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Sample records for diazotroph gluconacetobacter diazotrophicus

  1. Monitoring the colonization of sugarcane and rice plants by the endophytic diazotrophic bacterium Gluconacetobacter diazotrophicus marked with gfp and gusA reporter genes.

    Science.gov (United States)

    Rouws, L F M; Meneses, C H S G; Guedes, H V; Vidal, M S; Baldani, J I; Schwab, S

    2010-09-01

    To evaluate the colonization process of sugarcane plantlets and hydroponically grown rice seedlings by Gluconacetobacter diazotrophicus strain PAL5 marked with the gusA and gfp reporter genes. Sugarcane plantlets inoculated in vitro with PAL5 carrying the gfp::gusA plasmid pHRGFPGUS did not present green fluorescence, but beta-glucuronidase (GUS)-stained bacteria could be observed inside sugarcane roots. To complement this existing inoculation methodology for micropropagated sugarcane with a more rapid colonization assay, we employed hydroponically grown gnotobiotic rice seedlings to study PAL5-plant interaction. PAL5 could be isolated from the root surface (10(8) CFU g(-1)) and from surface-disinfected root and stem tissues (10(4) CFU g(-1)) of inoculated plants, suggesting that PAL5 colonized the internal plant tissues. Light microscopy confirmed the presence of bacteria inside the root tissue. After inoculation of rice plantlets with PAL5 marked with the gfp plasmid pHRGFPTC, bright green fluorescent bacteria could be seen colonizing the rice root surface, mainly at the sites of lateral root emergence, at root caps and on root hairs. The plasmids pHRGFPGUS and pHRGFPTC are valid tools to mark PAL5 and monitor the colonization of micropropagated sugarcane and hydroponic rice seedlings. These tools are of use to: (i) study PAL5 mutants affected in bacteria-plant interactions, (ii) monitor plant colonization in real time and (iii) distinguish PAL5 from other bacteria during the study of mixed inoculants.

  2. Drought Tolerance Conferred to Sugarcane by Association with Gluconacetobacter diazotrophicus: A Transcriptomic View of Hormone Pathways

    Science.gov (United States)

    Vargas, Lívia; Santa Brígida, Ailton B.; Mota Filho, José P.; de Carvalho, Thais G.; Rojas, Cristian A.; Vaneechoutte, Dries; Van Bel, Michiel; Farrinelli, Laurent; Ferreira, Paulo C. G.; Vandepoele, Klaas; Hemerly, Adriana S.

    2014-01-01

    Sugarcane interacts with particular types of beneficial nitrogen-fixing bacteria that provide fixed-nitrogen and plant growth hormones to host plants, promoting an increase in plant biomass. Other benefits, as enhanced tolerance to abiotic stresses have been reported to some diazotrophs. Here we aim to study the effects of the association between the diazotroph Gluconacetobacter diazotrophicus PAL5 and sugarcane cv. SP70-1143 during water depletion by characterizing differential transcriptome profiles of sugarcane. RNA-seq libraries were generated from roots and shoots of sugarcane plants free of endophytes that were inoculated with G. diazotrophicus and subjected to water depletion for 3 days. A sugarcane reference transcriptome was constructed and used for the identification of differentially expressed transcripts. The differential profile of non-inoculated SP70-1143 suggests that it responds to water deficit stress by the activation of drought-responsive markers and hormone pathways, as ABA and Ethylene. qRT-PCR revealed that root samples had higher levels of G. diazotrophicus 3 days after water deficit, compared to roots of inoculated plants watered normally. With prolonged drought only inoculated plants survived, indicating that SP70-1143 plants colonized with G. diazotrophicus become more tolerant to drought stress than non-inoculated plants. Strengthening this hypothesis, several gene expression responses to drought were inactivated or regulated in an opposite manner, especially in roots, when plants were colonized by the bacteria. The data suggests that colonized roots would not be suffering from stress in the same way as non-inoculated plants. On the other hand, shoots specifically activate ABA-dependent signaling genes, which could act as key elements in the drought resistance conferred by G. diazotrophicus to SP70-1143. This work reports for the first time the involvement of G. diazotrophicus in the promotion of drought-tolerance to sugarcane cv. SP70

  3. Drought tolerance conferred to sugarcane by association with Gluconacetobacter diazotrophicus: a transcriptomic view of hormone pathways.

    Directory of Open Access Journals (Sweden)

    Lívia Vargas

    Full Text Available Sugarcane interacts with particular types of beneficial nitrogen-fixing bacteria that provide fixed-nitrogen and plant growth hormones to host plants, promoting an increase in plant biomass. Other benefits, as enhanced tolerance to abiotic stresses have been reported to some diazotrophs. Here we aim to study the effects of the association between the diazotroph Gluconacetobacter diazotrophicus PAL5 and sugarcane cv. SP70-1143 during water depletion by characterizing differential transcriptome profiles of sugarcane. RNA-seq libraries were generated from roots and shoots of sugarcane plants free of endophytes that were inoculated with G. diazotrophicus and subjected to water depletion for 3 days. A sugarcane reference transcriptome was constructed and used for the identification of differentially expressed transcripts. The differential profile of non-inoculated SP70-1143 suggests that it responds to water deficit stress by the activation of drought-responsive markers and hormone pathways, as ABA and Ethylene. qRT-PCR revealed that root samples had higher levels of G. diazotrophicus 3 days after water deficit, compared to roots of inoculated plants watered normally. With prolonged drought only inoculated plants survived, indicating that SP70-1143 plants colonized with G. diazotrophicus become more tolerant to drought stress than non-inoculated plants. Strengthening this hypothesis, several gene expression responses to drought were inactivated or regulated in an opposite manner, especially in roots, when plants were colonized by the bacteria. The data suggests that colonized roots would not be suffering from stress in the same way as non-inoculated plants. On the other hand, shoots specifically activate ABA-dependent signaling genes, which could act as key elements in the drought resistance conferred by G. diazotrophicus to SP70-1143. This work reports for the first time the involvement of G. diazotrophicus in the promotion of drought-tolerance to

  4. Drought tolerance conferred to sugarcane by association with Gluconacetobacter diazotrophicus: a transcriptomic view of hormone pathways.

    Science.gov (United States)

    Vargas, Lívia; Santa Brígida, Ailton B; Mota Filho, José P; de Carvalho, Thais G; Rojas, Cristian A; Vaneechoutte, Dries; Van Bel, Michiel; Farrinelli, Laurent; Ferreira, Paulo C G; Vandepoele, Klaas; Hemerly, Adriana S

    2014-01-01

    Sugarcane interacts with particular types of beneficial nitrogen-fixing bacteria that provide fixed-nitrogen and plant growth hormones to host plants, promoting an increase in plant biomass. Other benefits, as enhanced tolerance to abiotic stresses have been reported to some diazotrophs. Here we aim to study the effects of the association between the diazotroph Gluconacetobacter diazotrophicus PAL5 and sugarcane cv. SP70-1143 during water depletion by characterizing differential transcriptome profiles of sugarcane. RNA-seq libraries were generated from roots and shoots of sugarcane plants free of endophytes that were inoculated with G. diazotrophicus and subjected to water depletion for 3 days. A sugarcane reference transcriptome was constructed and used for the identification of differentially expressed transcripts. The differential profile of non-inoculated SP70-1143 suggests that it responds to water deficit stress by the activation of drought-responsive markers and hormone pathways, as ABA and Ethylene. qRT-PCR revealed that root samples had higher levels of G. diazotrophicus 3 days after water deficit, compared to roots of inoculated plants watered normally. With prolonged drought only inoculated plants survived, indicating that SP70-1143 plants colonized with G. diazotrophicus become more tolerant to drought stress than non-inoculated plants. Strengthening this hypothesis, several gene expression responses to drought were inactivated or regulated in an opposite manner, especially in roots, when plants were colonized by the bacteria. The data suggests that colonized roots would not be suffering from stress in the same way as non-inoculated plants. On the other hand, shoots specifically activate ABA-dependent signaling genes, which could act as key elements in the drought resistance conferred by G. diazotrophicus to SP70-1143. This work reports for the first time the involvement of G. diazotrophicus in the promotion of drought-tolerance to sugarcane cv. SP70

  5. INTERACCIÓN DE LA BACTERIA Gluconacetobacter diazotrophicus Y HORTALIZAS DE RAÍZ

    OpenAIRE

    Yoania Ríos Rocafull; Bernardo Dibut Álvarez; Marcia Rojas Badía; Marisel Ortega García; Noel Arozarena Daza; Janet Rodríguez Sánchez

    2016-01-01

    Gluconacetobacter diazotrophicus es una bacteria endófita que presenta dentro de sus características metabólicas, mecanismos directos e indirectos de estimulación del crecimiento vegetal. Se han obtenido resultados positivos por su aplicación, fundamentalmente, en gramíneas y viandas tropicales. Sin embargo, en hortalizas, las investigaciones de esta interacción plantamicroorganismo son escasas. El presente trabajo tuvo como objetivo evaluar el efecto de cuatro cepas de G. diazotrophicus aisl...

  6. Quantification of natural populations of Gluconacetobacter diazotrophicus and Herbaspirillum spp. In sugar cane (Saccharum spp.) Using differente polyclonal antibodies

    Science.gov (United States)

    da Silva-Froufe, Lúcia Gracinda; Boddey, Robert Michael; Reis, Veronica Massena

    2009-01-01

    The species Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN) Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay) can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 105 cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques. PMID:24031435

  7. Quantification of natural populations of Gluconacetobacter diazotrophicus and Herbaspirillum spp. In sugar cane (Saccharum spp.) Using differente polyclonal antibodies.

    Science.gov (United States)

    da Silva-Froufe, Lúcia Gracinda; Boddey, Robert Michael; Reis, Veronica Massena

    2009-10-01

    The species Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN) Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay) can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 10(5) cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques.

  8. Diversidade de Gluconacetobacter diazotrophicus isolada de plantas de cana-de-açúcar cultivadas no Brasil Diversity of Gluconacetobacter diazotrophicus isolated from sugarcane plants cultivated in Brazil

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    Liamara Perin

    2004-08-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade da população de Gluconacetobacter diazotrophicus oriunda de cultivares de cana-de-açúcar (Saccharum spp. de diferentes regiões e bancos de germoplasma. O estudo foi realizado com 123 isolados, obtidos de folhas, colmos e raízes de 80 espécies e híbridos de cana-de-açúcar, procedentes de diferentes países e mantidos em coleções de germoplasma nos Estados da Bahia e Rio de Janeiro. Foram utilizados cinco isolados obtidos de plantas de café (Coffea arabica, dois de abacaxi (Ananas comosus e um de Pennisetum purpureum e mais 10 estirpes com padrões eletroforéticos distintos, após o uso de enzimas comuns do metabolismo microbiano (MLEE. O agrupamento obtido por meio da técnica de imunoadsorção com enzima acoplada (ELISA sugere que as variações expressas pelos isolados não estão relacionadas com a espécie de planta, a variedade de cana-de-açúcar, a origem geográfica, a parte da planta de onde os isolados foram obtidos e o tempo de amostragem. Altas doses de nitrogênio levaram à diminuição da diversidade de G. diazotrophicus.The objective of this study was to evaluate the diversity of Gluconacetobacter diazotrophicus isolates from different sugarcane (Saccharum spp. varieties. This study was developed using 123 strains from the internal tissues of stems and roots, isolated from 80 species and hybrids of sugarcane, originated from different countries and maintained in two germoplasm collections localized in Bahia and Rio de Janeiro States. Five isolates obtained from coffee plants (Coffea arabica, two from pineapple (Ananas comosus, one from Pennisetum purpureum and 10 strains, which present different electrophoretic patterns, were used in the comparison. Cluster analysis of enzyme-linked immunosorbent assay (ELISA results suggested that variations were not correlated with the plant species, sugarcane variety, geographic origin, parts of plants nor with sampling time

  9. Phosphate enhances levan production in the endophytic bacterium Gluconacetobacter diazotrophicus Pal5

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    Idogawa, Nao; Amamoto, Ryuta; Murata, Kousaku; Kawai, Shigeyuki

    2014-01-01

    Gluconacetobacter diazotrophicus is a gram-negative and endophytic nitrogen-fixing bacterium that has several beneficial effects in host plants; thus, utilization of this bacterium as a biofertilizer in agriculture may be possible. G. diazotrophicus synthesizes levan, a D-fructofuranosyl polymer with β-(2→6) linkages, as an exopolysaccharide and the synthesized levan improves the stress tolerance of the bacterium. In this study, we found that phosphate enhances levan production by G. diazotrophicus Pal5, a wild type strain that showed a stronger mucous phenotype on solid medium containing 28 mM phosphate than on solid medium containing 7 mM phosphate. A G. diazotrophicus Pal5 levansucrase disruptant showed only a weak mucous phenotype regardless of the phosphate concentration, indicating that the mucous phenotype observed on 28 mM phosphate medium was caused by levan. To our knowledge, this is the first report of the effect of a high concentration of phosphate on exopolysaccharide production. PMID:24717418

  10. Gluconacetobacter diazotrophicus: UN MICROORGANISMO PROMISORIO EN LA ELABORACIÓN DE BIOPREPARADOS

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    Yoania Ríos; B. Dibut

    2007-01-01

    Gluconacetobacter diazotrophicus, bacteria descubierta recientemente, se asocia a especies vegetales con altos contenidos de azúcares. Ello permite el incremento del nitrógeno disponible para la planta a partir del proceso de fijación biológica. El microorganismo libera al medio de hasta el 50 % de este elemento químico transformado. Además, posibilita el incremento de los rendimientos por la producción de sustancias fisiológicamente activas. A pesar de las diversas teorías en cuanto a la dis...

  11. Ecological occurrence of Gluconacetobacter diazotrophicus and nitrogen-fixing Acetobacteraceae members: their possible role in plant growth promotion.

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    Saravanan, V S; Madhaiyan, M; Osborne, Jabez; Thangaraju, M; Sa, T M

    2008-01-01

    Gluconacetobacter diazotrophicus has a long-standing history of bacterial-plant interrelationship as a symbiotic endophyte capable of fixing atmospheric nitrogen. In low nitrogen fertilized sugarcane fields it plays a significant role and its occurrence was realised in most of the sugarcane growing countries. In this mini review, the association of G. diazotrophicus with sugarcane, other crop plants and with various hosts is discussed. The factors affecting survival in the rhizosphere and the putative soil mode of transmission are emphasized. In addition, other N(2)-fixing Acetobacteraceae members, including Gluconacetobacter azotocaptans, Gluconacetobacter johannae and Swaminathania salitolerans, occurring in coffee, corn and rice plants are also covered. Lastly, the plant-growth-promoting traits identified in this group of bacteria, including N(2) fixation, phytohormone synthesis, P and Zn solubilization and biocontrol, are analysed.

  12. Solubilization of insoluble zinc compounds by Gluconacetobacter diazotrophicus and the detrimental action of zinc ion (Zn2+) and zinc chelates on root knot nematode Meloidogyne incognita.

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    Saravanan, V S; Kalaiarasan, P; Madhaiyan, M; Thangaraju, M

    2007-03-01

    To examine the zinc (Zn) solubilization potential and nematicidal properties of Gluconacetobacter diazotrophicus. Atomic Absorption Spectrophotometer, Differential Pulse Polarography and Gas Chromatography Coupled Mass Spectrometry were used to estimate the total Zn and Zn(2+) ions and identify the organic acids present in the culture supernatants. The effect of culture filtrate of Zn-amended G. diazotrophicus PAl5 on Meloidogyne incognita in tomato was examined under gnotobiotic conditions. Gluconacetobacter diazotrophicus PAl5 effectively solubilized the Zn compounds tested and 5-ketogluconic acid was identified as the major organic acid aiding the solubilization of zinc oxide. The presence of Zn compounds in the culture filtrates of G. diazotrophicus enhanced the mortality and reduced the root penetration of M. incognita under in vitro conditions. 5-ketogluconic acid produced by G. diazotrophicus mediated the solubilization process and the available Zn(2+) ions enhanced the nematicidal activity of G. diazotrophicus against M. incognita. Zn solubilization and enhanced nematicidal activity of Zn-amended G. diazotrophicus provides the possibility of exploiting it as a plant growth promoting bacteria.

  13. Uso de diferentes fontes de carbono por estirpes de Gluconacetobacter diazotrophicus isoladas de cana-de-açúcar Use of different carbon sources by strains of Gluconacetobacter diazotrophicus isolated from sugarcane

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    Edilene Alves Barbosa

    2006-05-01

    Full Text Available O objetivo deste trabalho foi quantificar a população de Gluconactobacter diazotrophicus presente em plântulas de 34 variedades de cana-de-açúcar, e caracterizar os isolados quanto à capacidade de utilizar diversas fontes de carbono e formar pigmento róseo em presença de luz. A população de G. diazotrophicus foi quantificada pela técnica do número mais provável e isolada em meios de cultura semi-seletivos. A população de G. diazotrophicus foi maior no sistema radicular do que na parte aérea, e maior nas variedades brasileiras de cana-de-açúcar. Os isolados utilizaram poucas fontes de carbono, e aqueles obtidos do sistema radicular utilizaram maior número de fontes, em comparação aos da parte aérea. Apenas a estirpe padrão Ppe-4 e quatro outros isolados formam pigmento róseo em presença de luz. A baixa variação observada pode estar relacionada com o habitat ou com a adaptação dessa população às condições específicas do interior das plantas e, por isso, a estrutura genética da população dessas bactérias é limitada e conservada.The objective of this work was to quantify the population of Gluconacetobacter diazotrophicus present in plantlets of 34 sugarcane varieties and to characterize the isolates according to their capacity to use carbon sources and produce pink pigments under light. G. diazotrophicus population was estimated using the most probable number technique, and then, isolated in semi-selective culture media. The highest population number of G. diazotrophicus was found in the roots; Brazilian varieties presented the highest population. The isolates used little carbon sources, and those strains obtained from the root used the highest number of carbon sources. Only the strain Ppe-4 and four other ones produce pink pigments under light. The low variation observed may be related to the habitat or to the adaptation of this population to specific internal plant conditions, and for this reason, the

  14. Inoculation of Burkholderia cepacia and Gluconacetobacter diazotrophicus on phenotype and biomass of Triticum aestivum var. Nana-F2007 at 50% of nitrogen fertilizer

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    Jesús Jaime Hernández-Escareño

    2015-03-01

    Full Text Available Wheat (Triticum aestivum L consuming requires of nitrogen fertilizer (NF, as ammonium nitrate (NH4NO3, which one in excess causes lost soil productivity. An alternative to reduce and optimize NF to wheat is to inoculate with endophytic promoting growth bacteria (EPGB, as genus Burkholderia cepacia and Gluconacetobacter diazotrophicus able to improve radical uptake of NF, its suggesting by inducing synthesis of growth promoting vegetal substances (GPVS. The aim of this research was to evaluate the inoculation of Burkholderia cepacia and Gluconacetobacter diazotrophicus on phenology and biomass of T.aestivum at 50% dose of NF. A trial in greenhouse condition wasconducted inoculating seed T.aestivum´s with both EPGB by measuring its phenology: (PH plant height, (RL root length and biomass: total fresh weight (TFW and dry (TDW at seedling and flowering stages. Results showed a positive effect of B. cepacia in wheat on its TDW with 0.61g value statistically significant compared to 0.53g TDW of wheat used as relative control fed with NF 100% dose (RC. B. cepacia and G. diazotrophicus inoculated to wheat had a positive increased on its TDW with 4.23 g value statistically significant compared to 1.13 g TDW of wheat used as RC. Conclusion suggested that B. cepacia and G. diazotrophicus by synthetized GPVS had a positive effect on wheat growth at reduced dose of NF.

  15. The decrease in the population of Gluconacetobacter diazotrophicus in sugarcane after nitrogen fertilization is related to plant physiology in split root experiments.

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    Rodríguez-Andrade, Osvaldo; Fuentes-Ramírez, Luis E; Morales-García, Yolanda E; Molina-Romero, Dalia; Bustillos-Cristales, María R; Martínez-Contreras, Rebeca D; Muñoz-Rojas, Jesús

    2015-01-01

    It has been established that a decrease in the population of Gluconacetobacter diazotrophicus associated with sugarcane occurs after nitrogen fertilization. This fact could be due to a direct influence of NH(4)NO(3) on bacterial cells or to changes in plant physiology after fertilizer addition, affecting bacterial establishment. In this work, we observed that survival of G. diazotrophicus was directly influenced when 44.8mM of NH(4)NO(3) (640mgN/plant) was used for in vitro experiments. Furthermore, micropropagated sugarcane plantlets were inoculated with G. diazotrophicus and used for split root experiments, in which both ends of the system were fertilized with a basal level of NH(4)NO(3) (0.35mM; 10mgN/plant). Twenty days post inoculation (dpi) one half of the plants were fertilized with a high dose of NH(4)NO(3) (6.3mM; 180 mgN/plant) on one end of the system. This nitrogen level was lower than that directly affecting G. diazotrophicus cells; however, it caused a decrease in the bacterial population in comparison with control plants fertilized with basal nitrogen levels. The decrease in the population of G. diazotrophicus was higher in pots fertilized with a basal nitrogen level when compared with the corresponding end supplied with high levels of NH4NO3 (100dpi; 80 days post fertilization) of the same plant system. These observations suggest that the high nitrogen level added to the plants induce systemic physiological changes that affect the establishment of G. diazotrophicus. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  16. Quantification of natural populations of Gluconacetobacter diazotrophicus and Herbaspirillum spp. In sugar cane (Saccharum spp. Using differente polyclonal antibodies

    Directory of Open Access Journals (Sweden)

    Lúcia Gracinda da Silva-Froufe

    2009-12-01

    Full Text Available The species Gluconacetobacterdiazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 10(5 cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques.

  17. The Oxidative Fermentation of Ethanol in Gluconacetobacter diazotrophicus Is a Two-Step Pathway Catalyzed by a Single Enzyme: Alcohol-Aldehyde Dehydrogenase (ADHa

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    Saúl Gómez-Manzo

    2015-01-01

    Full Text Available Gluconacetobacter diazotrophicus is a N2-fixing bacterium endophyte from sugar cane. The oxidation of ethanol to acetic acid of this organism takes place in the periplasmic space, and this reaction is catalyzed by two membrane-bound enzymes complexes: the alcohol dehydrogenase (ADH and the aldehyde dehydrogenase (ALDH. We present strong evidence showing that the well-known membrane-bound Alcohol dehydrogenase (ADHa of Ga. diazotrophicus is indeed a double function enzyme, which is able to use primary alcohols (C2–C6 and its respective aldehydes as alternate substrates. Moreover, the enzyme utilizes ethanol as a substrate in a reaction mechanism where this is subjected to a two-step oxidation process to produce acetic acid without releasing the acetaldehyde intermediary to the media. Moreover, we propose a mechanism that, under physiological conditions, might permit a massive conversion of ethanol to acetic acid, as usually occurs in the acetic acid bacteria, but without the transient accumulation of the highly toxic acetaldehyde.

  18. Development and nitrate reductase activity of sugarcane inoculated with five diazotrophic strains.

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    Dos Santos, Silvana Gomes; da Silva Ribeiro, Flaviane; da Fonseca, Camila Sousa; Pereira, Willian; Santos, Leandro Azevedo; Reis, Veronica Massena

    2017-08-01

    Diazotrophs are able to stimulate plant growth. This study aimed at evaluating the effect of inoculation of five diazotrophic strains on growth promotion and nitrate reductase (NR, EC 1.7.1.1) activity in sugarcane. An experiment was carried out from three stages of cultivation: sprouting, tubes, and in hydroponics. On the first two stages, seven treatments were adopted: uninoculated control; mixed inoculation with five strains; and individual inoculation with Gluconacetobacter diazotrophicus (Gd), Herbaspirillum rubrisubalbicans (Hr), Herbaspirillum seropedicae (Hs), Nitrospirillum amazonense (Na), and Paraburkholderia tropica (Pt). The four treatments showing the best performance were transferred to the hydroponic system for analysis of NR activity. Hs, Pt, and the mixture of all strains led to the highest seedling biomass in tubes, followed by Hr. In hydroponics, the mixture and the strain Hr had the highest growth-promoting effect. NR activity was influenced by inoculation only under low N supply conditions, with positive effect of Hr, Pt, and the mixture.

  19. Use of endophytic diazotrophic bacteria as a vector to express the cry3A gene from Bacillus thuringiensis

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    Salles Joana Falcão

    2000-01-01

    Full Text Available The goal of this study was to evaluate the potential of endophytic diazotrophic bacteria as a vector to express a cry gene from Bacillus thuringiensis, envisaging the control of pests that attack sugarcane plants. The endophytic nitrogen-fixing bacteria Gluconacetobacter diazotrophicus strain BR11281 and Herbaspirillum seropedicae strain BR11335 were used as models. The cry3A gene was transferred by conjugation using a suicide plasmid and the recombinant strains were selected by their ability to fix nitrogen in semi-solid N-free medium. The presence of the cry gene was detected by Southern-blot using an internal fragment of 1.0 kb as a probe. The production of delta-endotoxin by the recombinant H. seropedicae strain was detected by dot blot while for G. diazotrophicus the Western-blot technique was used. In both cases, a specific antibody raised against the B. thuringiensis toxin was applied. The delta-endotoxin production showed by the G. diazotrophicus recombinant strain was dependent on the nitrogen fixing conditions since the cry3A gene was fused to a nif promoter. In the case of H. seropedicae the delta-endotoxin expression was not affected by the promoter (rhi used. These results suggest that endophytic diazotrophic bacteria can be used as vectors to express entomopathogenic genes envisaging control of sugarcane pests.

  20. Relación de la fijación de nitrógeno y la producción de auxinas en cepas de Gluconacetobacter diazotrophicus procedentes de diferentes cultivos

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    Marcia M. Rojas

    2009-01-01

    Full Text Available Se determinó la capacidad de fijar nitrógeno mediante la actividad reductora de acetileno para 8 cepas de G. diazotrophicus aisladas de diferentes ecosistemas empleando el medio LGI-P. Además, se determinó la producción de auxinas a través del método de Salkowski y se analizó la influencia del aminoácido triptófano y del AIA en la actividad de la nitrogenasa. El triptófano, al igual que otros aminoácidos y las diferentes concentraciones de AIA, inhiben en distinta medida la actividad de la enzima solo parcialmente. Se demostró que las condiciones de nitrofijación no afectan la producción de AIA en esta bacteria. Esta relación entre ambas capacidades fisiológicas beneficiosas para los cultivos agrícolas pudiera tener gran importancia ya que pueden desarrollarse paralelamente, y potenciar la acción beneficiosa hacia la planta, basada en la dinitrofijación y la producción de auxinas estimuladoras del crecimiento vegetal. The capacity to fix nitrogen of 8 strains of G. diazotrophicus from different ecosystems was determined by acetylene reduction assay using LGIP medium. Moreover, auxins production was determined by Salkowski’s method and the influence of triptophan and indoleacetic acid (IAA in the nitrogenase activity were analyzed. The triptophan as other aminoacids and different concentrations of IAA, inhibit at different levels the nitrogen fixation only partially. There were demonstrated that the nitrogen fixation conditions do not affect the auxins production of this bacteria. This relationship between both crop beneficial physiological capacities should be a great importance since they may be parallely developed, and enhance the beneficial action to the plant, based on dinitrogen fixation and stimulating plant growth auxins production.

  1. NCBI nr-aa BLAST: CBRC-MMUR-01-0729 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0729 ref|YP_001601963.1| hypothetical protein GDI_1718 [Gluconacetobacter diazo...trophicus PAl 5] ref|YP_002277838.1| hypothetical protein Gdia_3499 [Gluconacetobacter diazotrophic...us PAl 5] emb|CAP55661.1| putative membrane protein [Gluconacetobacter diazotrophicus PAl 5] gb|ACI53223.1| ...conserved hypothetical protein [Gluconacetobacter diazotrophicus PAl 5] YP_001601963.1 0.11 24% ...

  2. Natural Endophytic Occurrence of Acetobacter diazotrophicus in Pineapple Plants.

    Science.gov (United States)

    Tapia-Hernández; Bustillos-Cristales; Jiménez-Salgado; Caballero-Mellado; Fuentes-Ramírez

    2000-01-01

    The presence of endophytic Acetobacter diazotrophicus was tested for pineapple plants (Ananas comosus [L.] Merr.) grown in the field. Diazotrophic bacteria were isolated from the inner tissues of surface sterilized roots, stems, and leaves of pineapple plants. Phenotypic tests permitted the selection of presumptive nitrogen-fixing A. diazotrophicus isolates. Restriction fragment length polymorphisms (RFLPs) of small subunit (SSU) rDNA using total DNA digested with endonuclease SphI and with endonuclease NcoI, hybridizations of RNA with an A. diazotrophicus large subunit (LSU) rRNA specific probe, as well as patterns in denaturing protein electrophoresis (SDS-PAGE) and multilocus enzyme tests allowed the identification of A. diazotrophicus isolates. High frequencies of isolation were obtained from propagative buds that had not been nitrogen-fertilized, and lower frequencies from 3-month-old plants that had been nitrogen-fertilized. No isolates were recovered from 5- to 7-month-old nitrogen-fertilized plants. All the A. diazotrophicus isolates recovered from pineapple plants belonged to the multilocus genotype which shows the most extensive distribution among all host species previously analyzed.

  3. In silico differential display of defense-related expressed sequence tags from sugarcane tissues infected with diazotrophic endophytes

    Directory of Open Access Journals (Sweden)

    Lambais Marcio R.

    2001-01-01

    Full Text Available The expression patterns of 277 sugarcane expressed sequence tags (EST-contigs encoding putative defense-related (DR proteins were evaluated using the Sugarcane EST database. The DR proteins evaluated included chitinases, beta-1,3-glucanases, phenylalanine ammonia-lyases, chalcone synthases, chalcone isomerases, isoflavone reductases, hydroxyproline-rich glycoproteins, proline-rich glycoproteins, peroxidases, catalases, superoxide dismutases, WRKY-like transcription factors and proteins involved in cell death control. Putative sugarcane WRKY proteins were compared and their phylogenetic relationships determined. A hierarchical clustering approach was used to identify DR ESTs with similar expression profiles in representative cDNA libraries. To identify DR ESTs differentially expressed in sugarcane tissues infected with Gluconacetobacter diazotrophicus or Herbaspirillum rubrisubalbicans, 179 putative DR EST-contigs expressed in non-infected tissues (leaves and roots and/or infected tissues were selected and arrayed by similarity of their expression profiles. Changes in the expression levels of 124 putative DR EST-contigs, expressed in non-infected tissues, were evaluated in infected tissues. Approximately 42% of these EST-contigs showed no expression in infected tissues, whereas 15% and 3% showed more than 2-fold suppression in tissues infected with G. diazotrophicus or H. rubrisubalbicans, respectively. Approximately 14 and 8% of the DR EST-contigs evaluated showed more than 2-fold induction in tissues infected with G. diazotrophicus or H. rubrisubalbicans, respectively. The differential expression of clusters of DR genes may be important in the establishment of a compatible interaction between sugarcane and diazotrophic endophytes. It is suggested that the hierarchical clustering approach can be used on a genome-wide scale to identify genes likely involved in controlling plant-microorganism interactions.

  4. Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen-fixing acetobacteria.

    Science.gov (United States)

    Jimenez-Salgado, T; Fuentes-Ramirez, L E; Tapia-Hernandez, A; Mascarua-Esparza, M A; Martinez-Romero, E; Caballero-Mellado, J

    1997-09-01

    Acetobacter diazotrophicus was isolated from coffee plant tissues and from rhizosphere soils. Isolation frequencies ranged from 15 to 40% and were dependent on soil pH. Attempts to isolate this bacterial species from coffee fruit, from inside vesicular-arbuscular mycorrhizal fungi spores, or from mealybugs (Planococcus citri) associated with coffee plants were not successful. Other acid-producing diazotrophic bacteria were recovered with frequencies of 20% from the coffee rhizosphere. These N2-fixing isolates had some features in common with the genus Acetobacter but should not be assigned to the species Acetobacter diazotrophicus because they differed from A. diazotrophicus in morphological and biochemical traits and were largely divergent in electrophoretic mobility patterns of metabolic enzymes at coefficients of genetic distance as high as 0.950. In addition, these N2-fixing acetobacteria differed in the small-subunit rRNA restriction fragment length polymorphism patterns obtained with EcoRI, and they exhibited very low DNA-DNA homology levels, ranging from 11 to 15% with the A. diazotrophicus reference strain PAI 5T. Thus, some of the diazotrophic acetobacteria recovered from the rhizosphere of coffee plants may be regarded as N2-fixing species of the genus Acetobacter other than A. diazotrophicus. Endophytic diazotrophic bacteria may be more prevalent than previously thought, and perhaps there are many more potentially beneficial N2-fixing bacteria which can be isolated from other agronomically important crops.

  5. Effects of the inoculation of Burkholderia vietnamensis and related endophytic diazotrophic bacteria on grain yield of rice.

    Science.gov (United States)

    Govindarajan, Munusamy; Balandreau, Jacques; Kwon, Soon-Wo; Weon, Hang-Yeon; Lakshminarasimhan, Cunthipuram

    2008-01-01

    During a survey of endophytic diazotrophic bacteria associated with different rice varieties in Tamilnadu, some "endophytes" were obtained. Thirteen bacterial isolates from surface-sterilized roots and shoots were obtained in pure culture, which produced indole acetic acid (IAA) and reduced acetylene to ethylene. Polymerase chain reaction (PCR) amplification confirmed the presence of nif-H gene in all the isolates. Morphological, biochemical, and molecular characteristics indicated that all of them belonged to the genus Burkholderia One of them, MGK3, was consistently more active in reducing acetylene, and 16S rDNA sequences of isolate MGK3 confirmed its identification as Burkholderia vietnamiensis. Colonization of rice root was confirmed by strain MGK3 marked with gusA gene. The inoculated roots showed a blue color, which was most intense at the points of lateral root emergence and at the root tip. Transverse sections of roots, 15 days after inoculation, revealed beta-glucuronidase (GUS) activity within many of the cortical intercellular spaces next to the stele and within the aerenchyma. Nitrogen fixation was quantified by using (15)N isotope dilution method with two different cultivars grown in pot and field experiments. Higher nitrogen fixation was observed in variety Ponni than in ADT-43, where nearly 42% (field) and 40% (pot) of the nitrogen was derived from the atmosphere (% Ndfa). Isolate MGK3 was used to inoculate rice seedlings in a comparison with four other diazotrophs, viz., Gluconacetobacter diazotrophicus LMG7603, Herbaspirillum seropedicae LMG6513, Azospirillum lipoferum 4B LMG4348, and B. vietnamiensis LMG10929. They were used to conduct two pot and four field inoculation experiments. MGK3 alone, and combined with other diazotrophs, performed best under both pot and field conditions: combined inoculation produced yield increases between 9.5 and 23.6%, while MGK3 alone increased yield by 5.6 to 12.16% over the uninoculated control treatment.

  6. Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen-fixing acetobacteria.

    OpenAIRE

    Jimenez-Salgado, T; Fuentes-Ramirez, L E; Tapia-Hernandez, A; Mascarua-Esparza, M A; Martinez-Romero, E; Caballero-Mellado, J

    1997-01-01

    Acetobacter diazotrophicus was isolated from coffee plant tissues and from rhizosphere soils. Isolation frequencies ranged from 15 to 40% and were dependent on soil pH. Attempts to isolate this bacterial species from coffee fruit, from inside vesicular-arbuscular mycorrhizal fungi spores, or from mealybugs (Planococcus citri) associated with coffee plants were not successful. Other acid-producing diazotrophic bacteria were recovered with frequencies of 20% from the coffee rhizosphere. These N...

  7. Lipopolysaccharides in diazotrophic bacteria

    OpenAIRE

    Serrato, Rodrigo V.

    2014-01-01

    Biological nitrogen fixation (BNF) is a process in which the atmospheric nitrogen (N2) is transformed into ammonia (NH3) by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are...

  8. Unraveling the molecular mechanisms of nitrogenase conformational protection against oxygen in diazotrophic bacteria.

    Science.gov (United States)

    Lery, Letícia M S; Bitar, Mainá; Costa, Mauricio G S; Rössle, Shaila C S; Bisch, Paulo M

    2010-12-22

    G. diazotrophicus and A. vinelandii are aerobic nitrogen-fixing bacteria. Although oxygen is essential for the survival of these organisms, it irreversibly inhibits nitrogenase, the complex responsible for nitrogen fixation. Both microorganisms deal with this paradox through compensatory mechanisms. In A. vinelandii a conformational protection mechanism occurs through the interaction between the nitrogenase complex and the FeSII protein. Previous studies suggested the existence of a similar system in G. diazotrophicus, but the putative protein involved was not yet described. This study intends to identify the protein coding gene in the recently sequenced genome of G. diazotrophicus and also provide detailed structural information of nitrogenase conformational protection in both organisms. Genomic analysis of G. diazotrophicus sequences revealed a protein coding ORF (Gdia0615) enclosing a conserved "fer2" domain, typical of the ferredoxin family and found in A. vinelandii FeSII. Comparative models of both FeSII and Gdia0615 disclosed a conserved beta-grasp fold. Cysteine residues that coordinate the 2[Fe-S] cluster are in conserved positions towards the metallocluster. Analysis of solvent accessible residues and electrostatic surfaces unveiled an hydrophobic dimerization interface. Dimers assembled by molecular docking presented a stable behaviour and a proper accommodation of regions possibly involved in binding of FeSII to nitrogenase throughout molecular dynamics simulations in aqueous solution. Molecular modeling of the nitrogenase complex of G. diazotrophicus was performed and models were compared to the crystal structure of A. vinelandii nitrogenase. Docking experiments of FeSII and Gdia0615 with its corresponding nitrogenase complex pointed out in both systems a putative binding site presenting shape and charge complementarities at the Fe-protein/MoFe-protein complex interface. The identification of the putative FeSII coding gene in G. diazotrophicus genome

  9. Lipopolysaccharides in diazotrophic bacteria.

    Science.gov (United States)

    Serrato, Rodrigo V

    2014-01-01

    Biological nitrogen fixation (BNF) is a process in which the atmospheric nitrogen (N2) is transformed into ammonia (NH3) by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are found between associative diazotrophs and plants, resulting in plant infection and colonization of internal tissues. Independent of the type of ecological interaction, glycans, and glycoconjugates produced by these bacteria play an important role in the molecular communication prior and during colonization. Even though exopolysaccharides (EPS) and lipochitooligosaccharides (LCO) produced by diazotrophic bacteria and released onto the environment have their importance in the microbe-plant interaction, it is the lipopolysaccharides (LPS), anchored on the external membrane of these bacteria, that mediates the direct contact of the diazotroph with the host cells. These molecules are extremely variable among the several species of nitrogen fixing-bacteria, and there are evidences of the mechanisms of infection being closely related to their structure.

  10. Quantification of diazotrophs bacteria isolated from cocoa soils (Theobroma cacao L., by the technique of Most Probable Number (MPN

    Directory of Open Access Journals (Sweden)

    Adriana Zulay Argüello Navarro

    2016-07-01

    Full Text Available The objective of this research was to quantify diazotrophic bacteria and compare physicochemically rhizospheric soils of three cocoa plantations (Theobroma cacao L. in Norte de Santander Department, Colombia; for which they were characterized, differing in cultivated area, agronomic management and crop age. From serial dilutions of the samples and using the technique of Most Probable Number (MPN, In semisolid culture media (NFb, JMV, LGI, JNFb, the diazotrophs were quantified, evaluating as positive the formation of a subsurface film in the medium contained in sealed vials; equal samples were sent to the Bioambiental laboratory (UNET for physicochemical analyzes. As a result, the evaluated samples showed deficiencies in the percentage of organic matter and elements such as Potassium, Phosphorus and Magnesium. Statistically highly significant differences in MPN were reported. The highest quantification of diazotrophs was reported in the Florilandia farm, which was characterized by drip irrigation. The highest quantification of diazotrophs was recorded in the media NFb and JMV, demonstrating a greater presence of the presumed genera Azospirillum sp. and Burkholderia sp. which are easily isolated from rhizospheric soils, unlike the genera Herbaspirillum sp. and Gluconacetobacter sp. which by their endophytic character tend to be less predominant in this type of samples. It is also concluded that the physicochemical characteristics of the soil, humidity and climatic relationships at the moment of sampling, condition the amount of root exudates and therefore are factors that conditioned the presence of diazotrophs in the samples.

  11. Strain improvement of Gluconacetobacter xylinus NCIM 2526 for ...

    African Journals Online (AJOL)

    The present investigation demonstrates the effectiveness of ultraviolet (UV) radiation and ethyl methanesulfonate (EMS) in strain improvement for enhanced cellulose production by Gluconacetobacter xylinus NCIM 2526. The mutants were compared with wild type for cellulose production. UV mutants GHUV3, GHUV4, and ...

  12. Occurrence of diazotrophic bacteria in Araucaria angustifolia

    OpenAIRE

    Neroni,Rafaela de Fátima; Cardoso,Elke Jurandy Bran Nogueira

    2007-01-01

    Araucaria angustifolia is an environmentally threatened tree and the whole biota of the Araucaria Forest should be investigated with the aim of its preservation. Diazotrophic bacteria are extremely important for the maintenance of ecosystems, but they have never been studied in Araucaria Forests. In this study, diazotrophic bacteria were isolated from Araucaria roots and soil, when grown in semi-specific, semi-solid media. The diazotrophic character of some recovered isolates could be confirm...

  13. Genome Sequence of Gluconacetobacter sp. Strain SXCC-1, Isolated from Chinese Vinegar Fermentation Starter▿

    OpenAIRE

    Du, Xin-jun; Jia, Shi-ru; Yang, Yue; Wang, Shuo

    2011-01-01

    Gluconacetobacter strains are prominent bacteria during traditional vinegar fermentation. Here, we report a draft genome sequence of Gluconacetobacter sp. strain SXCC-1. This strain was isolated from a fermentation starter (Daqu) used for commercial production of Shanxi vinegar, the best-known vinegar of China.

  14. Differentiation of species of the family Acetobacteraceae by AFLP DNA fingerprinting: Gluconacetobacter kombuchae is a later heterotypic synonym of Gluconacetobacter hansenii.

    Science.gov (United States)

    Cleenwerck, Ilse; De Wachter, Marjan; González, Angel; De Vuyst, Luc; De Vos, Paul

    2009-07-01

    Amplified fragment length polymorphism (AFLP) DNA fingerprinting was investigated as a tool for fast and accurate identification of acetic acid bacteria (AAB) to the species level. One hundred and thirty five reference strains and 15 additional strains, representing 50 recognized species of the family Acetobacteraceae, were subjected to AFLP analysis using the restriction enzyme combination ApaI/TaqI and the primer combination A03/T03. The reference strains had been previously subjected to either DNA-DNA hybridization or 16S-23S rRNA spacer region gene sequence analysis and were regarded as being accurately classified at the species level. The present study revealed that six of these strains should be reclassified, namely Gluconacetobacter europaeus LMG 1518 and Gluconacetobacter xylinus LMG 1510 as Gluconacetobacter xylinus and Gluconacetobacter europaeus, respectively; Gluconacetobacter kombuchae LMG 23726(T) as Gluconacetobacter hansenii; and Acetobacter orleanensis strains LMG 1545, LMG 1592 and LMG 1608 as Acetobacter cerevisiae. Cluster analysis of the AFLP DNA fingerprints of the reference strains revealed one cluster for each species, showing a linkage level below 50 % with other clusters, except for Acetobacter pasteurianus, Acetobacter indonesiensis and Acetobacter cerevisiae. These three species were separated into two, two, and three clusters, respectively. At present, confusion exists regarding the taxonomic status of Gluconacetobacter oboediens and Gluconacetobacter intermedius; the AFLP data from this study supported their classification as separate taxa. The 15 additional strains could all be identified at the species level. AFLP analysis further revealed that some species harboured genetically diverse strains, whereas other species consisted of strains showing similar banding patterns, indicating a more limited genetic diversity. It can be concluded that AFLP DNA fingerprinting is suitable for accurate identification and classification of a broad

  15. Ocorrência de bactérias diazotróficas em diferentes genótipos de cana-de-açúcar Occurrence of diazotrophic bacteria in different sugar cane genotypes

    Directory of Open Access Journals (Sweden)

    FÁBIO BUENO DOS REIS JUNIOR

    2000-05-01

    Full Text Available O objetivo deste trabalho foi avaliar a localização e o número de bactérias endofíticas em quatro genótipos de cana-de-açúcar e investigar sobre a possível existência de correlação com os resultados apresentados em trabalhos de quantificação da fixação biológica de nitrogênio (FBN. Fez-se um levantamento das bactérias diazotróficas presentes, e quantificou-se a população de Herbaspirillum spp. e Acetobacter diazotrophicus, em genótipos de cana-de-açúcar contrastantes quanto à capacidade de obter N da FBN. De acordo com o levantamento realizado neste trabalho, as bactérias estudadas (Azospirillum lipoferum, A. brasilense, A. amazonense, Herbaspirillum spp. e Acetobacter diazotrophicus estavam presentes nos quatro genótipos avaliados e em todas as partes da planta, exceto A. amazonense, que não foi isolado de amostras de folhas. A quantificação das bactérias Herbaspirillum spp. e A. diazotrophicus mostrou não haver diferenças significativas entre os genótipos, e que, geralmente, elas estão presentes em maior número nas raízes. Enquanto Herbaspirillum spp. mantém-se mais estável ao longo do ciclo da cultura, a população de A. diazotrophicus decresce com a aproximação do final do ciclo comercial. Pode-se sugerir que as diferenças entre as taxas de FBN encontradas nos diversos genótipos não é causada por diferenças na presença ou no número das bactérias aqui estudadas.The objective of this work was to find out the localization and number of endophytic bacteria in four sugar cane genotypes and investigate upon the possible existence of correlation to the results obtained in some studies about quantification of biological nitrogen fixation (BNF. A survey of the diazotrophic bacteria present in sugar cane genotypes differing in their capacity to obtain nitrogen through BNF was performed, and population of Herbaspirillum spp. and Acetobacter diazotrophicus was quantified. The bacteria tested in the

  16. Diazotrophic Bacterial Community of Degraded Pastures

    OpenAIRE

    João Tiago Correia Oliveira; Everthon Fernandes Figueredo; Williane Patrícia da Silva Diniz; Lucianne Ferreira Paes de Oliveira; Pedro Avelino Maia de Andrade; Fernando Dini Andreote; Júlia Kuklinsky-Sobral; Danúbia Ramos de Lima; Fernando José Freire

    2017-01-01

    Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady’s table was used to obtain the Most Probable Number (MPN) of bacteria per gram ...

  17. Diazotrophic Bacterial Community of Degraded Pastures

    Directory of Open Access Journals (Sweden)

    João Tiago Correia Oliveira

    2017-01-01

    Full Text Available Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady’s table was used to obtain the Most Probable Number (MPN of bacteria per gram of sample, in order to determine population density and calculate the Shannon-Weaver diversity index. The diversity of total diazotrophic bacterial community was determined by the technique of Denaturing Gradient Gel Electrophoresis (DGGE of the nifH gene, while the diversity of the culturable diazotrophic bacteria was determined by the Polymerase Chain Reaction (BOX-PCR technique. The increase in the degradation stage of the B. decumbens Stapf. pasture did not reduce the population density of the cultivated diazotrophic bacterial community, suggesting that the degradation at any degree of severity was highly harmful to the bacteria. The structure of the total diazotrophic bacterial community associated with B. decumbens Stapf. was altered by the pasture degradation stage, suggesting a high adaptive capacity of the bacteria to altered environments.

  18. Gluconacetobacter kakiaceti sp. nov., an acetic acid bacterium isolated from a traditional Japanese fruit vinegar.

    Science.gov (United States)

    Iino, Takao; Suzuki, Rei; Tanaka, Naoto; Kosako, Yoshimasa; Ohkuma, Moriya; Komagata, Kazuo; Uchimura, Tai

    2012-07-01

    Two novel acetic acid bacteria, strains G5-1(T) and I5-1, were isolated from traditional kaki vinegar (produced from fruits of kaki, Diospyros kaki Thunb.), collected in Kumamoto Prefecture, Japan. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains G5-1(T) and I5-1 formed a distinct subline in the genus Gluconacetobacter and were closely related to Gluconacetobacter swingsii DST GL01(T) (99.3% 16S rRNA gene sequence similarity). The isolates showed 96-100% DNA-DNA relatedness with each other, but genus Gluconacetobacter. The isolates could be distinguished from closely related members of the genus Gluconacetobacter by not producing 2- and 5-ketogluconic acids from glucose, producing cellulose, growing without acetic acid and with 30% (w/v) d-glucose, and producing acid from sugars and alcohols. Furthermore, the genomic DNA G+C contents of strains G5-1(T) and I5-1 were a little higher than those of their closest phylogenetic neighbours. On the basis of the phenotypic characteristics and phylogenetic position, strains G5-1(T) and I5-1 are assigned to a novel species, for which the name Gluconacetobacter kakiaceti sp. nov. is proposed; the type strain is G5-1(T) (=JCM 25156(T)=NRIC 0798(T)=LMG 26206(T)).

  19. Inoculation of sugarcane with diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Nivaldo Schultz

    2014-04-01

    Full Text Available The sugarcane industry, a strategic crop in Brazil, requires technological improvements in production efficiency to increase the crop energy balance. Among the various currently studied alternatives, inoculation with diazotrophic bacteria proved to be a technology with great potential. In this context, the efficiency of a mixture of bacterial inoculant was evaluated with regard to the agronomic performance and N nutrition of sugarcane. The experiment was carried out on an experimental field of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, using a randomized block, 2 × 3 factorial design (two varieties and three treatments with four replications, totaling 24 plots. The varieties RB867515 and RB72454 were tested in treatments consisting of: inoculation with diazotrophic bacteria, N-fertilized control with 120 kg ha-1 N and absolute control (no inoculation and no N fertilizer. The inoculum was composed of five strains of five diazotrophic species. The yield, dry matter accumulation, total N in the shoot dry matter and the contribution of N by biological fixation were evaluated, using the natural 15N abundance in non-inoculated sugarcane as reference. The bacterial inoculant increased the stalk yield of variety RB72454 similarly to fertilization with 120 kg ha-1 N in the harvests of plant-cane and first ratoon crops, however the contribution of biological N fixation was unchanged by inoculation, indicating that the benefits of the inoculant in sugarcane may have resulted from plant growth promotion.

  20. Nitrogen-fixing and cellulose-producing Gluconacetobacter kombuchae sp. nov., isolated from Kombucha tea.

    Science.gov (United States)

    Dutta, Debasree; Gachhui, Ratan

    2007-02-01

    A few members of the family Acetobacteraceae are cellulose-producers, while only six members fix nitrogen. Bacterial strain RG3T, isolated from Kombucha tea, displays both of these characteristics. A high bootstrap value in the 16S rRNA gene sequence-based phylogenetic analysis supported the position of this strain within the genus Gluconacetobacter, with Gluconacetobacter hansenii LMG 1527T as its nearest neighbour (99.1 % sequence similarity). It could utilize ethanol, fructose, arabinose, glycerol, sorbitol and mannitol, but not galactose or xylose, as sole sources of carbon. Single amino acids such as L-alanine, L-cysteine and L-threonine served as carbon and nitrogen sources for growth of strain RG3T. Strain RG3T produced cellulose in both nitrogen-free broth and enriched medium. The ubiquinone present was Q-10 and the DNA base composition was 55.8 mol% G+C. It exhibited low values of 5.2-27.77 % DNA-DNA relatedness to the type strains of related gluconacetobacters, which placed it within a separate taxon, for which the name Gluconacetobacter kombuchae sp. nov. is proposed, with the type strain RG3T (=LMG 23726T=MTCC 6913T).

  1. History on the biological nitrogen fixation research in graminaceous plants: special emphasis on the Brazilian experience.

    Science.gov (United States)

    Baldani, José I; Baldani, Vera L D

    2005-09-01

    This review covers the history on Biological Nitrogen Fixation (BNF) in Graminaceous plants grown in Brazil, and describes research progress made over the last 40 years, most of which was coordinated by Johanna Döbereiner. One notable accomplishment during this period was the discovery of several nitrogen-fixing bacteria such as the rhizospheric (Beijerinckia fluminensis and Azotobacter paspali), associative (Azospirillum lipoferum, A. brasilense, A. amazonense) and the endophytic (Herbaspirillum seropedicae, H. rubrisubalbicans, Gluconacetobacter diazotrophicus, Burkholderia brasilensis and B. tropica). The role of these diazotrophs in association with grasses, mainly with cereal plants, has been studied and a lot of progress has been achieved in the ecological, physiological, biochemical, and genetic aspects. The mechanisms of colonization and infection of the plant tissues are better understood, and the BNF contribution to the soil/plant system has been determined. Inoculation studies with diazotrophs showed that endophytic bacteria have a much higher BNF contribution potential than associative diazotrophs. In addition, it was found that the plant genotype influences the plant/bacteria association. Recent data suggest that more studies should be conducted on the endophytic association to strengthen the BNF potential. The ongoing genome sequencing programs: RIOGENE (Gluconacetobacter diazotrophicus) and GENOPAR (Herbaspirillum seropedicae) reflect the commitment to the BNF study in Brazil and should allow the country to continue in the forefront of research related to the BNF process in Graminaceous plants.

  2. Ocorrência de micorrizas arbusculares e da bactéria diazotrófica Acetobacter diazotrophicus em cana-de-açúcar Occurrence of arbuscular mycorrhizae and bacterium Acetobacter diazotrophicus in sugar cane

    Directory of Open Access Journals (Sweden)

    Veronica Massena Reis

    1999-10-01

    Full Text Available Foi avaliada a ocorrência e a distribuição de espécies de fungos micorrízicos arbusculares (FMAs e Acetobacter diazotrophicus em plantios de cana-de-açúcar em diferentes tipos de manejo nos Estados do Rio de Janeiro e Pernambuco. Foram feitas 35 coletas de amostras de solo da rizosfera e de raízes de 14 variedades de cana-de-açúcar para extração de esporos e isolamento da bactéria. O número de esporos variou de 18 a 2.070/100 mL de solo, e os maiores número e diversidade de espécies foram verificados nos canaviais de Campos, RJ, especialmente naqueles que não adotam a queima do palhiço. As espécies predominantes nas três localidades amostradas foram: Acaulospora sp., Scutellospora heterogama, Glomus etunicatum, Glomus occultum e Gigaspora margarita. A. diazotrophicus estava presente nas amostras de raízes colhidas em canaviais de Campos, com exceção de uma coleta de cana-de-açúcar plantada num solo usado como bacia de sedimentação de vinhaça. Não foi possível isolar essa bactéria a partir de esporos desinfestados dos FMAs nativos, apenas dos esporos lavados com água estéril.The occurrence and distribution of species of arbuscular mycorrhizae fungi and Acetobacter diazotrophicus in sugar cane (Saccharum officinarum grown in different regimes of crop management in the States of Rio de Janeiro and Pernambuco, Brazil, were studied. Thirty five samples of the rhizosphere soil and roots were collected from 14 varieties of sugar cane for the extraction of spores and isolation of the bacterium. The number of spores varied from 18 to 2,070 per 100 mL of soil, and the greatest diversity of fungal species was found in the sugarcane fields of Campos (Rio de Janeiro State, especially in those where the sugarcane trash was not burned at harvest. The predominant species found in the three localities sampled were: Scutellospora heterogama, Glomus etunicatum, Glomus occultum, Acaulospora sp. and Gigaspora margarita. A

  3. Agrobacterium tumefaciens is a diazotrophic bacterium

    International Nuclear Information System (INIS)

    Kanvinde, L.; Sastry, G.R.K.

    1990-01-01

    This is the first report that Agrobacterium tumefaciens can fix nitrogen in a free-living condition as shown by its abilities to grown on nitrogen-free medium, reduce acetylene to ethylene, and incorporate 15 N supplied as 15 N 2 . As with most other well-characterized diazotrophic bacteria, the presence of NH 4 + in the medium and aerobic conditions repress nitrogen fixation by A. tumefaciens. The system requires molybdenum. No evidence for nodulation was found with pea, peanut, or soybean plants. Further understanding of the nitrogen-fixing ability of this bacterium, which has always been considered a pathogen, should cast new light on the evolution of a pathogenic versus symbiotic relationship

  4. Agrobacterium tumefaciens is a diazotrophic bacterium

    Energy Technology Data Exchange (ETDEWEB)

    Kanvinde, L.; Sastry, G.R.K. (Univ. of Leeds (England))

    1990-07-01

    This is the first report that Agrobacterium tumefaciens can fix nitrogen in a free-living condition as shown by its abilities to grown on nitrogen-free medium, reduce acetylene to ethylene, and incorporate {sup 15}N supplied as {sup 15}N{sub 2}. As with most other well-characterized diazotrophic bacteria, the presence of NH{sub 4}{sup +} in the medium and aerobic conditions repress nitrogen fixation by A. tumefaciens. The system requires molybdenum. No evidence for nodulation was found with pea, peanut, or soybean plants. Further understanding of the nitrogen-fixing ability of this bacterium, which has always been considered a pathogen, should cast new light on the evolution of a pathogenic versus symbiotic relationship.

  5. Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments

    Science.gov (United States)

    Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Molybdenum-independent nitrogenases from aquatic environments...

  6. NCBI nr-aa BLAST: CBRC-PVAM-01-1081 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PVAM-01-1081 ref|YP_001603021.1| putative dehydrogenase [Gluconacetobacter diazo...trophicus PAl 5] emb|CAP56727.1| putative dehydrogenase [Gluconacetobacter diazotrophicus PAl 5] YP_001603021.1 0.42 27% ...

  7. NCBI nr-aa BLAST: CBRC-MLUC-01-1006 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MLUC-01-1006 ref|YP_001603545.1| hypothetical protein GDI_3314 [Gluconacetobacter diazo...trophicus PAl 5] emb|CAP57257.1| putative membrane protein [Gluconacetobacter diazotrophicus PAl 5] YP_001603545.1 0.065 35% ...

  8. Culture medium pH influence on Gluconacetobacter physiology: Cellulose production rate and yield enhancement in presence of multiple carbon sources.

    Science.gov (United States)

    Yassine, Fatima; Bassil, Nathalie; Flouty, Roula; Chokr, Ali; Samrani, Antoine El; Boiteux, Gisèle; Tahchi, Mario El

    2016-08-01

    Gluconacetobacter genera are valued for bacterial cellulose (BC) and acetic acid production. BC is produced at optimal yields in classical microbiological media that are expensive for a large scale of production. In addition, BC usage for industrial purposes is limited due to low conversion rate into cellulose and to long incubation duration. In this paper, Gluconacetobacter isolated from apple vinegar was kinetically studied to evaluate cellulose production in presence of different carbon sources. Acetic and citric acid effect on Gluconacetobacter metabolism is clarified. It was shown that Gluconacetobacter uses glucose as a primary carbon source for cells growth and products formation. Acetic acid employment as a co-carbon source in Hestrin Schramm medium showed an increase of 17% in BC yield with a moderate decrease in the crystallite size of the resulting polymer. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Technetium and diazotrophic organisms: toxicity, localization, transfer factors

    International Nuclear Information System (INIS)

    Vandecasteele, C.M.; Delmotte, A.; Roucoux, P.; Hove, C. van

    1982-01-01

    Three diazotrophic organisms, together with one leguminous organism in symbiosis with one of them, were cultivated in the presence of various quantities of technetium, of which the localization, transfer factors and toxicity were studied in relation to the age of the organisms and their type of metabolism. The paper discusses the biochemical aspects of the results. (author)

  10. Isolation of heterotrophic diazotrophic bacteria from estuarine surface waters.

    Science.gov (United States)

    Farnelid, Hanna; Harder, Jens; Bentzon-Tilia, Mikkel; Riemann, Lasse

    2014-10-01

    The wide distribution of diverse nitrogenase (nifH) genes affiliated with those of heterotrophic bacteria in marine and estuarine waters indicates ubiquity and an ecologically relevant role for heterotrophic N2 -fixers (diazotrophs) in aquatic nitrogen (N) cycling. However, the lack of cultivated representatives currently precludes an evaluation of their N2 -fixing capacity. In this study, microoxic or anoxic N-free media were inoculated with estuarine Baltic Sea surface water to select for N2 -fixers. After visible growth and isolation of single colonies on oxic plates or in anoxic agar tubes, nifH gene amplicons were obtained from 64 strains and nitrogenase activity, applying the acetylene reduction assay, was confirmed for 40 strains. Two strains, one Gammaproteobacterium affiliated with Pseudomonas and one Alphaproteobacterium affiliated with Rhodopseudomonas were shown to represent established members of the indigenous diazotrophic community in the Baltic Sea, with abundances of up to 7.9 × 10(4) and 4.7 × 10(4)  nifH copies l(-1) respectively. This study reports media for successful isolation of heterotrophic diazotrophs. The applied methodology and the obtained strains will facilitate future identification of factors controlling heterotrophic diazotrophic activity in aquatic environments, which is a prerequisite for understanding and evaluating their ecology and contribution to N cycling at local and regional scales. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  11. Thin stillage supplementation greatly enhances bacterial cellulose production by Gluconacetobacter xylinus.

    Science.gov (United States)

    Wu, Jyh-Ming; Liu, Ren-Han

    2012-09-01

    Thin stillage (TS), a wastewater from rice wine distillery can well sustain the growth of Gluconacetobacter xylinus for production of bacterial cellulose (BC). When used as a supplement to the traditional BC production medium (Hestrin and Schramm medium), the enhancement of BC production increased with the amount of TS supplemented in a static culture of G. xylinus. When TS was employed to replace distilled water for preparing HS medium (100%TS-HS medium), the BC production in this 100%TS-HS medium was enhanced 2.5-fold to a concentration of 10.38 g/l with sugar to BC conversion yield of 57% after 7 days cultivation. The cost-free TS as a supplement in BC production medium not only can greatly enhance the BC production, but also can effectively dispose the nuisance wastewater of rice wine distillery. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Biochemical and cellular properties of Gluconacetobacter xylinus cultures exposed to different modes of rotating magnetic field

    Directory of Open Access Journals (Sweden)

    Fijałkowski Karol

    2017-06-01

    Full Text Available The aim of the present study was to evaluate the impact of a rotating magnetic field (RMF on cellular and biochemical properties of Gluconacetobacter xylinus during the process of cellulose synthesis by these bacteria. The application of the RMF during bacterial cellulose (BC production intensified the biochemical processes in G. xylinus as compared to the RMF-unexposed cultures. Moreover, the RMF had a positive impact on the growth of cellulose-producing bacteria. Furthermore, the application of RMF did not increase the number of mutants unable to produce cellulose. In terms of BC production efficacy, the most favorable properties were found in the setting where RMF generator was switched off for the first 72 h of cultivation and switched on for the further 72 h. The results obtained can be used in subsequent studies concerning the optimization of BC production using different types of magnetic fields including RMF, especially.

  13. Performance of pineapple slips inoculated with diazotrophic phosphate-solubilizing bacteria and rock phosphate

    OpenAIRE

    Lílian Estrela Borges Baldotto; Marihus Altoé Baldotto; Fábio Lopes Olivares; Adriane Nunes de Souza

    2014-01-01

    Besides fixing N2, some diazotrophic bacteria or diazotrophs, also synthesize organic acids and are able to solubilize rock phosphates, increasing the availability of P for plants. The application of these bacteria to pineapple leaf axils in combination with rock phosphate could increase N and P availability for the crop, due to the bacterial activity of biological nitrogen fixation and phosphate solubilization. The objectives of this study were: (i) to select and characterize diazotrophs abl...

  14. Transfer of diazotroph-derived nitrogen towards non-diazotrophic planktonic communities: a comparative study between Trichodesmium erythraeum, Crocosphaera watsonii and Cyanothece sp.

    Science.gov (United States)

    Berthelot, Hugo; Bonnet, Sophie; Grosso, Olivier; Cornet, Véronique; Barani, Aude

    2016-07-01

    Biological dinitrogen (N2) fixation is the major source of new nitrogen (N) for the open ocean, and thus promotes marine productivity, in particular in the vast N-depleted regions of the surface ocean. Yet, the fate of the diazotroph-derived N (DDN) in marine ecosystems is poorly understood, and its transfer to auto- and heterotrophic surrounding plankton communities is rarely measured due to technical limitations. Moreover, the different diazotrophs involved in N2 fixation (Trichodesmium spp. vs. UCYN) exhibit distinct patterns of N2 fixation and inhabit different ecological niches, thus having potentially different fates in the marine food webs that remain to be explored. Here we used nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling and flow cytometry cell sorting to examine the DDN transfer to specific groups of natural phytoplankton and bacteria during artificially induced diazotroph blooms in New Caledonia (southwestern Pacific). The fate of the DDN was compared according to the three diazotrophs: the filamentous and colony-forming Trichodesmium erythraeum (IMS101), and the unicellular strains Crocosphaera watsonii WH8501 and Cyanothece ATCC51142. After 48 h, 7-17 % of the N2 fixed during the experiment was transferred to the dissolved pool and 6-12 % was transferred to non-diazotrophic plankton. The transfer was twice as high in the T. erythraeum bloom than in the C. watsonii and Cyanothece blooms, which shows that filamentous diazotrophs blooms are more efficient at promoting non-diazotrophic production in N-depleted areas. The amount of DDN released in the dissolved pool did not appear to be a good indicator of the DDN transfer efficiency towards the non-diazotrophic plankton. In contrast, the 15N-enrichment of the extracellular ammonium (NH4+) pool was a good indicator of the DDN transfer efficiency: it was significantly higher in the T. erythraeum than in unicellular diazotroph blooms, leading to a DDN

  15. Production of Bacterial Cellulose by Gluconacetobacter hansenii Using Corn Steep Liquor As Nutrient Sources

    Directory of Open Access Journals (Sweden)

    Andrea F. S. Costa

    2017-10-01

    Full Text Available Cellulose is mainly produced by plants, although many bacteria, especially those belonging to the genus Gluconacetobacter, produce a very peculiar form of cellulose with mechanical and structural properties that can be exploited in numerous applications. However, the production cost of bacterial cellulose (BC is very high to the use of expensive culture media, poor yields, downstream processing, and operating costs. Thus, the purpose of this work was to evaluate the use of industrial residues as nutrients for the production of BC by Gluconacetobacter hansenii UCP1619. BC pellicles were synthesized using the Hestrin–Schramm (HS medium and alternative media formulated with different carbon (sugarcane molasses and acetylated glucose and nitrogen sources [yeast extract, peptone, and corn steep liquor (CSL]. A jeans laundry was also tested. None of the tested sources (beside CSL worked as carbon and nutrient substitute. The alternative medium formulated with 1.5% glucose and 2.5% CSL led to the highest yield in terms of dry and hydrated mass. The BC mass produced in the alternative culture medium corresponded to 73% of that achieved with the HS culture medium. The BC pellicles demonstrated a high concentration of microfibrils and nanofibrils forming a homogenous, compact, and three-dimensional structure. The biopolymer produced in the alternative medium had greater thermal stability, as degradation began at 240°C, while degradation of the biopolymer produced in the HS medium began at 195°C. Both biopolymers exhibited high crystallinity. The mechanical tensile test revealed the maximum breaking strength and the elongation of the break of hydrated and dry pellicles. The dry BC film supported up to 48 MPa of the breaking strength and exhibited greater than 96.98% stiffness in comparison with the hydrated film. The dry film supported up to 48 MPa of the breaking strength and exhibited greater than 96.98% stiffness in comparison with the hydrated film

  16. Anthropogenic impact on diazotrophic diversity in the mangrove rhizosphere revealed by nifH pyrosequencing.

    Science.gov (United States)

    Jing, Hongmei; Xia, Xiaomin; Liu, Hongbin; Zhou, Zhi; Wu, Chen; Nagarajan, Sanjay

    2015-01-01

    Diazotrophs in the mangrove rhizosphere play a major role in providing new nitrogen to the mangrove ecosystem and their composition and activity are strongly influenced by anthropogenic activity and ecological conditions. In this study, the diversity of the diazotroph communities in the rhizosphere sediment of five tropical mangrove sites with different levels of pollution along the north and south coastline of Singapore were studied by pyrosequencing of the nifH gene. Bioinformatics analysis revealed that in all the studied locations, the diazotroph communities comprised mainly of members of the diazotrophic cluster I and cluster III. The detected cluster III diazotrophs, which were composed entirely of sulfate-reducing bacteria, were more abundant in the less polluted locations. The metabolic capacities of these diazotrophs indicate the potential for bioremediation and resiliency of the ecosystem to anthropogenic impact. In heavily polluted locations, the diazotrophic community structures were markedly different and the diversity of species was significantly reduced when compared with those in a pristine location. This, together with the increased abundance of Marinobacterium, which is a bioindicator of pollution, suggests that anthropogenic activity has a negative impact on the genetic diversity of diazotrophs in the mangrove rhizosphere.

  17. Production of green biocellulose nanofibers by Gluconacetobacter xylinus through utilizing the renewable resources of agriculture residues.

    Science.gov (United States)

    Al-Abdallah, Wahib; Dahman, Yaser

    2013-11-01

    The present study demonstrates the ability to produce green biocellulose nanofibers using the renewable resources of agriculture residues. Locally grown wheat straws (WS) were hydrolyzed under different conditions. Their hydrolysates were utilized to produce the nanofibers in separate hydrolysis fermentation process by Gluconacetobacter xylinus strain bacterium. Highest biocellulose production of ~10.6 g/L was achieved with samples that were enzymatically hydrolyzed. Moreover, acidic hydrolyzed WS produced up to 9.7 g/L, with total sugar concentrations in culture media of 43 g/L. Generally, enzymatic hydrolysis of WS resulted in more total sugar concentration than the acidic hydrolysis (i.e., 52.12 g/L), while water hydrolysis produced the least. This can be related to utilizing Xylanase in addition to Cellulase and Beta-glucosidase that helps to hydrolyse WS dry basis of cellulose and hemicelluloses. Sugar mixtures produced under all hydrolysis conditions were mainly composed of glucose and xylose with average percentages of 56 and 28 %, respectively. Acidic hydrolysis at higher acid concentration, as well as soaking WS in the acidic solution for longer time, improved the total sugar concentration in the culture media by 18 %. Conducting thermal treatment at more intense conditions of higher temperature or heating time improved the total sugar produced with acidic hydrolysis. These conditions, however, resulted in further production of furfural, which considerably affected bacterial cells proliferation. This resulted in lowest sugar consumption in the range of 62-64 % that affected final BC production.

  18. Bacterial cellulose production by Gluconacetobacter xylinus by employing alternative culture media.

    Science.gov (United States)

    Jozala, Angela Faustino; Pértile, Renata Aparecida Nedel; dos Santos, Carolina Alves; de Carvalho Santos-Ebinuma, Valéria; Seckler, Marcelo Martins; Gama, Francisco Miguel; Pessoa, Adalberto

    2015-02-01

    Bacterial cellulose (BC) is used in different fields as a biological material due to its unique properties. Despite there being many BC applications, there still remain many problems associated with bioprocess technology, such as increasing productivity and decreasing production cost. New technologies that use waste from the food industry as raw materials for culture media promote economic advantages because they reduce environmental pollution and stimulate new research for science sustainability. For this reason, BC production requires optimized conditions to increase its application. The main objective of this study was to evaluate BC production by Gluconacetobacter xylinus using industry waste, namely, rotten fruits and milk whey, as culture media. Furthermore, the structure of BC produced at different conditions was also determined. The culture media employed in this study were composed of rotten fruit collected from the disposal of free markets, milk whey from a local industrial disposal, and their combination, and Hestrin and Schramm media was used as standard culture media. Although all culture media studied produced BC, the highest BC yield-60 mg/mL-was achieved with the rotten fruit culture. Thus, the results showed that rotten fruit can be used for BC production. This culture media can be considered as a profitable alternative to generate high-value products. In addition, it combines environmental concern with sustainable processes that can promote also the reduction of production cost.

  19. Morphological structure of Gluconacetobacter xylinus cellulose and cellulose-based organic-inorganic composite materials

    Science.gov (United States)

    Smyslov, R. Yu; Ezdakova, K. V.; Kopitsa, G. P.; Khripunov, A. K.; Bugrov, A. N.; Tkachenko, A. A.; Angelov, B.; Pipich, V.; Szekely, N. K.; Baranchikov, A. E.; Latysheva, E.; Chetverikov, Yu O.; Haramus, V.

    2017-05-01

    Scanning electron microscopy, ultra-small-angle neutron scattering (USANS), small-angle neutron and X-ray scattering (SANS and SAXS), as well as low-temperature nitrogen adsorption, were used in the studies of micro- and mesostructure of polymer matrix prepared from air-dry preliminarily disintegrated cellulose nano-gel film (synthesized by Gluconacetobacter xylinus) and the composites based on this bacterial cellulose. The composites included ZrO2 nanoparticles, Tb3+ in the form of low molecular weight salt and of metal-polymer complex with poly(vinylpyrrolydone)-poly(methacryloyl-o-aminobenzoic acid) copolymer. The combined analysis of the data obtained allowed revealing three levels of fractal organization in mesostructure of G. xylinus cellulose and its composites. It was shown that both the composition and an aggregation state of dopants have a significant impact on the structural characteristics of the organic-inorganic composites. The composites containing Tb3+ ions demonstrate efficient luminescence; its intensity is an order of magnitude higher in the case of the composites with the metal-polymer complex. It was found that there is the optimal content of ZrO2 nanoparticles in composites resulting in increased Tb3+ luminescence.

  20. Isolation and Characterization of Diazotrophic Rhizobacteria of Oil Palm Roots

    Directory of Open Access Journals (Sweden)

    Azlin, C. O.

    2005-01-01

    Full Text Available Beneficial rhizobacteria were isolated from two different compartments of oil palm roots; the rhizosphere or rhizoplane and the inner root tissues. The root samples were collected from oil palm plantation at Felda Lepar 9, Temerloh Pahang (Block 17, Square 6 (soil pH 4.30; 10:25 0.01M CaCl2. Identification of the isolates was conducted by classical biochemical and physiological tests. Acetylene Reduction Assay (ARA test was also conducted to quantify the ability of the isolates to fix atmospheric N2. Twenty-nine strains of rhizobacteria were isolated from root samples and were maintained aerobically on N-free solid media. Seven of the isolates were identified as Gram negative while the rest were Gram positive. The isolates were successfully identified as Paenibacillus durus (formerly P. azotofixans, Paenibacillus polymyxa, Azospirillum lipoferum, Herbaspirillum seropedicae and Acetobacter diazotrophicus. The N2 fixation capacities of the isolates ranged from 7.0 x 10-12 to 1.0 x 10-8 mol C2H4/cfu/hour.

  1. Identification and Characterization of Non-Cellulose-Producing Mutants of Gluconacetobacter hansenii Generated by Tn5 Transposon Mutagenesis

    Science.gov (United States)

    Deng, Ying; Nagachar, Nivedita; Xiao, Chaowen; Tien, Ming

    2013-01-01

    The acs operon of Gluconacetobacter is thought to encode AcsA, AcsB, AcsC, and AcsD proteins that constitute the cellulose synthase complex, required for the synthesis and secretion of crystalline cellulose microfibrils. A few other genes have been shown to be involved in this process, but their precise role is unclear. We report here the use of Tn5 transposon insertion mutagenesis to identify and characterize six non-cellulose-producing (Cel−) mutants of Gluconacetobacter hansenii ATCC 23769. The genes disrupted were acsA, acsC, ccpAx (encoding cellulose-complementing protein [the subscript “Ax” indicates genes from organisms formerly classified as Acetobacter xylinum]), dgc1 (encoding guanylate dicyclase), and crp-fnr (encoding a cyclic AMP receptor protein/fumarate nitrate reductase transcriptional regulator). Protein blot analysis revealed that (i) AcsB and AcsC were absent in the acsA mutant, (ii) the levels of AcsB and AcsC were significantly reduced in the ccpAx mutant, and (iii) the level of AcsD was not affected in any of the Cel− mutants. Promoter analysis showed that the acs operon does not include acsD, unlike the organization of the acs operon of several strains of closely related Gluconacetobacter xylinus. Complementation experiments confirmed that the gene disrupted in each Cel− mutant was responsible for the phenotype. Quantitative real-time PCR and protein blotting results suggest that the transcription of bglAx (encoding β-glucosidase and located immediately downstream from acsD) was strongly dependent on Crp/Fnr. A bglAx knockout mutant, generated via homologous recombination, produced only ∼16% of the wild-type cellulose level. Since the crp-fnr mutant did not produce any cellulose, Crp/Fnr may regulate the expression of other gene(s) involved in cellulose biosynthesis. PMID:24013627

  2. Diversity and Structure of Diazotrophic Communities in Mangrove Rhizosphere, Revealed by High-Throughput Sequencing.

    Science.gov (United States)

    Zhang, Yanying; Yang, Qingsong; Ling, Juan; Van Nostrand, Joy D; Shi, Zhou; Zhou, Jizhong; Dong, Junde

    2017-01-01

    Diazotrophic communities make an essential contribution to the productivity through providing new nitrogen. However, knowledge of the roles that both mangrove tree species and geochemical parameters play in shaping mangove rhizosphere diazotrophic communities is still elusive. Here, a comprehensive examination of the diversity and structure of microbial communities in the rhizospheres of three mangrove species, Rhizophora apiculata , Avicennia marina , and Ceriops tagal , was undertaken using high - throughput sequencing of the 16S rRNA and nifH genes. Our results revealed a great diversity of both the total microbial composition and the diazotrophic composition specifically in the mangrove rhizosphere. Deltaproteobacteria and Gammaproteobacteria were both ubiquitous and dominant, comprising an average of 45.87 and 86.66% of total microbial and diazotrophic communities, respectively. Sulfate-reducing bacteria belonging to the Desulfobacteraceae and Desulfovibrionaceae were the dominant diazotrophs. Community statistical analyses suggested that both mangrove tree species and additional environmental variables played important roles in shaping total microbial and potential diazotroph communities in mangrove rhizospheres. In contrast to the total microbial community investigated by analysis of 16S rRNA gene sequences, most of the dominant diazotrophic groups identified by nifH gene sequences were significantly different among mangrove species. The dominant diazotrophs of the family Desulfobacteraceae were positively correlated with total phosphorus, but negatively correlated with the nitrogen to phosphorus ratio. The Pseudomonadaceae were positively correlated with the concentration of available potassium, suggesting that diazotrophs potentially play an important role in biogeochemical cycles, such as those of nitrogen, phosphorus, sulfur, and potassium, in the mangrove ecosystem.

  3. Diversity and Structure of Diazotrophic Communities in Mangrove Rhizosphere, Revealed by High-Throughput Sequencing

    Directory of Open Access Journals (Sweden)

    Yanying Zhang

    2017-10-01

    Full Text Available Diazotrophic communities make an essential contribution to the productivity through providing new nitrogen. However, knowledge of the roles that both mangrove tree species and geochemical parameters play in shaping mangove rhizosphere diazotrophic communities is still elusive. Here, a comprehensive examination of the diversity and structure of microbial communities in the rhizospheres of three mangrove species, Rhizophora apiculata, Avicennia marina, and Ceriops tagal, was undertaken using high-throughput sequencing of the 16S rRNA and nifH genes. Our results revealed a great diversity of both the total microbial composition and the diazotrophic composition specifically in the mangrove rhizosphere. Deltaproteobacteria and Gammaproteobacteria were both ubiquitous and dominant, comprising an average of 45.87 and 86.66% of total microbial and diazotrophic communities, respectively. Sulfate-reducing bacteria belonging to the Desulfobacteraceae and Desulfovibrionaceae were the dominant diazotrophs. Community statistical analyses suggested that both mangrove tree species and additional environmental variables played important roles in shaping total microbial and potential diazotroph communities in mangrove rhizospheres. In contrast to the total microbial community investigated by analysis of 16S rRNA gene sequences, most of the dominant diazotrophic groups identified by nifH gene sequences were significantly different among mangrove species. The dominant diazotrophs of the family Desulfobacteraceae were positively correlated with total phosphorus, but negatively correlated with the nitrogen to phosphorus ratio. The Pseudomonadaceae were positively correlated with the concentration of available potassium, suggesting that diazotrophs potentially play an important role in biogeochemical cycles, such as those of nitrogen, phosphorus, sulfur, and potassium, in the mangrove ecosystem.

  4. Understanding predicted shifts in diazotroph biogeography using resource competition theory

    Directory of Open Access Journals (Sweden)

    S. Dutkiewicz

    2014-10-01

    Full Text Available We examine the sensitivity of the biogeography of nitrogen fixers to a warming climate and increased aeolian iron deposition in the context of a global earth system model. We employ concepts from the resource-ratio theory to provide a simplifying and transparent interpretation of the results. First we demonstrate that a set of clearly defined, easily diagnosed provinces are consistent with the theory. Using this framework we show that the regions most vulnerable to province shifts and changes in diazotroph biogeography are the equatorial and South Pacific, and central Atlantic. Warmer and dustier climates favor diazotrophs due to an increase in the ratio of supply rate of iron to fixed nitrogen. We suggest that the emergent provinces could be a standard diagnostic for global change models, allowing for rapid and transparent interpretation and comparison of model predictions and the underlying mechanisms. The analysis suggests that monitoring of real world province boundaries, indicated by transitions in surface nutrient concentrations, would provide a clear and easily interpreted indicator of ongoing global change.

  5. Structure determination and functional analysis of a chromate reductase from Gluconacetobacter hansenii.

    Directory of Open Access Journals (Sweden)

    Hongjun Jin

    Full Text Available Environmental protection through biological mechanisms that aid in the reductive immobilization of toxic metals (e.g., chromate and uranyl has been identified to involve specific NADH-dependent flavoproteins that promote cell viability. To understand the enzyme mechanisms responsible for metal reduction, the enzyme kinetics of a putative chromate reductase from Gluconacetobacter hansenii (Gh-ChrR was measured and the crystal structure of the protein determined at 2.25 Å resolution. Gh-ChrR catalyzes the NADH-dependent reduction of chromate, ferricyanide, and uranyl anions under aerobic conditions. Kinetic measurements indicate that NADH acts as a substrate inhibitor; catalysis requires chromate binding prior to NADH association. The crystal structure of Gh-ChrR shows the protein is a homotetramer with one bound flavin mononucleotide (FMN per subunit. A bound anion is visualized proximal to the FMN at the interface between adjacent subunits within a cationic pocket, which is positioned at an optimal distance for hydride transfer. Site-directed substitutions of residues proposed to involve in both NADH and metal anion binding (N85A or R101A result in 90-95% reductions in enzyme efficiencies for NADH-dependent chromate reduction. In comparison site-directed substitution of a residue (S118A participating in the coordination of FMN in the active site results in only modest (50% reductions in catalytic efficiencies, consistent with the presence of a multitude of side chains that position the FMN in the active site. The proposed proximity relationships between metal anion binding site and enzyme cofactors is discussed in terms of rational design principles for the use of enzymes in chromate and uranyl bioremediation.

  6. Specificity of salt marsh diazotrophs for vegetation zones and plant hosts

    Directory of Open Access Journals (Sweden)

    Debra Aline Davis

    2012-03-01

    Full Text Available Salt marshes located on the east coast of temperate North America are highly productive, typically nitrogen-limited, and support diverse assemblages of nitrogen fixing (diazotrophic bacteria. The distributions of these diazotrophs are strongly influenced by plant host and abiotic environmental parameters. Crab Haul Creek Basin, North Inlet, SC, USA is a tidally dominated marsh that displays discrete plant zones distributed along an elevation gradient from the tidal creek bank to the terrestrial forest. These zones are defined by gradients of abiotic environmental variables, particularly salinity and sulfide. DGGE fingerprinting and phylogenetic analyses of recovered sequences demonstrated that the distributions of some diazotrophs indicate plant host specificity and that diazotroph assemblages across the marsh gradient are heavily influenced by edaphic conditions. Broadly distributed diazotrophs capable of maintaining populations in all environmental conditions across the gradient are also present in these assemblages. Parsimony test results confirm that diazotroph assemblages in different plant zones are significantly (p<0.01 different across the marsh landscape. Results also indicated that diazotroph assemblages associated with different plant hosts growing in the same area of the marsh were structurally similar confirming the influence of edaphic parameters on these assemblages. Principal Component Analysis of DGGE gel banding patterns confirmed these results. This article reviews and analyzes data from North Inlet Estuary, addressing diazotroph assemblage structure and the influence of plant host and environmental conditions. New data demonstrate the heterogeneity of salt marsh rhizosphere microenvironments, and corroborate previous findings from different plant hosts growing at several locations within this estuary. These data support the hypothesis that the biogeography of microorganisms is non-random and is partially driven by

  7. Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

    OpenAIRE

    Silva,Aurilena de Aviz; Carvalho,Almy Junior Cordeiro de; Freitas,Flávia Paiva de; Pessanha,Patrícia Gomes de Oliveira; Santos,Paulo Cesar dos; Silva,Mírian Peixoto Soares da; Vasconcelos,Tábatha de Souza; Olivares,Fábio Lopes

    2016-01-01

    ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholder...

  8. Evidence for the functional significance of diazotroph community structure in soil.

    Science.gov (United States)

    Hsu, Shi-Fang; Buckley, Daniel H

    2009-01-01

    Microbial ecologists continue to seek a greater understanding of the factors that govern the ecological significance of microbial community structure. Changes in community structure have been shown to have functional significance for processes that are mediated by a narrow spectrum of organisms, such as nitrification and denitrification, but in some cases, functional redundancy in the community seems to buffer microbial ecosystem processes. The functional significance of microbial community structure is frequently obscured by environmental variation and is hard to detect in short-term experiments. We examine the functional significance of free-living diazotrophs in a replicated long-term tillage experiment in which extraneous variation is minimized and N-fixation rates can be related to soil characteristics and diazotroph community structure. Soil characteristics were found to be primarily impacted by tillage management, whereas N-fixation rates and diazotroph community structure were impacted by both biomass management practices and interactions between tillage and biomass management. The data suggest that the variation in diazotroph community structure has a greater impact on N-fixation rates than do soil characteristics at the site. N-fixation rates displayed a saturating response to increases in diazotroph community diversity. These results show that the changes in the community structure of free-living diazotrophs in soils can have ecological significance and suggest that this response is related to a change in community diversity.

  9. Diversity and Activity of Diazotrophs in Great Barrier Reef Surface Waters.

    Science.gov (United States)

    Messer, Lauren F; Brown, Mark V; Furnas, Miles J; Carney, Richard L; McKinnon, A D; Seymour, Justin R

    2017-01-01

    Discrepancies between bioavailable nitrogen (N) concentrations and phytoplankton growth rates in the oligotrophic waters of the Great Barrier Reef (GBR) suggest that undetermined N sources must play a significant role in supporting primary productivity. One such source could be biological dinitrogen (N 2 ) fixation through the activity of "diazotrophic" bacterioplankton. Here, we investigated N 2 fixation and diazotroph community composition over 10° S of latitude within GBR surface waters. Qualitative N 2 fixation rates were found to be variable across the GBR but were relatively high in coastal, inner and outer GBR waters, reaching 68 nmol L -1 d -1 . Diazotroph assemblages, identified by amplicon sequencing of the nifH gene, were dominated by the cyanobacterium Trichodesmium erythraeum , γ-proteobacteria from the Gamma A clade, and δ-proteobacterial phylotypes related to sulfate-reducing genera. However, diazotroph communities exhibited significant spatial heterogeneity, correlated with shifts in dissolved inorganic nutrient concentrations. Specifically, heterotrophic diazotrophs generally increased in relative abundance with increasing concentrations of phosphate and N, while Trichodesmium was proportionally more abundant when concentrations of these nutrients were low. This study provides the first in-depth characterization of diazotroph community composition and N 2 fixation dynamics within the oligotrophic, N-limited surface waters of the GBR. Our observations highlight the need to re-evaluate N cycling dynamics within oligotrophic coral reef systems, to include diverse N 2 fixing assemblages as a potentially significant source of dissolved N within the water column.

  10. Impact of copper on the diazotroph abundance and community composition during swine manure composting.

    Science.gov (United States)

    Yin, Yanan; Gu, Jie; Wang, Xiaojuan; Zhang, Kaiyu; Hu, Ting; Ma, Jiyue; Wang, Qianzhi

    2018-05-01

    Biological nitrogen fixation is a major pathway in ecosystems. This study investigated the effects of adding Cu at different levels (0, 200, and 2000 mg kg -1 ) on the diazotroph community during swine manure composting. Quantitative PCR and high-throughput sequencing were used to analyze the abundances of diazotrophs and the community composition based on the nifH gene. The nifH gene copy number was relatively high in the early stage of composting and Cu had a significant inhibitory effect on the nifH copy number. Furthermore, Cu decreased the diversity of nifH and changed the microbial community structure in the early stage. The nifH genes from members of Firmicutes and Clostridium were most abundant. Co-occurrence ecological network analysis showed that the Cu treatments affected the co-occurrence patterns of diazotroph communities and reduced the associations between different diazotrophs. Interestingly, Cu may weaken symbiotic diazotrophic interactions and enhance the roles of free-living diazotrophs. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Effects of aromatic compounds on the production of bacterial nanocellulose by Gluconacetobacter xylinus.

    Science.gov (United States)

    Zhang, Shuo; Winestrand, Sandra; Guo, Xiang; Chen, Lin; Hong, Feng; Jönsson, Leif J

    2014-04-30

    Bacterial cellulose (BC) is a polymeric nanostructured fibrillar network produced by certain microorganisms, principally Gluconacetobacter xylinus. BC has a great potential of application in many fields. Lignocellulosic biomass has been investigated as a cost-effective feedstock for BC production through pretreatment and hydrolysis. It is well known that detoxification of lignocellulosic hydrolysates may be required to achieve efficient production of BC. Recent results suggest that phenolic compounds contribute to the inhibition of G. xylinus. However, very little is known about the effect on G. xylinus of specific lignocellulose-derived inhibitors. In this study, the inhibitory effects of four phenolic model compounds (coniferyl aldehyde, ferulic acid, vanillin and 4-hydroxybenzoic acid) on the growth of G. xylinus, the pH of the culture medium, and the production of BC were investigated in detail. The stability of the phenolics in the bacterial cultures was investigated and the main bioconversion products were identified and quantified. Coniferyl aldehyde was the most potent inhibitor, followed by vanillin, ferulic acid, and 4-hydroxybenzoic acid. There was no BC produced even with coniferyl aldehyde concentrations as low as 2 mM. Vanillin displayed a negative effect on the bacteria and when the vanillin concentration was raised to 2.5 mM the volumetric yield of BC decreased to ~40% of that obtained in control medium without inhibitors. The phenolic acids, ferulic acid and 4-hydroxybenzoic acid, showed almost no toxic effects when less than 2.5 mM. The bacterial cultures oxidized coniferyl aldehyde to ferulic acid with a yield of up to 81%. Vanillin was reduced to vanillyl alcohol with a yield of up to 80%. This is the first investigation of the effect of specific phenolics on the production of BC by G. xylinus, and is also the first demonstration of the ability of G. xylinus to convert phenolic compounds. This study gives a better understanding of how

  12. Enhancement of crystallinity of cellulose produced by Escherichia coli through heterologous expression of bcsD gene from Gluconacetobacter xylinus.

    Science.gov (United States)

    Sajadi, Elaheh; Babaipour, Valiollah; Deldar, Ali Asghar; Yakhchali, Bagher; Fatemi, Seyed Safa-Ali

    2017-09-01

    To evaluate the crystallinity index of the cellulose produced by Escherichia coli Nissle 1917 after heterologous expression of the cellulose synthase subunit D (bcsD) gene of Gluconacetobacter xylinus BPR2001. The bcsD gene of G. xylinus BPR2001 was expressed in E. coli and its protein product was visualized using SDS-PAGE. FTIR analysis showed that the crystallinity index of the cellulose produced by the recombinants was 0.84, which is 17% more than that of the wild type strain. The increased crystallinity index was also confirmed by X-ray diffraction analysis. The cellulose content was not changed significantly after over-expressing the bcsD. The bcsD gene can improve the crystalline structure of the bacterial cellulose but there is not any significant difference between the amounts of cellulose produced by the recombinant and wild type E. coli Nissle 1917.

  13. Occurrence of Cellulose-Producing Gluconacetobacter spp. in Fruit Samples and Kombucha Tea, and Production of the Biopolymer.

    Science.gov (United States)

    Neera; Ramana, Karna Venkata; Batra, Harsh Vardhan

    2015-06-01

    Cellulose producing bacteria were isolated from fruit samples and kombucha tea (a fermented beverage) using CuSO4 solution in modified Watanabe and Yamanaka medium to inhibit yeasts and molds. Six bacterial strains showing cellulose production were isolated and identified by 16S rRNA gene sequencing as Gluconacetobacter xylinus strain DFBT, Ga. xylinus strain dfr-1, Gluconobacter oxydans strain dfr-2, G. oxydans strain dfr-3, Acetobacter orientalis strain dfr-4, and Gluconacetobacter intermedius strain dfr-5. All the cellulose-producing bacteria were checked for the cellulose yield. A potent cellulose-producing bacterium, i.e., Ga. xylinus strain DFBT based on yield (cellulose yield 5.6 g/L) was selected for further studies. Cellulose was also produced in non- conventional media such as pineapple juice medium and hydrolysed corn starch medium. A very high yield of 9.1 g/L cellulose was obtained in pineapple juice medium. Fourier transform infrared spectrometer (FT-IR) analysis of the bacterial cellulose showed the characteristic peaks. Soft cellulose with a very high water holding capacity was produced using limited aeration. Scanning electron microscopy (SEM) was used to analyze the surface characteristics of normal bacterial cellulose and soft cellulose. The structural analysis of the polymer was performed using (13)C solid-state nuclear magnetic resonance (NMR). More interfibrillar space was observed in the case of soft cellulose as compared to normal cellulose. This soft cellulose can find potential applications in the food industry as it can be swallowed easily without chewing.

  14. History on the biological nitrogen fixation research in graminaceous plants: special emphasis on the Brazilian experience

    Directory of Open Access Journals (Sweden)

    José I. Baldani

    2005-09-01

    Full Text Available This review covers the history on Biological Nitrogen Fixation (BNF in Graminaceous plants grown in Brazil, and describes research progress made over the last 40 years, most of whichwas coordinated by Johanna Döbereiner. One notable accomplishment during this period was the discovery of several nitrogen-fixing bacteria such as the rhizospheric (Beijerinckia fluminensis and Azotobacter paspali, associative (Azospirillum lipoferum, A. brasilense, A. amazonense and the endophytic (Herbaspirillum seropedicae, H. rubrisubalbicans, Gluconacetobacter diazotrophicus, Burkholderia brasilensis and B. tropica. The role of these diazotrophs in association with grasses, mainly with cereal plants, has been studied and a lot of progress has been achieved in the ecological, physiological, biochemical, and genetic aspects. The mechanisms of colonization and infection of the plant tissues are better understood, and the BNF contribution to the soil/plant system has been determined. Inoculation studies with diazotrophs showed that endophytic bacteria have a much higher BNF contribution potential than associative diazotrophs. In addition, it was found that the plant genotype influences the plant/bacteria association. Recent data suggest that more studies should be conducted on the endophytic association to strengthen the BNF potential. The ongoing genome sequencing programs: RIOGENE (Gluconacetobacter diazotrophicus and GENOPAR (Herbaspirillum seropedicae reflect the commitment to the BNF study in Brazil and should allow the country to continue in the forefront of research related to the BNF process in Graminaceous plants.A presente revisão aborda a história da Fixação Biológica de Nitrogênio (FBN em Gramíneas no Brasil, procurando mostrar a evolução da pesquisa na área iniciada a mais de 40 anos sob a liderança da pesquisadora Johanna Döbereiner. Um aspecto marcante deste período foi a descoberta de diversas bactérias fixadoras de nitrogênio atmosf

  15. Diazotrophic diversity in the rhizosphere of two exotic weed plants, Prosopis juliflora and Parthenium hysterophorus.

    Science.gov (United States)

    Cibichakravarthy, B; Preetha, R; Sundaram, S P; Kumar, K; Balachandar, D

    2012-02-01

    This study is aimed at assessing culturable diazotrophic bacterial diversity in the rhizosphere of Prosopis juliflora and Parthenium hysterophorus, which grow profusely in nutritionally-poor soils and environmentally-stress conditions so as to identify some novel strains for bioinoculant technology. Diazotrophic isolates from Prosopis and Parthenium rhizosphere were characterized for nitrogenase activity by Acetylene Reduction Assay (ARA) and 16S rRNA gene sequencing. Further, the culture-independent quantitative PCR (qPCR) was performed to compare the abundance of diazotrophs in rhizosphere with bulk soils. The proportion of diazotrophs in total heterotrophs was higher in rhizosphere than bulk soils and 32 putative diazotrophs from rhizosphere of two plants were identified by nifH gene amplification. The ARA activity of the isolates ranged from 40 to 95 nmol ethylene h(-1) mg protein(-1). The 16S rRNA gene analysis identified the isolates to be members of alpha, beta and gamma Proteobacteria and firmicutes. The qPCR assay also confirmed that abundance of nif gene in rhizosphere of these two plants was 10-fold higher than bulk soil.

  16. Sheep manure vermicompost supplemented with a native diazotrophic bacteria and mycorrhizas for maize cultivation.

    Science.gov (United States)

    Gutiérrez-Miceli, F A; Moguel-Zamudio, B; Abud-Archila, M; Gutiérrez-Oliva, V F; Dendooven, L

    2008-10-01

    An orthogonal experimental design L9 (3(4)) with 10 repetitions was used to investigate the effect of Glomus claroideum (0, 1 or 2g(-1) plant), G. fasciculatum (0, 1 or 2g plant(-1)), native diazotrophic bacteria (0, 10(3) and 10(5) UFC ml(-1)) and sheep manure vermicompost (0%, 5% and 10% v/v) on maize plant growth, N and P in leaves and mycorrhization percent. Vermicompost explained most of the variation found for leaf number, wet weight, stem height, and diameter. Both mycorrhizas increased the plant wet weight but G. fasciculatum the most. Mycorrhization increased the P content, but not the N content. Mycorrhizal colonization increased when diazotrophic bacteria and vermicompost were added. It was found that weight of maize plants cultivated in peat moss amended with vermicompost increased when supplemented with G. fasciculatum and diazotrophic bacteria.

  17. Divergent Responses of the Diazotrophic Microbiome to Elevated CO2 in Two Rice Cultivars

    Directory of Open Access Journals (Sweden)

    Yongjie Yu

    2018-06-01

    Full Text Available The species-specific responses of plant growth to elevated atmospheric CO2 concentration (eCO2 could lead to N limitation and potentially influence the sustainability of ecosystem. Questions remain unanswered with regards to the response of soil N2-fixing community to eCO2 when developing high-yielding agroecosystem to dampen the future rate of increase in CO2 levels and associated climate warming. This study demonstrates the divergent eCO2 influences on the paddy diazotrophic community between weak- and strong-responsive rice cultivars. In response to eCO2, the diazotrophic abundance increased more for the strong-responsive cultivar treatments than for the weak-responsive ones. Only the strong-responsive cultivars decreased the alpha diversity and separated the composition of diazotrophic communities in response to eCO2. The topological indices of the ecological networks further highlighted the different co-occurrence patterns of the diazotrophic microbiome in rice cultivars under eCO2. Strong-responsive cultivars destabilized the diazotrophic community by complicating and centralizing the co-occurrence network as well as by shifting the hub species from Bradyrhizobium to Dechloromonas in response to eCO2. On the contrary, the network pattern of the weak-responsive cultivars was simplified and decentralized in response to eCO2, with the hub species shifting from Halorhodospira under aCO2 to Sideroxydans under eCO2. Collectively, the above information indicates that the strong-responsive cultivars could potentially undermine the belowground ecosystem from the diazotrophs perspective in response to eCO2. This information highlights that more attention should be paid to the stability of the belowground ecosystem when developing agricultural strategies to adapt prospective climatic scenarios by growing high-yielding crop cultivars under eCO2.

  18. Grazing by Zooplankton on Diazotrophs in the Amazon River Plume and Western Tropical North Atlantic

    Science.gov (United States)

    Conroy, B.; Steinberg, D. K.; Song, B.; Foster, R.

    2016-02-01

    Organisms capable of fixing di-nitrogen (N2), known as diazotrophs, are important primary producers and a potentially significant source for new nitrogen entering the planktonic food web. However, limited evidence exists for zooplankton grazing on diazotrophs compared to other primary producers. In the western tropical North Atlantic Ocean (WTNA), the Amazon River plume creates a niche for symbiotic diatom-diazotroph associations (DDAs) which can form large blooms. In adjacent non-plume-influenced waters, the colonial cyanobacterium Trichodesmium is abundant. In order to reveal zooplankton-diazotroph grazing interactions and determine the fate of newly fixed nitrogen, gut contents of zooplankton captured in these two regions were compared based on quantitative PCR (qPCR) assay of nitrogenase genes (nifH), and their microbiomes compared using next generation sequencing (NGS) analysis of 16S rRNA genes. We sampled individual copepods from discrete depth intervals (0-25m and 25-50m) and in two size classes (0.5-1mm and 1-2mm) for analysis. A modified DNA extraction protocol was developed and 54 extracts were used as templates in nifH qPCR assays for the larger size fraction diazotrophs (>10µm): Trichodesmium, and Hemiaulus or Rhizosolenia (diatoms)-Richelia (diazotroph) associations. Copepod gut content nifH copies ranged from 1.6 to 13.6 copies individual-1 for the assay targeting the Hemiaulus-Richelia DDA and from 1.1 to 3.0 copies individual-1 for Trichodesmium. 16S NGS conducted on 35 extracts with an Ion Torrent PGM and mothur revealed that cyanobacteria sequences accounted for up to 20% of sequences per extract. Our results show that both DDAs and Trichodesmium are prey for zooplankton, and that new nitrogen moves through the food web via these grazing interactions. These interactions should be considered in future explorations of the global ocean nitrogen cycle.

  19. Assimilation of diazotrophic nitrogen into pelagic food webs.

    Directory of Open Access Journals (Sweden)

    Ryan J Woodland

    Full Text Available The fate of diazotrophic nitrogen (N(D fixed by planktonic cyanobacteria in pelagic food webs remains unresolved, particularly for toxic cyanophytes that are selectively avoided by most herbivorous zooplankton. Current theory suggests that N(D fixed during cyanobacterial blooms can enter planktonic food webs contemporaneously with peak bloom biomass via direct grazing of zooplankton on cyanobacteria or via the uptake of bioavailable N(D (exuded from viable cyanobacterial cells by palatable phytoplankton or microbial consortia. Alternatively, N(D can enter planktonic food webs post-bloom following the remineralization of bloom detritus. Although the relative contribution of these processes to planktonic nutrient cycles is unknown, we hypothesized that assimilation of bioavailable N(D (e.g., nitrate, ammonium by palatable phytoplankton and subsequent grazing by zooplankton (either during or after the cyanobacterial bloom would be the primary pathway by which N(D was incorporated into the planktonic food web. Instead, in situ stable isotope measurements and grazing experiments clearly documented that the assimilation of N(D by zooplankton outpaced assimilation by palatable phytoplankton during a bloom of toxic Nodularia spumigena Mertens. We identified two distinct temporal phases in the trophic transfer of N(D from N. spumigena to the plankton community. The first phase was a highly dynamic transfer of N(D to zooplankton with rates that covaried with bloom biomass while bypassing other phytoplankton taxa; a trophic transfer that we infer was routed through bloom-associated bacteria. The second phase was a slowly accelerating assimilation of the dissolved-N(D pool by phytoplankton that was decoupled from contemporaneous variability in N. spumigena concentrations. These findings provide empirical evidence that N(D can be assimilated and transferred rapidly throughout natural plankton communities and yield insights into the specific processes

  20. Attachment of associative diazotroph alcaligenes faecalis to rice roots

    International Nuclear Information System (INIS)

    Lin Min; Fang Xuanjun; You Chongbiao

    1993-01-01

    The process of attachment of diazotroph Alcaligenes faecalis to host plant rice was studied by using 15 N-labelled bacteria and Tn5-induced mutants. A three-step attachment mechanism of A. faecalis to rice root surface is proposed on the basis of experimental data. Adsorption is the first step. The number of adsorbed bacteria reaches maximal level after 3 h of inoculation, it consists 3.7% of the total number of bacteria inoculated. Adsorbed bacteria could be removed from rice root surface quantitatively by shaking in water. Therefore, the adsorption forces are weak. Anchoring is the second step. It begins only after 9h of inoculation and reaches a maximal level (21%) after 16 h. Anchored bacteria could not be removed by shaking. Colonization is the third step. After 20 h of inoculation. part of anchored bacteria colonizes on rice root surface tightly, and it can not be removed by vortex. At this time, the pectolytic activity of bacteria appears. Chemotaxis and exopolysaccharide (EPS) play important roles in the attachment of A. faecalis to rice root surface. EPS mutants (Exo - , Exo ++ ) showed less anchoring-capability in comparison with wild type of bacterium, but they remained the adsorption capability. While chemotaxis (Che - ) mutants are defective in adsorption, but not in anchoring. Che - , Exo - mutant lost both adsorption and anchoring capabilities. A. faecalis absorbed on all part of rice root, but the anchoring and colonization of bacteria were occurred mainly on root hairs, particularly on the joint area of main root and lateral root

  1. Assimilation of diazotrophic nitrogen into pelagic food webs.

    Science.gov (United States)

    Woodland, Ryan J; Holland, Daryl P; Beardall, John; Smith, Jonathan; Scicluna, Todd; Cook, Perran L M

    2013-01-01

    The fate of diazotrophic nitrogen (N(D)) fixed by planktonic cyanobacteria in pelagic food webs remains unresolved, particularly for toxic cyanophytes that are selectively avoided by most herbivorous zooplankton. Current theory suggests that N(D) fixed during cyanobacterial blooms can enter planktonic food webs contemporaneously with peak bloom biomass via direct grazing of zooplankton on cyanobacteria or via the uptake of bioavailable N(D) (exuded from viable cyanobacterial cells) by palatable phytoplankton or microbial consortia. Alternatively, N(D) can enter planktonic food webs post-bloom following the remineralization of bloom detritus. Although the relative contribution of these processes to planktonic nutrient cycles is unknown, we hypothesized that assimilation of bioavailable N(D) (e.g., nitrate, ammonium) by palatable phytoplankton and subsequent grazing by zooplankton (either during or after the cyanobacterial bloom) would be the primary pathway by which N(D) was incorporated into the planktonic food web. Instead, in situ stable isotope measurements and grazing experiments clearly documented that the assimilation of N(D) by zooplankton outpaced assimilation by palatable phytoplankton during a bloom of toxic Nodularia spumigena Mertens. We identified two distinct temporal phases in the trophic transfer of N(D) from N. spumigena to the plankton community. The first phase was a highly dynamic transfer of N(D) to zooplankton with rates that covaried with bloom biomass while bypassing other phytoplankton taxa; a trophic transfer that we infer was routed through bloom-associated bacteria. The second phase was a slowly accelerating assimilation of the dissolved-N(D) pool by phytoplankton that was decoupled from contemporaneous variability in N. spumigena concentrations. These findings provide empirical evidence that N(D) can be assimilated and transferred rapidly throughout natural plankton communities and yield insights into the specific processes underlying

  2. Performance of pineapple slips inoculated with diazotrophic phosphate-solubilizing bacteria and rock phosphate

    Directory of Open Access Journals (Sweden)

    Lílian Estrela Borges Baldotto

    2014-06-01

    Full Text Available Besides fixing N2, some diazotrophic bacteria or diazotrophs, also synthesize organic acids and are able to solubilize rock phosphates, increasing the availability of P for plants. The application of these bacteria to pineapple leaf axils in combination with rock phosphate could increase N and P availability for the crop, due to the bacterial activity of biological nitrogen fixation and phosphate solubilization. The objectives of this study were: (i to select and characterize diazotrophs able to solubilize phosphates in vitro and (ii evaluate the initial performance of the pineapple cultivars Imperial and Pérola in response to inoculation with selected bacteria in combination with rock phosphate. The experiments were conducted at Universidade Estadual do Norte Fluminense Darcy Ribeiro, in 2009. In the treatments with bacteria the leaf contents of N, P and K were higher than those of the controls, followed by an increase in plant growth. These results indicate that the combined application of diazotrophic phosphate-solubilizing bacteria Burkholderia together with Araxá rock phosphate can be used to improve the initial performance of pineapple slips.

  3. Dissolved organic nitrogen and carbon release by a marine unicellular diazotrophic cyanobacterium

    NARCIS (Netherlands)

    Benavides, M.; Agawin, N.S.R.; Aristegui, J.; Peene, J.; Stal, L.J.

    2013-01-01

    Dinitrogen (N-2) fixation rates may be underestimated when recently fixed N2 is released as dissolved organic nitrogen (DON). DON release (DONr) is substantial in the filamentous cyanobacterium Trichodesmium but has never been reported in unicellular diazotrophic cyanobacteria. We used axenic

  4. Phylogenetic changes in soil microbial and diazotrophic diversity with application of butachlor.

    Science.gov (United States)

    Yen, Jui-Hung; Wang, Yei-Shung; Hsu, Wey-Shin; Chen, Wen-Ching

    2013-01-01

    We investigated changes in population and taxonomic distribution of cultivable bacteria and diazotrophs with butachlor application in rice paddy soils. Population changes were measured by the traditional plate-count method, and taxonomic distribution was studied by 16S rDNA sequencing, then maximum parsimony phylogenic analysis with bootstrapping (1,000 replications). The bacterial population was higher after 39 than 7 days of rice cultivation, which indicated the augmentation of soil microbes by rice root exudates. The application of butachlor increased the diazotrophic population in both upper (0-3 cm) and lower (3-15 cm) layers of soils. Especially at day 39, the population of diazotrophs was 1.8 and 1.6 times that of the control in upper and lower layer soils, respectively. We found several bacterial strains only with butachlor application; examples are strains closest to Bacillus arsenicus, B. marisflavi, B. luciferensis, B. pumilus, and Pseudomonas alvei. Among diazotrophs, three strains closely related to Streptomyces sp. or Rhrizobium sp. were found only with butachlor application. The population of cultivable bacteria and the species composition were both changed with butachlor application, which explains in part the contribution of butachlor to augmenting soil nitrogen-fixing ability.

  5. The effects of nitrogen fixation and plant growth-promoting in rice-diazotroph association

    International Nuclear Information System (INIS)

    Lin Fan; Wang Lu

    1999-05-01

    This is a review of studies on applications of the genetic engineered ammonium-tolerant diazotroph as an inoculum with the effects of nitrogen-fixation, plant growth-promoting and yield-increasing on rice and some crops by using 15 N tracer in mini-plot and field experiments in resent years

  6. Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments▿

    Science.gov (United States)

    Betancourt, Doris A.; Loveless, Telisa M.; Brown, James W.; Bishop, Paul E.

    2008-01-01

    Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Mo-independent nitrogenases from aquatic environments. In the present study, we extend these results to include diazotrophs isolated from wood chip mulch, soil, “paraffin dirt,” and sediments from mangrove swamps. Mo-deficient, N-free media under both aerobic and anaerobic conditions were used for the isolations. A total of 26 isolates were genetically and physiologically characterized. Their phylogenetic placement was determined using 16S rRNA gene sequence analysis. Most of the isolates are members of the gamma subdivision of the class Proteobacteria and appear to be specifically related to fluorescent pseudomonads and azotobacteria. Two other isolates, AN1 and LPF4, are closely related to Enterobacter spp. and Paenibacillus spp., respectively. PCR and/or Southern hybridization were used to detect the presence of nitrogenase genes in the isolates. PCR amplification of vnfG and anfG was used to detect the genetic potential for the expression of the vanadium-containing nitrogenase and the iron-only nitrogenase in the isolates. This study demonstrates that diazotrophs with Mo-independent nitrogenases can be readily isolated from diverse natural environments. PMID:18378646

  7. Seasonal variations in the diversity and abundance of diazotrophic communities across soils

    NARCIS (Netherlands)

    Pereira e Silva, Michele C.; Semenov, Alexander V.; van Elsas, Jan Dirk; Salles, Joana Falcao

    The nitrogen (N)-fixing community is a key functional community in soil, as it replenishes the pool of biologically available N that is lost to the atmosphere via anaerobic ammonium oxidation and denitrification. We characterized the structure and dynamic changes in diazotrophic communities, based

  8. Dissolved organic nitrogen and carbon release by a marine unicellular diazotrophic cyanobacterium

    NARCIS (Netherlands)

    Benavides, M.; Agawin, N.S.R.; Aristegui, J.; Peene, J.; Stal, L.J.

    2013-01-01

    Dinitrogen (N2) fixation rates may be underestimated when recently fixed N2 is released as dissolved organic nitrogen (DON). DON release (DONr) is substantial in the filamentous cyanobacterium Trichodesmium but has never been reported in unicellular diazotrophic cyanobacteria. We used axenic

  9. [Dynamics of diazotrophic bacteria number in the root zone of wheat Vrn lines isogenic by genes].

    Science.gov (United States)

    Samoĭlov, A M; Zhmurko, V V

    2012-01-01

    The number of diazotrophic bacteria and nitrogenase activity in the root zone of isogenic monogene-dominant Vrn lines were measured in the field experiments throughout their vegetation from tillering to heading. The total number of diazotrophic bacteria and nitrogenase activity in the root zone of these lines during this period were increased irrespective of their genotypes. The above indices of the winter cultivar (Vrn loci bottom recessive) were lower than those of the spring lines--Vrn-A1, Vrn-B1 and Vrn-D1. Plants of Vrn-B1 line have the lowest indices among the spring lines with the exception of some indices. This line plants flowered later than those of Vrn-A1 and Vrn-D1 lines. We hypothesized the differences between plants of these lines as to nitrogen fixation activity and the number of diazotrophic bacteria are mediately determined by Vrn loci through their effects on metabolism intensity and assimilate reflux in the form of root exudates, therefore the total number of diazotrophic bacteria and nitrogenase activity increases.

  10. Dominance of unicellular cyanobacteria in the diazotrophic community in the Atlantic Ocean

    NARCIS (Netherlands)

    Agawin, N.S.R.; Benavides, M.; Busquets, A.; Ferriol, P.; Stal, L.J.; Aristegui, J.

    2014-01-01

    ABSTRACT: The horizontal and vertical distribution of representatives of diazotrophic unicellular cyanobacteria was investigated in the subtropical northeast Atlantic Ocean (28.87 to 42.00°N; 9.01 to 20.02°W). Samples from stations encompassing different water conditions (from oceanic oligotrophic

  11. Dominance of unicellular cyanobacteria in the diazotrophic community in the Atlantic Ocean

    NARCIS (Netherlands)

    Agawin, N.S.R.; Benavides, M.; Busquets, A.; Ferriol, P.; Stal, L.J.; Arístegui, J.

    2014-01-01

    The horizontal and vertical distribution of representatives of diazotrophic unicellular cyanobacteria was investigated in the subtropical northeast Atlantic Ocean (28.87 to 42.00°N; 9.01 to 20.02°W). Samples from stations encompassing different water conditions (from oceanic oligotrophic waters to

  12. Glycerol as a Cheaper Carbon Source in Bacterial Cellulose (BC) Production by Gluconacetobacter Xylinus DSM46604 in Batch Fermentation System

    International Nuclear Information System (INIS)

    Azila Adnan; Nair, G.R.; Roslan Umar; Roslan Umar

    2015-01-01

    Bacterial cellulose (BC) is a polymer of glucose monomers, which has unique properties including high crystallinity and high strength. It has potential to be used in biomedical applications such as making artificial blood vessel, wound dressings, and in the paper making industry. Extensive study on BC aimed to improve BC production such as by using glycerol as a cheaper carbon source. BC was produced in shake flask culture using five different concentrations of glycerol (10, 20, 30, 40 and 50 g/ L). Using concentration of glycerol above 20 g/ L inhibited culture growth and BC production. Further experiments were performed in batch culture (3-L bioreactor) using 20 g/ L glycerol. It produced yield and productivity of 0.15 g/ g and 0.29 g/ L/ day BC, respectively. This is compared with the control medium, 50 g/ L glucose, which only gave yield and productivity of 0.05 g/ g and 0.23 g/ L/ day, respectively. Twenty g/ L of glycerol enhanced BC production by Gluconacetobacter xylinus DSM46604 in batch fermentation system. (author)

  13. Bio-cellulose Production by Beijerinckia fluminensis WAUPM53 and Gluconacetobacter xylinus 0416 in Sago By-product Medium.

    Science.gov (United States)

    Voon, W W Y; Muhialdin, B J; Yusof, N L; Rukayadi, Y; Meor Hussin, A S

    2018-06-19

    Bio-cellulose is the microbial extracellular cellulose that is produced by growing several microorganisms on agriculture by-products, and it is used in several food applications. This study aims to utilize sago by-product, coconut water, and the standard medium Hestrin-Schramm as the carbon sources in the culture medium for bio-cellulose production. The bacteria Beijerinkia fluminensis WAUPM53 and Gluconacetobacter xylinus 0416 were selected based on their bio-cellulose production activity. The structure was determined by Fourier transform infrared spectroscopy and scanning electron microscopy, while the toxicity safety was evaluated by brine shrimp lethality test. The results of Fourier transform infrared spectroscopy showed that the bio-cellulose produced by B. fluminensis cultivated in sago by-products was of high quality. The bio-cellulose production by B. fluminensis in the sago by-product medium was slightly higher than that in the coconut water medium and was comparable with the production in the Hestrin-Schramm medium. Brine shrimp lethality test confirmed that the bio-cellulose produced by B. fluminensis in the sago by-product medium has no toxicity, which is safe for applications in the food industry. This is the first study to determine the high potential of sago by-product to be used as a new carbon source for the bio-cellulose production.

  14. Analogous nutrient limitations in unicellular diazotrophs and Prochlorococcus in the South Pacific Ocean.

    Science.gov (United States)

    Moisander, Pia H; Zhang, Ruifeng; Boyle, Edward A; Hewson, Ian; Montoya, Joseph P; Zehr, Jonathan P

    2012-04-01

    Growth limitation of phytoplankton and unicellular nitrogen (N(2)) fixers (diazotrophs) were investigated in the oligotrophic Western South Pacific Ocean. Based on change in abundances of nifH or 23S rRNA gene copies during nutrient-enrichment experiments, the factors limiting net growth of the unicellular diazotrophs UCYN-A (Group A), Crocosphaera watsonii, γ-Proteobacterium 24774A11, and the non-diazotrophic picocyanobacterium Prochlorococcus, varied within the region. At the westernmost stations, numbers were enhanced by organic carbon added as simple sugars, a combination of iron and an organic chelator, or iron added with phosphate. At stations nearest the equator, the nutrient-limiting growth was not apparent. Maximum net growth rates for UCYN-A, C. watsonii and γ-24774A11 were 0.19, 0.61 and 0.52 d(-1), respectively, which are the first known empirical growth rates reported for the uncultivated UCYN-A and the γ-24774A11. The addition of N enhanced total phytoplankton biomass up to 5-fold, and the non-N(2)-fixing Synechococcus was among the groups that responded favorably to N addition. Nitrogen was the major nutrient-limiting phytoplankton biomass in the Western South Pacific Ocean, while availability of organic carbon or iron and organic chelator appear to limit abundances of unicellular diazotrophs. Lack of phytoplankton response to nutrient additions in the Pacific warm pool waters suggests diazotroph growth in this area is controlled by different factors than in the higher latitudes, which may partially explain previously observed variability in community composition in the region.

  15. Nitrogen signalling in plant interactions with associative and endophytic diazotrophic bacteria.

    Science.gov (United States)

    Carvalho, T L G; Balsemão-Pires, E; Saraiva, R M; Ferreira, P C G; Hemerly, A S

    2014-10-01

    Some beneficial plant-interacting bacteria can biologically fix N2 to plant-available ammonium. Biological nitrogen fixation (BNF) is an important source of nitrogen (N) input in agriculture and represents a promising substitute for chemical N fertilizers. Diazotrophic bacteria have the ability to develop different types of root associations with different plant species. Among the highest rates of BNF are those measured in legumes nodulated by endosymbionts, an already very well documented model of plant-diazotrophic bacterial association. However, it has also been shown that economically important crops, especially monocots, can obtain a substantial part of their N needs from BNF by interacting with associative and endophytic diazotrophic bacteria, that either live near the root surface or endophytically colonize intercellular spaces and vascular tissues of host plants. One of the best reported outcomes of this association is the promotion of plant growth by direct and indirect mechanisms. Besides fixing N, these bacteria can also produce plant growth hormones, and some species are reported to improve nutrient uptake and increase plant tolerance against biotic and abiotic stresses. Thus, this particular type of plant-bacteria association consists of a natural beneficial system to be explored; however, the regulatory mechanisms involved are still not clear. Plant N status might act as a key signal, regulating and integrating various metabolic processes that occur during association with diazotrophic bacteria. This review will focus on the recent progress in understanding plant association with associative and endophytic diazotrophic bacteria, particularly on the knowledge of the N networks involved in BNF and in the promotion of plant growth. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  16. The Assimilation of Diazotroph-Derived Nitrogen by Scleractinian Corals Depends on Their Metabolic Status

    Directory of Open Access Journals (Sweden)

    Vanessa N. Bednarz

    2017-01-01

    Full Text Available Tropical corals are associated with a diverse community of dinitrogen (N2-fixing prokaryotes (diazotrophs providing the coral an additional source of bioavailable nitrogen (N in oligotrophic waters. The overall activity of these diazotrophs changes depending on the current environmental conditions, but to what extent it affects the assimilation of diazotroph-derived N (DDN by corals is still unknown. Here, in a series of 15N2 tracer experiments, we directly quantified DDN assimilation by scleractinian corals from the Red Sea exposed to different environmental conditions. We show that DDN assimilation strongly varied with the corals’ metabolic status or with phosphate availability in the water. The very autotrophic shallow-water (~5 m corals showed low or no DDN assimilation, which significantly increased under elevated phosphate availability (3 µM. Corals that depended more on heterotrophy (i.e., bleached and deep-water [~45 m] corals assimilated significantly more DDN, which contributed up to 15% of the corals’ N demand (compared to 1% in shallow corals. Furthermore, we demonstrate that a substantial part of the DDN assimilated by deep corals was likely obtained from heterotrophic feeding on fixed N compounds and/or diazotrophic cells in the mucus. Conversely, in shallow corals, the net release of mucus, rich in organic carbon compounds, likely enhanced diazotroph abundance and activity and thereby the release of fixed N to the pelagic and benthic reef community. Overall, our results suggest that DDN assimilation by corals varies according to the environmental conditions and is likely linked to the capacity of the coral to acquire nutrients from seawater.

  17. Evaluation of the biological nitrogen fixation contribution in sugarcane plants originated from seeds and inoculated with nitrogen-fixing endophytes Avaliação da contribuição da fixação biológica de nitrogênio em cana-de-açúcar originada de sementes e inoculada com endófitos fixadores de nitrogênio

    Directory of Open Access Journals (Sweden)

    Erineudo de Lima Canuto

    2003-11-01

    Full Text Available The inoculation technique with endophytic diazotrophic bacteria in sugarcane has been shown as an alternative practice to plant growth promotion. The aim of this work was to evaluate the biological nitrogen fixation (BNF contribution by different strains of Herbaspirillum seropedicae and Gluconacetobacter diazotrophicus in sugarcane plant inoculated from seeds. The seeds were planted in pots filled with non-sterile soil, inoculated with the bacterial strains and grown 10 months outside of the greenhouse. The BNF contribution of the inoculated bacteria varied depending on the plant species used as a control. The highest BNF contribution as well as the highest populations of reisolated bacteria was observed with inoculation of H. seropedicae strains. The roots appeared to be the preferential tissues for the establishment of the inoculated species.A técnica de inoculação com bactérias diazotróficas endofíticas na cana-de-açúcar apresenta-se como uma prática alternativa para promover o crescimento vegetal menos dependente da adubação nitrogenada. Este trabalho teve como objetivo avaliar a contribuição da fixação biológica de nitrogênio (FBN por diferentes estirpes de Herbaspirillum seropedicae e Gluconacetobacter diazotrophicus inoculadas em plantas de cana-de-açúcar originadas de semente. As sementes foram plantadas em vasos com solo não estéril, inoculadas com as diferentes bactérias e mantidas por 10 meses ao ar livre. As maiores contribuições da FBN ocorreram com a inoculação de estirpes Herbaspirillum seropedicae, e dependeram da espécie vegetal utilizada como testemunha. As raízes apresentaram-se como o órgão vegetal preferencial para o estabelecimento das espécies inoculadas.

  18. Enumeration and characterization of arsenic-tolerant diazotrophic bacteria in a long-term heavy-metal-contaminated soil

    OpenAIRE

    Oliveira, A.; Pampulha, M.E.; Neto, M.M.; Almeida, A.C.

    2009-01-01

    The abundance of arsenic-tolerant diazotrophic bacteria was compared in a long-term contaminated soil versus a non-contaminated one. In addition, the characterization of tolerant diazotrophic bacteria was carried out. Differences in the number of heterotrophic N2 fixers were found between soils. Contaminated soil showed a decrease in the microbial population size of about 80%, confirming the great sensitivity of this group of soil bacteria to metals. However, quantitat...

  19. Characterization of diazotrophic bacteria non-symbiotic associated with eucalyptus (eucalyptus sp.) in Codazzi, Cesar (Colombia)

    International Nuclear Information System (INIS)

    Obando Castellanos, Dolly Melissa; Burgos Zabala, Ludy Beatriz; Rivera Botia, Diego Mauricio; Rubiano Garrido, Maria Fernanda; Divan Baldini, Vera Lucia; Bonilla Buitrago, Ruth Rebeca

    2010-01-01

    The effect of climatic seasons (rainy and dry) and the stratum sample (rhizospheric soil, roots and leaves) the population of the genera Azotobacter, Beijerinckia, Derxia, Azospirillum, Herbaspirillum, Gluconacetobacter and Burkholderia in soil rhizosphere, roots and leaves of eucalyptus (eucalyptus sp.). It also assesses their ability to produce indoles compounds as plant growth promoters and their acetylene reduction activity as an indicator of biological fixation of nitrogen. The results showed no statistically significant differences in the Duncan test (p ≤ 0.05) in the population with respect to the climate epoch, suggesting that these bacteria are able to tolerate stress conditions by different physiological mechanisms. With respect to the stratum sample isolates attempts of Herbaspirillum sp. and Azospirillum sp. significant differences in rhizospheric soil and roots. we obtained 44 isolates of which were grouped by phenotypic characterization as 14 suspected of Beijerinckia sp., 12 Azotobacter sp., 8 Derxia sp., 4 Herbaspirillum sp., 5 Azospirillum sp., 1 Gluconacetobacter sp. and 1 Burkholderia sp. due to their high potential were selected isolates C27, C26 and C25. These four strains present the best values of efficiency in vitro, exceeding production values of the reference strains used (A. chroococcum (AC1) and a. brasilense (SP7)).

  20. Revealing differences in metabolic flux distributions between a mutant strain and its parent strain Gluconacetobacter xylinus CGMCC 2955.

    Directory of Open Access Journals (Sweden)

    Cheng Zhong

    Full Text Available A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955 using DEC (diethyl sulfate and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA cycle was obtained in mutant strain (57.0% compared with parent strain (17.0%. It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH, which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53-6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain.

  1. Isolation and Characterization of Two Cellulose Morphology Mutants of Gluconacetobacter hansenii ATCC23769 Producing Cellulose with Lower Crystallinity

    Science.gov (United States)

    Deng, Ying; Nagachar, Nivedita; Fang, Lin; Luan, Xin; Catchmark, Jeffrey M.; Tien, Ming; Kao, Teh-hui

    2015-01-01

    Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC). These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of peptidoglycan in the

  2. Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity.

    Directory of Open Access Journals (Sweden)

    Ying Deng

    Full Text Available Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC. These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of

  3. Characteristics of nitrogen fixation of mixed diazotrophs associated with rice plant

    International Nuclear Information System (INIS)

    Ling Fan; Wang Zhengfang; Wang Yaodong; Song Wei

    1997-01-01

    Characteristics of N 2 fixation of diazotrophs associated with rice plant in paddy soils in Nanjing was studied by 15 N tracing technique. The results showed that amount of N fixed by rice plant was 1.03 kg/666.7 m 2 and the rate of fixed N was 6.7%. The maximum N fixed was occurred during jointing-complete heading stage. The daily average amount of N fixed reached to 24.31 mg/m 2 ·day. The fixed N of jointing-complete heading stage was 40.9% of that whole rice growth stage. The amount of fixed N during jointing-maturing stage was over 70% of whole rice growth stage. The economic benefits for fertilizer saving was 13.3 kg/666.7 m 2 of ammonium sulphate. The yield of rice grain was increased by 4.14% after inoculation with the mixed diazotrophs

  4. GENETIC VARIABILITY OF SUGARCANE-ASSOCIATED DIAZOTROPHIC BACTERIA CAPABLE OF INORGANIC PHOSPHATE SOLUBILIZING

    OpenAIRE

    Lira-Cadete, Luana; Barbosa de Farias, Andreza Raquel; de Souza Ramos, Andresa Priscila; da Costa, Diogo Paes; Freire, Fernando Jose; Kuklinsky-Sobral, Julia

    2013-01-01

    The sugarcane is a culture of great importance for the Brazilian agriculture. Every year this culture consumes great amounts of nitrogen and phosphate fertilizers. However, the use of plant growth-promoting bacteria can reduce the use of the chemical fertilizers, contributing to the economy and the environment conservation. So, the goal of this study was to select sugarcane-associated diazotrophic bacteria able to solubilize inorganic phosphate and to evaluate the genetic diversity of these b...

  5. Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima

    Directory of Open Access Journals (Sweden)

    Fan eZhang

    2014-10-01

    Full Text Available Sponges that harbor microalgal or cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM. nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem.

  6. Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima.

    Science.gov (United States)

    Zhang, Fan; Vicente, Jan; Hill, Russell T

    2014-01-01

    Sponges that harbor microalgal or, cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM). nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem.

  7. Associative diazotrophic bacteria in grass roots and soils from heavy metal contaminated sites.

    Science.gov (United States)

    Moreira, Fátima M S; Lange, Anderson; Klauberg-Filho, Osmar; Siqueira, José O; Nóbrega, Rafaela S A; Lima, Adriana S

    2008-12-01

    This work aimed to evaluate density of associative diazotrophic bacteria populations in soil and grass root samples from heavy metal contaminated sites, and to characterize isolates from these populations, both, phenotypically (Zinc, Cadmium and NaCl tolerance in vitro, and protein profiles) and genotypically (16S rDNA sequencing), as compared to type strains of known diazotrophic species. Densities were evaluated by using NFb, Fam and JNFb media, commonly used for enrichment cultures of diazotrophic bacteria. Bacterial densities found in soil and grass root samples from contaminated sites were similar to those reported for agricultural soils. Azospirillum spp. isolates from contaminated sites and type strains from non-contaminated sites varied substantially in their in vitro tolerance to Zn+2 and Cd+2, being Cd+2 more toxic than Zn+2. Among the most tolerant isolates (UFLA 1S, 1R, S181, S34 and S22), some (1R, S34 and S22) were more tolerant to heavy metals than rhizobia from tropical and temperate soils. The majority of the isolates tolerant to heavy metals were also tolerant to salt stress as indicated by their ability to grow in solid medium supplemented with 30 g L(-1) NaCl. Five isolates exhibited high dissimilarity in protein profiles, and the 16S rDNA sequence analysis of two of them revealed new sequences for Azospirillum.

  8. "Cold" Fixation: Reconciliation of Nitrogen Fixation Rates and Diazotroph Assemblages in the Arctic Ocean

    Science.gov (United States)

    Fong, A. A.; Waite, A.; Rost, B.; Richter, K. U.

    2016-02-01

    Measurements of biological nitrogen fixation are typically conducted in oligotrophic subtropical and tropical marine environments where concentrations of fixed inorganic nitrogen are low. To date, only a handful of nitrogen fixation studies have been conducted in high latitude marine environments, but further investigation is needed to resolve the distribution of cold ocean diazotrophic assemblages. Nitrogen fixation rates and nifH gene distributions were measured at seven stations from 5°E to 20°E, north of 81°N in the Arctic Ocean at the onset of summer 2015. Discrete water samples in ice-covered regions were collected from the sea surface to 200 m for 15N2-tracer additions and targeted nifH gene and transcript analyses. Previous work suggests that heterotrophic bacteria dominate diazotrophic communities in the Arctic Ocean. Therefore, additional nifH gene surveys of sinking particles were conducted to test for enrichment on organic matter-rich microenvironments. Together, these measurements aim to reconcile diazotrophic activity with microbial community composition, further elucidating how nitrogen fixers could impact current concepts in polar carbon and nutrient cycling.

  9. Strong shift in the diazotrophic endophytic bacterial community inhabiting rice (Oryza sativa) plants after flooding.

    Science.gov (United States)

    Ferrando, Lucía; Fernández Scavino, Ana

    2015-09-01

    Flooding impacts soil microbial communities, but its effect on endophytic communities has rarely been explored. This work addresses the effect of flooding on the abundance and diversity of endophytic diazotrophic communities on rice plants established in a greenhouse experiment. The nifH gene was significantly more abundant in roots after flooding, whereas the nifH gene copy numbers in leaves were unaffected and remained low. The PCA (principal component analysis) of T-RFLP (terminal restriction fragment length polymorphism) profiles indicated that root communities of replicate plots were more similar and diverse after flooding than before flooding. The nifH libraries obtained by cloning and 454 pyrosequencing consistently showed a remarkable shift in the diazotrophic community composition after flooding. Gammaproteobacteria (66-98%), mainly of the genus Stenotrophomonas, prevailed in roots before flooding, whereas Betaproteobacteria was the dominant class (26-34%) after flooding. A wide variety of aerotolerant and anaerobic diazotrophic bacteria (e.g. Dechloromonas, Rhodopseudomonas, Desulfovibrio, Geobacter, Chlorobium, Spirochaeta, Selenomonas and Dehalobacter) with diverse metabolic traits were retrieved from flooded rice roots. These findings suggest that endophytic communities could be significantly impacted by changes in plant-soil conditions derived from flooding during rice cropping. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  10. Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

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    Aurilena de Aviz Silva

    Full Text Available ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholderia silvatlantica UENF 11711, and Herbaspirillum seropedicae HRC 54, five levels of urea (0, 2, 5, 7, and 10g L-1, and five evaluation periods (30, 60, 90, 120, and 150 days following the planting of plantlets in trays, and with four replications of each treatment. An increase in the dried weight of shoots and roots of micropropagated pineapple cv. 'Vitória' plants depended on inoculation with diazotrophic bacteria, and plantlets acclimatization could be reduced by up to 57 days, depending on the concentration of urea fertilizer.

  11. Isolation and molecular identification of endophytic diazotrophs from seeds and stems of three cereal crops.

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    Huawei Liu

    Full Text Available Ten strains of endophytic diazotroph were isolated and identified from the plants collected from three different agricultural crop species, wheat, rice and maize, using the nitrogen-free selective isolation conditions. The nitrogen-fixing ability of endophytic diazotroph was verified by the nifH-PCR assay that showed positive nitrogen fixation ability. These identified strains were classified by 879F-RAPD and 16S rRNA sequence analysis. RAPD analyses revealed that the 10 strains were clustered into seven 879F-RAPD groups, suggesting a clonal origin. 16S rRNA sequencing analyses allowed the assignment of the 10 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Paenibacillus, Enterobacter, Klebsiella and Pantoea. These representative genus are not endophytic diazotrophs in the conventional sense. They may have obtained nitrogen fixation ability through lateral gene transfer, however, the evolutionary forces of lateral gene transfer are not well known. Molecular identification results from 16S rRNA analyses were also confirmed by morphological and biochemical data. The test strains SH6A and MZB showed positive effect on the growth of plants.

  12. Diazotroph Diversity in the Sea Ice, Melt Ponds, and Surface Waters of the Eurasian Basin of the Central Arctic Ocean.

    Science.gov (United States)

    Fernández-Méndez, Mar; Turk-Kubo, Kendra A; Buttigieg, Pier L; Rapp, Josephine Z; Krumpen, Thomas; Zehr, Jonathan P; Boetius, Antje

    2016-01-01

    The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing toward a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77°N. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed.

  13. Diazotroph diversity in the sea ice, melt ponds and surface waters of the Eurasian Basin of the Central Arctic Ocean

    Directory of Open Access Journals (Sweden)

    Mar Fernández-Méndez

    2016-11-01

    Full Text Available The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing towards a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77ºN. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed.

  14. Endophytic colonization of plant roots by nitrogen-fixing bacteria

    International Nuclear Information System (INIS)

    Cocking, Edward C.

    2001-01-01

    Nitrogen-fixing bacteria are able to enter into roots from the rhizosphere, particularly at the base of emerging lateral roots, between epidermal cells and through root hairs. In the rhizosphere growing root hairs play an important role in symbiotic recognition in legume crops. Nodulated legumes in endosymbiosis with rhizobia are amongst the most prominent nitrogen-fixing systems in agriculture. The inoculation of non-legumes, especially cereals, with various non-rhizobial diazotrophic bacteria has been undertaken with the expectation that they would establish themselves intercellularly within the root system, fixing nitrogen endophytic ally and providing combined nitrogen for enhanced crop production. However, in most instances bacteria colonize only the surface of the roots and remain vulnerable to competition from other rhizosphere micro-organisms, even when the nitrogen-fixing bacteria are endophytic, benefits to the plant may result from better uptake of soil nutrients rather than from endophytic nitrogen fixation. Azorhizobium caulinodans is known to enter the root system of cereals, other nonlegume crops and Arabidopsis, by intercellular invasion between epidermal cells and to internally colonize the plant intercellularly, including the xylem. This raises the possibility that xylem colonization might provide a nonnodular niche for endosymbiotic nitrogen fixation in rice, wheat, maize, sorghum and other non-legume crops. A particularly interesting, naturally occurring, non-qodular xylem colonising endophytic diazotrophic interaction with evidence for endophytic nitrogen fixation is that of Gluconacetobacter diazotrophicus in sugarcane. Could this beneficial endophytic colonization of sugarcane by G. diazotrophicus be extended to other members of the Gramineae, including the major cereals, and to other major non-legume crops of the World? (author)

  15. Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae in the Brazilian Amazon

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    Paulo Ivan Fernandes Júnior

    2013-06-01

    Full Text Available The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg’s medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH+ bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty- one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus

  16. Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon.

    Science.gov (United States)

    Júnior, Paulo Ivan Fernandes; Pereira, Gilmara Maria Duarte; Perin, Liamara; da Silva, Luana Mesquita; Baraúna, Alexandre Cardoso; Alvess, Francilene Muniz; Passos, Samuel Ribeiro; Zilli, Jerri Edson

    2013-06-01

    The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg's medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty-one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360 bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growth

  17. Effects of different fermentation methods on bacterial cellulose and acid production by Gluconacetobacter xylinus in Cantonese-style rice vinegar.

    Science.gov (United States)

    Fu, Liang; Chen, Siqian; Yi, Jiulong; Hou, Zongxia

    2014-07-01

    A strain of acidogenic bacterium was isolated from the fermentation liquid of Cantonese-style rice vinegar produced by traditional surface fermentation. 16S rDNA identification confirmed the bacterium as Gluconacetobacter xylinus, which synthesizes bacterial cellulose, and the acid productivity of the strain was investigated. In the study, the effects of the membrane integrity and the comparison of the air-liquid interface membrane with immerged membrane on total acidity, cellulose production, alcohol dehydrogenase (ADH) activity and number of bacteria were investigated. The cellulose membrane and the bacteria were observed under SEM for discussing their relationship. The correlations between oxygen consumption and total acid production rate were compared in surface and shake flask fermentation. The results showed the average acid productivity of the strain was 0.02g/(100mL/h), and the integrity of cellulose membrane in surface fermentation had an important effect on total acidity and cellulose production. With a higher membrane integrity, the total acidity after 144 h of fermentation was 3.75 g/100 mL, and the cellulose production was 1.71 g/100 mL after 360 h of fermentation. However, when the membrane was crushed by mechanical force, the total acidity and the cellulose production were as low as 0.36 g/100 mL and 0.14 g/100 mL, respectively. When the cellulose membrane was forced under the surface of fermentation liquid, the total acid production rate was extremely low, but the activity of ADH in the cellulose membrane was basically the same with the one above the liquid surface. The bacteria were mainly distributed in the cellulose membrane during the fermentation. The bacterial counts in surface fermentation were more than in the shake flask fermentation and G. xylinus consumed the substrate faster, in surface fermentation than in shake flask fermentation. The oxygen consumption rate and total acid production rate of surface fermentation were respectively 26

  18. Influence of nitrogen fertilization on diazotrophic communities in the rhizosphere of the Jerusalem artichoke (Helianthus tuberosus L.).

    Science.gov (United States)

    Meng, Xianfa; Wang, Lin; Long, Xiaohua; Liu, Zhaopu; Zhang, Zhenhua; Zed, Rengel

    2012-06-01

    Diazotrophs in the soil may be influenced by plant factors as well as nitrogen (N) fertilization. In this study, we investigated potential diazotrophic communities in the rhizosphere of the Jerusalem artichoke (Helianthus tuberosus L.) supplied with differing amounts of N. The community structure of N(2)-fixing bacteria was profiled using the length heterogeneity polymerase chain reaction (LH-PCR) and terminal restriction fragment length polymorphism (T-RFLP) based on a variation in the nifH gene. Higher numbers of diazotrophs were detected by T-RFLP compared to LH-PCR. The lowest number of N(2)-fixing bacteria was observed in the rhizosphere soil with high N fertilization. T-RFLP was a better method than LH-PCR for profiling microbial diversity of diazotrophs using multidimensional scaling (MDS) and analysis of similarity (ANOSIM) of fingerprints as well as diversity measures. The supply of N fertilizer appeared to negatively influence the abundance of diazotrophs in the rhizophere of the Jerusalem artichoke. Copyright © 2012 Institut Pasteur. All rights reserved.

  19. Comparative study of endophytic and endophytic diazotrophic bacterial communities across rice landraces grown in the highlands of northern Thailand.

    Science.gov (United States)

    Rangjaroen, Chakrapong; Rerkasem, Benjavan; Teaumroong, Neung; Sungthong, Rungroch; Lumyong, Saisamorn

    2014-01-01

    Communities of bacterial endophytes within the rice landraces cultivated in the highlands of northern Thailand were studied using fingerprinting data of 16S rRNA and nifH genes profiling by polymerase chain reaction-denaturing gradient gel electrophoresis. The bacterial communities' richness, diversity index, evenness, and stability were varied depending on the plant tissues, stages of growth, and rice cultivars. These indices for the endophytic diazotrophic bacteria within the landrace rice Bue Wah Bo were significantly the lowest. The endophytic bacteria revealed greater diversity by cluster analysis with seven clusters compared to the endophytic diazotrophic bacteria (three clusters). Principal component analysis suggested that the endophytic bacteria showed that the community structures across the rice landraces had a higher stability than those of the endophytic diazotrophic bacteria. Uncultured bacteria were found dominantly in both bacterial communities, while higher generic varieties were observed in the endophytic diazotrophic bacterial community. These differences in bacterial communities might be influenced either by genetic variation in the rice landraces or the rice cultivation system, where the nitrogen input affects the endophytic diazotrophic bacterial community.

  20. Database of diazotrophs in global ocean: abundance, biomass and nitrogen fixation rates

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    Y.-W. Luo

    2012-08-01

    Full Text Available Marine N2 fixing microorganisms, termed diazotrophs, are a key functional group in marine pelagic ecosystems. The biological fixation of dinitrogen (N2 to bioavailable nitrogen provides an important new source of nitrogen for pelagic marine ecosystems and influences primary productivity and organic matter export to the deep ocean. As one of a series of efforts to collect biomass and rates specific to different phytoplankton functional groups, we have constructed a database on diazotrophic organisms in the global pelagic upper ocean by compiling about 12 000 direct field measurements of cyanobacterial diazotroph abundances (based on microscopic cell counts or qPCR assays targeting the nifH genes and N2 fixation rates. Biomass conversion factors are estimated based on cell sizes to convert abundance data to diazotrophic biomass. The database is limited spatially, lacking large regions of the ocean especially in the Indian Ocean. The data are approximately log-normal distributed, and large variances exist in most sub-databases with non-zero values differing 5 to 8 orders of magnitude. Reporting the geometric mean and the range of one geometric standard error below and above the geometric mean, the pelagic N2 fixation rate in the global ocean is estimated to be 62 (52–73 Tg N yr−1 and the pelagic diazotrophic biomass in the global ocean is estimated to be 2.1 (1.4–3.1 Tg C from cell counts and to 89 (43–150 Tg C from nifH-based abundances. Reporting the arithmetic mean and one standard error instead, these three global estimates are 140 ± 9.2 Tg N yr−1, 18 ± 1.8 Tg C and 590 ± 70 Tg C, respectively. Uncertainties related to biomass conversion factors can change the estimate of geometric mean pelagic diazotrophic biomass in the global ocean by about ±70%. It was recently established that the most commonly applied method used to measure N2

  1. An OmpA Family Protein, a Target of the GinI/GinR Quorum-Sensing System in Gluconacetobacter intermedius, Controls Acetic Acid Fermentation▿ †

    Science.gov (United States)

    Iida, Aya; Ohnishi, Yasuo; Horinouchi, Sueharu

    2008-01-01

    Via N-acylhomoserine lactones, the GinI/GinR quorum-sensing system in Gluconacetobacter intermedius NCI1051, a gram-negative acetic acid bacterium, represses acetic acid and gluconic acid fermentation. Two-dimensional polyacrylamide gel electrophoretic analysis of protein profiles of strain NCI1051 and ginI and ginR mutants identified a protein that was produced in response to the GinI/GinR regulatory system. Cloning and nucleotide sequencing of the gene encoding this protein revealed that it encoded an OmpA family protein, named GmpA. gmpA was a member of the gene cluster containing three adjacent homologous genes, gmpA to gmpC, the organization of which appeared to be unique to vinegar producers, including “Gluconacetobacter polyoxogenes.” In addition, GmpA was unique among the OmpA family proteins in that its N-terminal membrane domain forming eight antiparallel transmembrane β-strands contained an extra sequence in one of the surface-exposed loops. Transcriptional analysis showed that only gmpA of the three adjacent gmp genes was activated by the GinI/GinR quorum-sensing system. However, gmpA was not controlled directly by GinR but was controlled by an 89-amino-acid protein, GinA, a target of this quorum-sensing system. A gmpA mutant grew more rapidly in the presence of 2% (vol/vol) ethanol and accumulated acetic acid and gluconic acid in greater final yields than strain NCI1051. Thus, GmpA plays a role in repressing oxidative fermentation, including acetic acid fermentation, which is unique to acetic acid bacteria and allows ATP synthesis via ethanol oxidation. Consistent with the involvement of gmpA in oxidative fermentation, its transcription was also enhanced by ethanol and acetic acid. PMID:18487322

  2. An OmpA family protein, a target of the GinI/GinR quorum-sensing system in Gluconacetobacter intermedius, controls acetic acid fermentation.

    Science.gov (United States)

    Iida, Aya; Ohnishi, Yasuo; Horinouchi, Sueharu

    2008-07-01

    Via N-acylhomoserine lactones, the GinI/GinR quorum-sensing system in Gluconacetobacter intermedius NCI1051, a gram-negative acetic acid bacterium, represses acetic acid and gluconic acid fermentation. Two-dimensional polyacrylamide gel electrophoretic analysis of protein profiles of strain NCI1051 and ginI and ginR mutants identified a protein that was produced in response to the GinI/GinR regulatory system. Cloning and nucleotide sequencing of the gene encoding this protein revealed that it encoded an OmpA family protein, named GmpA. gmpA was a member of the gene cluster containing three adjacent homologous genes, gmpA to gmpC, the organization of which appeared to be unique to vinegar producers, including "Gluconacetobacter polyoxogenes." In addition, GmpA was unique among the OmpA family proteins in that its N-terminal membrane domain forming eight antiparallel transmembrane beta-strands contained an extra sequence in one of the surface-exposed loops. Transcriptional analysis showed that only gmpA of the three adjacent gmp genes was activated by the GinI/GinR quorum-sensing system. However, gmpA was not controlled directly by GinR but was controlled by an 89-amino-acid protein, GinA, a target of this quorum-sensing system. A gmpA mutant grew more rapidly in the presence of 2% (vol/vol) ethanol and accumulated acetic acid and gluconic acid in greater final yields than strain NCI1051. Thus, GmpA plays a role in repressing oxidative fermentation, including acetic acid fermentation, which is unique to acetic acid bacteria and allows ATP synthesis via ethanol oxidation. Consistent with the involvement of gmpA in oxidative fermentation, its transcription was also enhanced by ethanol and acetic acid.

  3. Effects of long-term elevated CO2, warming, and prolonged drought on Pleurozium-associated diazotrophic activity and abundance

    Science.gov (United States)

    Dyrnum, Kristine; Priemé, Anders; Michelsen, Anders

    2014-05-01

    Nitrogen (N2) fixation is the primary natural influx of N to terrestrial ecosystems, and changes in N2 fixation may have consequences for primary productivity and thus ecosystem function. We studied the activity and abundance of diazotrophs associated with the feather moss Pleurozium schreberi in a temperate heathland, after seven years of global change manipulations, including elevated atmospheric CO2 (510 ppm), increased temperature (0.5-1.5 ° C), and prolonged pre-summer droughts (4-6 weeks /year). Acetylene reduction assay was carried out monthly to monitor N2 fixation rates throughout one year, while nif H copy abundance, serving as a diazotroph abundance estimate, was measured by quantitative polymerase chain reaction (q-PCR). Prolonged summer droughts significantly increased both N2 fixation and nif H copy abundance, contrasting previous studies that demonstrate a direct negative correlation between N2 fixation and water availability. A shift in the relative abundance of N2-fixing bacteria from the green, upper parts of the moss stem to the lower, brown parts was observed. This shift could make diazotrophs less sensitive to desiccation, enabling N2 fixation to be upheld for longer during drought and thus causing higher abundance. Increased temperature likewise had a positive effect on the diazotroph abundance, although this did not translate into increased activity. Possibly, warming protects diazotrophs during extreme cold events, while actual N2 fixation is limited by water, disregarding a rise in potential N2 fixation caused by higher abundance. Increased CO2 caused no significant diazotroph response. Our study showed that long-term increase in temperature and recurrent drought events cause higher diazotroph abundance in Pleurozium schreberi and thus enhance the potential N2 fixations rate. Furthermore, our results indicate that diazotrophs may alter colonization patterns and thereby actively remain in the moss fraction less likely affected by

  4. Bacterial N2-fixation in mangrove ecosystems: insights from a diazotroph-mangrove interaction.

    Science.gov (United States)

    Alfaro-Espinoza, Gabriela; Ullrich, Matthias S

    2015-01-01

    Mangrove forests are highly productive ecosystems but represent low nutrient environments. Nitrogen availability is one of the main factors limiting mangrove growth. Diazotrophs have been identified as key organisms that provide nitrogen to these environments. N2-fixation by such organisms was found to be higher in the mangrove roots than in surrounding rhizosphere. Moreover, previous studies showed that mangroves grew better in the presence of N2-fixers indicating a potentially mutualistic relationship. However, the molecular signals and mechanisms that govern these interactions are still poorly understood. Here we present novel insights in the interaction of a diazotroph with a mangrove species to improve our understanding of the molecular and ecophysiological relationship between these two organisms under controlled conditions. Our results showed that Marinobacterium mangrovicola is a versatile organism capable of competing with other organisms to survive for long periods in mangrove soils. N2-fixation by this bacterium was up-regulated in the presence of mangrove roots, indicating a possible beneficial interaction. The increase in N2-fixation was limited to cells of the exponential growth phase suggesting that N2-fixation differs over the bacterial growth cycle. Bacterial transformants harboring a transcriptional nifH::gusA fusion showed that M. mangrovicola successfully colonized mangrove roots and simultaneously conducted N2-fixation. The colonization process was stimulated by the lack of an external carbon source suggesting a possible mutualistic relationship. M. mangrovicola represents an interesting genetically accessible diazotroph, which colonize mangrove roots and exhibit higher N2-fixation in the presence of mangrove roots. Consequently, we propose this microorganism as a tool to study molecular interactions between N2-fixers and mangrove plants and to better understand how changes in the environment could impact these important and relatively unknown

  5. Diversity of diazotrophic gut inhabitants of pikas (Ochotonidae) revealed by PCR-DGGE analysis.

    Science.gov (United States)

    Kizilova, A K; Kravchenko, I K

    2014-01-01

    Diazotrophic gut symbionts are considered to act as nitrogen providers for their hosts, as was shown for various termite species. Although the diet of lagomorphs, like pikas or rabbits, is very poor in nitrogen and energy, their fecal matter contains 30-40% of protein. Since our hypothesis was that pikas maintained a diazotrophic consortium in their gastrointestinal tract, we conducted the first investigation of microbial diversity in pika guts. We obtained gut samples from animals of several Ochotona species, O. hyperborea (Northern pika), O. mantchurica (Manchurian pika), and O. dauurica (Daurian pika), in order to retrieve and compare the nitrogen-fixing communities of different pika species. The age and gender of the animals were taken into consideration. We amplified 320-bp long fragments of the nifH gene using the DNA extracted directly from the colon and cecum samples of pika's gut, resolved them by DGGE, and performed phylogenetic reconstruction of 51 sequences obtained from excised bands. No significant difference was detected between the nitrogen-fixing gut inhabitants of different pika species. NifH sequences fell into two clusters. The first cluster contained the sequences affiliated with NifH Cluster I (Zehr et al., 2003) with similarity to Sphingomonas sp., Bradyrhizobium sp., and various uncultured bacteria from soil and rhizosphere. Sequences from the second group were related to Treponema sp., Fibrobacter succinogenes, and uncultured clones from the guts of various termites and belonged to NifH Cluster III. We suggest that diazotrophic organisms from the second cluster are genuine endosymbionts of pikas and provide nitrogen for further synthesis processes thus allowing these animals not to be short of protein.

  6. Genome sequence of Bradyrhizobium sp. LMTR 3, a diazotrophic symbiont of Lima bean (Phaseolus lunatus

    Directory of Open Access Journals (Sweden)

    Ernesto Ormeño-Orrillo

    2017-09-01

    Full Text Available Bradyrhizobium sp. LMTR 3 is a representative strain of one of the geno(species of diazotrophic symbionts associated with Lima bean (Phaseolus lunatus in Peru. Its 7.83 Mb genome was sequenced using the Illumina technology and found to encode a complete set of genes required for nodulation and nitrogen fixation, and additional genes putatively involved in root colonization. Its draft genome sequence and annotation have been deposited at GenBank under the accession number MAXC00000000.

  7. Enhancement of Chlorella vulgaris growth and bioremediation ability of aquarium wastewater using diazotrophs.

    Science.gov (United States)

    Ali, Sayeda Mohammed; Nasr, Hoda Shafeek; Abbas, Wafaa Tawfik

    2012-08-15

    Treatment of aquarium wastewater represents an important process to clean and recycle wastewater to be safely returned to the environment, used for cultivation or to minimize the multiple renewal of water. Chlorella vulgaris was an important freshwater microalgae which used in wastewater treatment, and increasing its potential of treatment can be achieved with existence of N2-fixing bacteria. Co-culturing of Chlorella vulgaris with the diazotrophs, Azospirillum brasilense or Azotobacter chroococcum in three different media; aquarium wastewater (AWW), sterile enriched natural aquarium wastewater (GPM) and synthetic wastewater media (SWW) were studied. Biomass yield of the microalgae was estimated by determination of chlorophylls (a and b), total carotenoid and the dry weight of C. vulgaris. Also determination of ammonia, nitrite, phosphate and nitrate in the culture were done. The presence of diazotrophs significantly increased the biomass of C. vulgaris by increasing its microalgae pigments (chlorophylls a and b, and total carotenoids). The highest pigments percentage was reported due to addition of A. brasilense to C. vulgaris (18.3-133.5%) compared to A. chroococcum (23.9-56.9%). As well as increased dry weight from 12 to 50%. There was also improved removal of nitrate, nitrite, ammonia and phosphate; where, the highest removal percentage was reported due to addition of A. chroococcum to C. vulgaris (0.0-52%) compared to A. brasilense (0.6-16.4%). A. brasilense and A. chroococcum can support C. vulgaris biomass production and bioremediation activity in the aquarium to minimize the periodical water renewal.

  8. Seasonal resource conditions favor a summertime increase in North Pacific diatom-diazotroph associations.

    Science.gov (United States)

    Follett, Christopher L; Dutkiewicz, Stephanie; Karl, David M; Inomura, Keisuke; Follows, Michael J

    2018-02-15

    In the North Pacific Subtropical Gyre (NPSG), an annual pulse of sinking organic carbon is observed at 4000 m between July and August, driven by large diatoms found in association with nitrogen fixing, heterocystous, cyanobacteria: Diatom-Diazotroph Associations (DDAs). Here we ask what drives the bloom of DDAs and present a simplified trait-based model of subtropical phototroph populations driven by observed, monthly averaged, environmental characteristics. The ratio of resource supply rates favors nitrogen fixation year round. The relative fitness of DDA traits is most competitive in early summer when the mixed layer is shallow, solar irradiance is high, and phosphorus and iron are relatively abundant. Later in the season, as light intensity drops and phosphorus is depleted, the traits of small unicellular diazotrophs become more competitive. The competitive transition happens in August, at the time when the DDA export event occurs. This seasonal dynamic is maintained when embedded in a more complex, global-scale, ecological model, and provides predictions for the extent of the North Pacific DDA bloom. The model provides a parsimonious and testable hypothesis for the stimulation of DDA blooms.

  9. Spatial and temporal distribution of two diazotrophic bacteria in the Chesapeake Bay.

    Science.gov (United States)

    Short, Steven M; Jenkins, Bethany D; Zehr, Jonathan P

    2004-04-01

    The aim of this study was to initiate autecological studies on uncultivated natural populations of diazotrophic bacteria by examining the distribution of specific diazotrophs in the Chesapeake Bay. By use of quantitative PCR, the abundance of two nifH sequences (907h22 and 912h4) was quantified in water samples collected along a transect from the head to the mouth of the Chesapeake Bay during cruises in April and October 2001 and 2002. Standard curves for the quantitative PCR assays demonstrated that the relationship between gene copies and cycle threshold was linear and highly reproducible from 1 to 10(7) gene copies. The maximum number of 907h22 gene copies detected was approximately 140 ml(-1) and the maximum number of 912h4 gene copies detected was approximately 340 ml(-1). Sequence 912h4 was most abundant at the mouth of the Chesapeake Bay, and in general, its abundance increased with increasing salinity, with the highest abundances observed in April 2002. Overall, the 907h22 phylotype was most abundant at the mid-bay station. Additionally, 907h22 was most abundant in the April samples from the mid-bay and mouth of the Chesapeake Bay. Despite the fact that the Chesapeake Bay is rarely nitrogen limited, our results show that individual nitrogen-fixing bacteria have distinct nonrandom spatial and seasonal distributions in the Chesapeake Bay and are either distributed by specific physical processes or adapted to different environmental niches.

  10. Characterization and crop production efficiency of diazotrophic bacterial isolates from coastal saline soils.

    Science.gov (United States)

    Barua, Shilajit; Tripathi, Sudipta; Chakraborty, Ashis; Ghosh, Sagarmoy; Chakrabarti, Kalyan

    2012-01-20

    Use of eco-friendly area specific salt tolerant bioinoculants is better alternatives to chemical fertilizer for sustainable agriculture in coastal saline soils. We isolated diverse groups of diazotrophic bacteria from coastal saline soils of different forest and agricultural lands in the Sundarbans, West Bengal, India, to study their effect on crop productivity in saline soils. Phenotypic, biochemical and molecular identifications of the isolates were performed. The isolates produced indole acetic acid, phosphatase, and solubilized insoluble phosphates. Sequence analysis of 16S rDNA identified the SUND_BDU1 strain as Agrobacterium and the strains SUND_LM2, Can4 and Can6 belonging to the genus Bacillus. The ARA activity, dinitrogen fixation and presence of nifH genes indicated they were diazotrophs. Field trials with these strains as bioinoculants were carried out during 2007-2009, with rice during August-December followed by Lady's finger during April-June. Microplots, amended with FYM inoculated with four bioinoculants individually were compared against sole FYM (5 t ha(-1)) and a sole chemical fertilizer (60:30:30 kg ha(-1) NPK) treated plot. The strain Can6 was by far the best performer in respect of yield attributes and productivity of studied crops. Copyright © 2011 Elsevier GmbH. All rights reserved.

  11. Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars.

    Science.gov (United States)

    Ji, Sang Hye; Gururani, Mayank Anand; Chun, Se-Chul

    2014-01-20

    We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants. Copyright © 2013 Elsevier GmbH. All rights reserved.

  12. Quantification and characterization of putative diazotrophic bacteria from forage palm under saline water irrigation

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    Gabiane dos Reis Antunes

    2017-09-01

    Full Text Available The aim of this study was to evaluate the density and phenotypical diversity of diazotrophic endophytic bacteria from the forage palm irrigated with different saline water depths. Opuntia stricta (IPA-200016 received five depths of saline water (L1: 80%. ETo; L2: 60%.ETo; L3: 40%; ETo; L4: 20%; ETo and, L5: 0% ETo, where ETo is the reference evapotranspiration. The roots were collected in the field, disinfected, grounded and serial diluted from 10-1 to 10-4. The total concentration of diazotrophic bacteria was determined by the most probable number method (MPN and the isolated bacteria were characterized phenotipically. The concentration of bacteria found in forage palm roots ranged from 0.36 x 104 to 109.89 104 cells per gram of root, with highest occurrence on the 60 and 80% ETo. In the dendrogram of similarity it was possible to observe the formation of 24 phenotypic groups with 100% similarity. All bacteria presented similarity superior to 40%. Among these groups, 14 are rare groups, formed by only a single bacterial isolate. In the Semi-Arid conditions, the forage palm that receives the highest amount of saline water, presents a higher density of putative nitrogen-fixing endophytic bacteria with high phenotypic diversity.

  13. Diazotrophic potential among bacterial communities associated with wild and cultivated Agave species.

    Science.gov (United States)

    Desgarennes, Damaris; Garrido, Etzel; Torres-Gomez, Miryam J; Peña-Cabriales, Juan J; Partida-Martinez, Laila P

    2014-12-01

    Agaves are major biotic resources in arid and semi-arid ecosystems. Despite their ecological, economical and cultural relevance, many aspects of the microbial communities associated with agaves are still unknown. Here, we investigated the bacterial communities associated with two Agave species by 16S rRNA- Denaturing gradient gel electrophoresis fingerprinting and sequencing. We also evaluated the effects of biotic and abiotic factors in the structure of the bacterial communities. In parallel, we isolated and characterized diazotrophic bacteria associated with agaves, as Agave soils are characterized by their low nitrogen content. Our results demonstrate that in Agave, the structure of prokaryotic assemblages was mostly influenced by the community group, where the soil, episphere, and endosphere were clearly distinct. Proteobacteria (γ and α), Actinobacteria, and Acidobacteria were the dominant phyla. Bacterial communities in the episphere of agaves were mainly influenced by the host species, whereas in the endosphere were affected by the season. Fifteen bacterial taxa were common and abundant in the endosphere of both Agave species during the dry season. Notably, some of the confirmed diazotrophic strains belonged to this group, suggesting a possible beneficial role in planta. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  14. Toxicity of herbicides used in the sugarcane crop to diazotrophic bacterium Herbaspirillum seropedicae

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    Sergio de Oliveira Procópio

    2014-10-01

    Full Text Available The objective of this work was to identify herbicides used in the sugarcane crop that affects neither the growth, the development, of nor the process of biological nitrogen fixation (BNF by the diazotrophic bacterium Herbaspirillum seropedicae. Eighteen herbicides (paraquat, ametryne, tebuthiuron, amicarbazone, diuron, metribuzin, [hexazinone + diuron], [hexazinone + clomazone], clomazone, isoxaflutole, sulfentrazone, oxyfluorfen, imazapic, imazapyr, [trifloxysulfuron sodium + ametryne], glyphosate, MSMA e 2,4-D were tested in their respective commercial doses regarding their impact on the growth of the bacteria in liquid medium DIGs. For this, we determined the duration of lag phase, generation time and maximum cell density of H. seropedicae, calculated from optical density data obtained at regular intervals during the incubation of cultures for 33 h at 32oC. We also evaluated the impact of herbicides on nitrogenase activity of H. seropedicae grown in semi-solid N-free JNFb medium. The effects of herbicides on the growth variables and the ARA were compared with the untreated control by Dunnett test. A completely randomized design was used. The herbicides paraquat, imazapyr, ametryne, glyphosate and oxyfluorfen inhibited the growth of H. seropedicae in vitro. Ametryne, oxyfluorfen and glyphosate caused a small reduction in the duration of the lag phase of diazotrophic bacteria H. seropedicae. Oxyfluorfen, ametryne and imazapyr resulted in increased the generation time by H. seropedicae. Glyphosate promoted drastic reduction in biological nitrogen fixation in vitro by H. seropedicae. The other tested herbicides did not affect the growth or the same BNF by H. seropedicae.

  15. Differential distribution and abundance of diazotrophic bacterial communities across different soil niches using a gene-targeted clone library approach.

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    Yousuf, Basit; Kumar, Raghawendra; Mishra, Avinash; Jha, Bhavanath

    2014-11-01

    Diazotrophs are key players of the globally important biogeochemical nitrogen cycle, having a significant role in maintaining ecosystem sustainability. Saline soils are pristine and unexplored habitats representing intriguing ecosystems expected to harbour potential diazotrophs capable of adapting in extreme conditions, and these implicated organisms are largely obscure. Differential occurrence of diazotrophs was studied by the nifH gene-targeted clone library approach. Four nifH gene clone libraries were constructed from different soil niches, that is saline soils (low and high salinity; EC 3.8 and 7.1 ds m(-1) ), and agricultural and rhizosphere soil. Additionally, the abundance of diazotrophic community members was assessed using quantitative PCR. Results showed environment-dependent metabolic versatility and the presence of nitrogen-fixing bacteria affiliated with a range of taxa, encompassing members of the Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Gammaproteobacteria, Cyanobacteria and Firmicutes. The analyses unveiled the dominance of Alphaproteobacteria and Gammaproteobacteria (Pseudomonas, Halorhodospira, Ectothiorhodospira, Bradyrhizobium, Agrobacterium, Amorphomonas) as nitrogen fixers in coastal-saline soil ecosystems, and Alphaproteobacteria and Betaproteobacteria (Bradyrhizobium, Azohydromonas, Azospirillum, Ideonella) in agricultural/rhizosphere ecosystems. The results revealed a repertoire of novel nitrogen-fixing bacterial guilds particularly in saline soil ecosystems. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  16. Effect of salinity on nitrogenase activity and composition of the active diazotrophic community in intertidal microbial mats

    NARCIS (Netherlands)

    Severin, I.; Confurius-Guns, V.; Stal, L.J.

    2012-01-01

    Microbial mats are often found in intertidal areas experiencing a large range of salinities. This study investigated the effect of changing salinities on nitrogenase activity and on the composition of the active diazotrophic community ( transcript libraries) of three types of microbial mats situated

  17. Isolation and Screening of Bacteria for Their Diazotrophic Potential and Their Influence on Growth Promotion of Maize Seedlings in Greenhouses.

    Science.gov (United States)

    Kifle, Medhin H; Laing, Mark D

    2015-01-01

    Poor soil fertility is one of the major constraints for crop production. Nitrogen is the most limiting nutrient for increasing crop productivity. Therefore, there is a need to identify diazotrophic inoculants as an alternative or supplement to N-fertilizers for sustainable agriculture. In the current study, a number of free-living diazotrophic bacteria were isolated from soils collected from maize rhizosphere and from leaves and roots of maize within the KwaZulu-Natal Province, Republic of South Africa. Ninety-two isolates were selected for further screening because they were able to grow on N-free media containing different carbon sources. Isolates that were very slow to grow on N-free media were discarded. The isolates were screened in vitro for diazotrophic potential tests for ammonia production and acetylene reduction. Ethylene (C2H4) production was quantified and ranged from 4 to 73 nmoles of C2H4h(-1) culture(-1). The top 20 isolates were re-screened on maize seedlings, and eight isolates significantly (P = 0.001) enhanced some growth parameters of maize above the un-inoculated control. Isolates that showed significant effect on at least two growth parameters were identified at species or genera level. In conclusion, selected diazotrophic isolates may be potentially beneficial but they should be tested more in greenhouse and field conditions with maize to confirm their potential for application as biofertilizers.

  18. Isolation and screening of bacteria for their diazotrophic potential and their influence on growth promotion of maize seedlings in greenhouses

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    Medhin Hadish Kifle

    2016-01-01

    Full Text Available Poor soil fertility is one of the major constraints for crop production. Nitrogen is the most limiting nutrient for increasing crop productivity. Therefore, there is a need to identify diazotrophic inoculants as an alternative or supplement to N-fertilizers for sustainable agriculture. In the current study, a number of free-living diazotrophic bacteria were isolated from soils collected from maize rhizosphere and from leaves and roots of maize within the KwaZulu-Natal Province, Republic of South Africa. Ninety two isolates were selected for further screening because they were able to grow on N-free media containing different carbon sources. Isolates that were very slow to grow on N-free media were discarded. The isolates were screened in vitro for diazotrophic potential tests for ammonia production and acetylene reduction. Ethylene (C2H4 production was quantified and ranged from 4 to 73 nmoles of C2H4 h-1 culture-1. The top twenty isolates were re-screened on maize seedlings, and eight isolates significantly (P=0.001 enhanced some growth parameters of maize above the un-inoculated control. Isolates that showed significant effect on at least two growth parameters were identified at species or genera level. In conclusion, selected diazotrophic isolates may be potentially beneficial but they should be tested more in greenhouse and field conditions with maize to confirm their potential for application as biofertilizers.

  19. Distribution and drivers of symbiotic and free-living diazotrophic cyanobacteria in the western tropical South Pacific

    Science.gov (United States)

    Stenegren, Marcus; Caputo, Andrea; Berg, Carlo; Bonnet, Sophie; Foster, Rachel A.

    2018-03-01

    The abundance and distribution of cyanobacterial diazotrophs were quantified in two regions (Melanesian archipelago, MA; and subtropical gyre, SG) of the western tropical South Pacific using nifH quantitative polymerase chain reaction (qPCR) assays. UCYN-A1 and A2 host populations were quantified using 18S rRNA qPCR assays including one newly developed assay. All phylotypes were detected in the upper photic zone (0-50 m), with higher abundances in the MA region. Trichodesmium and UCYN-B dominated and ranged from 2.18 × 102 to 9.41 × 106 and 1.10 × 102 to 2.78 × 106 nifH copies L-1, respectively. Het-1 (symbiont of Rhizosolenia diatoms) was the next most abundant (1.40 × 101-1.74 × 105 nifH copies L-1) and co-occurred with het-2 and het-3. UCYN-A1 and A2 were the least abundant diazotrophs and were below detection (bd) in 63 and 79, respectively, of 120 samples. In addition, in up to 39 % of samples in which UCYN-A1 and A2 were detected, their respective hosts were bd. Pairwise comparisons of the nifH abundances and various environmental parameters supported two groups: a deep-dwelling group (45 m) comprised of UCYN-A1 and A2 and a surface group (0-15 m) comprised of Trichodesmium, het-1 and het-2. Temperature and photosynthetically active radiation were positively correlated with the surface group, while UCYN-A1 and A2 were positively correlated with depth, salinity, and oxygen. Similarly, in a meta-analysis of 11 external datasets, all diazotrophs, except UCYN-A were correlated with temperature. Combined, our results indicate that conditions favoring the UCYN-A symbiosis differ from those of diatom diazotroph associations and free-living cyanobacterial diazotrophs.

  20. Efecto de la sacarosa en la producción de celulosa por Gluconacetobacter xylinus en cultivo estático

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    Rubén Jaramillo L.

    2012-08-01

    Full Text Available Objetivo. Determinar el efecto de sacarosa en la productividad de BC por Gluconacetobacter xylinus IFO 13693 en condición estática. Materiales y métodos. La síntesis de celulosa bacteriana (BC por Gluconacetobacter xylinus se llevo a cabo en un cultivo estático discontinuo a temperatura ambiente, en presencia de sacarosa como la principal fuente de carbono a concentraciones iniciales de 0.8 a 7.6 % (p/v. Las concentraciones remanentes de BC, sacarosa, glucosa y fructosa se determinaron cada semana. Para la cinética de la hidrólisis de la sacarosa y formación de celulosa y el coeficiente de rendimiento del producto se utilizo el software Microcal Origin 6.0®. Resultados. En la cuarta semana los valores de BC se encontraron entre 32.5 a 39.5 g/L para las diferentes concentraciones de sacarosa. La cinética para la hidrólisis de sacarosa se ajusta al modelo de Michaelis-Menten, con una Vmax de 0.0002 mol L-1 h-1 y Km de 0.018 M. La producción de BC se ajusta al modelo propuesto por Marx-Figini y Pion, con un valor de la pendiente (kc, entre 0.0018 y 0.0024 h-1 para las diferentes concentraciones iniciales de sacarosa. Los coeficientes de rendimiento tienen valores de 0.8 a 2.4 g de BC producida/g de sacarosa consumida. Conclusiones. La hidrólisis de sacarosa, el consumo de glucosa y fructosa se refleja en la síntesis de celulosa. La hidrólisis de sacarosa y la producción de BC se ajustan a los modelos de Michaelis-Menten y al propuesto por Marx-Figini y Pion, respectivamente. Finalmente, el rendimiento depende de la concentración de sacarosa.

  1. Diversity of bacteria nesting the plant cover of north Sinai deserts, Egypt

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    Amira L. Hanna

    2013-01-01

    Full Text Available North Sinai deserts were surveyed for the predominant plant cover and for the culturable bacteria nesting their roots and shoots. Among 43 plant species reported, 13 are perennial (e.g. Fagonia spp., Pancratium spp. and 30 annuals (e.g. Bromus spp., Erodium spp.. Eleven species possessed rhizo-sheath, e.g. Cyperus capitatus, Panicum turgidum and Trisetaria koelerioides. Microbiological analyses demonstrated: the great diversity and richness of associated culturable bacteria, in particular nitrogen-fixing bacteria (diazotrophs; the majority of bacterial residents were of true and/or putative diazotrophic nature; the bacterial populations followed an increasing density gradient towards the root surfaces; sizeable populations were able to reside inside the root (endorhizosphere and shoot (endophyllosphere tissues. Three hundred bacterial isolates were secured from studied spheres. The majority of nitrogen-fixing bacilli isolates belonged to Bacillus megaterium, Bacillus pumilus, Bacillus polymexa, Bacillus macerans, Bacillus circulans and Bacillus licheniformis. The family Enterobacteriaceae represented by Enterobacter agglomerans, Enterobacter sackazakii, Enterobacter cloacae, Serratia adorifera, Serratia liquefaciens and Klebsiella oxytoca. The non-Enterobacteriaceae population was rich in Pantoae spp., Agrobacterium rdiobacter, Pseudomonas vesicularis, Pseudomonas putida, Stenotrophomonas maltophilia, Ochrobactrum anthropi, Sphingomonas paucimobilis and Chrysemonas luteola. Gluconacetobacter diazotrophicus were reported inside root and shoot tissues of a number of tested plants. The dense bacterial populations reported speak well to the very possible significant role played by the endophytic bacterial populations in the survival, in respect of nutrition and health, of existing plants. Such groups of diazotrophs are good candidates, as bio-preparates, to support the growth of future field crops grown in deserts of north Sinai and irrigated by the

  2. Basin scale variability of active diazotrophs and nitrogen fixation in the North Pacific, from the tropics to the subarctic Bering Sea

    Science.gov (United States)

    Shiozaki, Takuhei; Bombar, Deniz; Riemann, Lasse; Hashihama, Fuminori; Takeda, Shigenobu; Yamaguchi, Tamaha; Ehama, Makoto; Hamasaki, Koji; Furuya, Ken

    2017-06-01

    Nitrogen-fixing microorganisms (diazotrophs) provide biologically available nitrogen to plankton communities and thereby greatly influence the productivity in many marine regions. Various cyanobacterial groups have traditionally been considered the major oceanic diazotrophs, but later noncyanobacterial and presumably heterotrophic diazotrophs were also found to be widespread and potentially important in nitrogen fixation. However, the distribution and activity of different diazotroph groups is still poorly constrained for most oceanic ecosystems. Here we examined diazotroph community structure and activity along a 7500 km south-north transect between the central equatorial Pacific and the Bering Sea. Nitrogen fixation contributed up to 84% of new production in the upper waters of the subtropical gyre, where the diazotroph community included the gammaproteobacterium γ-24774A11 and highly active cyanobacterial phylotypes (>50% of total nifH transcript abundance). Nitrogen fixation was sometimes detectable down to 150 m depth and extended horizontally to the edge of the gyre at around 35°N. Nitrogen fixation was even detected far north on the Bering Sea shelf. In the Alaskan Coastal Waters on the Bering Sea shelf, low nitrate together with high dissolved iron concentrations seemed to foster diazotroph growth, including a prominent role of UCYN-A2, which was abundant near the surface (1.2×105 nifH gene copies L-1). Our study provides evidence for nitrogen fixation in the Bering Sea and suggests a clear contrast in the composition of diazotrophs between the tropical/subtropical gyre and the separate waters in the cold northern regions of the North Pacific.

  3. Response of the unicellular diazotrophic cyanobacterium Crocosphaera watsonii to iron limitation.

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    Violaine Jacq

    Full Text Available Iron (Fe is widely suspected as a key controlling factor of N2 fixation due to the high Fe content of nitrogenase and photosynthetic enzymes complex, and to its low concentrations in oceanic surface seawaters. The influence of Fe limitation on the recently discovered unicellular diazotrophic cyanobacteria (UCYN is poorly understood despite their biogeochemical importance in the carbon and nitrogen cycles. To address this knowledge gap, we conducted culture experiments on Crocosphaera watsonii WH8501 growing under a range of dissolved Fe concentrations (from 3.3 to 403 nM. Overall, severe Fe limitation led to significant decreases in growth rate (2.6-fold, C, N and chlorophyll a contents per cell (up to 4.1-fold, N2 and CO2 fixation rates per cell (17- and 7-fold as well as biovolume (2.2-fold. We highlighted a two phased response depending on the degree of limitation: (i under a moderate Fe limitation, the biovolume of C. watsonii was strongly reduced, allowing the cells to keep sufficient energy to maintain an optimal growth, volume-normalized contents and N2 and CO2 fixation rates; (ii with increasing Fe deprivation, biovolume remained unchanged but the entire cell metabolism was affected, as shown by a strong decrease in the growth rate, volume-normalized contents and N2 and CO2 fixation rates. The half-saturation constant for growth of C. watsonii with respect to Fe is twice as low as that of the filamentous Trichodesmium indicating a better adaptation of C. watsonii to poor Fe environments than filamentous diazotrophs. The physiological response of C. watsonii to Fe limitation was different from that previously shown on the UCYN Cyanothece sp, suggesting potential differences in Fe requirements and/or Fe acquisition within the UCYN community. These results contribute to a better understanding of how Fe bioavailability can control the activity of UCYN and explain the biogeography of diverse N2 fixers in ocean.

  4. Non-symbiotic bacterial diazotrophs from of agricultural crops of San Carlos. Córdoba, Colombia

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    Cecilia Lara Mantilla

    2007-07-01

    Full Text Available Diazotrophic Azotobacter sp and Azospirrillum sp bacteria were isolated from the rhizosphere of plantain, (Musa paradisiaca, corn, wheat and fallow areas (uncultivated, but covered by fallen leaves in the San Carlos region (in the Sinú valley in the Córdoba department, Colombia. Microbial populations were identified by biochemical tests; physiological characters were examined using Gram reaction in different mediums: Burk’s, Ashby and Jensen’s (Azotobacter sp and Burk’s, NFB and Congo-red medium (Azospirillum sp. The study was aimed at determining ion ammonium production from Azotobacter sp and Azospirrillum sp isolates; ion ammonium was quantified according to Berthelot’s colorimetric technique (phenol-hypochlorite. This method was modified and standardised in line with using a Perkin-Elmer Lamba 11 UV-Vis spectrometer. As a result of this study, 14 isolates have been shown to be ammonium-producers at concentrations ranging from 0.9 mg/l to 5.2 mg/l; A16PG (5.1545 mg/l and A26M1P (5.1743 mg/l yielded the highest ammonium concentrations for Azotobacter sp and A5M1G (4.6741 mg/l for Azospirrillum sp. Biological N2 fixation (BNF by associative diazotrophic bacteria has contributed towards increasing harvest yield, thereby reducing the need for nitrogenised fertilisers and the emission of greenhouse gases (such as N2O and obtaining economic and environmental benefits for farming.

  5. Nitrogen fixation dynamics of two diazotrophic communities in Mono Lake, California

    Science.gov (United States)

    Oremland, R.S.

    1990-01-01

    Two types of diazotrophic microbial communities were found in the littoral zone of alkaline hypersaline Mono Lake, California. One consisted of anaerobic bacteria inhabiting the flocculent surface layers of sediments. Nitrogen fixation (acetylene reduction) by flocculent surface layers occurred under anaerobic conditions, was not stimulated by light or by additions of organic substrates, and was inhibited by O2, nitrate, and ammonia. The second community consisted of a ball-shaped association of a filamentous chlorophyte (Ctenocladus circinnatus) with diazotrophic, nonheterocystous cyanobacteria, as well as anaerobic bacteria (Ctenocladus balls). Nitrogen fixation by Ctenocladus balls was usually, but not always, stimulated by light. Rates of anaerobic dark fixation equaled those in the light under air. Fixation in the light was stimulated by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and by propanil [N-(3,4-dichlorophenyl)propanamide]. 3-(3,4-Dichlorophenyl)-1,1-dimethyl urea-elicited nitrogenase activity was inhibited by ammonia (96%) and nitrate (65%). Fixation was greatest when Ctenocladus balls were incubated anaerobically in the light with sulfide. Dark anaerobic fixation was not stimulated by organic substrates in short-term (4-h) incubations, but was in long-term (67-h) ones. Areal estimates of benthic N2 fixation were measured seasonally, using chambers. Highest rates (~29.3 ??mol of C2H4 m-2 h-1) occurred under normal diel regimens of light and dark. These estimates indicate that benthic N2 fixation has the potential to be a significant nitrogen source in Mono Lake.

  6. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing.

    Science.gov (United States)

    Li, Ou; Xiao, Rong; Sun, Lihua; Guan, Chenglin; Kong, Dedong; Hu, Xiufang

    2017-01-01

    As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ) to 96.08% (YN), and Delftia (10.39-38.42%), Burkholderia (2.71-15.98%), Escherichia/Shigella (4.90-25.12%), Pseudomonas (2.68-30.72%) and Sphingomonas (1.83-2.05%) dominated in four planting bases. Pseudomonas (17.94-22.06%), Escherichia/Shigella (6.59-11.59%), Delftia (9.65-22.14%) and Burkholderia (3.12-11.05%) dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples.

  7. Effect of diazotrophic bacteria as phosphate solubilizing and indolic compound producers on maize plants

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    Mónica Del Pilar López Ortega

    2013-07-01

    Full Text Available Phosphorus is limiting for growth of maize plants, and because of that use of fertilizers like Rock Phosphate has been proposed. However, direct use of Rock Phosphate is not recommended because of its low availability, so it is necessary to improve it. In this study, a group of diazotrophic bacteria were evaluated as phosphate-solubilizing bacteria, for their production of indolic compounds and for their effects on growth of maize plants. Strains of the genera Azosporillum, Azotobacter, Rhizobium and Klebsiella, were quantitatively evaluated for solubilization of Ca3(PO42 and rock phosphate as a single source of phosphorous in SRS culture media. Additionally, the phosphatase enzyme activity was quantified at pH 5.0, 7.0 and 8.0 using p-nitrophenyl phosphate, and production of indolic compound was determined by colorimetric quantification. The effect of inoculation of bacteria on maize was determined in a completely randomized greenhouse experiment where root and shoot dry weights and phosphorus content were assessed. Results showed that strain C50 produced 107.2 mg .L-1 of available-P after 12 days of fermentation, and AC10 strain had the highest phosphatase activity at pH 8 with 12.7 mg of p-nitrophenol mL .h-1. All strains synthetized indolic compounds, and strain AV5 strain produced the most at 63.03 µg .mL-1. These diazotrophic bacteria increased plant biomass up to 39 % and accumulation of phosphorus by 10%. Hence, use of diazotrphic phosphate-solubilizing bacteria may represent an alternative technology for fertilization systems in maize plants.

  8. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing.

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    Ou Li

    Full Text Available As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ to 96.08% (YN, and Delftia (10.39-38.42%, Burkholderia (2.71-15.98%, Escherichia/Shigella (4.90-25.12%, Pseudomonas (2.68-30.72% and Sphingomonas (1.83-2.05% dominated in four planting bases. Pseudomonas (17.94-22.06%, Escherichia/Shigella (6.59-11.59%, Delftia (9.65-22.14% and Burkholderia (3.12-11.05% dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples.

  9. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing

    Science.gov (United States)

    Li, Ou; Sun, Lihua; Guan, Chenglin; Kong, Dedong

    2017-01-01

    As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ) to 96.08% (YN), and Delftia (10.39–38.42%), Burkholderia (2.71–15.98%), Escherichia/Shigella (4.90–25.12%), Pseudomonas (2.68–30.72%) and Sphingomonas (1.83–2.05%) dominated in four planting bases. Pseudomonas (17.94–22.06%), Escherichia/Shigella (6.59–11.59%), Delftia (9.65–22.14%) and Burkholderia (3.12–11.05%) dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples. PMID:28931073

  10. Permanent Draft Genome of Strain ESFC-1: Ecological Genomics of a Newly Discovered Lineage of Filamentous Diazotrophic Cyanobacteria

    Science.gov (United States)

    Everroad, R. Craig; Stuart, Rhona K.; Bebout, Brad M.; Detweiler, Angela M.; Lee, Jackson Zan; Woebken, Dagmar; Bebout, Leslie E.; Pett-Ridge, Jennifer

    2016-01-01

    The nonheterocystous filamentous cyanobacterium, strain ESFC-1, is a recently described member of the order Oscillatoriales within the Cyanobacteria. ESFC-1 has been shown to be a major diazotroph in the intertidal microbial mat system at Elkhorn Slough, CA, USA. Based on phylogenetic analyses of the 16S RNA gene, ESFC-1 appears to belong to a unique, genus-level divergence; the draft genome sequence of this strain has now been determined. Here we report features of this genome as they relate to the ecological functions and capabilities of strain ESFC-1. The 5,632,035 bp genome sequence encodes 4914 protein-coding genes and 92 RNA genes. One striking feature of this cyanobacterium is the apparent lack of either uptake or bi-directional hydrogenases typically expected within a diazotroph. Additionally, a large genomic island is found that contains numerous low GC-content genes and genes related to extracellular polysaccharide production and cell wall synthesis and maintenance.

  11. Biofilm comprising phototrophic, diazotrophic, and hydrocarbon-utilizing bacteria: a promising consortium in the bioremediation of aquatic hydrocarbon pollutants.

    Science.gov (United States)

    Al-Bader, Dhia; Kansour, Mayada K; Rayan, Rehab; Radwan, Samir S

    2013-05-01

    Biofilms harboring simultaneously anoxygenic and oxygenic phototrophic bacteria, diazotrophic bacteria, and hydrocarbon-utilizing bacteria were established on glass slides suspended in pristine and oily seawater. Via denaturing gradient gel electrophoresis analysis on PCR-amplified rRNA gene sequence fragments from the extracted DNA from biofilms, followed by band amplification, biofilm composition was determined. The biofilms contained anoxygenic phototrophs belonging to alphaproteobacteria; pico- and filamentous cyanobacteria (oxygenic phototrophs); two species of the diazotroph Azospirillum; and two hydrocarbon-utilizing gammaproteobacterial genera, Cycloclasticus and Oleibacter. The coexistence of all these microbial taxa with different physiologies in the biofilm makes the whole community nutritionally self-sufficient and adequately aerated, a condition quite suitable for the microbial biodegradation of aquatic pollutant hydrocarbons.

  12. Comparative study of endophytic and endophytic diazotrophic bacterial communities across rice landraces grown in the highlands of northern Thailand

    OpenAIRE

    Rangjaroen, C.; Rerkasem, B.; Teaumroong, N.; Sungthong, R.; Lumyong, S.

    2014-01-01

    Communities of bacterial endophytes within the rice landraces cultivated in the highlands of northern Thailand were studied using fingerprinting data of 16S rRNA and nifH genes profiling by polymerase chain reaction-denaturing gradient gel electrophoresis. The bacterial communities' richness, diversity index, evenness, and stability were varied depending on the plant tissues, stages of growth, and rice cultivars. These indices for the endophytic diazotrophic bacteria within the landrace rice ...

  13. Genome-wide identification of microRNA and siRNA responsive to endophytic beneficial diazotrophic bacteria in maize.

    Science.gov (United States)

    Thiebaut, Flávia; Rojas, Cristian A; Grativol, Clícia; Motta, Mariana Romeiro; Vieira, Tauan; Regulski, Michael; Martienssen, Robert A; Farinelli, Laurent; Hemerly, Adriana S; Ferreira, Paulo C G

    2014-09-06

    Small RNA (sRNA) has been described as a regulator of gene expression. In order to understand the role of maize sRNA (Zea mays-hybrid UENF 506-8) during association with endophytic nitrogen-fixing bacteria, we analyzed the sRNA regulated by its association with two diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. Deep sequencing analysis was done with RNA extracted from plants inoculated with H. seropedicae, allowing the identification of miRNA and siRNA. A total of 25 conserved miRNA families and 15 novel miRNAs were identified. A dynamic regulation in response to inoculation was also observed. A hypothetical model involving copper-miRNA is proposed, emphasizing the fact that the up-regulation of miR397, miR398, miR408 and miR528, which is followed by inhibition of their targets, can facilitate association with diazotrophic bacteria. Similar expression patterns were observed in samples inoculated with A. brasilense. Moreover, novel miRNA and siRNA were classified in the Transposable Elements (TE) database, and an enrichment of siRNA aligned with TE was observed in the inoculated samples. In addition, an increase in 24-nt siRNA mapping to genes was observed, which was correlated with an increase in methylation of the coding regions and a subsequent reduction in transcription. Our results show that maize has RNA-based silencing mechanisms that can trigger specific responses when plants interact with beneficial endophytic diazotrophic bacteria. Our findings suggest important roles for sRNA regulation in maize, and probably in other plants, during association with diazotrophic bacteria, emphasizing the up-regulation of Cu-miRNA.

  14. Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer.

    Science.gov (United States)

    Singh, Surendra; Datta, Pallavi

    2006-01-01

    Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain.

  15. Comparisons of diazotrophic communities in native and agricultural desert ecosystems reveal plants as important drivers in diversity.

    Science.gov (United States)

    Köberl, Martina; Erlacher, Armin; Ramadan, Elshahat M; El-Arabi, Tarek F; Müller, Henry; Bragina, Anastasia; Berg, Gabriele

    2016-02-01

    Diazotrophs provide the only biological source of fixed atmospheric nitrogen in the biosphere. Although they are the key player for plant-available nitrogen, less is known about their diversity and potential importance in arid ecosystems. We investigated the nitrogenase gene diversity in native and agricultural desert soil as well as within root-associated microbiota of medicinal plants grown in Egypt through the combination of nifH-specific qPCR, fingerprints, amplicon pyrosequencing and fluorescence in situ hybridization-confocal laser scanning microscopy. Although the diazotrophic microbiota were characterized by generally high abundances and diversity, statistically significant differences were found between both soils, the different microhabitats, and between the investigated plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn.). We observed a considerable community shift from desert to agriculturally used soil that demonstrated a higher abundance and diversity in the agro-ecosystem. The endorhiza was characterized by lower abundances and only a subset of species when compared to the rhizosphere. While the microbiomes of the Asteraceae were similar and dominated by potential root-nodulating rhizobia acquired primarily from soil, the perennial S. distichum generally formed associations with free-living nitrogen fixers. These results underline the importance of diazotrophs in desert ecosystems and additionally identify plants as important drivers in functional gene pool diversity. © FEMS 2015.

  16. Characterization of plant-growth promoting diazotrophic bacteria isolated from field grown Chinese cabbage under different fertilization conditions.

    Science.gov (United States)

    Yim, Woo-Jong; Poonguzhali, Selvaraj; Madhaiyan, Munusamy; Palaniappan, Pitchai; Siddikee, M A; Sa, Tongmin

    2009-04-01

    Diazotrophic bacteria isolated from the rhizosphere of Chinese cabbage were assessed for other plant growth promoting characteristics viz., production of IAA, ethylene, ACC deaminase, phosphate solubilization, and gnotobiotic root elongation. Their effect on inoculation to Chinese cabbage was also observed under growth chamber conditions. A total of 19 strains that showed higher nitrogenase activity identified by 16S rRNA gene sequence analysis were found to be the members of the genera Pseudomonas and Agrobacterium belonging to alpha- and gamma-Proteobacteria groups. These strains were also efficient in producing IAA and ACC deaminase though they produced low levels of ethylene and no phosphate solubilization. In addition, inoculation of selected diazotrophic bacterial strains significantly increased seedling length, dry weight, and total nitrogen when compared to uninoculated control. The colonization of crop plants by diazotrophic bacteria can be affected by many biotic and abiotic factors, and further studies are oriented towards investigating the factors that could influence the establishment of a selected bacterial community.

  17. Associative diazotrophic bacteria in grass roots and soils from heavy metal contaminated sites

    Directory of Open Access Journals (Sweden)

    Fátima M.S. Moreira

    2008-12-01

    Full Text Available This work aimed to evaluate density of associative diazotrophic bacteria populations in soil and grass root samples from heavy metal contaminated sites, and to characterize isolates from these populations, both, phenotypically (Zinc, Cadmium and NaCl tolerance in vitro, and protein profiles and genotypically (16S rDNA sequencing, as compared to type strains of known diazotrophic species. Densities were evaluated by using NFb, Fam and JNFb media, commonly used for enrichment cultures of diazotrophic bacteria. Bacterial densities found in soil and grass root samples from contaminated sites were similar to those reported for agricultural soils. Azospirillum spp. isolates from contaminated sites and type strains from non-contaminated sites varied substantially in their in vitro tolerance to Zn+2 and Cd+2, being Cd+2 more toxic than Zn+2. Among the most tolerant isolates (UFLA 1S, 1R, S181, S34 and S22, some (1R, S34 and S22 were more tolerant to heavy metals than rhizobia from tropical and temperate soils. The majority of the isolates tolerant to heavy metals were also tolerant to salt stress as indicated by their ability to grow in solid medium supplemented with 30 g L-1 NaCl. Five isolates exhibited high dissimilarity in protein profiles, and the 16S rDNA sequence analysis of two of them revealed new sequences for Azospirillum.Objetivou-se avaliar a densidade de populações de bactérias diazotróficas associativas em amostras de solos e de raízes de gramíneas oriundas de sítios contaminados com metais pesados, e caracterizar isolados destas populações através da análise fenotípica (tolerância aos metais pesados zinco e cádmio e à NaCl in vitro, perfis protéicos, e genotípica (seqüenciamento de 16S rDNA, comparados às estirpes tipo das mesmas espécies. As densidades foram avaliadas nos meios NFb, Fam e LGI, comumente utilizados para culturas de enriquecimento de populações de bactérias diazotróficas associativas. As densidades

  18. Diversity analysis of diazotrophic bacteria associated with the roots of tea (Camellia sinensis (L.) O. Kuntze).

    Science.gov (United States)

    Gulati, Arvind; Sood, Swati; Rahi, Praveen; Thakur, Rishu; Chauhan, Sunita; Chawla, Isha

    2011-06-01

    The diversity elucidation by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of 96 associative diazotrophs, isolated from the feeder roots of tea on enriched nitrogen-free semisolid media, revealed the predominance of Gram-positive over Gram-negative bacteria within the Kangra valley in Himachal Pradesh, India. The Gram-positive bacteria observed belong to two taxonomic groupings; Firmicutes, including the genera Bacillus and Paenibacillus; and Actinobacteria, represented by the genus Microbacterium. The Gram-negative bacteria included alpha-Proteobacteria genera Brevundimonas, Rhizobium, and Mesorhizobium; gamma-Proteobacteria genera Pseudomonas and Stenotrophomonas; and beta-Proteobacteria genera Azospira, Burkholderia, Delftia, Herbaspirillum and Ralstonia. The low level of similarity of two isolates, with the type strains Paenibacillus xinjiangensis and Mesorhizobium albiziae, suggests the possibility of raising species novum. The bacterial strains of different phylogenetic groups exhibited distinct carbon-source utilization patterns and fatty acid methyl ester profiles. The strains differed in their nitrogenase activities with relatively high activity seen in the Gramnegative strains exhibiting the highest similarity to Azospira oryzae, Delftia lacustris and Herbaspirillum huttiense.

  19. Association with an ammonium-excreting bacterium allows diazotrophic culture of oil-rich eukaryotic microalgae.

    Science.gov (United States)

    Ortiz-Marquez, Juan Cesar Federico; Do Nascimento, Mauro; Dublan, Maria de Los Angeles; Curatti, Leonardo

    2012-04-01

    Concerns regarding the depletion of the world's reserves of oil and global climate change have promoted an intensification of research and development toward the production of biofuels and other alternative sources of energy during the last years. There is currently much interest in developing the technology for third-generation biofuels from microalgal biomass mainly because of its potential for high yields and reduced land use changes in comparison with biofuels derived from plant feedstocks. Regardless of the nature of the feedstock, the use of fertilizers, especially nitrogen, entails a potential economic and environmental drawback for the sustainability of biofuel production. In this work, we have studied the possibility of nitrogen biofertilization by diazotrophic bacteria applied to cultured microalgae as a promising feedstock for next-generation biofuels. We have obtained an Azotobacter vinelandii mutant strain that accumulates several times more ammonium in culture medium than wild-type cells. The ammonium excreted by the mutant cells is bioavailable to promote the growth of nondiazotrophic microalgae. Moreover, this synthetic symbiosis was able to produce an oil-rich microalgal biomass using both carbon and nitrogen from the air. This work provides a proof of concept that artificial symbiosis may be considered an alternative strategy for the low-N-intensive cultivation of microalgae for the sustainable production of next-generation biofuels and other bioproducts.

  20. Endophytic colonization and in planta nitrogen fixation by a diazotrophic Serratia sp. in rice.

    Science.gov (United States)

    Sandhiya, G S; Sugitha, T C K; Balachandar, D; Kumar, K

    2005-09-01

    Nitrogen fixing endophytic Serratia sp. was isolated from rice and characterized. Re-colonization ability of Serratia sp. in the rice seedlings as endophyte was studied under laboratory condition. For detecting the re-colonization potential in the rice seedlings, Serratia sp. was marked with reporter genes (egfp and Kmr) using transposon mutagenesis. The conjugants were screened for re-colonization ability and presence of nif genes using PCR. Further, the influence of flavonoids and growth hormones on the endophytic colonization and in planta nitrogen fixation of Serratia was also investigated. The flavonoids, quercetin (3 microg/ml) and diadzein (2 microg/ml) significantly increased the re-colonization ability of the endophytic Serratia, whereas the growth hormones like IAA and NAA (5 microg/ml) reduced the endophytic colonization ability of Serratia sp. Similarly, the in planta nitrogen fixation by Serratia sp. in rice was significantly increased due to flavonoids. The inoculation of endophytic diazotrophs increased the plant biomass and biochemical constituents.

  1. Genome of Herbaspirillum seropedicae strain SmR1, a specialized diazotrophic endophyte of tropical grasses.

    Science.gov (United States)

    Pedrosa, Fábio O; Monteiro, Rose Adele; Wassem, Roseli; Cruz, Leonardo M; Ayub, Ricardo A; Colauto, Nelson B; Fernandez, Maria Aparecida; Fungaro, Maria Helena P; Grisard, Edmundo C; Hungria, Mariangela; Madeira, Humberto M F; Nodari, Rubens O; Osaku, Clarice A; Petzl-Erler, Maria Luiza; Terenzi, Hernán; Vieira, Luiz G E; Steffens, Maria Berenice R; Weiss, Vinicius A; Pereira, Luiz F P; Almeida, Marina I M; Alves, Lysangela R; Marin, Anelis; Araujo, Luiza Maria; Balsanelli, Eduardo; Baura, Valter A; Chubatsu, Leda S; Faoro, Helisson; Favetti, Augusto; Friedermann, Geraldo; Glienke, Chirlei; Karp, Susan; Kava-Cordeiro, Vanessa; Raittz, Roberto T; Ramos, Humberto J O; Ribeiro, Enilze Maria S F; Rigo, Liu Un; Rocha, Saul N; Schwab, Stefan; Silva, Anilda G; Souza, Eliel M; Tadra-Sfeir, Michelle Z; Torres, Rodrigo A; Dabul, Audrei N G; Soares, Maria Albertina M; Gasques, Luciano S; Gimenes, Ciela C T; Valle, Juliana S; Ciferri, Ricardo R; Correa, Luiz C; Murace, Norma K; Pamphile, João A; Patussi, Eliana Valéria; Prioli, Alberto J; Prioli, Sonia Maria A; Rocha, Carmem Lúcia M S C; Arantes, Olívia Márcia N; Furlaneto, Márcia Cristina; Godoy, Leandro P; Oliveira, Carlos E C; Satori, Daniele; Vilas-Boas, Laurival A; Watanabe, Maria Angélica E; Dambros, Bibiana Paula; Guerra, Miguel P; Mathioni, Sandra Marisa; Santos, Karine Louise; Steindel, Mario; Vernal, Javier; Barcellos, Fernando G; Campo, Rubens J; Chueire, Ligia Maria O; Nicolás, Marisa Fabiana; Pereira-Ferrari, Lilian; Silva, José L da Conceição; Gioppo, Nereida M R; Margarido, Vladimir P; Menck-Soares, Maria Amélia; Pinto, Fabiana Gisele S; Simão, Rita de Cássia G; Takahashi, Elizabete K; Yates, Marshall G; Souza, Emanuel M

    2011-05-01

    The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

  2. Evidence for the endophytic colonization of Phaseolus vulgaris (common bean) roots by the diazotroph Herbaspirillum seropedicae.

    Science.gov (United States)

    Schmidt, M A; Souza, E M; Baura, V; Wassem, R; Yates, M G; Pedrosa, F O; Monteiro, R A

    2011-03-01

    Herbaspirillum seropedicae is an endophytic diazotrophic bacterium, which associates with important agricultural plants. In the present study, we have investigated the attachment to and internal colonization of Phaseolus vulgaris roots by the H. seropedicae wild-type strain SMR1 and by a strain of H. seropedicae expressing a red fluorescent protein (DsRed) to track the bacterium in the plant tissues. Two-day-old P. vulgaris roots were incubated at 30°C for 15 min with 6 x 10(8) CFU/mL H. seropedicae SMR1 or RAM4. Three days after inoculation, 4 x 10(4) cells of endophytic H. seropedicae SMR1 were recovered per gram of fresh root, and 9 days after inoculation the number of endophytes increased to 4 x 10(6) CFU/g. The identity of the recovered bacteria was confirmed by amplification and sequencing of the 16SrRNA gene. Furthermore, confocal microscopy of P. vulgaris roots inoculated with H. seropedicae RAM4 showed that the bacterial cells were attached to the root surface 15 min after inoculation; fluorescent bacteria were visible in the internal tissues after 24 h and were found in the central cylinder after 72 h, showing that H. seropedicae RAM4 is capable of colonizing the roots of the dicotyledon P. vulgaris. Determination of dry weight of common bean inoculated with H. seropedicae SMR1 suggested that this bacterium has a negative effect on the growth of P. vulgaris.

  3. Genome of Herbaspirillum seropedicae strain SmR1, a specialized diazotrophic endophyte of tropical grasses.

    Directory of Open Access Journals (Sweden)

    Fábio O Pedrosa

    2011-05-01

    Full Text Available The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

  4. INOCULATION OF DIAZOTROPHIC BACTERIA AND NITROGEN FERTILIZATION IN TOPDRESSING IN IRRIGATED CORN

    Directory of Open Access Journals (Sweden)

    VANESSA ZIRONDI LONGHINI

    2016-01-01

    Full Text Available Corn is a nitrogen-intensive crop, and the use of management practices such as inoculation of the seed with diazotrophic bacteria, which can maximize crop productivity and reduce the need of nitrogen fertilizers, may result in lower production costs. The present study aimed to evaluate the effect of inoculation of corn seed with Azospirillum brasilense and controlled addition of nitrogen to topdressing on the nutrition, production components, and productivity of crop grain. The experimental design was a randomized block design, with four replications in a 2 × 5 factorial scheme. The treatments consisted of inoculation or not of corn seed with A. brasilense (at 100 mL per 25 kg of seed and five nitrogen (N levels in topdressing (0, 30, 60, 90, and 120 kg N ha-1 from urea [45% N] were applied when the corn was in the phenological growth stage V6. Foliar macronutrients, foliar chlorophyll index (FCI, production components, and yield of corn grain were valuated. Inoculation of corn seeds with A. brasilense increased plant height and grain yield. Fertilization in topdressing, with N levels up to 120 kg ha-1, linearly increased the foliar nutrients and productivity of corn cultivated in the spring/summer in the low-altitude Cerrado region of Brazil.

  5. Dynamics of soil diazotrophic community structure, diversity, and functioning during the cropping period of cotton (Gossypium hirsutum).

    Science.gov (United States)

    Rai, Sandhya; Singh, Dileep Kumar; Annapurna, Kannepalli

    2015-01-01

    The soil sampled at different growth stages along the cropping period of cotton were analyzed using various molecular tools: restriction fragment length polymorphism (RFLP), terminal restriction length polymorphism (T-RFLP), and cloning-sequencing. The cluster analysis of the diazotrophic community structure of early sampled soil (0, 15, and 30 days) was found to be more closely related to each other than the later sampled one. Phylogenetic and diversity analysis of sequences obtained from the first (0 Day; C0) and last soil sample (180 day; C180) confirmed the data. The phylogenetic analysis revealed that C0 was having more unique sequences than C180 (presence of γ-Proteobacteria exclusively in C0). A relatively higher richness of diazotrophic community sequences was observed in C0 (S(ACE) : 30.76; S(Chao1) : 20.94) than C180 (S(ACE) : 18.00; S(Chao1) : 18.00) while the evenness component of Shannon diversity index increased from C0 (0.97) to C180 (1.15). The impact of routine agricultural activities was more evident based on diazotrophic activity (measured by acetylene reduction assay) than its structure and diversity. The nitrogenase activity of C0 (1264.85 ± 35.7 ηmol of ethylene production g(-1) dry soil h(-1) ) was statistically higher when compared to all other values (p structure/diversity and N2 fixation rates. Thus, considerable functional redundancy of nifH was concluded to be existing at the experimental site. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Single and mixed formulations of inoculants with diazotrophic bacteria, under different nitrogen rates and on the paddy rice crop

    Directory of Open Access Journals (Sweden)

    Paula Bianchet

    2013-12-01

    Full Text Available The use of diazotrophic bacteria as a biological input for the production of paddy rice can reduce nitrogen fertilizer applications and contribute to plant development. The use of mixed inoculants’ formulations can increase the efficiency of nitrogen fixation biological process. The objective of this study was to evaluate the effect of single and mixed formulations of inoculants with diazotrophic bacteria on the initial growth of paddy rice plants under different levels of N. The experiment was set in a greenhouse. Treatments consisted of four types of inoculation (no inoculation, inoculation with the isolated AI UDESC 27, inoculation with the isolated FE UDESC 22, and inoculation with the mixed formulation of isolated AI UDESC UDESC 27 and FE UDESC 22; and two levels of mineral nitrogen (30 and 60 mg kg-1 of N. The cultivar used was Epagri 109, which presents late maturity (over 140 days and high yield potential. Treatments were arranged in a factorial design (4 x 2 with five replicates. The experimental design was completely randomized. Inoculation with diazotrophic bacteria reduced by 18% and 26% shoot and root dry matter of rice plants, respectively. Plants also presented lower root area and volume when they were inoculated. There was no significant effect of inoculation and nitrogen rates on the number of leaves and tillers produced per plant or shoot nitrogen accumulation. The results showed that the isolated used in this work were not effective to stimulate shoot and root growth of cv Epagri 109, regardless of formulation type and rate of N.

  7. The Green Berry Consortia of the Sippewissett Salt Marsh: Millimeter-Sized Aggregates of Diazotrophic Unicellular Cyanobacteria.

    Science.gov (United States)

    Wilbanks, Elizabeth G; Salman-Carvalho, Verena; Jaekel, Ulrike; Humphrey, Parris T; Eisen, Jonathan A; Buckley, Daniel H; Zinder, Stephen H

    2017-01-01

    Microbial interactions driving key biogeochemical fluxes often occur within multispecies consortia that form spatially heterogeneous microenvironments. Here, we describe the "green berry" consortia of the Sippewissett salt marsh (Falmouth, MA, United States): millimeter-sized aggregates dominated by an uncultured, diazotrophic unicellular cyanobacterium of the order Chroococcales (termed GB-CYN1). We show that GB-CYN1 is closely related to Crocosphaera watsonii (UCYN-B) and " Candidatus Atelocyanobacterium thalassa" (UCYN-A), two groups of unicellular diazotrophic cyanobacteria that play an important role in marine primary production. Other green berry consortium members include pennate diatoms and putative heterotrophic bacteria from the Alphaproteobacteria and Bacteroidetes . Tight coupling was observed between photosynthetic oxygen production and heterotrophic respiration. When illuminated, the green berries became supersaturated with oxygen. From the metagenome, we observed that GB-CYN1 encodes photosystem II genes and thus has the metabolic potential for oxygen production unlike UCYN-A. In darkness, respiratory activity rapidly depleted oxygen creating anoxia within the aggregates. Metagenomic data revealed a suite of nitrogen fixation genes encoded by GB-CYN1, and nitrogenase activity was confirmed at the whole-aggregate level by acetylene reduction assays. Metagenome reads homologous to marker genes for denitrification were observed and suggest that heterotrophic denitrifiers might co-occur in the green berries, although the physiology and activity of facultative anaerobes in these aggregates remains uncharacterized. Nitrogen fixation in the surface ocean was long thought to be driven by filamentous cyanobacterial aggregates, though recent work has demonstrated the importance of unicellular diazotrophic cyanobacteria (UCYN) from the order Chroococcales. The green berries serve as a useful contrast to studies of open ocean UCYN and may provide a tractable

  8. Estimation of Nitrogenase Enzyme Activities and Plant Growth of Legume and Non-legume Inoculated with Diazotrophic Bacteria

    Directory of Open Access Journals (Sweden)

    Salwani S.

    2012-06-01

    Full Text Available Biological Nitrogen Fixation (BNF process benefits the agriculture sector especially for reducing cost of nitrogenfertilizer. In the process, the diazotrophs convert N2 into ammonia (NH3 which is useable by plants. The BNF process iscatalysed by nitrogenase enzyme that involved protons and electrons together with evolution of H2 therefore, theassessment of N2 fixation is also available via H2 production and electron allocation analysis. Thus, the aims of thisexperiment were to estimate the nitrogenase enzyme activities and observe the influence of diazothrophs on growth oflegume (soybean and non legume (rice plants. Host plants were inoculated with respective inocula; Bradyrhizobiumjaponicum (strain 532C for soybean while Azospirillum brasilense (Sp7 and locally isolated diazotroph (isolate 5 forrice. At harvest, the plants were observed for plant growth parameters, H2 evolution, N2 fixation and electron allocationcoefficient (EAC values. The experiment recorded N2 fixation activities of inoculated soybean plants at 141.2 μmol N2 h-1g-1 dry weight nodule, and the evolution of H2 at 144.4 μmol H2 h-1 g-1 dry weight nodule. The electron allocationcoefficient (EAC of soybean was recorded at 0.982. For inoculated rice plants, none of the observations was successfully recorded. However, results for chlorophyll contents and plant dry weight of both plants inoculated with respective inocula were similar to the control treatments supplied with full nitrogen fertilization (+N. The experiment clearly showed that inoculation of diazotrophic bacteria could enhance growth of the host plants similar to plants treated with nitrogenous fertilizer due to efficient N2 fixation process

  9. Selection of diazotrophic bacteria isolated from wastewater treatment plant sludge at a poultry slaughterhouse for their effect on maize plants

    Directory of Open Access Journals (Sweden)

    Jorge Avelino Rodriguez Lozada

    Full Text Available ABSTRACT The economic and environmental costs of nitrogen fertilization have intensified the search for technologies that reduce mineral fertilization, for example atmospheric nitrogen-fixing (diazotrophic bacteria inoculation. In this context, the present study addressed the isolation and quantification of diazotrophic bacteria in the sludge from treated wastewater of a poultry slaughterhouse; a description of the bacteria, based on cell and colony morphology; and an assessment of growth and N content of maize plants in response to inoculation. Sixteen morphotypes of bacteria were isolated in six N-free culture media (JMV, JMVL, NFb, JNFb, LGI, and LGI-P. The bacteria stained gram-positive, with 10 rod- and six coccoid-shaped isolates. To evaluate the potential of bacteria to promote plant growth, maize seeds were inoculated. The experiment consisted of 17 treatments (control plus 16 bacterial isolates and was carried out in a completely randomized design with six replicates. The experimental units consisted of one pot containing two maize plants in a greenhouse. Forty-five days after planting, the variables plant height, leaf number, stem diameter, root and shoot fresh and dry weight, and N content were measured. The highest values were obtained with isolate UFV L-162, which produced 0.68 g total dry matter per plant and increased N content to 22.14 mg/plant, representing increments of 74 and 133%, respectively, compared with the control. Diazotrophs inhabit sludge from treated wastewater of poultry slaughterhouses and can potentially be used to stimulate plant development and enrich inoculants.

  10. Rates of Dinitrogen Fixation and the Abundance of Diazotrophs in North American Coastal Waters Between Cape Hatteras and Georges Bank

    Science.gov (United States)

    Mulholland, M.R.; Bernhardt, P. W.; Blanco-Garcia, J. L.; Mannino, A.; Hyde, K.; Mondragon, E.; Turk, K.; Moisander, P. H.; Zehr, J. P.

    2012-01-01

    We coupled dinitrogen (N2) fixation rate estimates with molecular biological methods to determine the activity and abundance of diazotrophs in coastal waters along the temperate North American Mid-Atlantic continental shelf during multiple seasons and cruises. Volumetric rates of N2 fixation were as high as 49.8 nmol N L(sup -1) d(sup -1) and areal rates as high as 837.9 micromol N m(sup -2) d(sup -1) in our study area. Our results suggest that N2 fixation occurs at high rates in coastal shelf waters that were previously thought to be unimportant sites of N2 fixation and so were excluded from calculations of pelagic marine N2 fixation. Unicellular N2-fixing group A cyanobacteria were the most abundant diazotrophs in the Atlantic coastal waters and their abundance was comparable to, or higher than, that measured in oceanic regimes where they were discovered. High rates of N2 fixation and the high abundance of diazotrophs along the North American Mid-Atlantic continental shelf highlight the need to revise marine N budgets to include coastal N2 fixation. Integrating areal rates of N2 fixation over the continental shelf area between Cape Hatteras and Nova Scotia, the estimated N2 fixation in this temperate shelf system is about 0.02 Tmol N yr(sup -1), the amount previously calculated for the entire North Atlantic continental shelf. Additional studies should provide spatially, temporally, and seasonally resolved rate estimates from coastal systems to better constrain N inputs via N2 fixation from the neritic zone.

  11. Effects of the Atrazine herbicide over the diazotrophic microorganisms associated with corn in a Saldana soil (Tolima)

    International Nuclear Information System (INIS)

    Ordonez, Alba R; Lozano de Yunda, A; Fuentes, C

    1999-01-01

    In this study, it was evaluated, at greenhouse in soil pots, the effect of the herbicide atrazine in different dose (corresponding to o, 0.75, 1.5 and 3.0 kg of soil hectare) over time (15, 30, 45, 60 and 90 days after the application) on the diazotrophic micro-organisms associated with rhizosphere soil and root of a corn cultivation in a clay loam soil of Saldana - Tolima. It was determined the population of diazotrophic microorganisms in soil by the most probable number technique; also it was observed the characteristic growth in the specific cultivation semisolid Nfb-malic acid medium and was evaluated the nitrogenase activity by means of the acetylene reduction technique. Of the same form, it was observed the characteristic growth and it was evaluated the nitrogenase activity of direct fragment sowings of root in the Nfb-malic acid medium. The 60.8% of soil isolates presented characteristic cultural growth in the form of whitish pellicle under the surface of the medium. in the root 83.3% showed this type of growth. The population of diazotrophic microorganisms of soil did not present significant differences (α = 0.05) between treatments neither over time the nitrogenase activity of soil dilutions demonstrated that there is no a significant effect (α = 0.05) of the treatments (dose) but if there is significant effect of the time. The highest values were presented to 45 days of application of the herbicide, with the application of the highest dose. the nitrogenase activity in the root to what is long of the time was different from rhizosphere soil. The treatment without application of atrazine presented greater values to what is long of the time, however, also in this case were not presented significant differences (α = 0.05) between the treatments (dose) neither to what is long of the time. For so much, as a conclusion, they were not presented effects on the population and the nitrogenase activity of the diazotrophic microorganisms until 90 days of after the

  12. Nitrogenase (nifH gene expression in diazotrophic cyanobacteria in the Tropical North Atlantic in response to nutrient amendments

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    Kendra A Turk-Kubo

    2012-11-01

    Full Text Available The Tropical North Atlantic (TNAtl plays a critical role in the marine nitrogen cycle, as it supports high rates of biological nitrogen (N2 fixation, yet it is unclear whether this process is limited by the availability of iron (Fe, phosphate (P or is co-limited by both. In order to investigate the impact of nutrient limitation on the N2-fixing microorganisms (diazotrophs in the TNAtl, trace metal clean nutrient amendment experiments were conducted, and the expression of nitrogenase (nifH in cyanobacterial diazotrophs in response to the addition of Fe, P, or Fe+P was measured using quantitative PCR. To provide context, N2 fixation rates associated with the <10 μm community and diel nifH expression in natural cyanobacterial populations were measured. In the western TNAtl, nifH expression in Crocosphaera, Trichodesmium, and Richelia was stimulated by Fe and Fe+P additions, but not by P, implying that diazotrophs may be Fe-limited in this region. In the eastern TNAtl, nifH expression in unicellular cyanobacteria UCYN-A and Crocosphaera was stimulated by P, implying P-limitation. In equatorial waters, nifH expression in Trichodesmium was highest in Fe+P treatments, implying co-limitation in this region. Nutrient additions did not measurably stimulate N2 fixation rates in the <10 μm fraction in most of the experiments, even when upregulation of nifH expression was evident. These results demonstrate the utility of using gene expression to investigate the physiological state of natural populations of microorganisms, while underscoring the complexity of nutrient limitation on diazotrophy, and providing evidence that diazotroph populations are slow to respond to the addition of limiting nutrients and may be limited by different nutrients on basin-wide spatial scales. This has important implications for our current understanding of controls on N2 fixation in the TNAtl and may partially explain why it appears to be intermittently limited by Fe, P, or

  13. Analysis of carbohydrate storage granules in the diazotrophic cyanobacterium Cyanothece sp. PCC 7822

    Energy Technology Data Exchange (ETDEWEB)

    Welkie, David G. [Purdue Univ., West Lafayette, IN (United States); Sherman, Debra M. [Purdue Univ., West Lafayette, IN (United States); Chrisler, William B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Orr, Galya [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sherman, Louis A. [Purdue Univ., West Lafayette, IN (United States)

    2013-10-19

    The unicellular diazotrophic cyanobacteria of the genus Cyanothece demonstrate oscillations in nitrogenase activity and H2 production when grown under 12h light-12h dark cycles. We established that Cyanothece sp. PCC 7822 allows for the construction of knock-out mutants and our objective was to improve the growth characteristics of this strain and to identify the nature of the intracellular storage granules. We report the physiological and morphological effects of reduction in nitrate and phosphate concentrations in BG-11 media on this strain. We developed a series of BG-11-derived growth media and monitored batch culture growth, nitrogenase activity and nitrogenase-mediated hydrogen production, culture synchronicity, and intracellular storage content. Reduction in NaNO3 and K2HPO4 concentrations from 17.6 and 0.23 mM to 4.41 and 0.06 mM, respectively, improved growth characteristics such as cell size and uniformity, and enhanced the rate of cell division. Cells grown in this low NP BG-11 were less complex, a parameter that related to the composition of the intracellular storage granules. Cells grown in low NP BG-11 had less polyphosphate, fewer polyhydroxybutyrate granules and many smaller granules became evident. Biochemical analysis and transmission electron microscopy using the histocytochemical PATO technique demonstrated that these small granules contained glycogen. The glycogen levels and the number of granules per cell correlated nicely with a 2.3 to 3.3-fold change from the minimum at L0 to the maximum at D0. The differences in granule morphology and enzymes between Cyanothece ATCC 51142 and Cyanothece PCC 7822 provide insights into the formation of large starch-like granules in some cyanobacteria.

  14. Metabolic engineering of a diazotrophic bacterium improves ammonium release and biofertilization of plants and microalgae.

    Science.gov (United States)

    Ambrosio, Rafael; Ortiz-Marquez, Juan Cesar Federico; Curatti, Leonardo

    2017-03-01

    The biological nitrogen fixation carried out by some Bacteria and Archaea is one of the most attractive alternatives to synthetic nitrogen fertilizers. However, with the exception of the symbiotic rhizobia-legumes system, progress towards a more extensive realization of this goal has been slow. In this study we manipulated the endogenous regulation of both nitrogen fixation and assimilation in the aerobic bacterium Azotobacter vinelandii. Substituting an exogenously inducible promoter for the native promoter of glutamine synthetase produced conditional lethal mutant strains unable to grow diazotrophically in the absence of the inducer. This mutant phenotype could be reverted in a double mutant strain bearing a deletion in the nifL gene that resulted in constitutive expression of nif genes and increased production of ammonium. Under GS non-inducing conditions both the single and the double mutant strains consistently released very high levels of ammonium (>20mM) into the growth medium. The double mutant strain grew and excreted high levels of ammonium under a wider range of concentrations of the inducer than the single mutant strain. Induced mutant cells could be loaded with glutamine synthetase at different levels, which resulted in different patterns of extracellular ammonium accumulation afterwards. Inoculation of the engineered bacteria into a microalgal culture in the absence of sources of C and N other than N 2 and CO 2 from the air, resulted in a strong proliferation of microalgae that was suppressed upon addition of the inducer. Both single and double mutant strains also promoted growth of cucumber plants in the absence of added N-fertilizer, while this property was only marginal in the parental strain. This study provides a simple synthetic genetic circuit that might inspire engineering of optimized inoculants that efficiently channel N 2 from the air into crops. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All

  15. DRIVERS OF THE DYNAMICS OF DIAZOTROPHS AND DENITRIFIERS IN NORTH SEA BOTTOM WATERS AND SEDIMENTS

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    Lucas eStal

    2015-07-01

    Full Text Available The fixation of dinitrogen (N2 and denitrification are two opposite processes in the nitrogen cycle. The former transfers atmospheric dinitrogen gas into bound nitrogen in the biosphere, while the latter returns this bound nitrogen back to atmospheric dinitrogen. It is unclear whether or not these processes are intimately connected in any microbial ecosystem or that they are spatially and/or temporally separated. Here, we measured seafloor nitrogen fixation and denitrification as well as pelagic nitrogen fixation by using the stable isotope technique. Alongside, we measured the diversity, abundance, and activity of nitrogen-fixing and denitrifying microorganisms at three stations in the southern North Sea. Nitrogen fixation ranged from undetectable to 2.4 nmol N L-1 d-1 and from undetectable to 8.2 nmol N g-1 d-1 in the water column and seafloor, respectively. The highest rates were measured in August at Doggersbank, both for the water column and for the seafloor. Denitrification ranged from 1.7 to 208.8 µmol m-2 d-1 and the highest rates were measured in May at the Oyster Grounds. DNA sequence analysis showed sequences of nifH, a structural gene for nitrogenase, related to sequences from anaerobic sulfur/iron reducers and sulfate reducers. Sequences of the structural gene for nitrite reductase, nirS, were related to environmental clones from marine sediments. Quantitative polymerase chain reaction (qPCR data revealed the highest abundance of nifH and nirS genes at the Oyster Grounds. Quantitative reverse transcription polymerase chain reaction (qRT-PCR data revealed the highest nifH expression at Doggersbank and the highest nirS expression at the Oyster Grounds. The distribution of the diazotrophic and denitrifying communities seems to be subject to different selecting factors, leading to spatial and temporal separation of nitrogen fixation and denitrification. These selecting factors include temperature, organic matter availability, and

  16. Endophytic nitrogen fixation in sugarcane: Present knowledge and future applications

    International Nuclear Information System (INIS)

    Boddey, Robert M.; Urquiaga, Segundo; Alves, Bruno J.R.; Reis, Veronica

    2001-01-01

    In Brazil the long-term continuous cultivation of sugarcane with low N fertiliser inputs, without apparent depletion of soil-N reserves, led to the suggestion that N 2 -fixing bacteria associated with the plants may be the source of agronomically significant N inputs to this crop. From the 1950s to 1970s, considerable numbers of N 2 -fixing bacteria were found to be associated with the crop, but it was not until the late 1980s that evidence from N balance and 15 N dilution experiments showed that some Brazilian varieties of sugarcane were able to obtain significant contributions from this source. The results of these studies renewed the efforts to search for N 2 -fixing bacteria, but this time the emphasis was on those diazotrophs that infected the interior of the plants. Within a few years several species of such 'endophytic diazotrophs' were discovered including Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, H. rubrisubalbicans and Burkholderia sp. Work has continued on these endophytes within sugarcane plants, but to date little success has been attained in elucidating which endophyte is responsible for the observed BNF and in what site, or sites, within the cane plants the N 2 fixation mainly occurs. Until such important questions are answered further developments or extension of this novel N 2 -fixing system to other economically important non-legumes (e.g. cereals) will be seriously hindered. As far as application of present knowledge to maximise BNF with sugarcane is concerned, molybdenum is an essential micronutrient. An abundant water supply favours high BNF inputs, and the best medium term strategy to increase BNF would appear to be based on cultivar selection on irrigated N deficient soils fertilised with Mo. (author)

  17. Azolla along a phosphorus gradient: biphasic growth response linked to diazotroph traits and phosphorus-induced iron chlorosis.

    Science.gov (United States)

    Temmink, Ralph J M; Harpenslager, Sarah F; Smolders, Alfons J P; van Dijk, Gijs; Peters, Roy C J H; Lamers, Leon P M; van Kempen, Monique M L

    2018-03-13

    Azolla spp., a water fern often used for phytoremediation, is a strong phosphorus (P) accumulator due to its high growth rate and N 2 fixing symbionts (diazotrophs). It is known that plant growth is stimulated by P, but the nature of the interactive response of both symbionts along a P gradient, and related changes in growth-limiting factors, are unclear. We determined growth, and N and P sequestration rates of Azolla filiculoides in N-free water at different P concentrations. The growth response appeared to be biphasic and highest at levels ≥10 P µmol l -1 . Diazotrophic N sequestration increased upon P addition, and rates were three times higher at high P than at low P. At 10 µmol P l -1 , N sequestration rates reached its maximum and A. filiculoides growth became saturated. Due to luxury consumption, P sequestration rates increased until 50 µmol P l -1 . At higher P concentrations (≥50 µmol l -1 ), however, chlorosis occurred that seems to be caused by iron- (Fe-), and not by N-deficiency. We demonstrate that traits of the complete symbiosis in relation to P and Fe availability determine plant performance, stressing the role of nutrient stoichiometry. The results are discussed regarding Azolla's potential use in a bio-based economy.

  18. Macroalgal blooms favor heterotrophic diazotrophic bacteria in nitrogen-rich and phosphorus-limited coastal surface waters in the Yellow Sea

    Science.gov (United States)

    Zhang, Xiaoli; Song, Yanjing; Liu, Dongyan; Keesing, John K.; Gong, Jun

    2015-09-01

    Macroalgal blooms may lead to dramatic changes in physicochemical variables and biogeochemical cycling in affected waters. However, little is known about the effects of macroalgal blooms on marine bacteria, especially those functioning in nutrient cycles. We measured environmental factors and investigated bacterial diazotrophs in two niches, surface waters that were covered (CC) and non-covered (CF) with massive macroalgal canopies of Ulva prolifera, in the Yellow Sea in the summer of 2011 using real-time PCR and clone library analysis of nifH genes. We found that heterotrophic diazotrophs (Gammaproteobacteria) dominated the communities and were mostly represented by Vibrio-related phylotypes in both CC and CF. Desulfovibrio-related phylotypes were only detected in CC. There were significant differences in community composition in these two environments (p diazotrophic abundance and community composition and that vibrios and Desulfovibrio-related heterotrophic diazotrophs adapt well to the (N-rich but P-limited) environment during blooming. Potential ecological and microbiological mechanisms behind this scenario are discussed.

  19. Visualizing Single Cell Biology: Nanosims Studies of Carbon and Nitrogen Metabolism in Diazotrophic Cyanobacteria

    Science.gov (United States)

    Pett-Ridge, J.; Finzi, J. A.; Capone, D. G.; Popa, R.; Nealson, K. H.; Ng, W.; Spormann, A. M.; Hutcheon, I. D.; Weber, P. K.

    2007-12-01

    Filamentous nitrogen fixing (diazotrophic) cyanobacteria are key players in global nutrient cycling, but the relationship between CO2- and N2-fixation and intercellular exchange of these elements remains poorly understood in many genera. These bacteria are faced with the challenge of isolating regions of N-fixation (O2 inhibited) and photosynthetic (O2 producing) activity. We used isotope labeling in conjunction with a high-resolution isotope and elemental mapping technique (NanoSIMS) to quantitatively describe 13C and 15N uptake and transport in two aquatic cyanobacteria grown on NaH13CO3 and 15N2. The technical challenges of tracing isotopes within individual bacteria can be overcome with high resolution Secondary Ion Mass Spectrometry (NanoSIMS). In NanoSIMS analysis, samples are sputtered with an energetic primary beam (Cs+, O-) liberating secondary ions that are separated by the mass spectrometer and detected in a suite of electron multipliers. Five isotopic species may be analyzed concurrently with spatial resolution as fine as 50nm. A high sensitivity isotope ratio 'map' can then be generated for the analyzed area. Using sequentially harvested cyanobacteria in conjunction with enriched H13CO3 and 15N2 incubations, we measured temporal enrichment patterns that evolve over the course of a day's growth and suggest tightly regulated changes in fixation kinetics. With a combination of TEM, SEM and NanoSIMS analyses, we also mapped the distribution of C, N and Mo (a critical nitrogenase co-factor) isotopes in intact cells. Our results suggest that NanoSIMS mapping of metal enzyme co-factors may be a powerful method of identifying physiological and morphological characteristics within individual bacterial cells, and could be used to provide a 3-dimensional context for more traditional analyses such as immunogold labeling. Finally, we resolved patterns of isotope enrichment at multiple spatial scales: sub-cellular variation, cell-cell differences along filaments

  20. Analysis of diversity of diazotrophic bacteria associated with the rhizosphere of a tropical Arbor, Melastoma malabathricum L.

    Science.gov (United States)

    Sato, Atsuya; Watanabe, Toshihiro; Unno, Yusuke; Purnomo, Erry; Osaki, Mitsuru; Shinano, Takuro

    2009-01-01

    The diversity of diazotrophic bacteria in the rhizosphere of Melastoma malabathricum L. was investigated by cloning-sequencing of the nifH gene directly amplified from DNA extracted from soil. Samples were obtained from the rhizosphere and bulk soil of M. malabathricum growing in three different soil types (acid sulfate, peat and sandy clay soils) located very close to each other in south Kalimantan, Indonesia. Six clone libraries were constructed, generated from bulk and rhizosphere soil samples, and 300 nifH clones were produced, then assembled into 29 operational taxonomic units (OTUs) based on percent identity values. Our results suggested that nifH gene diversity is mainly dependent on soil properties, and did not differ remarkably between the rhizosphere and bulk soil of M. malabathricum except in acid sulfate soil. In acid sulfate soil, as the Shannon diversity index was lower in rhizosphere than in bulk soil, it is suggested that particular bacterial species might accumulate in the rhizosphere.

  1. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

    Directory of Open Access Journals (Sweden)

    L.P.S. Alves

    Full Text Available The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i cell permeabilization, ii Nile red staining, and iii analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99 compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

  2. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry.

    Science.gov (United States)

    Alves, L P S; Almeida, A T; Cruz, L M; Pedrosa, F O; de Souza, E M; Chubatsu, L S; Müller-Santos, M; Valdameri, G

    2017-01-16

    The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

  3. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

    Science.gov (United States)

    Alves, L.P.S.; Almeida, A.T.; Cruz, L.M.; Pedrosa, F.O.; de Souza, E.M.; Chubatsu, L.S.; Müller-Santos, M.; Valdameri, G.

    2017-01-01

    The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots. PMID:28099582

  4. Response of biological soil crust diazotrophs to season, altered summer precipitation and year-round increased temperature in an arid grassland of the Colorado Plateau, USA

    Directory of Open Access Journals (Sweden)

    Chris M Yeager

    2012-10-01

    Full Text Available Biological soil crusts (biocrusts, which supply significant amounts of fixed nitrogen into terrestrial ecosystems worldwide (~33 Tg y-1, are likely to respond to changes in temperature and precipitation associated with climate change. Using nifH gene-based surveys, we explored variation in the diazotrophic community of biocrusts of the Colorado Plateau, USA in response to season (autumn vs. spring, as well as field manipulations that increased the frequency of small-volume precipitation events and year-round soil temperature. Abundance of nifH genes in biocrusts ranged from 3x106 – 1x108 g-1 soil, and nifH from heterocystous cyanobacteria closely related to Scytonema hyalinum, Spirirestis rafaelensis, and Nostoc commune comprised > 98% of the total. Although there was no apparent seasonal effect on total nifH gene abundance in the biocrusts, T-RFLP analysis revealed a strong seasonal pattern in nifH composition. Spirirestis nifH abundance was estimated to oscillate 1 to >2 orders of magnitude between autumn (low and spring (high. A year-round increase of soil temperature (2 − 3 °C had little effect on the diazotroph community structure over 2 years. Altered summer precipitation had little impact on diazotroph community structure over the first 1.5 years of the study, when natural background patterns across years and seasons superseded any treatment effects. However, after the second summer of treatments, nifH abundance was 2.6 fold lower in biocrusts receiving altered precipitation. Heterocystous cyanobacteria were apparently more resilient to altered precipitation than other cyanobacteria. The results demonstrate that diazotrophic community composition of biocrusts in this semi-arid grassland undergoes strong seasonal shifts and that the abundance of its dominant members decreased in response to more frequent, small-volume precipitation events.

  5. Response of biological soil crust diazotrophs to season, altered summer precipitation, and year-round increased temperature in an arid grassland of the Colorado Plateau, USA

    Science.gov (United States)

    Yeager, Chris M.; Kuske, Cheryl R.; Carney, Travis D.; Johnson, Shannon L.; Ticknor, Lawrence O.; Belnap, Jayne

    2012-01-01

    Biological soil crusts (biocrusts), which supply significant amounts of fixed nitrogen into terrestrial ecosystems worldwide (~33Tg y-1), are likely to respond to changes in temperature and precipitation associated with climate change. Using nifH gene-based surveys, we explored variation in the diazotrophic community of biocrusts of the Colorado Plateau, USA in response to season (autumn vs. spring), as well as field manipulations that increased the frequency of small volume precipitation events and year-round soil temperature. Abundance of nifH genes in biocrusts ranged from 3×106 to 1×8 g-1 soil, and nifH from heterocystous cyanobacteria closely related to Scytonema hyalinum, Spirirestis rafaelensis, and Nostoc commune comprised >98% of the total. Although there was no apparent seasonal effect on total nifH gene abundance in the biocrusts, T-RFLP analysis revealed a strong seasonal pattern in nifH composition. Spirirestis nifH abundance was estimated to oscillate 1 to >2 orders of magnitude between autumn (low) and spring (high). A year-round increase of soil temperature (2–3°C) had little effect on the diazotroph community structure over 2 years. Altered summer precipitation had little impact on diazotroph community structure over the first 1.5years of the study, when natural background patterns across years and seasons superseded any treatment effects. However, after the second summer of treatments, nifH abundance was 2.6-fold lower in biocrusts receiving altered precipitation. Heterocystous cyanobacteria were apparently more resilient to altered precipitation than other cyanobacteria. The results demonstrate that diazotrophic community composition of biocrusts in this semi-arid grassland undergoes strong seasonal shifts and that the abundance of its dominant members decreased in response to more frequent, small volume precipitation events.

  6. On the Relationship Between Hydrogen Saturation in the Tropical Atlantic Ocean and Nitrogen Fixation by the Symbiotic Diazotroph UCYN-A

    Science.gov (United States)

    Moore, R. M.; Grefe, I.; Zorz, J.; Shan, S.; Thompson, K.; Ratten, J.; LaRoche, J.

    2018-04-01

    Dissolved hydrogen measurements were made at high resolution in surface waters along a tropical north Atlantic transect between Guadeloupe and Cape Verde in 2015 (Meteor 116). Parallel water samples acquired to assess the relative abundance of the nifH gene from several types of diazotrophs, indicated that Trichodesmium and UCYN-A were dominant in this region. We show that a high degree of correlation exists between the hydrogen saturations and UCYN-A nifH abundance, and a weak correlation with Trichodesmium. The findings suggest that nitrogen fixation by UCYN-A is a major contributor to hydrogen supersaturations in this region of the ocean. The ratio of hydrogen released to nitrogen fixed has not been determined for this symbiont, but the indications are that it may be high in comparison with the small number of diazotrophs for which the ratio has been measured in laboratory cultures. We speculate that this would be consistent with the diazotroph being an exosymbiont on its haptophyte host. Our high resolution measurements of hydrogen concentrations are capable of illustrating the time and space scales of inferred activity of diazotrophs in near real-time in a way that cannot be achieved by biological sampling and rate measurements requiring incubations with 15N2. Direct measurement of high resolution spatial variability would be relatively challenging through collection and analysis of biological samples by qPCR, and extremely challenging by 15N-uptake techniques, neither of which methods yields real-time data. Nonetheless, determination of fixation rates still firmly depends on the established procedure of incubations in the presence of 15N2.

  7. Biochemical studies of amylase, lipase and protease in Callosobruchus maculatus (Coleoptera: Chrysomelidae) populations fed with Vigna unguiculata grain cultivated with diazotrophic bacteria strains.

    Science.gov (United States)

    Silva, L B; Torres, É B; Nóbrega, R A S; Lopes, G N; Vogado, R F; Pavan, B E; Fernandes-Junior, P I

    2017-12-01

    The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: 'BRS Acauã', 'BRS Carijó', 'BRS Pujante', and 'BRS Tapaihum' grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03-11B, 03-84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of 'BRS Carijó' inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

  8. Significance of diazotrophic plant growth-promoting Herbaspirillum sp. GW103 on phytoextraction of Pband Zn by Zea mays L.

    Science.gov (United States)

    Praburaman, Loganathan; Park, Sung-Hee; Cho, Min; Lee, Kui-Jae; Ko, Jeong-Ae; Han, Sang-Sub; Lee, Sang-Hyun; Kamala-Kannan, Seralathan; Oh, Byung-Taek

    2017-01-01

    Microbe-assisted phytoremediation has been considered a promising measure for the remediation of heavy metal-polluted soil. The aim of this study was to assess the effect of diazotrophic plant growth-promoting Herbaspirillum sp. GW103 on growth and lead (Pb) and zinc (Zn) accumulation in Zea mays L. The strain GW103 exhibited plant growth-promoting traits such as indole-3-acetic acid, siderophores, and 1-aminocyclopropane-1-carboxylic deaminase. Treatment of Z. mays L. plants with GW103 significantly increased 19, 31, and 52% of plant biomass and 10, 50, and 126% of chlorophyll a contents in Pb, Zn, and Pb + Zn-amended soils, respectively. Similarly, the strain GW103 significantly increased Pb and Zn accumulation in shoots and roots of Z. mays L., which were 77 and 25% in Pb-amended soil, 42 and 73% in Zn-amended soil, and 27 and 84% in Pb + Zn-amended soil. Furthermore, addition of GW103 increased 8, 12, and 7% of total protein content, catalase, and superoxide dismutase levels, respectively, in Z. mays L. plants. The results pointed out that isolate GW103 could potentially reduce the phytotoxicity of metals and increase Pb and Zn accumulation in Z. mays L. plant.

  9. Effect of salinity on diazotrophic activity and microbial composition of phototrophic communities from Bitter-1 soda lake (Kulunda Steppe, Russia).

    Science.gov (United States)

    Namsaraev, Zorigto; Samylina, Olga; Sukhacheva, Marina; Borisenko, Gennadii; Sorokin, Dimitry Y; Tourova, Tatiana

    2018-04-16

    Bitter-1 is a shallow hypersaline soda lake in Kulunda Steppe (Altai region, Russia). During a study period between 2005 and 2016, the salinity in the littoral area of the lake fluctuated within the range from 85 to 400 g/L (in July of each year). Light-dependent nitrogen fixation occurred in this lake up to the salt-saturating conditions. The rates increased with a decrease in salinity, both under environmental conditions and in laboratory simulations. The salinities below 100 g/L were favorable for light-dependent nitrogen fixation, while the process was dramatically inhibited above 200 g/L salts. The analysis of nifH genes in environmental samples and in enrichment cultures of diazotrophic phototrophs suggested that anaerobic fermenting and sulfate-reducing bacteria could participate in the dark nitrogen fixation process up to soda-saturating conditions. However, we cannot exclude the possibility that haloalkaliphilic nonheterocystous cyanobacteria (Euhalothece sp. and Geitlerinema sp.) and anoxygenic purple sulfur bacteria (Ectothiorhodospira sp.) might also play a role in the process at light conditions. The heterocystous cyanobacterium Nodularia sp. develops at low salinity (below 80 g/L) that is not characteristic for Bitter-1 Lake and thus does not make a significant contribution to the nitrogen fixation in this lake.

  10. Analysis of a diverse assemblage of diazotrophic bacteria from Spartina alterniflora using DGGE and clone library screening.

    Science.gov (United States)

    Lovell, Charles R; Decker, Peter V; Bagwell, Christopher E; Thompson, Shelly; Matsui, George Y

    2008-05-01

    Methods to assess the diversity of the diazotroph assemblage in the rhizosphere of the salt marsh cordgrass, Spartina alterniflora were examined. The effectiveness of nifH PCR-denaturing gradient gel electrophoresis (DGGE) was compared to that of nifH clone library analysis. Seventeen DGGE gel bands were sequenced and yielded 58 nonidentical nifH sequences from a total of 67 sequences determined. A clone library constructed using the GC-clamp nifH primers that were employed in the PCR-DGGE (designated the GC-Library) yielded 83 nonidentical sequences from a total of 257 nifH sequences. A second library constructed using an alternate set of nifH primers (N-Library) yielded 83 nonidentical sequences from a total of 138 nifH sequences. Rarefaction curves for the libraries did not reach saturation, although the GC-Library curve was substantially dampened and appeared to be closer to saturation than the N-Library curve. Phylogenetic analyses showed that DGGE gel band sequencing recovered nifH sequences that were frequently sampled in the GC-Library, as well as sequences that were infrequently sampled, and provided a species composition assessment that was robust, efficient, and relatively inexpensive to obtain. Further, the DGGE method permits a large number of samples to be examined for differences in banding patterns, after which bands of interest can be sampled for sequence determination.

  11. Potential for hydrogen-oxidizing chemolithoautotrophic and diazotrophic populations to initiate biofilm formation in oligotrophic, deep terrestrial subsurface waters.

    Science.gov (United States)

    Wu, Xiaofen; Pedersen, Karsten; Edlund, Johanna; Eriksson, Lena; Åström, Mats; Andersson, Anders F; Bertilsson, Stefan; Dopson, Mark

    2017-03-23

    Deep terrestrial biosphere waters are separated from the light-driven surface by the time required to percolate to the subsurface. Despite biofilms being the dominant form of microbial life in many natural environments, they have received little attention in the oligotrophic and anaerobic waters found in deep bedrock fractures. This study is the first to use community DNA sequencing to describe biofilm formation under in situ conditions in the deep terrestrial biosphere. In this study, flow cells were attached to boreholes containing either "modern marine" or "old saline" waters of different origin and degree of isolation from the light-driven surface of the earth. Using 16S rRNA gene sequencing, we showed that planktonic and attached populations were dissimilar while gene frequencies in the metagenomes suggested that hydrogen-fed, carbon dioxide- and nitrogen-fixing populations were responsible for biofilm formation across the two aquifers. Metagenome analyses further suggested that only a subset of the populations were able to attach and produce an extracellular polysaccharide matrix. Initial biofilm formation is thus likely to be mediated by a few bacterial populations which were similar to Epsilonproteobacteria, Deltaproteobacteria, Betaproteobacteria, Verrucomicrobia, and unclassified bacteria. Populations potentially capable of attaching to a surface and to produce extracellular polysaccharide matrix for attachment were identified in the terrestrial deep biosphere. Our results suggest that the biofilm populations were taxonomically distinct from the planktonic community and were enriched in populations with a chemolithoautotrophic and diazotrophic metabolism coupling hydrogen oxidation to energy conservation under oligotrophic conditions.

  12. Most hydrocarbonoclastic bacteria in the total environment are diazotrophic, which highlights their value in the bioremediation of hydrocarbon contaminants.

    Science.gov (United States)

    Dashti, Narjes; Ali, Nedaa; Eliyas, Mohamed; Khanafer, Majida; Sorkhoh, Naser A; Radwan, Samir S

    2015-01-01

    Eighty-two out of the 100 hydrocarbonoclastic bacterial species that have been already isolated from oil-contaminated Kuwaiti sites, characterized by 16S rRNA nucleotide sequencing, and preserved in our private culture collection, grew successfully in a mineral medium free of any nitrogenous compounds with oil vapor as the sole carbon source. Fifteen out of these 82 species were selected for further study based on the predominance of most of the isolates in their specific sites. All of these species tested positive for nitrogenase using the acetylene reduction reaction. They belonged to the genera Agrobacterium, Sphingomonas, and Pseudomonas from oily desert soil and Nesiotobacter, Nitratireductor, Acinetobacter, Alcanivorax, Arthrobacter, Marinobacter, Pseudoalteromonas, Vibrio, Diatzia, Mycobacterium, and Microbacterium from the Arabian/Persian Gulf water body. A PCR-DGGE-based sequencing analysis of nifH genes revealed the common occurrence of the corresponding genes among all the strains tested. The tested species also grew well and consumed crude oil effectively in NaNO3 -containing medium with and without nitrogen gas in the top space. On the other hand, these bacteria only grew and consumed crude oil in the NaNO3 -free medium when the top space gas contained nitrogen. We concluded that most hydrocarbonoclastic bacteria are diazotrophic, which allows for their wide distribution in the total environment. Therefore, these bacteria are useful for the cost-effective, environmentally friendly bioremediation of hydrocarbon contaminants.

  13. Oil-bioremediation potential of two hydrocarbonoclastic, diazotrophic Marinobacter strains from hypersaline areas along the Arabian Gulf coasts.

    Science.gov (United States)

    Al-Mailem, D M; Eliyas, M; Radwan, S S

    2013-05-01

    Two halophilic, hydrocarbonoclastics bacteria, Marinobacter sedimentarum and M. flavimaris, with diazotrophic potential occured in hypersaline waters and soils in southern and northern coasts of Kuwait. Their numbers were in the magnitude of 10(3) colony forming units g(-1). The ambient salinity in the hypersaline environments was between 3.2 and 3.5 M NaCl. The partial 16S rRNA gene sequences of the two strains showed, respectively, 99 and 100% similarities to the sequences in the GenBank. The two strains failed to grow in the absence of NaCl, exhibited best growth and hydrocarbon biodegradation in the presence of 1 to 1.5 M NaCl, and still grew and maintained their hydrocarbonoclastic activity at salinities up to 5 M NaCl. Both species utilized Tween 80, a wide range of individual aliphatic hydrocarbons (C9-C40) and the aromatics benzene, biphenyl, phenanthrene, anthracene and naphthalene as sole sources of carbon and energy. Experimental evidence was provided for their nitrogen-fixation potential. The two halophilic Marinobacter strains successfully mineralized crude oil in nutrient media as well as in hypersaline soil and water microcosms without the use of any nitrogen fertilizers.

  14. Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets 'VitóRia'.

    Science.gov (United States)

    Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

    2011-01-01

    The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

  15. Evidence for the endophytic colonization of Phaseolus vulgaris(common bean roots by the diazotroph Herbaspirillum seropedicae

    Directory of Open Access Journals (Sweden)

    M.A. Schmidt

    2011-03-01

    Full Text Available Herbaspirillum seropedicae is an endophytic diazotrophic bacterium, which associates with important agricultural plants. In the present study, we have investigated the attachment to and internal colonization of Phaseolus vulgaris roots by the H. seropedicae wild-type strain SMR1 and by a strain of H. seropedicae expressing a red fluorescent protein (DsRed to track the bacterium in the plant tissues. Two-day-old P. vulgaris roots were incubated at 30°C for 15 min with 6 x 10(8 CFU/mL H. seropedicae SMR1 or RAM4. Three days after inoculation, 4 x 10(4 cells of endophytic H. seropedicae SMR1 were recovered per gram of fresh root, and 9 days after inoculation the number of endophytes increased to 4 x 10(6 CFU/g. The identity of the recovered bacteria was confirmed by amplification and sequencing of the 16SrRNA gene. Furthermore, confocal microscopy of P. vulgaris roots inoculated with H. seropedicae RAM4 showed that the bacterial cells were attached to the root surface 15 min after inoculation; fluorescent bacteria were visible in the internal tissues after 24 h and were found in the central cylinder after 72 h, showing that H. seropedicae RAM4 is capable of colonizing the roots of the dicotyledon P. vulgaris. Determination of dry weight of common bean inoculated with H. seropedicae SMR1 suggested that this bacterium has a negative effect on the growth of P. vulgaris.

  16. Structural interaction between GFP-labeled diazotrophic endophytic bacterium Herbaspirillum seropedicae RAM10 and pineapple plantlets 'Vitória'

    Directory of Open Access Journals (Sweden)

    Lílian Estrela Borges Baldotto

    2011-03-01

    Full Text Available The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

  17. Occurrence of diazotrophic bacteria in Araucaria angustifolia Ocorrência de bactérias diazotróficas em Araucaria angustifolia

    Directory of Open Access Journals (Sweden)

    Rafaela de Fátima Neroni

    2007-06-01

    Full Text Available Araucaria angustifolia is an environmentally threatened tree and the whole biota of the Araucaria Forest should be investigated with the aim of its preservation. Diazotrophic bacteria are extremely important for the maintenance of ecosystems, but they have never been studied in Araucaria Forests. In this study, diazotrophic bacteria were isolated from Araucaria roots and soil, when grown in semi-specific, semi-solid media. The diazotrophic character of some recovered isolates could be confirmed using the acetylene reduction assay. According to their 16S rRNA sequences, most of these isolates belong to the genus Burkholderia.Araucaria angustifolia é uma espécie arbórea ameaçada de extinção e toda a comunidade viva das Florestas de Araucária deve ser estudada com a finalidade de sua preservação. Bactérias diazotróficas apresentam grande importância para a manutenção de ecossistemas, mas sua ocorrência nunca foi pesquisada em araucária. Neste estudo foi demonstrada a associação de bactérias fixadoras de nitrogênio com A. angustifolia. Amostras de solo e raízes de araucária apresentaram a ocorrência de bactérias diazotróficas, quando inoculadas em meios semi-sólidos semi-específícos. O caráter diazotrófico de alguns isolados recuperados foi confirmado através da análise de redução do acetileno. De acordo com a análise das seqüências do 16S rRNA, estes isolados são na sua maioria pertencentes ao gênero Burkholderia.

  18. Studies on the process of attachment of diazotroph alcaligenes faecalis and its Tn5 mutants to rice roots using 15N-labelling technique

    International Nuclear Information System (INIS)

    Fang Xuanjun; Lin Min; You Chongbiao

    1993-09-01

    By using 15 N-labelling technique and Tn5-induced mutants the attachment of associative diazotroph Alcaligenes faecalis to intact rice plants was examined in vitro. Three distinguished modes of attachment of Alcaligenes faecalis: adsorption, anchoring and colonization were proposed by using 15 N-labelling bacterial cells and Tn5-induced mutants. Che - mutants affected on adsorption, but not on anchoring. Exo - Che - mutant is defective in both adsorption and anchoring. Exo - or exo ++ mutants are only defective in anchoring. Effective colonization is benefit for establishment on the associative system. The data also indicated that EPS (exopolysaccharide) play rather important roles in the association between the host plant and bacteria

  19. Growth Kinetics of Diazotrophic Bacillus sphaericus UPMB10 Cultured Using Different Types and Concentrations of Carbon and Nitrogen Sources

    Directory of Open Access Journals (Sweden)

    Ooi, T. C.

    2008-01-01

    Full Text Available Growth kinetics of newly isolated diazotrophic Bacillus sphaericus UPMB10 grown in various carbon (lactate, acetate, glycerol, malate, fructose, xylose and sucrose and nitrogen (glutamate, yeast extract, arginine, hystadine, glycine, polypeptone, tryptophan, lysine, NH4Cl and urea sources was investigated using 2 L stirred tank fermenter. The highest growth was obtained in a medium containing lactate as a carbon source, which gave the highest maximum cell concentration of 2.30 g/L, which is corresponding to maximum viable cell count of 4.60 x 10^9 cfu/mL. However, the highest cell yield (1.06 g cell/g carbon consumed was obtained in cultivation using glycerol though slightly lower maximum viable cell count was obtained (3.22 x 10^9 cfu/mL. In addition, cost for the production of live cell using glycerol was about 15 times lower than the cost using lactate. Growth performance of this bacterium when yeast extract was used as a nitrogen source was comparable to the use of pure amino acid. The medium containing 1.8 g/L glycerol and 2 g/L yeast extract was suggested as optimal for growth of this bacterium, which gave carbon to nitrogen ratio (C/N of 10:1. The maximum viable cell count obtained in cultivation using optimised medium in 2 L stirred tank fermenter was 3.34 x 10^9 cfu/mL and the cells maintained its capacity for N2 fixation at 18 nmol C2H2/h.mL.

  20. Transcriptional analysis of the unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC 51142 grown under short day/night cycles

    Energy Technology Data Exchange (ETDEWEB)

    Toepel, Jorg; McDermott, Jason E.; Summerfield, Tina; Sherman, Louis A.

    2009-06-01

    Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium that demonstrates extensive metabolic periodicities of photosynthesis, respiration and nitrogen fixation when grown under N2-fixing conditions. We have performed a global transcription analysis of this organism using 6 h light/dark cycles in order to determine the response of the cell to these conditions and to differentiate between diurnal and circadian regulated genes. In addition, we used a context-likelihood of relatedness (CLR) analysis with this data and those from two-day light/dark and light-dark plus continuous light experiments to better differentiate between diurnal and circadian regulated genes. Cyanothece sp. adapted in several ways to growth under short light/dark conditions. Nitrogen was fixed in every second dark period and only once in each 24 h period. Nitrogen fixation was strongly correlated to the energy status of the cells and glycogen breakdown and high respiration rates were necessary to provide appropriate energy and anoxic conditions for this process. We conclude that glycogen breakdown is a key regulatory step within these complex processes. Our results demonstrated that the main metabolic genes involved in photosynthesis, respiration, nitrogen fixation and central carbohydrate metabolism have strong (or total) circadian-regulated components. The short light/dark cycles enable us to identify transcriptional differences among the family of psbA genes, as well as the differing patterns of the hup genes, which follow the same pattern as nitrogenase genes, relative to the hox genes which displayed a diurnal, dark-dependent gene expression.

  1. Nice to meet you: genetic, epigenetic and metabolic controls of plant perception of beneficial associative and endophytic diazotrophic bacteria in non-leguminous plants.

    Science.gov (United States)

    Carvalho, T L G; Ballesteros, H G F; Thiebaut, F; Ferreira, P C G; Hemerly, A S

    2016-04-01

    A wide range of rhizosphere diazotrophic bacteria are able to establish beneficial associations with plants, being able to associate to root surfaces or even endophytically colonize plant tissues. In common, both associative and endophytic types of colonization can result in beneficial outcomes to the plant leading to plant growth promotion, as well as increase in tolerance against biotic and abiotic stresses. An intriguing question in such associations is how plant cell surface perceives signals from other living organisms, thus sorting pathogens from beneficial ones, to transduce this information and activate proper responses that will finally culminate in plant adaptations to optimize their growth rates. This review focuses on the recent advances in the understanding of genetic and epigenetic controls of plant-bacteria signaling and recognition during beneficial associations with associative and endophytic diazotrophic bacteria. Finally, we propose that "soil-rhizosphere-rhizoplane-endophytes-plant" could be considered as a single coordinated unit with dynamic components that integrate the plant with the environment to generate adaptive responses in plants to improve growth. The homeostasis of the whole system should recruit different levels of regulation, and recognition between the parties in a given environment might be one of the crucial factors coordinating these adaptive plant responses.

  2. Effects of tungsten and titanium oxide nanoparticles on the diazotrophic growth and metals acquisition by Azotobacter vinelandii under molybdenum limiting condition.

    Science.gov (United States)

    Allard, Patrick; Darnajoux, Romain; Phalyvong, Karine; Bellenger, Jean-Philippe

    2013-02-19

    The acquisition of essential metals, such as the metal cofactors (molybdenum (Mo) and iron (Fe)) of the nitrogenase, the enzyme responsible for the reduction of dinitrogen (N(2)) to ammonium, is critical to N(2) fixing bacteria in soil. The release of metal nanoparticles (MNPs) to the environment could be detrimental to N(2) fixing bacteria by introducing a new source of toxic metals and by interfering with the acquisition of essential metals such as Mo. Since Mo has been reported to limit nonsymbiotic N(2) fixation in many ecosystems from tropical to cold temperate, this question is particularly acute in the context of Mo limitation. Using a combination of microbiology and analytical chemistry techniques, we have evaluated the effect of titanium (Ti) and tungsten (W) oxide nanoparticles on the diazotrophic growth and metals acquisition in pure culture of the ubiquitous N(2) fixing bacterium Azotobacter vinelandii under Mo replete and Mo limiting conditions. We report that under our conditions (≤10 mg·L(-1)) TiO(2) NPs have no effects on the diazotrophic growth of A. vinelandii while WO(3) NPs are highly detrimental to the growth especially under Mo limiting conditions. Our results show that the toxicity of WO(3) NPs to A. vinelandii is due to an interference with the catechol-metalophores assisted uptake of Mo.

  3. Seagrass (Zostera marina) Colonization Promotes the Accumulation of Diazotrophic Bacteria and Alters the Relative Abundances of Specific Bacterial Lineages Involved in Benthic Carbon and Sulfur Cycling.

    Science.gov (United States)

    Sun, Feifei; Zhang, Xiaoli; Zhang, Qianqian; Liu, Fanghua; Zhang, Jianping; Gong, Jun

    2015-10-01

    Seagrass colonization changes the chemistry and biogeochemical cycles mediated by microbes in coastal sediments. In this study, we molecularly characterized the diazotrophic assemblages and entire bacterial community in surface sediments of a Zostera marina-colonized coastal lagoon in northern China. Higher nitrogenase gene (nifH) copy numbers were detected in the sediments from the vegetated region than in the sediments from the unvegetated region nearby. The nifH phylotypes detected were mostly affiliated with the Geobacteraceae, Desulfobulbus, Desulfocapsa, and Pseudomonas. Redundancy analysis based on terminal restriction fragment length polymorphism analysis showed that the distribution of nifH genotypes was mostly shaped by the ratio of total organic carbon to total organic nitrogen, the concentration of cadmium in the sediments, and the pH of the overlying water. High-throughput sequencing and phylogenetic analyses of bacterial 16S rRNA genes also indicated the presence of Geobacteraceae and Desulfobulbaceae phylotypes in these samples. A comparison of these results with those of previous studies suggests the prevalence and predominance of iron(III)-reducing Geobacteraceae and sulfate-reducing Desulfobulbaceae diazotrophs in coastal sedimentary environments. Although the entire bacterial community structure was not significantly different between these two niches, Desulfococcus (Deltaproteobacteria) and Anaerolineae (Chloroflexi) presented with much higher proportions in the vegetated sediments, and Flavobacteriaceae (Bacteroidetes) occurred more frequently in the bare sediments. These data suggest that the high bioavailability of organic matter (indicated by relatively lower carbon-to-nitrogen ratios) and the less-reducing anaerobic condition in vegetated sediments may favor Desulfococcus and Anaerolineae lineages, which are potentially important populations in benthic carbon and sulfur cycling in the highly productive seagrass ecosystem. Copyright © 2015

  4. Characterization of plant growth-promoting traits of free-living diazotrophic bacteria and their inoculation effects on growth and nitrogen uptake of crop plants.

    Science.gov (United States)

    Islam, Md Rashedul; Madhaiyan, M; Deka Boruah, Hari P; Yim, Woojong; Lee, Gillseung; Saravanan, V S; Fu, Qingling; Hu, Hongqing; Sa, Tongmin

    2009-10-01

    The search for diverse plant growth-promoting (PGP) diazotrophic bacteria is gaining momentum as efforts are made to exploit them as biofertilizers for various economically important crops. In the present study, 17 diazotrophic strains belonging to eight different genera isolated from rice paddy fields were screened for multiple PGP traits and evaluated for their inoculation effects on canola and rice plants. All of the strains tested positive for 1- aminocyclopropane-1-carboxylate (ACC) deaminase activity and production of indole 3-acetic acid (IAA) and ammonia (NH3). Additionally, four of the strains were able to solubilize phosphorus (P), five tested positive for zinc (Zn) solubilization and sulfur (S) oxidation, and eight strains produced siderophores. Based on the presence of multiple PGP traits, 10 strains were selected for inoculation studies. Treatment with Herbaspirillum sp. RFNB26 resulted in maximum root length (54.3%), seedling vigor, and dry biomass in canola, whereas Paenibacillus sp. RFNB4 exhibited the lowest activity under gnotobiotic conditions. However, under pot culture conditions, Paenibacillus sp. RFNB4 significantly increased plant height and dry biomass production by 42.3% and 29.5%, respectively. Canola plants and rhizosphere soils inoculated with Bacillus sp. RFNB6 exhibited significantly higher nitrogenase activity. In greenhouse experiments, Serratia sp. RFNB18 increased rice plant height by 35.1%, Xanthomonas sp. RFNB24 enhanced biomass production by 84.6%, and rice rhizosphere soils inoculated with Herbaspirillum sp. RFNB26 exhibited the highest nitrogenase activity. Our findings indicate that most of the selected strains possess multiple PGP properties that significantly improve the growth parameters of the two plants when tested under controlled conditions.

  5. Diazotrophic microbial mats

    NARCIS (Netherlands)

    Severin, I.; Stal, L.J.; Seckbach, J.; Oren, A.

    2010-01-01

    Microbial mats have been the focus of scientific research for a few decades. These small-scale ecosystems are examples of versatile benthic communities of microorganisms, usually dominated by phototrophic bacteria (e.g., Krumbein et al., 1977; Jørgensen et al., 1983). They develop as vertically

  6. Analysis of the early heterocyst Cys-proteome in the multicellular cyanobacterium Nostoc punctiforme reveals novel insights into the division of labor within diazotrophic filaments.

    Science.gov (United States)

    Sandh, Gustaf; Ramström, Margareta; Stensjö, Karin

    2014-12-04

    In the filamentous cyanobacterium Nostoc punctiforme ATCC 29133, removal of combined nitrogen induces the differentiation of heterocysts, a cell-type specialized in N2 fixation. The differentiation involves genomic, structural and metabolic adaptations. In cyanobacteria, changes in the availability of carbon and nitrogen have also been linked to redox regulated posttranslational modifications of protein bound thiol groups. We have here employed a thiol targeting strategy to relatively quantify the putative redox proteome in heterocysts as compared to N2-fixing filaments, 24 hours after combined nitrogen depletion. The aim of the study was to expand the coverage of the cell-type specific proteome and metabolic landscape of heterocysts. Here we report the first cell-type specific proteome of newly formed heterocysts, compared to N2-fixing filaments, using the cysteine-specific selective ICAT methodology. The data set defined a good quantitative accuracy of the ICAT reagent in complex protein samples. The relative abundance levels of 511 proteins were determined and 74% showed a cell-type specific differential abundance. The majority of the identified proteins have not previously been quantified at the cell-type specific level. We have in addition analyzed the cell-type specific differential abundance of a large section of proteins quantified in both newly formed and steady-state diazotrophic cultures in N. punctiforme. The results describe a wide distribution of members of the putative redox regulated Cys-proteome in the central metabolism of both vegetative cells and heterocysts of N. punctiforme. The data set broadens our understanding of heterocysts and describes novel proteins involved in heterocyst physiology, including signaling and regulatory proteins as well as a large number of proteins with unknown function. Significant differences in cell-type specific abundance levels were present in the cell-type specific proteomes of newly formed diazotrophic filaments

  7. Higher production of C-phycocyanin by nitrogen-free (diazotrophic) cultivation of Nostoc sp. NK and simplified extraction by dark-cold shock.

    Science.gov (United States)

    Lee, Na Kyeong; Oh, Hee-Mock; Kim, Hee-Sik; Ahn, Chi-Yong

    2017-03-01

    Nostoc sp. NK (KCTC 12772BP) was isolated and cultivated in a BG11 medium and a nitrate-free BG11 medium (BG11 0 ). To enhance C-phycocyanin (C-PC) content in the cells, different fluorescent lamps (white, plant, and red) were used as light sources for complementary chromatic adaptation (CCA). The maximum biomass productivity was 0.42g/L/d and 0.32g/L/d under BG11 and BG11 0 conditions, respectively. The maximum C-PC contents were 8.4% (w/w) under white lamps, 13.6% (w/w) under plant lamps, and 18% (w/w) under BG11 0 and the red light condition. The maximum C-PC productivity was 57.4mg/L/d in BG11 0 under the red lamp condition. These results indicate that a higher C-PC content could be obtained under a diazotrophic condition and a CCA reaction. The C-PC could be released naturally from cells without any extraction processes, when Nostoc sp. NK was cultivated in the BG11 0 medium with CO 2 aeration and put in dark conditions at 5°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Organic matter inoculated with diazotrophic bacterium Beijerinckia indica and Cunninghamella elegans fungus containing chitosan on banana “Williams” in field

    Directory of Open Access Journals (Sweden)

    Newton Pereira Stamford

    2017-01-01

    Full Text Available The production of biofertilizers from rocks increases nutrients for plant nutrition without environmental pollution. The aim of this study was to evaluate the effectiveness of biofertilizers from phosphate and potassium rocks mixed with organic matter (earthworm compound inoculated with free living diazotrophic bacteria (NFB 10001 and Cunninghamella elegans (fungus with chitosan on yield, characteristics, and nutrient uptake of banana (cv. Williams, and attributes of a Red Yellow Argisoil of the rainforest Zone of Pernambuco, Brazil. The experimental design included two biofertilizers: (a PK rock biofertilizers plus organic matter (NPKB and (b bioprotector (NPKP applied at 50, 100 and 150% of the recommended rate for banana, which were compared with soluble mineral fertilizers (NPKF applied at the recommended rate, and earthworm compound (20 ton ha-1. The best results of the plant parameters were obtained with NPKB and NPKP applied at the highest rates (150% RR. A normal yield was produced when NPKB and NPKP were applied at the highest rates and NPKF at the recommended rate. The available P and K in the soil showed a significant fertilization effect, especially when NPKB and NPKP were applied at the highest rates. The biofertilizer and bioprotector may be alternatives to mineral soluble fertilizers.

  9. The Multiple Functions of Common Microbial Carbon Polymers, Glycogen and PHB, during Stress Responses in the Non-Diazotrophic Cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Damrow, Ramon; Maldener, Iris; Zilliges, Yvonne

    2016-01-01

    Classical microbial carbon polymers such as glycogen and polyhydroxybutyrate (PHB) have a crucial impact as both a sink and a reserve under macronutrient stress conditions. Most microbial species exclusively synthesize and degrade either glycogen or PHB. A few bacteria such as the phototrophic model organism Synechocystis sp. PCC 6803 surprisingly produce both physico-chemically different polymers under conditions of high C to N ratios. For the first time, the function and interrelation of both carbon polymers in non-diazotrophic cyanobacteria are analyzed in a comparative physiological study of single- and double-knockout mutants (ΔglgC; ΔphaC; ΔglgC/ΔphaC), respectively. Most of the observed phenotypes are explicitly related to the knockout of glycogen synthesis, highlighting the metabolic, energetic, and structural impact of this process whenever cells switch from an active, photosynthetic 'protein status' to a dormant 'glycogen status'. The carbon flux regulation into glycogen granules is apparently crucial for both phycobilisome degradation and thylakoid layer disassembly in the presence of light. In contrast, PHB synthesis is definitely not involved in this primary acclimation response. Moreover, the very weak interrelations between the two carbon-polymer syntheses indicate that the regulation and role of PHB synthesis in Synechocystis sp. PCC 6803 is different from glycogen synthesis.

  10. Identities of epilithic hydrocarbon-utilizing diazotrophic bacteria from the Arabian Gulf Coasts, and their potential for oil bioremediation without nitrogen supplementation.

    Science.gov (United States)

    Radwan, Samir; Mahmoud, Huda; Khanafer, Majida; Al-Habib, Aamar; Al-Hasan, Redha

    2010-08-01

    Gravel particles from four sites along the Arabian Gulf coast in autumn, winter, and spring were naturally colonized with microbial consortia containing between 7 and 400 × 10(2) cm(-2) of cultivable oil-utilizing bacteria. The 16S rRNA gene sequences of 70 representatives of oil-utilizing bacteria revealed that they were predominantly affiliated with the Gammaproteobacteria and the Actinobacteria. The Gammaproteobacteria comprised among others, the genera Pseudomonas, Pseudoalteromonas, Shewanella, Marinobacter, Psychrobacter, Idiomarina, Alcanivorax, Cobetia, and others. Actinobacteria comprised the genera Dietzia, Kocuria, Isoptericola, Rhodococcus, Microbacterium, and others. In autumn, Firmicutes members were isolated from bay and nonbay stations while Alphaproteobacteria were detected only during winter from Anjefa bay station. Fingerprinting by denaturing gradient gel electrophoresis of amplified 16S rRNA genes of whole microbial consortia confirmed the culture-based bacterial diversities in the various epilithons in various sites and seasons. Most of the representative oil-utilizing bacteria isolated from the epilithons were diazotrophic and could attenuate oil also in nitrogen-rich (7.9-62%) and nitrogen-free (4-54%) cultures, which, makes the microbial consortia suitable for oil bioremediation in situ, without need for nitrogen supplementation. This was confirmed in bench-scale experiments in which unfertilized oily seawater was bioremediated by epilithon-coated gravel particles.

  11. Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets ‘VitóRia’

    Science.gov (United States)

    Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

    2011-01-01

    The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets ‘Vitória’ were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae. PMID:24031612

  12. Effects of the Atrazine herbicide over the diazotrophic microorganisms associated with corn in a Saldana soil (Tolima); Efecto del herbicida Atrazina sobre los microorganismos diazotrofos asociados a un cultivo de maiz en un suelo de Saldana (Tolima)

    Energy Technology Data Exchange (ETDEWEB)

    Ordonez, Alba R; Lozano de Yunda, A; Fuentes, C

    1999-12-01

    In this study, it was evaluated, at greenhouse in soil pots, the effect of the herbicide atrazine in different dose (corresponding to o, 0.75, 1.5 and 3.0 kg of soil hectare) over time (15, 30, 45, 60 and 90 days after the application) on the diazotrophic micro-organisms associated with rhizosphere soil and root of a corn cultivation in a clay loam soil of Saldana - Tolima. It was determined the population of diazotrophic microorganisms in soil by the most probable number technique; also it was observed the characteristic growth in the specific cultivation semisolid Nfb-malic acid medium and was evaluated the nitrogenase activity by means of the acetylene reduction technique. Of the same form, it was observed the characteristic growth and it was evaluated the nitrogenase activity of direct fragment sowings of root in the Nfb-malic acid medium. The 60.8% of soil isolates presented characteristic cultural growth in the form of whitish pellicle under the surface of the medium. in the root 83.3% showed this type of growth. The population of diazotrophic microorganisms of soil did not present significant differences ({alpha} = 0.05) between treatments neither over time the nitrogenase activity of soil dilutions demonstrated that there is no a significant effect ({alpha} = 0.05) of the treatments (dose) but if there is significant effect of the time. The highest values were presented to 45 days of application of the herbicide, with the application of the highest dose. the nitrogenase activity in the root to what is long of the time was different from rhizosphere soil. The treatment without application of atrazine presented greater values to what is long of the time, however, also in this case were not presented significant differences ({alpha} = 0.05) between the treatments (dose) neither to what is long of the time. For so much, as a conclusion, they were not presented effects on the population and the nitrogenase activity of the diazotrophic microorganisms until 90

  13. Tolerância de bactérias diazotróficas simbióticas à salinidade in vitro Tolerance of diazotrophic symbiotic bacteria to salinity

    Directory of Open Access Journals (Sweden)

    Rafaela Simão Abrahão Nóbrega

    2004-08-01

    Full Text Available A busca de estirpes de bactérias diazotróficas simbióticas tolerantes à salinidade, em conjunto com a seleção de hospedeiros, tem por objetivo aumentar o desempenho da simbiose e manter a produtividade vegetal de forma sustentada em condições de estresse salino. Doze estirpes de bactérias diazotróficas de diferentes procedências foram estudadas quanto à tolerância a diferentes concentrações de NaCl em meios de cultura 79 e LB. As estirpes que apresentaram maior tolerância à salinidade (30 g L-1 em meio 79 foram: BR 6806 e BR 4007, ambas de crescimento rápido isoladas de solos do nordeste brasileiro, UFLA 03-51 e UFLA 03-65, de crescimento rápido e UFLA 03-84, de crescimento lento, sendo as três últimas oriundas da região Amazônica. Essas também estiveram entre as mais tolerantes em meio de cultura LB. A estirpe UFLA 03-84, selecionada em trabalhos prévios por sua alta eficiência para caupi (Vigna unguiculata, pode ser indicada para estudos de inoculação, visando a aumentar o rendimento dessa cultura em solos salinos. Os meios de cultura 79 e LB foram eficientes para avaliar a tolerância relativa de rizóbio a NaCl, mas o meio 79 é mais indicado, pois permite o crescimento de todas as estirpes. Pelos resultados, infere-se haver relação entre tolerância in vitro à salinidade e origem dos microssimbiontes, pois as estirpes BR 4007 e BR 6806, oriundas do Ceará, estiveram entre as mais tolerantes.Tolerance to salinity of diazotrophic bacteriaand their host species could increase plant growth and sustainability in salt stressed soils. Twelve diazotrophic bacteria strains were studied regarding their tolerance to different NaCl concentrations in 79 and LB media. More tolerant (30 g L-1 strains in 79 medium were: the fast growers BR 6806 and BR 4007, isolated from northeast Brazil, UFLA 03-51 and UFLA 03-65, and slow growing strain UFLA03-84, being these last three isolated from Amazon region. These strains were among

  14. A Key Marine Diazotroph in a Changing Ocean: The Interacting Effects of Temperature, CO2 and Light on the Growth of Trichodesmium erythraeum IMS101.

    Directory of Open Access Journals (Sweden)

    Tobias G Boatman

    Full Text Available Trichodesmium is a globally important marine diazotroph that accounts for approximately 60 - 80% of marine biological N2 fixation and as such plays a key role in marine N and C cycles. We undertook a comprehensive assessment of how the growth rate of Trichodesmium erythraeum IMS101 was directly affected by the combined interactions of temperature, pCO2 and light intensity. Our key findings were: low pCO2 affected the lower temperature tolerance limit (Tmin but had no effect on the optimum temperature (Topt at which growth was maximal or the maximum temperature tolerance limit (Tmax; low pCO2 had a greater effect on the thermal niche width than low-light; the effect of pCO2 on growth rate was more pronounced at suboptimal temperatures than at supraoptimal temperatures; temperature and light had a stronger effect on the photosynthetic efficiency (Fv/Fm than did CO2; and at Topt, the maximum growth rate increased with increasing CO2, but the initial slope of the growth-irradiance curve was not affected by CO2. In the context of environmental change, our results suggest that the (i nutrient replete growth rate of Trichodesmium IMS101 would have been severely limited by low pCO2 at the last glacial maximum (LGM, (ii future increases in pCO2 will increase growth rates in areas where temperature ranges between Tmin to Topt, but will have negligible effect at temperatures between Topt and Tmax, (iii areal increase of warm surface waters (> 18°C has allowed the geographic range to increase significantly from the LGM to present and that the range will continue to expand to higher latitudes with continued warming, but (iv continued global warming may exclude Trichodesmium spp. from some tropical regions by 2100 where temperature exceeds Topt.

  15. Differential Responses of Dinitrogen Fixation, Diazotrophic Cyanobacteria and Ammonia Oxidation Reveal a Potential Warming-Induced Imbalance of the N-Cycle in Biological Soil Crusts.

    Directory of Open Access Journals (Sweden)

    Xiaobing Zhou

    Full Text Available N2 fixation and ammonia oxidation (AO are the two most important processes in the nitrogen (N cycle of biological soil crusts (BSCs. We studied the short-term response of acetylene reduction assay (ARA rates, an indicator of potential N2 fixation, and AO rates to temperature (T, -5°C to 35°C in BSC of different successional stages along the BSC ecological succession and geographic origin (hot Chihuahuan and cooler Great Basin deserts. ARA in all BSCs increased with T until saturation occurred between 15 and 20°C, and declined at 30-35°C. Culture studies using cyanobacteria isolated from these crusts indicated that the saturating effect was traceable to their inability to grow well diazotrophically within the high temperature range. Below saturation, temperature response was exponential, with Q10 significantly different in the two areas (~ 5 for Great Basin BSCs; 2-3 for Chihuahuan BSCs, but similar between the two successional stages. However, in contrast to ARA, AO showed a steady increase to 30-35°C in Great Basin, and Chihuhuan BSCs showed no inhibition at any tested temperature. The T response of AO also differed significantly between Great Basin (Q10 of 4.5-4.8 and Chihuahuan (Q10 of 2.4-2.6 BSCs, but not between successional stages. Response of ARA rates to T did not differ from that of AO in either desert. Thus, while both processes scaled to T in unison until 20°C, they separated to an increasing degree at higher temperature. As future warming is likely to occur in the regions where BSCs are often the dominant living cover, this predicted decoupling is expected to result in higher proportion of nitrates in soil relative to ammonium. As nitrate is more easily lost as leachate or to be reduced to gaseous forms, this could mean a depletion of soil N over large landscapes globally.

  16. Dynamics of the diazotroph Bacillus polymyxa in the rhizosphere of wheat (Triticum aestivum L.) after inoculation and its effect on uptake of 15N-labelled fertilizer

    International Nuclear Information System (INIS)

    Gouzou, L.; Cheneby, D.; Nicolardot, B.; Heulin, T.

    1995-01-01

    An experiment under glasshouse conditions with a sandy soil was conducted to evaluate the effect of the inoculation of wheat (Triticum aestivum L.) roots with Bacillus polymyxa on N uptake by plants and to investigate the effect of N fertilization on the dynamics of this strain. Different treatments were considered: (i) amended (100 kg N ha −1 ) or not with 15 N labelled ammonium nitrate; (ii) inoculated (2.5 10 7 spores per seed and 0.5 10 9 spores kg −1 soil) or not with B. polymyxa CF43. No promoting effect on wheat growth (dry yield) or N uptake was observed after inoculation under these experimental conditions; 62 per cent of the N fertilizer was assimilated by plants during the first 33 days. Inoculation had no effect on the quantity of soil adhering to the roots. Soluble organic C and inorganic N concentrations were higher in the soil adhering to the roots than in bulk soil. However, the nitrate concentration in soil adhering to the roots was influenced by inoculation at the beginning of plant growth, nitrate supply to the root system being delayed by its probable retention in the rhizosphere. Furthermore, total bacteria in roots, soil adhering to the roots and bulk soil was not affected by N fertilization. The number of B. polymyxa cells measured by an AB-ELISA method increased first in the bulk soil and later in the soil adhering to the roots after inoculation. Fertilization with N decreased the B. polymyxa population in adhering soil, competitive suppression of diazotrophs by non-fixing bacteria probably being induced by the presence of large amounts of nutrients in the rhizosphere when N was added. (author)

  17. Differential responses of dinitrogen fixation, diazotrophic cyanobacteria and ammonia oxidation reveal a potential warming-induced imbalance of the N-cycle in biological soil crusts

    Science.gov (United States)

    Zhou, Xiaobing; Smith, Hilda J.; Giraldo Silva, Ana; Belnap, Jayne; Garcia-Pichel, Ferran

    2017-01-01

    N2 fixation and ammonia oxidation (AO) are the two most important processes in the nitrogen (N) cycle of biological soil crusts (BSCs). We studied the short-term response of acetylene reduction assay (ARA) rates, an indicator of potential N2 fixation, and AO rates to temperature (T, -5°C to 35°C) in BSC of different successional stages along the BSC ecological succession and geographic origin (hot Chihuahuan and cooler Great Basin deserts). ARA in all BSCs increased with T until saturation occurred between 15 and 20°C, and declined at 30–35°C. Culture studies using cyanobacteria isolated from these crusts indicated that the saturating effect was traceable to their inability to grow well diazotrophically within the high temperature range. Below saturation, temperature response was exponential, with Q10 significantly different in the two areas (~ 5 for Great Basin BSCs; 2–3 for Chihuahuan BSCs), but similar between the two successional stages. However, in contrast to ARA, AO showed a steady increase to 30–35°C in Great Basin, and Chihuhuan BSCs showed no inhibition at any tested temperature. The T response of AO also differed significantly between Great Basin (Q10 of 4.5–4.8) and Chihuahuan (Q10 of 2.4–2.6) BSCs, but not between successional stages. Response of ARA rates to T did not differ from that of AO in either desert. Thus, while both processes scaled to T in unison until 20°C, they separated to an increasing degree at higher temperature. As future warming is likely to occur in the regions where BSCs are often the dominant living cover, this predicted decoupling is expected to result in higher proportion of nitrates in soil relative to ammonium. As nitrate is more easily lost as leachate or to be reduced to gaseous forms, this could mean a depletion of soil N over large landscapes globally.

  18. Piecewise Structural Equation Model (SEM) Disentangles the Environmental Conditions Favoring Diatom Diazotroph Associations (DDAs) in the Western Tropical North Atlantic (WTNA).

    Science.gov (United States)

    Stenegren, Marcus; Berg, Carlo; Padilla, Cory C; David, Stefan-Sebastian; Montoya, Joseph P; Yager, Patricia L; Foster, Rachel A

    2017-01-01

    Diatom diazotroph associations (DDAs) are important components in the world's oceans, especially in the western tropical north Atlantic (WTNA), where blooms have a significant impact on carbon and nitrogen cycling. However, drivers of their abundances and distribution patterns remain unknown. Here, we examined abundance and distribution patterns for two DDA populations in relation to the Amazon River (AR) plume in the WTNA. Quantitative PCR assays, targeting two DDAs (het-1 and het-2) by their symbiont's nifH gene, served as input in a piecewise structural equation model (SEM). Collections were made during high (spring 2010) and low (fall 2011) flow discharges of the AR. The distributions of dissolved nutrients, chlorophyll- a , and DDAs showed coherent patterns indicative of areas influenced by the AR. A symbiotic Hemiaulus hauckii-Richelia (het-2) bloom (>10 6 cells L -1 ) occurred during higher discharge of the AR and was coincident with mesohaline to oceanic (30-35) sea surface salinities (SSS), and regions devoid of dissolved inorganic nitrogen (DIN), low concentrations of both DIP (>0.1 μmol L -1 ) and Si (>1.0 μmol L -1 ). The Richelia (het-1) associated with Rhizosolenia was only present in 2010 and at lower densities (10-1.76 × 10 5 nifH copies L -1 ) than het-2 and limited to regions of oceanic SSS (>36). The het-2 symbiont detected in 2011 was associated with H. membranaceus (>10 3 nifH copies L -1 ) and were restricted to regions with mesohaline SSS (31.8-34.3), immeasurable DIN, moderate DIP (0.1-0.60 μmol L -1 ) and higher Si (4.19-22.1 μmol L -1 ). The piecewise SEM identified a profound direct negative effect of turbidity on the het-2 abundance in spring 2010, while DIP and water turbidity had a more positive influence in fall 2011, corroborating our observations of DDAs at subsurface maximas. We also found a striking difference in the influence of salinity on DDA symbionts suggesting a niche differentiation and preferences in oceanic and

  19. Piecewise Structural Equation Model (SEM Disentangles the Environmental Conditions Favoring Diatom Diazotroph Associations (DDAs in the Western Tropical North Atlantic (WTNA

    Directory of Open Access Journals (Sweden)

    Marcus Stenegren

    2017-05-01

    Full Text Available Diatom diazotroph associations (DDAs are important components in the world’s oceans, especially in the western tropical north Atlantic (WTNA, where blooms have a significant impact on carbon and nitrogen cycling. However, drivers of their abundances and distribution patterns remain unknown. Here, we examined abundance and distribution patterns for two DDA populations in relation to the Amazon River (AR plume in the WTNA. Quantitative PCR assays, targeting two DDAs (het-1 and het-2 by their symbiont’s nifH gene, served as input in a piecewise structural equation model (SEM. Collections were made during high (spring 2010 and low (fall 2011 flow discharges of the AR. The distributions of dissolved nutrients, chlorophyll-a, and DDAs showed coherent patterns indicative of areas influenced by the AR. A symbiotic Hemiaulus hauckii-Richelia (het-2 bloom (>106 cells L-1 occurred during higher discharge of the AR and was coincident with mesohaline to oceanic (30–35 sea surface salinities (SSS, and regions devoid of dissolved inorganic nitrogen (DIN, low concentrations of both DIP (>0.1 μmol L-1 and Si (>1.0 μmol L-1. The Richelia (het-1 associated with Rhizosolenia was only present in 2010 and at lower densities (10-1.76 × 105nifH copies L-1 than het-2 and limited to regions of oceanic SSS (>36. The het-2 symbiont detected in 2011 was associated with H. membranaceus (>103nifH copies L-1 and were restricted to regions with mesohaline SSS (31.8–34.3, immeasurable DIN, moderate DIP (0.1–0.60 μmol L-1 and higher Si (4.19–22.1 μmol L-1. The piecewise SEM identified a profound direct negative effect of turbidity on the het-2 abundance in spring 2010, while DIP and water turbidity had a more positive influence in fall 2011, corroborating our observations of DDAs at subsurface maximas. We also found a striking difference in the influence of salinity on DDA symbionts suggesting a niche differentiation and preferences in oceanic and mesohaline

  20. GenBank blastx search result: AK243269 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243269 J100049H19 L41732.4 ABCLSDA Gluconacetobacter diazotrophicus levansucrase precursor (lsd...A), levanase precursor (lsdB), pseudopilin G precursor (lsdG), prepilin peptidase and N-methyl...transferase O (gspO), kinase E (lsdE), LsdF (lsdF), pseudopilin H precursor (lsdH), pseudopilin I precursor (lsd...I), pseudopilin J precursor (lsdJ), LsdL (lsdL), LsdM (lsdM), and LsdN (lsdN)... genes, complete cds; secretin D (lsdD) gene, partial cds; and unknown gene. BCT 6e-26 1 ...

  1. Klebsiella pneumoniae inoculants for enhancing plant growth

    Science.gov (United States)

    Triplett, Eric W [Middleton, WI; Kaeppler, Shawn M [Oregon, WI; Chelius, Marisa K [Greeley, CO

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  2. Diversidade de bactérias diazotróficas endofíticas associadas a plantas de milho Diversity of diazotrophic endophytic bacteria associated with maize plants

    Directory of Open Access Journals (Sweden)

    Luiz Fernando Wurdig Roesch

    2007-12-01

    capazes de colonizar o interior de plantas de milho e que as diferentes condições edafoclimáticas estão correlacionadas com a diversidade dos genes nifH.Endophytic diazotrophic bacteria are capable of promoting maize growth through biological nitrogen fixation (BNF or by the production of plant hormones. The aim of this study was to characterize diversity of endophytic bacteria in maize at sites with different climate and soil conditions in Rio Grande do Sul, Brazil. A PCR-RFLP approach and sequence analysis of nifH Cluster I clone libraries were used to assess diversity in maize plants. The Shannon-Weaver and Equitability indices were calculated to estimate the diazotroph diversity as well as the nucleotide diversity and the average sequence divergence to estimate genetic diversity. To evaluate the variability in populations we performed the F ST test. A greater variation in bacterial communities was detected between rather than within regions, particularly among communities of different soil types and varying water regimes and geographical regions. The Shannon-Weaver index indicated a high difference in terms of diversity of taxonomic units among the communities. The diversity of the communities in the northern region, with higher precipitation and clay content, tended to be higher than that in the south. The Equitability index indicated that there was a dominant taxonomic unit within each community. All clones grouped into nifH gene cluster I. The nifH sequence types from Alpha, Beta and Gammaproteobacteria were recovered. These results demonstrate that there is a large diversity of endophytic nitrogen-fixing bacteria able to colonize maize tissue and that nifH diversity is correlated with the different environmental conditions.

  3. Ocorrência e efeito de bactérias diazotróficas em genótipos de trigo Occurrence and effect of diazotrophic bacteria in wheat genotypes

    Directory of Open Access Journals (Sweden)

    Valéria Marino Rodrigues Sala

    2005-06-01

    Full Text Available A pesquisa sobre bactérias diazotróficas na cultura do trigo tem demonstrado a necessidade de associar bactérias eficientes a genótipos promissores, os quais se beneficiariam dessa associação. Em um experimento com parcelas subdivididas, instalado em condições de campo, em Mococa (SP, empregando os tratamentos: três doses de N (0, 60 e 120 kg ha-1 e três genótipos de trigo (IAC-24, ITD-19 e IAC-355, foi avaliada a ocorrência de microrganismos diazotróficos endofíticos em raízes desinfestadas superficialmente, utilizando-se três meios de cultivo distintos, NFb, JNFb e LGI-P. Somente para o genótipo IAC-355, houve um ajuste linear ascendente da quantidade de bactérias diazotróficas com o aumento na quantidade de N adicionada, apesar de o mesmo genótipo apresentar o menor número de bactérias diazotróficas endofíticas nos três meios de cultivo utilizados para quantificação. Foram obtidos oito isolados bacterianos do meio NFb com as características de Azospirillum e doze do meio JNFb com as características de Herbaspirillum. Esses isolados foram testados "in vitro", nos genótipos dos quais foram originalmente isolados, ou seja, ITD-19 e IAC-24. Todos os isolados testados no genótipo ITD-19 causaram maior crescimento radicular que a testemunha e apenas um isolado do meio JNFb propiciou aumento significativo do N acumulado na parte aérea. A interação planta-bactéria diazotrófica associativa indicou que é possível obter benefícios desta associação.Research on wheat root diazotrophic bacteria has demonstrated the need to associate effective bacteria to promising genotypes, which would benefit from this association. A field experiment was carried out in Mococa County, State of São Paulo, Brazil, using a split-plot design. Treatments consisted of 3 wheat genotypes (IAC-24, ITD-19 and IAC-355 under 3 nitrogen doses (0, 60 and 120 kg ha-1. The occurrence of diazotrophic bacteria was evaluated in three growth media

  4. Resposta de plantas micropropagadas de abacaxizeiro à inoculação de bactérias diazotróficas em casa de vegetação Response of pineapple plantlets to inoculation with diazotrophic bacteria in greenhouse

    Directory of Open Access Journals (Sweden)

    Olmar Baller Weber

    2003-12-01

    Full Text Available O objetivo deste trabalho foi selecionar e avaliar bactérias diazotróficas isoladas de abacaxizeiro (Ananas comosus (L. Merril no desenvolvimento de cultivares micropropagadas da mesma espécie em casa de vegetação. Plantas da cultivar Perolera foram submetidas à inoculação com Asaia bogorensis (AB219 e cultivadas em tubetes, durante 145 dias, com as misturas: casca de arroz carbonizada, folha de carnaubeira triturada e vermicomposto; casca de arroz carbonizada, pó da casca do coco maduro e vermicomposto; casca de arroz carbonizada, vermiculita e vermicomposto. Plantas da cultivar Primavera receberam inóculos com o AB219 e bactérias relacionadas a Burkholderia cepacia (AB202 e AB213, enquanto plantas das cultivares Pérola e Smooth Cayenne receberam AB219 e AB213, sendo cultivadas, por 140 dias, em tubetes com a mistura de vermicomposto e vermiculita. A colonização dos abacaxizeiros pelas bactérias diazotróficas foi confirmada. As plantas da cultivar Perolera cresceram melhor em casca de arroz carbonizada, vermiculita e vermicomposto e responderam positivamente ao AB219. Já as plantas da cultivar Primavera não apresentaram resposta significativa à inoculação com AB219, AB202 e AB213. Houve incremento de 23,1% a 38,5% na matéria seca de raízes das plantas da cultivar Pérola na presença de AB213 e AB219, respectivamente. A presença de AB213 incrementou em 15,2% a matéria seca da parte aérea das plantas da cultivar Smooth Cayenne. Os resultados revelam a eficiência de bactérias diazotróficas na promoção do crescimento de abacaxizeiros.The objective of this work was to select and to evaluate isolates of diazotrophic bacteria associated to pineapple (Ananas comosus (L. Merril plants in the development of micropropagated plantlets of cultivars pineapple in greenhouse. Plantlets from the Perolera cultivar have been submitted to inoculation with Asaia bogorensis (AB219 and were cultivated during 145 days on the following

  5. Recobrimento de sementes de milho com ácidos húmicos e bactérias diazotróficas endofíticas Corn seed coating with humic acids and endophytic diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Patrícia Marluci da Conceição

    2008-04-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito do recobrimento de sementes de milho com ácidos húmicos (AH, bactérias diazotróficas endofíticas e o uso em conjunto de AH e bactérias diazotróficas endofíticas, na estimulação do crescimento vegetal e na população de bactérias estabelecidas na planta hospedeira. A adição de AH, bactérias e o uso em conjunto estimularam o crescimento vegetal. Os AH utilizados no recobrimento de sementes de milho têm menor capacidade de estimular o crescimento radicular, em comparação ao uso em solução. O recobrimento de sementes é uma opção de inoculação de bactérias diazotróficas endofíticas da espécie Herbaspirillum seropedicae (Z67.The objective of this work was to evaluate the effect of seed coating of maize with humic acid (HA, endophytic diazotrophic bacteria, and the combination of both, on plant growth stimulation and bacteria population establishment in roots of inoculated plant host. The addition of HA, bacteria, and the combined use of bacteria and HA stimulated plant growth. Humic acids used in the coated seed formulation show diminished capacity for stimulation of root growth compared with its use in solution. Seed coat is an option for inoculation of endophytic diazotrophic bacteria like Herbaspirillum seropedicae (Z67.

  6. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia.

    Science.gov (United States)

    Kumar, Manoj; van Elsas, Jan Dirk; Nissinen, Riitta

    2017-01-01

    as well as the endosphere communities in the Mayrhofen region were all characterized by high relative abundances of nifH sequences related to Geobacter . In contrast, the endosphere and soil (bulk or rhizosphere soil) communities in the High Arctic were highly divergent: endosphere communities in the arctic regions were shaped by Clostridium spp., while nifH sequences representing δ- Proteobacteria , β- Proteobacteria, Cyanobacteria (in Ny-Ålesund), and Verrucomicrobia (in Kilpisjärvi) dominated the soil communities. Interestingly, the major PNFB genera identified in this study have been previously identified as members of conserved core microbiomes in the endospheres and seeds of these plants by 16S rRNA gene based analyses combined with bacterial isolation, suggesting a very tight interaction between diazotrophic bacteria and these arctic pioneer plants. Overall, anaerobic bacterial taxa dominated the PNFB communities of the endospheres and rhizospheres of the two plant species in all study sites. This could indicate anoxic conditions in and around plant roots at the time of sampling (early growth season), created by melting snow and underlying permafrost.

  7. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia

    Directory of Open Access Journals (Sweden)

    Manoj Kumar

    2017-10-01

    rhizosphere soil as well as the endosphere communities in the Mayrhofen region were all characterized by high relative abundances of nifH sequences related to Geobacter. In contrast, the endosphere and soil (bulk or rhizosphere soil communities in the High Arctic were highly divergent: endosphere communities in the arctic regions were shaped by Clostridium spp., while nifH sequences representing δ-Proteobacteria, β-Proteobacteria, Cyanobacteria (in Ny-Ålesund, and Verrucomicrobia (in Kilpisjärvi dominated the soil communities. Interestingly, the major PNFB genera identified in this study have been previously identified as members of conserved core microbiomes in the endospheres and seeds of these plants by 16S rRNA gene based analyses combined with bacterial isolation, suggesting a very tight interaction between diazotrophic bacteria and these arctic pioneer plants. Overall, anaerobic bacterial taxa dominated the PNFB communities of the endospheres and rhizospheres of the two plant species in all study sites. This could indicate anoxic conditions in and around plant roots at the time of sampling (early growth season, created by melting snow and underlying permafrost.

  8. Ocorrência de bactérias diazotróficas e fungos micorrízicos arbusculares na cultura da mandioca Occurrence of diazotrophic bacteria and arbuscular mycorrhizal fungi on the cassava crop

    Directory of Open Access Journals (Sweden)

    Elcio Liborio Balota

    1999-07-01

    Full Text Available Este trabalho teve como objetivo avaliar a ocorrência, isolar e identificar fungos micorrízicos arbusculares associados à cultura da mandioca (Manihot esculenta. Amostras de solo rizosférico e de várias partes da planta (raízes, tubérculos, manivas e folhas de locais nos Estados do Rio de Janeiro, São Paulo e Paraná, foram inoculadas nos meios LGI-P, NFb-malato e NFb-GOC, avaliando-se o número mais provável de células e a atividade de redução de acetileno. Bactérias diazotróficas foram isoladas de todas as partes da planta, com exceção das folhas, sendo identificadas como Klebsiella sp., Azospirillum lipoferum e uma bactéria denominada "E", provavelmente pertencente ao gênero Burkholderia. A Bactéria E acumulou de 7,63 mg a 14,84 mg de N/g de C em meio semi-sólido, isento de N, e conseguiu manter a capacidade de fixação biológica de N, mesmo após uma dezena de repicagens consecutivas. A colonização micorrízica variou de 31% a 69%, e a densidade de esporos de 10 a 384 esporos/100 mL de solo, predominando as espécies Entrophospora colombiana e Acaulospora scrobiculata no Rio de Janeiro, A. scrobiculata e Scutellospora heterogama no Paraná e em Piracicaba (São Paulo e A. appendicula e S. pellucida em Campinas (São Paulo.This study was performed to evaluate the occurrence and to isolate and identify diazotrophic bacteria and arbuscular mycorrhizal fungi associated with the cassava (Manihot esculenta crop. Samples from rhizospherical soil, roots, tubers, stems and leaves from several localities of the States of Rio de Janeiro, São Paulo and Paraná, in Brazil, were inoculated in three media specific for diazotrophic associative bacteria, LGI-P, NFb-malate and NFb-GOC, evaluating the most probable number of cells and the acetylene-reducing activity. Diazotrophic bacteria were detected in all plant parts except for the leaves, and were identified as Klebsiella sp., Azospirillum lipoferum and a bacterium called "E

  9. Bactérias diazotróficas associadas a coqueiros na região de baixada litorânea em Sergipe Diazotroph bacteria associated to coconut palms in a coastal lowland region in Sergipe State, Brazil

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    Marcelo Ferreira Fernandes

    2001-12-01

    Full Text Available Objetivou-se neste trabalho, isolar, identificar e quantificar bactérias diazotróficas existentes nas raízes e folhas de coqueiro (híbrido PB121 cultivados na baixada litorânea de Sergipe. As populações de bactérias diazotróficas nesses órgãos foram quantificadas pela técnica do número mais provável (NMP em meios semi-sólidos JNFb e NFb, e identificadas por meio de avaliações microscópicas, culturais e de testes bioquímicos dos sistemas API 20 - E e API 20 - NE. O conteúdo de N em meios semi-sólidos NFb e JNFb foi determinado colorimetricamente, após oito dias de incubação das bactérias, a 32ºC, para estimar a capacidade de fixação biológica dos isolados. Enterobactérias pertencentes ao gênero Enterobacter e bactérias com características semelhantes às do gênero Pseudomonas (pseudomonadas foram predominantes entre os isolados obtidos. As enterobactérias e pseudomonadas isoladas de folhas foram predominantemente endofíticas. Quanto às diazotróficas isoladas de raízes, observou-se predominância de enterobactérias na sua superfície, ao passo que as pseudomonadas ocorreram em proporções semelhantes na superfície e no interior desse órgão. Após oito dias de incubação, os conteúdos de N nos meios com inoculação das bactérias pseudomonadas foram maiores do que nos meios com inoculação das enterobactérias.The aim of this work was to isolate, identify and quantify diazotrophic bacteria existing in roots and leaves of coconut palms (hybrid PB121 grown in a coastal lowland of Sergipe, in Brazil. Diazotrophic populations in these organs were quantified by the most probable number (MNP technique on NFb and JNFb semi-solid media and identified by the evaluation of microscopic, cultural and biochemical characteristics of the isolates. Biochemical characteristics were evaluated using the API 20 - E and API 20 - NE galleries. Nitrogen content in NFb and JNFb semi-solid media inoculated with each of the

  10. CARACTERIZAÇÃO FENOTÍPICA DE BACTÉRIAS DIAZOTRÓFICAS ENDOFÍTICAS ISOLADAS DE CANA DE AÇÚCAR PHENOTYPIC CHARACTERIZATION OF ENDOPHYTIC DIAZOTROPHIC BACTERIAS ISOLATED OF SUGARCANE

    Directory of Open Access Journals (Sweden)

    Robson Cavalcante de Lima

    2011-06-01

    Full Text Available A utilização de inoculantes contendo microrganismos diazotróficos na cana-de-açúcar pode contribuir para a promoção do crescimento destas plantas e promover ganhos significativos na produtividade. Objetivou-se no presente trabalho caracterizar bactérias diazotróficas isoladas de colmos cana-de-açúcar. Foram utilizados os meios de cultivo semi-sólidos NFb e JNFb sem adição de nitrogênio. Foram isoladas cinco estirpes nativas de bactérias diazotróficas as quais foram avaliadas microscopicamente pela coloração de Gram e em relação à resistência intrínseca aos antibióticos; fungicida furadan (i.a. carbofuran e inseticida regente (i.a. fipronil, e à capacidade de fixar o nitrogênio em condições de casa-de-vegetação e campo. Os resultados obtidos permitiram observar que todos os isolados apresentaram características de bacilos Gram-negativos, e ampla resistência aos antibióticos. A estirpe nativa UCCBc5 apresentou resistência ao fungicida furadan e ao inseticida regente. As bactérias isoladas UCCBc1 e UCCBc5 apresentam capacidade e eficiência de fixar o nitrogênio atmosférico em condições de casa-de-vegetação. Verificou-se que a estirpe UCCBc5, resistente ao furadan e ao regente apresentou capacidade fixadora do nitrogênio atmosférico, em condições de campo. Na avaliação de doses de inoculante observou-se que existe relação dose de inoculante/ eficiência de fixação de nitrogênio. Estes resultados permitiram afirmar que as bactérias diazotróficas endofíticas podem ser utilizadas, em alguns casos, como substituto da adubação nitrogenada, na cultura de cana-de- açúcar.The use of inoculants containing diazotrophic microorganism in sugarcane can contribute to promoting the growth of these plants and tp promote significant gains in productivity. The objective of this study was to characterize diazotrophic bacteria isolated from stalks of sugarcane. We used the means of a semi-solid NFB and JNFb

  11. Diversidade de bactérias diazotróficas endofíticas dos gêneros Herbaspirillum e Burkholderia na cultura do arroz inundado Diversity of endophytic diazotrophic bacteria of the genus Herbaspirillum and Burkholderia in wetland rice

    Directory of Open Access Journals (Sweden)

    Luciana da Silva Rodrigues

    2006-02-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade de bactérias diazotróficas endofíticas, dos gêneros Herbaspirillum e Burkholderia, em duas variedades de arroz, consideradas de alta (IR 42 e baixa (IAC 4440 eficiência de fixação biológica de nitrogênio. Foram realizados dois experimentos em casa de vegetação, em vasos com dois tipos de solos, provenientes dos Estados de Goiás e do Rio de Janeiro. Foi feita a contagem do número de bactérias e o isolamento em diferentes partes e estágios de desenvolvimento das plantas, mediante o uso de meios de cultivo JNFb e JMV. Os isolados bacterianos foram caracterizados a partir de aspectos morfológicos das colônias, com o crescimento em meios de cultivo, e de testes fisiológicos (uso de fontes de carbono e atividade de redução de acetileno. A contagem revelou grande número de bactérias diazotróficas (10(6 células g-1 matéria fresca, presentes em ambas as variedades de arroz, principalmente nas amostras radiculares. Os dados, obtidos na matriz de similaridade, mostram a presença de representantes da espécie Herbaspirillum seropedicae, bem como a diversidade entre isolados pertencentes ao gênero Burkholderia.The objective of this work was to evaluate the diversity of endophytic diazotrophic bacteria of the genera Herbaspirillum and Burkholderia, in two rice varieties, considered of high (IR 42 and low (IAC 4440 contribution on BNF. Two experiments were conducted in greenhouse conditions, in order to study the association of endophytic diazotrophic bacteria with wetland rice varieties, which were planted in two types of soil: one from Rio de Janeiro State and another from Goiás State, Brazil. Bacterial population (in different parts and physiological stages of the plants were evaluated, followed by the both genera strains isolation using culture media. The isolated bacteria were characterized based on morphological and physiological aspects. High bacterial counts were detected

  12. Strain improvement of Gluconacetobacter xylinus NCIM 2526 for ...

    African Journals Online (AJOL)

    TUOYO

    2010-08-09

    Aug 9, 2010 ... UV mutants GHUV3, GHUV4, and GHUV5 of G. xylinus showed higher cellulose yield than the ... peptone, 0.5% yeast extract, 0.27% disodium phosphate and. 0.115% ... reported mutation induced enhanced lipase production.

  13. 块根块茎类作物内生固氮菌分离鉴定、系统发育与促生特性%Isolation, Identification, Phylogeny and Growth Promoting Characteristics of Endophytic Diazotrophs from Tuber and Root Crops

    Institute of Scientific and Technical Information of China (English)

    李艳星; 郭平毅; 孙建光

    2017-01-01

    [Objective]The objectives of this study are to isolate, identify and analyze phylogenetics of endophytic diazotrophs from tuber and root crops, test plant growth promoting (PGP) characteristics of the isolates, and to explore population property and host distributions of endophytic diazotrophs from tuber and root crops. [Method] Surface sterilization and low nitrogen medium were used to isolate endophytic diazotrophs. nifH detection based on PCR amplification to confirm isolates as nitrogen-fixing bacteria. 16S rRNA was amplified with PCR, blasted in EzTaxon after sequencing, and analyzed with Clustalx-MEGA to make phylogenetic tree. PGP characteristics were evaluated by testing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, indole acetic acid (IAA) production and antagonism to Fusarium spp. [Result] Total 219 endophytic bacterial isolates were obtained from 14 tuber and root samples including radish, carrot, potato, ginger, beet, lotus, yam, taro, cabbage, and sweet potato. All isolates were verified as nitrogen-fixing bacteria after nifH inspection and identified as 79 species of 24 genera based on 16S rRNA. The 79 species are Acinetobacter harbinensis, Arthrobacter arilaitensis, Ar. bergerei, Ar. nicotianae, Ar. nicotinovorans, Ar. nitroguajacolicus, Bacillus amyloliquefaciens, Ba. aryabhattai, Ba. circulans, Ba. fengqiuensis, Ba. firmus, Ba. flexus, Ba. halosaccharovorans, Ba. idriensis, Ba. licheniformis, Ba. litoralis, Ba. luciferensis, Ba. marisflavi, Ba. megaterium, Ba. methylotrophicus, Ba. oceanisediminis, Ba. safensis, Ba. simplex, Ba. sonorensis, Ba. stratosphericus, Ba. subterraneus, Ba. tequilensis, Ba. thaonhiensis, Ba. thioparans, Brevibacillus brevis, Br. formosus, Br. nitrificans, Br. frigoritolerans, Chryseobacterium indoltheticum, Ch. lactis, Citrobacter youngae, Delftia lacustris, Domibacillus indicus, Enterobacter asburiae, E. ludwigii, E. mori, E. xiangfangensis, Fictibacillus barbaricus, Fi. enclensis, Fi. nanhaiensis, Fi

  14. Novas bactérias diazotróficas endofíticas na cultura do trigo em interação com a adubação nitrogenada, no campo Interaction of new diazotrophic endophytic bacteria and nitrogen fertilization on wheat crop under field conditions

    Directory of Open Access Journals (Sweden)

    Valeria Marino Rodrigues Sala

    2008-06-01

    Full Text Available Bactérias diazotróficas pertencentes a diferentes espécies podem estar associadas a gramíneas, como o trigo, promovendo incrementos no crescimento e na produtividade de grãos. Foi realizado um experimento, em condições de campo, com o objetivo de avaliar os benefícios e a viabilidade econômica da inoculação de novos isolados homólogos de bactérias diazotróficas endofíticas em genótipos de trigo sob diferentes doses de N. Foram empregados três isolados de bactérias diazotróficas endofíticas (IAC-AT-8-Azospirillum brasiliense, IAC-HT-11-Achromobacter insolitus e IAC-HT-12-Zoogloea ramigera, dois genótipos de trigo (ITD- 19-Triticum durum L. e IAC-370-Triticum aestivum hard L. e três doses de N (0, 60 e 120 kg ha-1. Foram avaliados, no estádio de perfilhamento e de maturidade fisiológica, a matéria seca, o N acumulado e o índice de eficiência de utilização do N na parte aérea e na colheita; a produtividade de grãos, os componentes de rendimento e os índices de colheita para biomassa e N. No estádio de perfilhamento, somente houve aumento na produção de biomassa com a inoculação de Azospirillum brasiliense e Achromobacter insolitus no genótipo IAC-370, com a adição de N. Entretanto, não houve especificidade dos genótipos de trigo empregados e essas novas bactérias diazotróficas endofíticas para as demais caracaterísticas avaliadas. Houve aumento no acumulado de N na espiga com a inoculação de Azospirillum brasiliense. A inoculação de Achromobacter insolitus propiciou aumento da produtividade de grãos, na maior dose de N utilizada (120 kg ha-1. A inoculação, embora não tenha substituído a adubação nitrogenada, reverteu-se em lucro para o agricultor.Diazotrophic bacteria belonging to different species may be associated with grasses, such as wheat, promoting increases in plant growth and grain yield. A field experiment was carried out to evaluate the benefits and economic viability of inoculation

  15. Seleção de inoculantes à base de turfa contendo bactérias diazotróficas em duas variedades de arroz = Selection of peats inoculants with diazotrophic bacteria in two rice varieties

    Directory of Open Access Journals (Sweden)

    Vera Lúcia Divan Baldani

    2010-01-01

    inoculants, grown in pots containing soil and maintained outside a greenhouse. The dry mass accumulation, Npercent, N-total and yield were determined during the plant cycle. The humidity content varied with storage period. The survival measurement showed that only Burkholderia sp. M130 maintained the number of viable cells around 108 g-1 of peat, while there was a reduction in population of other strains. An increase of yield and total N of 13 and 19.4%, respectively in comparison to treatment fertilized with 40 kg N ha-1 was observed for variety IAC4440,inoculated with strain ZAE94. There was no difference in yield of the IR42 inoculated with either ZAE94 or M130, as compared to N control treatment. No significant difference in development of both rice varieties was observed for both peat used. The results suggest that peat can be used as a carrier for production of an inoculant based on diazotrophic bacteria, since it allowed maintenance of a bacterial population up to 108 cells g-1 peat during the storage period of up to100 days. The results obtained encourage the practice of inoculation of non-leguminous plants.

  16. Toxicidade de herbicidas utilizados na cultura da cana-de-açúcar à bactéria diazotrófica Azospirillum brasilense Toxicity of herbicides applied on sugarcane to the diazotrophic bacterium Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    S.O Procópio

    2011-01-01

    Full Text Available Objetivou-se neste trabalho identificar herbicidas utilizados na cultura da cana-de-açúcar que não alteram o crescimento ou a capacidade de fixação biológica de nitrogênio (FBN da bactéria diazotrófica Azospirillum brasi lense. Dezoito herbicidas - paraquat, ametryn, amicarbazone, diuron, metribuzin, [hexazinone + diuron], [hexazinone + clomazone], clomazone, isoxaflutole, sulfentrazone, oxyfluorfen, imazapic, imazapyr, [trifloxysulfuron-sodium + ametryn], S-metolachlor, glyphosate, MSMA e 2,4-D - foram testados em suas doses comerciais quanto ao impacto sobre o crescimento da bactéria em meio líquido DIGs. As variáveis capacidade de suporte de crescimento (carrying capacity do meio de cultura, duração da fase lag e tempo de geração de A. brasilense foram calculadas a partir de dados de densidade ótica obtidos, em intervalos regulares, durante a incubação de culturas por 55 h. O impacto dos herbicidas na atividade da nitrogenase de A. brasilense foi avaliado em meio semissólido NFb, sem N, pela técnica da atividade de redução do acetileno (ARA. Os efeitos dos herbicidas sobre as variáveis de crescimento e ARA foram comparados ao controle pelo teste de Dunnett. Paraquat, oxyfluorfen, [trifloxysulfuron-sodium + ametryn] e glyphosate reduziram a capacidade do meio DIGs em suportar o crescimento de A. brasilense. Esse efeito foi associado ao aumento da duração da fase lag e do tempo de geração para [trifloxysulfuron-sodium + ametryn] e ao aumento no tempo de geração para glyphosate. MSMA, paraquat e amicarbazone reduzem a FBN in vitro de A. brasilense, porém essa redução é mais severa na presença do paraquat. Os demais herbicidas não alteram o crescimento e a FBN de A. brasilense.The objective of this work was to identify the herbicides applied on sugarcane that do not affect the growth nor the process of biological nitrogen fixation (BNF of the diazotrophic bacterium Azospirillum brasilense. Commercial doses of

  17. Search for endophytic diazotrophs in barley seeds

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    Myriam S. Zawoznik

    2014-06-01

    Full Text Available Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR.

  18. Symbiosome-like intracellular colonization of cereals and other crop plants by nitrogen-fixing bacteria for reduced inputs of synthetic nitrogen fertilizers.

    Science.gov (United States)

    Cocking, Edward C; Stone, Philip J; Davey, Michael R

    2005-09-01

    It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems, and that the problem of environmental nitrogen enrichment is most likely to be solved by substituting synthetic nitrogen fertilizers by the creation of cereal crops that are able to fix nitrogen symbiotically as legumes do. In legumes, rhizobia present intracellularly in membrane-bound vesicular compartments in the cytoplasm of nodule cells fix nitrogen endosymbiotically. Within these symbiosomes, membrane-bound vesicular compartments, rhizobia are supplied with energy derived from plant photosynthates and in return supply the plant with biologically fixed nitrogen, usually as ammonia. This minimizes or eliminates the need for inputs of synthetic nitrogen fertilizers. Recently we have demonstrated, using novel inoculation conditions with very low numbers of bacteria, that cells of root meristems of maize, rice, wheat and other major non-legume crops, such as oilseed rape and tomato, can be intracellularly colonized by the non-rhizobial, non-nodulating, nitrogen fixing bacterium,Gluconacetobacter diazotrophicus that naturally occurs in sugarcane.G. diazotrophicus expressing nitrogen fixing (nifH) genes is present in symbiosome-like compartments in the cytoplasm of cells of the root meristems of the target cereals and non-legume crop species, somewhat similar to the intracellular symbiosome colonization of legume nodule cells by rhizobia. To obtain an indication of the likelihood of adequate growth and yield, of maize for example, with reduced inputs of synthetic nitrogen fertilizers, we are currently determining the extent to which nitrogen fixation, as assessed using various methods, is correlated with the extent of systemic intracellular colonization byG. diazotrophicus, with minimal or zero inputs.

  19. Ocorrência e diversidade de bactérias diazotróficas associadas a gramíneas forrageiras do Pantanal Sul Matogrossense Occurrence and diversity of diazotrophic bacteria associated to forage grasses of the Pantanal in the state of Mato Grosso do Sul

    Directory of Open Access Journals (Sweden)

    Marivaine da Silva Brasil

    2005-04-01

    Full Text Available Este trabalho teve por objetivo avaliar a ocorrência e a diversidade genética de bactérias fixadoras de N2 associadas às gramíneas nativas Elyonurus muticus (capim carona e Axonopus purpusii (capim mimoso e à gramínea exótica Brachiaria humidicola (braquiária que formam as pastagens na região da Nhecolândia. As coletas das plantas e solo foram feitas nos períodos de seca e de cheia para determinar a população de bactérias diazotróficas. Identificaram-se Azospirillum brasilense, A. lipoferum, A. amazonense, Herbaspirillum spp., Burkholderia spp. em amostras de solo, raízes e folhas das três espécies forrageiras. As populações dessas bactérias foram menores na época da cheia em comparação com a época da seca. A diversidade genética das bactérias isoladas foi avaliada por meio da técnica de análise de restrição do DNA ribossomal amplificado (ARDRA. Os isolados foram divididos em cinco grupos genotípicos distintos. Os isolados de A. brasilense e A. lipoferum apresentaram cerca de 50 % de similaridade, enquanto A. amazonense formou um grupo a parte, com apenas 25 % de similaridade em relação ao grupo das espécies do gênero. As bactérias do gênero Herbaspirillum formaram um grupo isolado com apenas 25 % de similaridade em relação ao gênero Azospirillum. O quinto grupo foi formado por apenas um isolado com 25 % de similaridade em relação aos demais.This study was carried out to verify the occurrence and genetic diversity of diazotrophic bacteria associated to the native graminaceus plants Elyonurus muticus (Carona grass and Axonopus purpusii (Mimoso grass and the exotic Brachiaria humidicola (Brachiaria that form the pastures in the Nhecolândia region of the Pantanal in the state of Mato Grosso do Sul, Brazil. The plants were harvested in the dry and rainy seasons and the diazotrophic bacteria populations in the soil and on roots and leaves of the three grasses were determined. Strains belonging to the

  20. Resposta do dendezeiro à adição de nitrogênio e sua influência na população de bactérias diazotróficas Response of African oil palm to nitrogen addition and its influence on the diazotrophic bacteria population

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    André Vieira de Carvalho

    2006-02-01

    Full Text Available O dendê (Elaeis guineensis, Jaquim pode produzir até sete toneladas de óleo por hectare por ano. O óleo vegetal é muito versátil e pode ser usado desde a indústria alimentícia até a produção de biocombustíveis. A planta é capaz de se associar com bactérias diazotróficas que colonizam raízes e caules. O objetivo deste trabalho foi avaliar a resposta à adubação nitrogenada de 17 genótipos de dendê no primeiro ano de cultivo e avaliar a influência da adição do N mineral sobre a população de bactérias diazotróficas, naturalmente presentes nas plântulas de dendezeiro. Foram utilizados potes de plástico completados com 50% de areia quartzoza e 50% de horizonte B de um Argissolo Vermelho-Amarelo, série Itaguaí, não esterilizado e extremamente pobre em nitrogênio. A uréia foi aplicada na dosagem de 33,68 kg ha-1 de N. Na presença do N, todos os genótipos melhoram os parâmetros biométricos, e houve aumento tanto do N total quanto do N acumulado. As populações de bactérias diazotróficas não foram influenciadas pela adição desse elemento. Dois genótipos foram selecionados, na presença e ausência de N, respectivamente, C-2001 e La Mé.African oil palm (Elaeis guineensis, Jaquim can produce up to 7 tons of oil per hectare per year. The vegetable oil is greatly versatile in its use, since food industry up to the production of fuels favorable to environmental protection. The plant has the potential to be associative with diazotrophic bacteria which colonize the roots and stem. The objective of this work was to evaluate 17 genotypes of E. guineensis in response to nitrogen addition and to verify the influence of this nitrogen on the diazotrophic population in an experiment using plastic pots filled with 50% quartz sand and 50% of non sterilized Horizon B; Red-Yellow Podzolic Soil series Itaguaí, extremely poor in nitrogen. Urea was used at a dose of 33.68 kg ha-1 de N. In the presence of the nitrogen, all

  1. Resposta de genótipos de trigo à inoculação de bactérias diazotróficas em condições de campo Wheat genotypes response to inoculation of diazotrophic bacteria in field conditions

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    Valéria Marino Rodrigues Sala

    2007-06-01

    Full Text Available O objetivo deste trabalho foi avaliar, em condições de campo, o efeito e a viabilidade econômica da inoculação de novos isolados homólogos de bactérias diazotróficas endofíticas, sob diferentes doses de nitrogênio, em dois genótipos de trigo, em duas localidades. Foram utilizados: três isolados de bactérias diazotróficas endofíticas (IAC-AT-8, Azospirillum brasilense; IAC-HT-11, Achromobacter insolitus; IAC-HT-12, Zoogloea ramigera, dois genótipos de trigo (ITD-19 e IAC-370 e três doses de N, na forma de uréia (0, 60 e 120 kg ha-1. No estádio de quatro folhas e no perfilhamento, foram avaliados a massa de matéria seca e o N acumulado na parte aérea. Na colheita, foram avaliados o teor de N, a massa de 1.000 sementes e a produtividade de grãos. A inoculação promoveu maior massa de matéria seca e N acumulado e aumentou a produtividade de grãos, principalmente na presença de adubo nitrogenado, com lucro para o agricultor. Entretanto, o maior aumento na produtividade de grãos foi obtido nas plantas do genótipo IAC-370, com o emprego do isolado IAC-HT-12, na ausência de N, que superou em 45% o tratamento testemunha. As respostas variaram em relação ao local de cultivo, o que sugere expressiva interação planta-bactéria-ambiente.The aim of this work was to evaluate, in field conditions, the effect and the economic viability of inoculation of new homologous strains endophytic diazotrophic bacteria, under different nitrogen doses on two wheat genotypes. Three strains of diazotrophic bacteria (IAC-AT-8, Azospirillum brasilense; IAC-HT-11, Achromobacter insolitus; IAC-HT-12, Zoogloea ramigera, two wheat genotypes (ITD-19 and IAC-370, and three levels of nitrogen fertilizer as urea (0, 60 e 120 kg ha-1 were tested. Shoot dry matter and total shoot nitrogen were evaluated, at four leaves and at tillering stages. Nitrogen concentration in the grain, 1,000 grains weight and yield were evaluated at harvest. Plants with

  2. Efeito de bactérias diazotróficas na produção de abacaxizeiro 'Cayenne Champac', sob irrigação, em dois níveis de adubação nitrogenada Contribution of diazotrophic bacteria on yield of pineapple 'Cayenne Champac', under irrigation, with two levels of nitrogen fertilization

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    Olmar Baller Weber

    2004-08-01

    Full Text Available Avaliou-se o efeito de bactérias diazotróficas na produção de abacaxizeiro cv. Cayenne Champac (Champaka, sob irrigação, em dois níveis de adubação nitrogenada. Os tratamentos constaram dos níveis de N de 180 kg ha-1 ano-1 e 300 kg ha-1 ano-1, nas parcelas e subparcelas de mudas micropropagadas inoculadas com bactéria relacionada a Burkholderia cepacia AB213, de bactéria Asaia bogorensis AB219 e controles sem inoculação bacteriana, utilizando-se de três repetições. A inoculação bacteriana foi realizada no laboratório (10(8 células planta-1 e a aclimatação das mudas, em casa de vegetação. Após 5 meses, as mudas foram transplantadas no campo, utilizando-se de 0,9 m entre as linhas e 0,3 m entre as plantas. A cultivar apresentou bom desempenho sob irrigação no solo arenoso de Pacajus, Estado do Ceará, quando adubado com o maior nível de N. Para cada quilograma do elemento fornecido, houve incremento de 124,3 kg ha-1 na massa fresca dos frutos com coroa, entre os níveis da adubação nitrogenada. Na dose maior de N, plantas inoculadas do AB219 produziram frutos maiores e massas frescas 19,4% e 17,3% superiores aos controles, respectivamente, para frutos sem e com coroa. O efeito do AB213 na produção foi menor (9,9% no nível mais baixo de N. A evidência do efeito de bactérias diazotróficas na cultivar Cayenne Champac foi demonstrada.The objective of this work was to evaluate the effect of diazotrophic bacteria on yield of pineapple cv. Cayenne Champac (Champaka, under irrigation, with two levels of N fertilization. Treatments consisted of two N levels (180 kg and 300 kg ha-1 year-1, on the plots, and micropropagated plantlets inoculated with bacteria related to Burkholderia cepacia AB213, with Asaia bogorensis AB219 and without bacterial inoculation, on the subplots, using three replications. The bacterial inoculation was performed in a laboratory (with 10(8 bacterial cells plant-1 and the plant acclimatization in

  3. Adição de molibdênio ao inoculante turfoso com bactérias diazotróficas usado em duas cultivares de arroz irrigado Addition of molybdenum in peat innoculum with diazotrophic bacteria used in two rice cultivars

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    Salomão Lima Guimarães

    2007-03-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da utilização de bactérias diazotróficas com ou sem molibdênio em duas cultivares de arroz com diferentes potenciais de fixação biológica de nitrogênio. Dois experimentos foram montados: o primeiro foi conduzido em condições gnotobióticas, com utilização das estirpes BR 11417 (ZAE94 - Herbaspirillum seropedicae e BR 11340 (M130 - Burkholderia sp., duas doses de molibdênio, 1,12 e 2,25 g, e duas fontes, molibdato de sódio e molibdato de amônio. As sementes foram peletizadas com inoculante turfoso com bactérias diazotróficas e o molibdênio. Em seguida as cultivares foram plantadas em tubos de ensaio com capacidade para 120 mL com 60 mL da solução de Hoagland sem nitrogênio por 30 dias. Foi avaliada a matéria seca das plantas. O segundo experimento foi conduzido em vasos em casa de vegetação, com as mesmas estirpes e dosagens de molibdênio. Foram avaliados a produção e o N total dos grãos. O delineamento estatístico foi inteiramente casualizado com quatro repetições e quatro plantas por vaso. Foram observados aumentos na produção de grãos nas cultivares IR42 e IAC4440 de 88 e 57%, respectivamente, com a adição ao inoculante turfoso, das estirpes ZAE94 e M130 com molibdênio.The objective of this work was to evaluate the effect of the inoculation of diazotrophic bacteria associated with the molybdenum in two rice cultivars. Two experiments were performed: the first one was lead under gnotobiotics conditions, using Herbaspirillum seropedicae, strain BR 11417 and Burkholderia sp., strain BR 11340, two doses of molybdenum (1.12 and 2.25 g and two different sources of Mo - sodium molybdate and ammonium molybdate. The seeds were pelleted with peat containing diazotrophic bacteria and molybdenum. After inoculations were seeds planted in to test tubes with capacity for 120 mL content 60 mL Hoagland solution without nitrogen. Seedlings were harvested after 30 days and plant

  4. Response of micropropagated sugarcane varieties to inoculation with endophytic diazotrophic bacteria Resposta da inoculação com bactérias diazotróficas endofíticas em duas variedades micropropagadas de cana-de-açúcar

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    André Luiz Martinez de Oliveira

    2003-11-01

    Full Text Available Previous studies estimated that sugarcane could obtain up to 60% of total nitrogen accumulated from BNF. Here a mixture of five endophytic diazotrophic strains was tested in a field trial, inoculated in two micropropagated sugarcane varieties and three locals, to determine the effects on commercial crop conditions. The sugarcane plantlets were inoculated in vitro, and after 17 months of growing in the field, the productivity and BNF contribution showed to be influenced by the plant genotype and soil type. The highest BNF contributions was observed in the poorest soil for both varieties. Smaller increases in productivity were observed for SP 701143 variety grown in soil with low or medium fertility. In contrast, a decrease in the stem productivity was observed in the SP 813250 variety grown in the three localities.Experimentos anteriores estimaram que a cana-de-açúcar pode obter até 60% do N acumulado via fixação biológica de nitrogênio (FBN. Neste trabalho, os efeitos da inoculação da mistura de cinco espécies de bactérias diazotróficas endofíticas foram testados em duas variedades de cana-de-açúcar micropropagadas, sob condições de campo. Após 17 meses de crescimento, a produtividade e a FBN apresentaram influência do genótipo vegetal e da localidade de cultivo. As maiores contribuições via FBN foram observadas no solo de menor fertilidade, para ambas variedades de cana-de-açúcar. Pequenos aumentos de produtividade foram observados para a variedade SP 701143 nos solos de baixa e média fertilidade. Por outro lado, a inoculação na variedade SP 813250 apresentou decréscimo de produtividade nos três tipos de solo testados.

  5. Characterization of cellulose production by a Gluconacetobacter xylinus strain from Kombucha.

    Science.gov (United States)

    Nguyen, Vu Tuan; Flanagan, Bernadine; Gidley, Michael J; Dykes, Gary A

    2008-11-01

    The aims of this work were to characterize and improve cellulose production by a Gluconoacetobacter xylinus strain isolated from Kombucha and determine the purity and some structural features of the cellulose from this strain. Cellulose yield in tea medium with both black tea and green tea and in Hestrin and Schramm (HS) medium under both static and agitated cultures was compared. In the tea medium, the highest cellulose yield was obtained with green tea (approximately 0.20 g/L) rather than black tea (approximately 0.14 g/L). Yield in HS was higher (approximately 0.28 g/L) but did not differ between static and agitated incubation. (1)H-NMR and (13)C-NMR spectroscopy indicated that the cellulose is pure (free of acetan) and has high crystallinity, respectively. Cellulose yield was improved by changing the type and level of carbon and nitrogen source in the HS medium. A high yield of approximately 2.64 g/L was obtained with mannitol at 20 g/L and corn steep liquor at 40 g/L in combination. In the tea medium, tea at a level of 3 g/L gave the highest cellulose yield and the addition of 3 g/L of tea to the HS medium increased cellulose yield to 3.34 g/L. In conclusion, the G. xylinus strain from Kombucha had different cellulose-producing characteristics than previous strains isolated from fruit. Cellulose was produced in a pure form and showed high potential applicability. Our studies extensively characterized cellulose production from a G. xylinus strain from Kombucha for the first time, indicating both similarities and differences to strains from different sources.

  6. Promoção de enraizamento de microtoletes de cana-de-açúcar pelo uso conjunto de substâncias húmicas e bactérias diazotróficas endofíticas Rooting of micro seed pieces by combined use of humic substances and endophytic diazotrophic bacteria in sugar cane

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    Roberto Batista Marques Júnior

    2008-06-01

    éficos da inoculação de estirpes selecionadas de H. seropedicae, já para ácidos húmicos, as respostas positivas independem da termoterapia. A promoção do crescimento radicular por AH e a fixação biológica de N2 podem representar uma estratégia inovadora para produção sustentada em sistemas agrícolas.Besides the direct nutritional effect of mineralization of organic matter or by biological nitrogen fixation activity, the humic substances and endophytic diazotrophic bacteria can directly influence plant metabolism, modifying plant growth and development patterns. The purpose of this study was to evaluate the effect of the combined use of humic acid (HA and Herbaspirillum seropedicae, an endophytic nitrogen-fixing bacteria, on the root growth of seed pieces - heat-treated or not - of the sugarcane variety RB 72 454. After hot water treatment, the cane cuttings were immersed for 12 hours as follows: in water (control plant, in HA solution (20 mg L-1 of C from vermicompost, in bacterial inoculant of H. seropedicae, strain HRC54 (10(8 cells mL-1, and in a combination of bacteria and HA. Root growth was improved by 60 to 118 % in length and from 33 % to 233 % in surface area on sugarcane plant treatments compared to control, with more pronounced effect in plants under heat treatment. Likewise, the positive effect of the combinded treatment (bacteria inoculation and humic acid on shoot and root biomass was significant compared to the control with heat-treated cuts. For non-heated seed pieces, bacteria inoculation did not result in a positive plant growth effect, but only in the presence of humic acid. The combined or isolated use of both HA and bacteria did not significantly modify the bacteria population in the root tissue of heat-untreated sugarcane. For heat-treated cuts, bacteria inoculation, combined or not with HA, increased the size of diazotrophic bacteria population on roots. The results highlight the importance of thermotherapy to increase the positive

  7. Efeito dos ácidos húmicos na inoculação de bactérias diazotróficas endofíticas em sementes de milho Effect of the humic acids in endophytic diazotrophic bacteria inoculation in corn seeds

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    Patrícia Marluci da Conceição

    2009-09-01

    Full Text Available Os ácidos húmicos podem atuar no aumento da população de bactérias diazotróficas introduzidas no interior da planta e, consequentemente, no incremento dos efeitos benéficos sobre a planta hospedeira. Com este trabalho, objetivou-se avaliar o efeito dos ácidos húmicos na inoculação de bactérias diazotróficas endofíticas, em sementes de milho. Foi utilizada a estirpe Herbaspirillum seropedicae Z67 BR 11175. A inoculação das sementes com as bactérias e a adição de ácidos húmicos foram realizadas pelo recobrimento das sementes de milho UENF 506-8, com a mistura de calcário, meio de cultura semisólido, água e cimentante. As sementes recobertas foram semeadas em vasos Leonard. Aos 40 dias após a semeadura, as plântulas foram coletadas, e foi realizada a contagem de bactérias nas raízes pela técnica do Número Mais Provável. Os resultados deste trabalho mostram que os ácidos húmicos não interferem negativamente no crescimento das bactérias e estimulam a colonização da microbiota nativa. No entanto, nas condições avaliadas, a aplicação conjunta de bactérias + ácidos húmicos não estimulou o crescimento da população de bactérias inoculadas nas plântulas de milho.The objective of this research was to evaluate the effect of the humic acids in the inoculation of endophytic diazotrophic bacteria in corn seeds. It was used the bacteria Herbaspirillum seropedicae Z67 BR 11175. The inoculation of the seeds with the bacteria and the addition of humic acids were accomplished by the coating of the corn seeds UENF 506-8. The coating was accomplished with a mixture of lime, semi-solid middle culture, water and cement. The seeds covered were sown in Leonard vases. Forty days after sowing the plants were collected and the bacteria couting was accomplished in the roots by the Most probable Number technique. The results showed that the humic acids doesn't interfere negatively in the bacteria growth of and they stimulate the

  8. GERMINATION OF GRASSES DUE TO INOCULATION DIAZOTROPHIC BACTERIA

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    C. D. A. Moreira

    2014-07-01

    Full Text Available The germination of forage grasses suffers from numbness and a natural tendency to low quality. The use of microorganisms inoculated in seeds with the purpose of increasing and meet the demand of some nutrient has been shown to be efficient, but the role of the microorganism in germination and rate of force is still unknown. Therefore the goal as study was to evaluate the germination rate of seeds of three cultivars of Brachiaria brizantha CV. Marandu, b., b. brizantha CV. Xaraés and b. humidícola cv Tupi and a cultivar of millet, P. hybrid cv Massai depending on the bacterium Azospirillum brasilense diazotrofic inoculation (nitrogen-fixing. Germination test was used in seed dispersal to assess the effect of first count (VPC in the treatments with and without inoculation. It was done also conducted further tests of electrical conductivity, weight of thousand seeds and water content. The delineation used was randomized entirely (DIC and the statistical analysis carried out through the analysis of variance and comparison of means using the Tukey test, the 5% probability. Massai grass seeds have the highest rate of force of first count in both treatments. Inoculation of bacterium Azospirillum brasilense did not affect the values of force of first count on seeds of the cultivars Marandu, Xaraés, Tupi and Massai. The seeds of the massai have higher germination speed relative the other cultivars evaluated when inoculated.

  9. Potential of diazotrophic bacteria associated with sugarcane for energycane production

    Science.gov (United States)

    Crosses between sugarcane and wild species of Saccharum and other closely related genera are made to introgress new genes from the wild species into sugarcane. Characteristics of the progeny from these crosses may include increased biomass and the ability to be grown in a broader geographical range ...

  10. PHYSIOLOGICAL QUALITY OF CORN SEEDS REINOCULATED WITH DIAZOTROPHIC BACTERIA

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    Ricardo Felipe Braga de Sousa

    2015-09-01

    Full Text Available O objetivo deste trabalho foi avaliar a qualidade fisiológica de sementes de milho provenientes do cultivo associado com Azospirillum brasilense e Herbaspirillum seropedicae e reinoculadas com A. brasilense. As sementes utilizadas foram provenientes de cultivo em condições de campo, sendo os tratamentos: T1 - testemunha; T2 - 60 kg hectare-1 de N; T3 – inoculação (Azospirillum brasilense - AbV5; T4 - inoculação (Azospirillum brasilense + 60 kg hectare-1 de N; T5 - inoculação (Herbaspirillum seropedicae - SmR1 e T6 - inoculação (Herbaspirillum seropedicae + 60 kg hectare-1 de N. Foram separados dois lotes dos tratamentos, sendo um reinoculado com A. brasilense (AbV5. O delineamento foi inteiramente ao acaso com esquema fatorial (6x2, sendo seis tratamentos; com inoculação e reinoculadas. As sementes reinoculadas apresentaram maiores comprimentos, volumes e matéria seca de raízes e parte aérea, concluindo que a reinoculação com sementes promove um incremento no desenvolvimento inicial das plântulas.

  11. Caracterização fenotípica e diversidade de bactérias diazotróficas associativas isoladas de solos em reabilitação após a mineração de bauxita Phenotypic characterization and diversity of diazotrophic associative bacteria isolated from soils rehabilitated after bauxite mining

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    R. S. A. Nóbrega

    2004-04-01

    eles.Grass species, which are frequently used for rehabilitation of degraded areas, can establish root associations with nitrogen fixing bacteria thus contributing to the ecosystem's sustainability. On the other hand, microbial diversity plays an important role in the resilience of biological processes, including N2 fixation. This study aimed at the phenotypic characterization of 72 diazotrophic Gram-negative bacteria. Samples were isolated from areas under different rehabilitation strategies after bauxite mining in Poços de Caldas, Minas Gerais, Brazil, and inoculated in NFb, Fam, and JNFb media. Type and reference strains of Herbaspirillum, Azospirillum and Burkholderia species were used for comparison as they are able to grow in such media. The similarity dendrogram based on seven cultural characteristics of the isolates in GNA medium presented a great diversity, as 50 groups were formed with 81 % similarity. NaCl tolerance in the potato/sucrose/acid malic medium varied from 0 to 50 g L-1 and allowed a separation of isolates and type strains into five groups. Cell diameters varied from 0.61 to 1.21 µm, and 13 isolates were not similar to the type strains. Fifteen groups with 75 % similarity were formed based on total proteins patterns obtained by polyacrylamide gel electrophoresis (SDS-PAGE. Neither was there any relationship among groups based on the different characteristics, nor between these and the areas of bacteria isolation. Fam and JNFb media detected the target species as well as other unidentified ones. The high phenotypic dissimilarity among isolates and type strains, mainly regarding total protein eletrophoresis profiles suggests that new species could be present within these populations.

  12. citohistochemistry, Biocatalytic Effectors (EBc©, source of infection, biological control, agrosystems

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    Marcia M. Rojas

    2015-11-01

    Full Text Available Among the efforts done in Cuba to the sustainability in the agricultural system, one of them is the use of bioproducts, which have a relevant economic, ecological and social impact. The sugarcane is one of main crops in our country and it has a great importance at world level. In the present work is demonstrated the effect of different carbon and nitrogen sources in the growth of 5 entophytic bacteria (three of Gluconacetobacter diazotrophicus, one of Bacillus licheniformis and one of Enterobacter agglomerans were demonstrated. As the same form are studied the influence of juices from five varieties, as well as, different concentrations of fitohormones indole3acetic acid and giberelic acid on the growth. Was demonstrated that asparagine and ammonium sulfate as nitrogen sources added to LGI medium enhance the growth a major growth of the studied endophytic bacteria. The LGI medium supplied with juices of sugarcane enhance the growth of microorganisms (p≤0,05 and don't exist any relationships among the origin of the juice and the strains. On the other hand, the fitohormones at low concentrations don't affect the growth but at high levels of these hormones inhibit the growth. It's necessary to study the factors that have influence on the interaction between the plant and endophytes to use their potentialities as plant growth promoters.

  13. Diel regulation of photosynthetic activity in the oceanic unicellular diazotrophic cyanobacterium Crocosphaera watsonii WH8501

    Czech Academy of Sciences Publication Activity Database

    Masuda, Takako; Bernát, Gábor; Bečková, Martina; Kotabová, Eva; Lawrenz, Evelyn; Lukeš, Martin; Komenda, Josef; Prášil, Ondřej

    2018-01-01

    Roč. 20, č. 2 (2018), s. 546-560 ISSN 1462-2912 R&D Projects: GA MŠk(CZ) LO1416; GA MŠk(CZ) ED2.1.00/19.0392; GA ČR GA16-15467S Institutional support: RVO:61388971 Keywords : NORTH PACIFIC-OCEAN * SYNECHOCYSTIS SP PCC-6803; * PHOTOSYSTEM-II Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 5.395, year: 2016

  14. Transcriptional patterns identify resource controls on the diazotroph Trichodesmium in the Atlantic and Pacific oceans.

    Science.gov (United States)

    Rouco, Mónica; Frischkorn, Kyle R; Haley, Sheean T; Alexander, Harriet; Dyhrman, Sonya T

    2018-02-28

    The N 2 -fixing cyanobacterium Trichodesmium is intensely studied because of the control this organism exerts over the cycling of carbon and nitrogen in the low nutrient ocean gyres. Although iron (Fe) and phosphorus (P) bioavailability are thought to be major drivers of Trichodesmium distributions and activities, identifying resource controls on Trichodesmium is challenging, as Fe and P are often organically complexed and their bioavailability to a single species in a mixed community is difficult to constrain. Further, Fe and P geochemistries are linked through the activities of metalloenzymes, such as the alkaline phosphatases (APs) PhoX and PhoA, which are used by microbes to access dissolved organic P (DOP). Here we identified significant correlations between Trichodesmium-specific transcriptional patterns in the North Atlantic (NASG) and North Pacific Subtropical Gyres (NPSG) and patterns in Fe and P biogeochemistry, with the relative enrichment of Fe stress markers in the NPSG, and P stress markers in the NASG. We also observed the differential enrichment of Fe-requiring PhoX transcripts in the NASG and Fe-insensitive PhoA transcripts in the NPSG, suggesting that metalloenzyme switching may be used to mitigate Fe limitation of DOP metabolism in Trichodesmium. This trait may underpin Trichodesmium success across disparate ecosystems.

  15. A highly asynchronous developmental program triggered during germination of dormant akinetes of filamentous diazotrophic cyanobacteria

    Czech Academy of Sciences Publication Activity Database

    Perez, Rebeca; Wörmer, L.; Sass, P.; Maldener, I.

    2018-01-01

    Roč. 94, č. 1 (2018), č. článku fix131. ISSN 0168-6496 Institutional support: RVO:61388971 Keywords : akinetes * germination * heterocyst Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 3.720, year: 2016

  16. Ultradian metabolic rhythm in the diazotrophic cyanobacterium Cyanothece sp. ATCC 51142

    Czech Academy of Sciences Publication Activity Database

    Červený, Jan; Sinětova, Maria A.; Valledor, Luis; Sherman, L. A.; Nedbal, Ladislav

    2013-01-01

    Roč. 110, č. 32 (2013), s. 13210-13215 ISSN 0027-8424 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073; GA MŠk(CZ) EE2.3.20.0256; GA ČR GA206/09/1284 Institutional support: RVO:67179843 Keywords : cyanobacteria * diurnal * metabolism * oscillation Subject RIV: EH - Ecology, Behaviour Impact factor: 9.809, year: 2013

  17. Genomic and evolutionary comparisons of diazotrophic and pathogenic bacteria of the order Rhizobiales

    Directory of Open Access Journals (Sweden)

    Vasconcelos Ana

    2010-02-01

    Full Text Available Abstract Background Species belonging to the Rhizobiales are intriguing and extensively researched for including both bacteria with the ability to fix nitrogen when in symbiosis with leguminous plants and pathogenic bacteria to animals and plants. Similarities between the strategies adopted by pathogenic and symbiotic Rhizobiales have been described, as well as high variability related to events of horizontal gene transfer. Although it is well known that chromosomal rearrangements, mutations and horizontal gene transfer influence the dynamics of bacterial genomes, in Rhizobiales, the scenario that determine pathogenic or symbiotic lifestyle are not clear and there are very few studies of comparative genomic between these classes of prokaryotic microorganisms trying to delineate the evolutionary characterization of symbiosis and pathogenesis. Results Non-symbiotic nitrogen-fixing bacteria and bacteria involved in bioremediation closer to symbionts and pathogens in study may assist in the origin and ancestry genes and the gene flow occurring in Rhizobiales. The genomic comparisons of 19 species of Rhizobiales, including nitrogen-fixing, bioremediators and pathogens resulted in 33 common clusters to biological nitrogen fixation and pathogenesis, 15 clusters exclusive to all nitrogen-fixing bacteria and bacteria involved in bioremediation, 13 clusters found in only some nitrogen-fixing and bioremediation bacteria, 01 cluster exclusive to some symbionts, and 01 cluster found only in some pathogens analyzed. In BBH performed to all strains studied, 77 common genes were obtained, 17 of which were related to biological nitrogen fixation and pathogenesis. Phylogenetic reconstructions for Fix, Nif, Nod, Vir, and Trb showed possible horizontal gene transfer events, grouping species of different phenotypes. Conclusions The presence of symbiotic and virulence genes in both pathogens and symbionts does not seem to be the only determinant factor for lifestyle evolution in these microorganisms, although they may act in common stages of host infection. The phylogenetic analysis for many distinct operons involved in these processes emphasizes the relevance of horizontal gene transfer events in the symbiotic and pathogenic similarity.

  18. Drivers of the dynamics of diazotrophs and denitrifiers in North Sea bottom waters and sediments

    NARCIS (Netherlands)

    Fan, H.; Bolhuis, H.; Stal, L.J.

    2015-01-01

    The fixation of dinitrogen (N2) and denitrification are two opposite processes in the nitrogen cycle. The former transfers atmospheric dinitrogen gas into bound nitrogen in the biosphere, while the latter returns this bound nitrogen back to atmospheric dinitrogen. It is unclear whether or not these

  19. Agronomic and environmental aspects of diazotrophic bacteria in irrigated rice fields

    Science.gov (United States)

    This article provides an overview of the free-living and plant-associated nitrogen-fixing bacterial communities in irrigated rice fields, with a focus on describing the drivers affecting community assemblages in this soil-water-plant-atmosphere system. Theoretical and technical advances in non-legu...

  20. The Co-Distribution of Nitrifying Archaea and Diazotrophic Bacteria in Geothermal Springs

    Science.gov (United States)

    Hamilton, T. L.; Jewell, T. N. M.; de la Torre, J. R.; Boyd, E. S.

    2014-12-01

    Microbial processes that regulate availability of nutrients play key roles in shaping community composition. All life requires fixed nitrogen (N), and its bioavailability is what often limits ecosystem productivity. Biological nitrogen fixation, or the reduction of dinitrogen (N2) to ammonia (NH3), is a keystone process in N limited ecosystems, providing nitrogen for members of the community. N2 fixing organisms likely represent a 'bottom up control' on the structure of communities that develop in N limited environments. N2 fixation is catalyzed by a limited number of metabolically diverse bacteria and some methanogenic archaea and occurs in a variety of physically and geochemically diverse environments. Nitrification, or the sequential oxidation of NH4+ to nitrite (NO2-) and ultimately nitrate (NO3-), is catalyzed by several lineages of Proteobacteria at temperatures of < 62°C and by members of the Thaumarcheota at temperatures up to 90°C. Nitrification can thus be considered a 'top down control' on the structure of communities that develop in N limited environments. Our research in Yellowstone National Park (YNP) reveals a strong correspondence between the distribution of ammonia oxidizing archaea (AOA) and nitrogen fixing aquificae (NFA) in nitrogen-limited geothermal hot springs over large environmental gradients. Based on the physiology of AOA and NFA, we propose that the strong co-distributional pattern results from interspecies interactions, namely competition for bioavailable ammonia. Our recent work has shown that in springs where the niche dimension of AOA and NFA overlap (e.g., Perpetual Spouter; pH 7.1, 86.4°C), the dissimilar affinities for NH4 result in AOA metabolism maintaining a low NH4(T) pool and selecting for inclusion of NFA during the assembly of these communities. Here, we examine in situ physiological interactions of AOA and NFA, tracking changes in transcript levels of key genes involved in nitrogen metabolism and carbon fixation of these organisms in springs where the niche dimension of AOA and NFA overlap (e.g., Perpetual Spouter). These data suggest affinity for substrate and electron donor use play key roles in structuring the biodiversity of this hydrothermal community, and likely influences the structure of other N limited hydrothermal and non-hydrothermal ecosystems.

  1. Co-inoculation with diazotrophic bacteria in soybeans associated to urea topdressing

    Directory of Open Access Journals (Sweden)

    Glauber Monçon Fipke

    Full Text Available ABSTRACT Increased grain yield can be obtained via an interaction between plants and growth-promoting microorganisms. The Bradyrhizobium spp. are capable of fixing atmospheric nitrogen in soybeans [Glycine max (L. Merril], and Azospirillum spp. induce the synthesis of phytohormones. The aim of this study was to evaluate inoculation with Bradyrhizobium and co-inoculation with Bradyrhizobium + Azospirillum brasilense in soybeans in combination with the application a topdressing of 0, 75 or 150 kg of N ha-1 of urea during the reproductive stage. Three soybean cultivars (BMX Ativa, TEC 6029 and BMX Potência, were tested in field experiments in Santa Maria, RS, Brazil, during two agricultural years (2013/2014 and 2014/2015 and two sowing times. Morphological, nodulation and yield components were evaluated. Co-inoculation increased the grain yield by 240 kg ha-1 compared with conventional inoculation. When co-inoculated, cultivars BMX Ativa, TEC 6029 and BMX Potência showed increased grain yields of 6, 4 and 12%, respectively. The application of 150 kg ha-1 of N as a topdressing increased the grain yield by 300 kg ha-1 in the co-inoculated cultivars TEC 6029 and BMX Potência, but without a financial return. When inoculated only with Bradyrhizobium, the cultivars did not respond positively to the application of urea.

  2. Chemical characterization of the gels produced by the diazotrophic bacteria Rhizobium tropici and Mesorhizobium sp

    International Nuclear Information System (INIS)

    Monteiro, Nilson Kobori; Aranda-Selverio, Gabriel; Exposti, Diego Tadeu Degli; Silva, Maria de Lourdes Corradi da; Lemos, Eliana Gertrudes Macedo; Campanharo, Joao Carlos; Silveira, Joana Lea Meira

    2012-01-01

    The exopolysaccharides with characteristics of gel produced by Rhizobium tropici (EPSRT) and Mesorhizobium sp (EPSMR) are acidic heteropolysaccharide composed mainly of glucose and galactose in a molar ratio of 4:1 and 5:1 respectively, with traces of mannose (∼ 1%). Chemical analysis showed the presence of uronic acid, pyruvate and acetyl-substituents in the structures of both polymers. Experiments of gel permeation chromatography and polyacrylamide gel electrophoresis showed that EPSRT and EPSMR are homogeneous molecules with low grade of polydispersity. The EPS were characterized using spectroscopic techniques of FT-IR, 1 H and 13 C-NMR. (author)

  3. Complete genome sequence of Paenibacillus riograndensis SBR5(T), a Gram-positive diazotrophic rhizobacterium.

    Science.gov (United States)

    Brito, Luciana Fernandes; Bach, Evelise; Kalinowski, Jörn; Rückert, Christian; Wibberg, Daniel; Passaglia, Luciane M; Wendisch, Volker F

    2015-08-10

    Paenibacillus riograndensis is a Gram-positive rhizobacterium which exhibits plant growth promoting activities. It was isolated from the rhizosphere of wheat grown in the state of Rio Grande do Sul, Brazil. Here we announce the complete genome sequence of P. riograndensis strain SBR5(T). The genome of P. riograndensis SBR5(T) consists of a circular chromosome of 7,893,056bps. The genome was finished and fully annotated, containing 6705 protein coding genes, 87 tRNAs and 27 rRNAs. The knowledge of the complete genome helped to explain why P. riograndensis SBR5(T) can grow with the carbon sources arabinose and mannitol, but not myo-inositol, and to explain physiological features such as biotin auxotrophy and antibiotic resistances. The genome sequence will be valuable for functional genomics and ecological studies as well as for application of P. riograndensis SBR5(T) as plant growth-promoting rhizobacterium. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Multilocus sequence analysis (MLSA) of Bradyrhizobium strains: revealing high diversity of tropical diazotrophic symbiotic bacteria.

    Science.gov (United States)

    Delamuta, Jakeline Renata Marçon; Ribeiro, Renan Augusto; Menna, Pâmela; Bangel, Eliane Villamil; Hungria, Mariangela

    2012-04-01

    Symbiotic association of several genera of bacteria collectively called as rhizobia and plants belonging to the family Leguminosae (=Fabaceae) results in the process of biological nitrogen fixation, playing a key role in global N cycling, and also bringing relevant contributions to the agriculture. Bradyrhizobium is considered as the ancestral of all nitrogen-fixing rhizobial species, probably originated in the tropics. The genus encompasses a variety of diverse bacteria, but the diversity captured in the analysis of the 16S rRNA is often low. In this study, we analyzed twelve Bradyrhizobium strains selected from previous studies performed by our group for showing high genetic diversity in relation to the described species. In addition to the 16S rRNA, five housekeeping genes (recA, atpD, glnII, gyrB and rpoB) were analyzed in the MLSA (multilocus sequence analysis) approach. Analysis of each gene and of the concatenated housekeeping genes captured a considerably higher level of genetic diversity, with indication of putative new species. The results highlight the high genetic variability associated with Bradyrhizobium microsymbionts of a variety of legumes. In addition, the MLSA approach has proved to represent a rapid and reliable method to be employed in phylogenetic and taxonomic studies, speeding the identification of the still poorly known diversity of nitrogen-fixing rhizobia in the tropics.

  5. The genome of Cyanothece 51142, a unicellular diazotrophic cyanobacterium important in the marine nitrogen cycle

    Energy Technology Data Exchange (ETDEWEB)

    Welsh, Eric A.; Liberton, Michelle L.; Stockel, Jana; Loh, Thomas; Elvitigala, Thanura R.; Wang, Chunyan; Wollam, Aye; Fulton, Robert S.; Clifton, Sandra W.; Jacobs, Jon M.; Aurora, Rajeev; Ghosh, Bijoy K.; Sherman, Louis A.; Smith, Richard D.; Wilson, Richard K.; Pakrasi, Himadri B.

    2008-09-30

    Cyanobacteria are oxygenic photosynthetic bacteria that have significant roles in global biological carbon sequestration and oxygen production. They occupy a diverse range of habitats, from open ocean, to hot springs, deserts, and arctic waters. Cyanobacteria are known as the progenitors of the chloroplasts of plants and algae, and are the simplest known organisms to exhibit circadian behavior4. Cyanothece sp. ATCC 51142 is a unicellular marine cyanobacterium capable of N2-fixation, a process that is biochemically incompatible with oxygenic photosynthesis. To resolve this problem, Cyanothece performs photosynthesis during the day and nitrogen fixation at night, thus temporally separating these processes in the same cell. The genome of Cyanothece 51142 was completely sequenced and found to contain a unique arrangement of one large circular chromosome, four small plasmids, and one linear chromosome, the first report of such a linear element in a photosynthetic bacterium. Annotation of the Cyanothece genome was aided by the use of highthroughput proteomics data, enabling the reclassification of 25% of the proteins with no informative sequence homology. Phylogenetic analysis suggests that nitrogen fixation is an ancient process that arose early in evolution and has subsequently been lost in many cyanobacterial strains. In cyanobacterial cells, the circadian clock influences numerous processes, including carbohydrate synthesis, nitrogen fixation, photosynthesis, respiration, and the cell division cycle. During a diurnal period, Cyanothece cells actively accumulate and degrade different storage inclusion bodies for the products of photosynthesis and N2-fixation. This ability to utilize metabolic compartmentalization and energy storage makes Cyanothece an ideal system for bioenergy research, as well as studies of how a unicellular organism balances multiple, often incompatible, processes in the same cell.

  6. Genomic and evolutionary comparisons of diazotrophic and pathogenic bacteria of the order Rhizobiales.

    Science.gov (United States)

    Carvalho, Fabíola M; Souza, Rangel C; Barcellos, Fernando G; Hungria, Mariangela; Vasconcelos, Ana Tereza R

    2010-02-08

    Species belonging to the Rhizobiales are intriguing and extensively researched for including both bacteria with the ability to fix nitrogen when in symbiosis with leguminous plants and pathogenic bacteria to animals and plants. Similarities between the strategies adopted by pathogenic and symbiotic Rhizobiales have been described, as well as high variability related to events of horizontal gene transfer. Although it is well known that chromosomal rearrangements, mutations and horizontal gene transfer influence the dynamics of bacterial genomes, in Rhizobiales, the scenario that determine pathogenic or symbiotic lifestyle are not clear and there are very few studies of comparative genomic between these classes of prokaryotic microorganisms trying to delineate the evolutionary characterization of symbiosis and pathogenesis. Non-symbiotic nitrogen-fixing bacteria and bacteria involved in bioremediation closer to symbionts and pathogens in study may assist in the origin and ancestry genes and the gene flow occurring in Rhizobiales. The genomic comparisons of 19 species of Rhizobiales, including nitrogen-fixing, bioremediators and pathogens resulted in 33 common clusters to biological nitrogen fixation and pathogenesis, 15 clusters exclusive to all nitrogen-fixing bacteria and bacteria involved in bioremediation, 13 clusters found in only some nitrogen-fixing and bioremediation bacteria, 01 cluster exclusive to some symbionts, and 01 cluster found only in some pathogens analyzed. In BBH performed to all strains studied, 77 common genes were obtained, 17 of which were related to biological nitrogen fixation and pathogenesis. Phylogenetic reconstructions for Fix, Nif, Nod, Vir, and Trb showed possible horizontal gene transfer events, grouping species of different phenotypes. The presence of symbiotic and virulence genes in both pathogens and symbionts does not seem to be the only determinant factor for lifestyle evolution in these microorganisms, although they may act in common stages of host infection. The phylogenetic analysis for many distinct operons involved in these processes emphasizes the relevance of horizontal gene transfer events in the symbiotic and pathogenic similarity.

  7. Natronobacillus azotifigens gen. nov., sp. nov., an anaerobic diazotrophic haloalkaliphile from soda-rich habitats

    NARCIS (Netherlands)

    Sorokin, I.D.; Zadorina, E.V.; Kravchenko, I.K.; Boulygina, E.S.; Tourova, T.P.; Sorokin, D.Y.

    2009-01-01

    Gram-positive bacteria capable of nitrogen fixation were obtained in microoxic enrichments from soda soils in south-western Siberia, north-eastern Mongolia, and the Lybian desert (Egypt). The same organisms were obtained in anoxic enrichments with glucose from soda lake sediments in the Kulunda

  8. Molecular characterization of diazotrophic and denitrifying bacteria associated with mangrove roots.

    Science.gov (United States)

    Flores-Mireles, Ana L; Winans, Stephen C; Holguin, Gina

    2007-11-01

    An analysis of the molecular diversity of N(2) fixers and denitrifiers associated with mangrove roots was performed using terminal restriction length polymorphism (T-RFLP) of nifH (N(2) fixation) and nirS and nirK (denitrification), and the compositions and structures of these communities among three sites were compared. The number of operational taxonomic units (OTU) for nifH was higher than that for nirK or nirS at all three sites. Site 3, which had the highest organic matter and sand content in the rhizosphere sediment, as well as the lowest pore water oxygen concentration, had the highest nifH diversity. Principal component analysis of biogeochemical parameters identified soil texture, organic matter content, pore water oxygen concentration, and salinity as the main variables that differentiated the sites. Nonmetric multidimensional scaling (MDS) analyses of the T-RFLP data using the Bray-Curtis coefficient, group analyses, and pairwise comparisons between the sites clearly separated the OTU of site 3 from those of sites 1 and 2. For nirS, there were statistically significant differences in the composition of OTU among the sites, but the variability was less than for nifH. OTU defined on the basis of nirK were highly similar, and the three sites were not clearly separated on the basis of these sequences. The phylogenetic trees of nifH, nirK, and nirS showed that most of the cloned sequences were more similar to sequences from the rhizosphere isolates than to those from known strains or from other environments.

  9. Diversity and antifungal activity of endophytic diazotrophic bacteria colonizing sugarcane in Egypt

    Science.gov (United States)

    The price of nitrogen continues to increase and is a major input in sugarcane production. Sugarcane grown in Egypt was screened for the presence of nitrogen-fixing bacteria. Nitrogen-free medium LGI-P was used to isolate bacteria from cane stalks. Among the 52 isolates subjected to acetylene redu...

  10. Molecular Characterization of Diazotrophic and Denitrifying Bacteria Associated with Mangrove Roots▿

    Science.gov (United States)

    Flores-Mireles, Ana L.; Winans, Stephen C.; Holguin, Gina

    2007-01-01

    An analysis of the molecular diversity of N2 fixers and denitrifiers associated with mangrove roots was performed using terminal restriction length polymorphism (T-RFLP) of nifH (N2 fixation) and nirS and nirK (denitrification), and the compositions and structures of these communities among three sites were compared. The number of operational taxonomic units (OTU) for nifH was higher than that for nirK or nirS at all three sites. Site 3, which had the highest organic matter and sand content in the rhizosphere sediment, as well as the lowest pore water oxygen concentration, had the highest nifH diversity. Principal component analysis of biogeochemical parameters identified soil texture, organic matter content, pore water oxygen concentration, and salinity as the main variables that differentiated the sites. Nonmetric multidimensional scaling (MDS) analyses of the T-RFLP data using the Bray-Curtis coefficient, group analyses, and pairwise comparisons between the sites clearly separated the OTU of site 3 from those of sites 1 and 2. For nirS, there were statistically significant differences in the composition of OTU among the sites, but the variability was less than for nifH. OTU defined on the basis of nirK were highly similar, and the three sites were not clearly separated on the basis of these sequences. The phylogenetic trees of nifH, nirK, and nirS showed that most of the cloned sequences were more similar to sequences from the rhizosphere isolates than to those from known strains or from other environments. PMID:17827324

  11. Effect of microwave irradiation on alfalfa seeds germination and nitrogenase activity of endophytic diazotrophs in seeds

    International Nuclear Information System (INIS)

    Zhang Shuqing; Li Jianfeng; Shi Shangli; Huo Pinghui; Zhu Xinqiang; Zhao Wenhan; Tao Rong

    2011-01-01

    Various microwave powers were used to irradiate alfalfa seeds with various time to study the effect of microwave irradiation on nitrogenase activity of endogenous azotobacter and germination of seeds. Germination rate, germination speed and nitrogenase activity of pure cultures that derived from seed-carried azotobacter were tested. The results indicate that : 800 W, 20 s and 500 W, 40 s are found with highest germination rate on the 1 st day, which is 122% and 88.9% times higher than the control group (P th day is 29.8% and 41.9% times longer than the control group, and more sensitive nitrogenase activity is found on condition of various time than various powers. Short time treatments on condition of the two irradiation powers can increase nitrogenase activity conspicuously, and the treatments that treated more than 32 s make nitrogenase activity lower than the control group, conspicuously. Nitrogenase activity is found 104.9% times higher than the control group on condition of 24 s. (authors)

  12. Endophytic Colonization of Rice by a Diazotrophic Strain of Serratia marcescens

    Science.gov (United States)

    Gyaneshwar, Prasad; James, Euan K.; Mathan, Natarajan; Reddy, Pallavolu M.; Reinhold-Hurek, Barbara; Ladha, Jagdish K.

    2001-01-01

    Six closely related N2-fixing bacterial strains were isolated from surface-sterilized roots and stems of four different rice varieties. The strains were identified as Serratia marcescens by 16S rRNA gene analysis. One strain, IRBG500, chosen for further analysis showed acetylene reduction activity (ARA) only when inoculated into media containing low levels of fixed nitrogen (yeast extract). Diazotrophy of IRBG500 was confirmed by measurement of 15N2 incorporation and by sequence analysis of the PCR-amplified fragment of nifH. To examine its interaction with rice, strain IRBG500 was marked with gusA fused to a constitutive promoter, and the marked strain was inoculated onto rice seedlings under axenic conditions. At 3 days after inoculation, the roots showed blue staining, which was most intense at the points of lateral root emergence and at the root tip. At 6 days, the blue precipitate also appeared in the leaves and stems. More detailed studies using light and transmission electron microscopy combined with immunogold labeling confirmed that IRBG500 was endophytically established within roots, stems, and leaves. Large numbers of bacteria were observed within intercellular spaces, senescing root cortical cells, aerenchyma, and xylem vessels. They were not observed within intact host cells. Inoculation of IRBG500 resulted in a significant increase in root length and root dry weight but not in total N content of rice variety IR72. The inoculated plants showed ARA, but only when external carbon (e.g., malate, succinate, or sucrose) was added to the rooting medium. PMID:11274124

  13. Draft genome sequence of Paraburkholderia tropica Ppe8 strain, a sugarcane endophytic diazotrophic bacterium.

    Science.gov (United States)

    Silva, Paula Renata Alves da; Simões-Araújo, Jean Luiz; Vidal, Márcia Soares; Cruz, Leonardo Magalhães; Souza, Emanuel Maltempi de; Baldani, José Ivo

    Paraburkholderia tropica (syn Burkholderia tropica) are nitrogen-fixing bacteria commonly found in sugarcane. The Paraburkholderia tropica strain Ppe8 is part of the sugarcane inoculant consortium that has a beneficial effect on yield. Here, we report a draft genome sequence of this strain elucidating the mechanisms involved in its interaction mainly with Poaceae. A genome size of approximately 8.75Mb containing 7844 protein coding genes distributed in 526 subsystems was de novo assembled with ABySS and annotated by RAST. Genes related to the nitrogen fixation process, the secretion systems (I, II, III, IV, and VI), and related to a variety of metabolic traits, such as metabolism of carbohydrates, amino acids, vitamins, and proteins, were detected, suggesting a broad metabolic capacity and possible adaptation to plant association. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  14. Compost biofortification with diazotrophic and P-solubilizing bacteria improves maturation process and P availability.

    Science.gov (United States)

    Busato, Jader G; Zandonadi, Daniel B; Mól, Alan R; Souza, Rafaela S; Aguiar, Kamilla P; Júnior, Fábio B Reis; Olivares, Fábio L

    2017-02-01

    Phosphorus-containing fertilizers play an important role in tropical agriculture owing to the well documented shortage of plant-available P in soils. Traditional P fertilizer production is based on chemical processing of insoluble rock phosphate (RP), which includes an acid treatment at high temperature. Processing the RP increases fertilizer costs, making it unavailable for undercapitalized and typically family-based farmers. Biotechnological methods have been proposed as an alternative to increase phosphate availability in RP. In this study, Burkholderia silvatlantica and Herbaspirillum seropedicae were co-inoculated into an RP-enriched compost with the aim of determining the effects of this technology on the levels of phosphatase activities and release of plant-available P. Inoculation of both microorganisms resulted in higher organic matter decomposition and higher humic acid formation in composting. Herbaspirillum seropedicae was the most promising microorganism for the production of acid and alkaline phosphatase enzymes. Both microorganisms presented potential to increase the supply of P from poorly soluble sources owing to increased levels of water-soluble P and citric acid P. Burkholderia silvatlantica and H. seropedicae in RP-enriched compost may represent an important biotechnological tool to reduce the overall time required for composting and increase the supply of P from poorly soluble sources. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  15. Naringenin degradation by the endophytic diazotroph Herbaspirillum seropedicae SmR1.

    Science.gov (United States)

    Marin, A M; Souza, E M; Pedrosa, F O; Souza, L M; Sassaki, G L; Baura, V A; Yates, M G; Wassem, R; Monteiro, R A

    2013-01-01

    Several bacteria are able to degrade flavonoids either to use them as carbon sources or as a detoxification mechanism. Degradation pathways have been proposed for several bacteria, but the genes responsible are not known. We identified in the genome of the endophyte Herbaspirillum seropedicae SmR1 an operon potentially associated with the degradation of aromatic compounds. We show that this operon is involved in naringenin degradation and that its expression is induced by naringenin and chrysin, two closely related flavonoids. Mutation of fdeA, the first gene of the operon, and fdeR, its transcriptional activator, abolished the ability of H. seropedicae to degrade naringenin.

  16. Characterisation of the effect of a simulated hydrocarbon spill on diazotrophs in mangrove sediment mesocosm

    NARCIS (Netherlands)

    Taketani, R.G.; dos Santos, H.F.; van Elsas, J.D.; Rosado, A.S.

    2009-01-01

    An analysis of the effect of an oil spill on mangrove sediments was carried out by contamination of mesocosms derived from two different mangroves, one with a history of contamination and one pristine. The association between N(2) fixers and hydrocarbon degradation was assessed using quantitative

  17. Desenvolvimento Inicial de Duas Variedades de Cana-de-açúcar Inoculadas com Bactérias Diazotróficas

    Directory of Open Access Journals (Sweden)

    Valfredo Almeida Chaves

    2015-12-01

    Full Text Available RESUMO A cana-de-açúcar é uma cultura de grande destaque na economia, em razão da produção de açúcar, etanol e energia. Tecnologias que possam contribuir para o aumento da produtividade e qualidade da cultura com mínimos danos ao meio ambiente são necessárias. Objetivou-se avaliar a produção de ácido indol acético de cinco estirpes de bactérias diazotróficas e o efeito da inoculação delas na brotação de duas variedades de cana-de-açúcar, RB867515 e IACSP95-5000. A produção de auxina foi determinada pelo teste colorimétrico, usando o reagente de Salkowski. Para avaliar a germinação, foi conduzido um experimento em casa de vegetação, utilizando-se delineamento experimental em blocos ao acaso com quatro repetições e sete tratamentos: controle não inoculado; inoculação mista com as cinco estirpes e inoculação individual com Gluconacetobacter diazotrophicus (Gd estirpe BR11281T(PAL-5T, Herbaspirillum seropedicae (Hs - BR11335 = HRC54, Herbaspirillum rubrisubalbicans (Hr - BR11504 = HCC103, Burkholderia tropica (Bt - BR11366T = PPe 8 T e Azospirillum amazonense (Aa - BR11145 = CBAMc. As bactérias mais eficientes na produção de auxina foram Hs e Hr, declinando 48 h após o crescimento. Hr, Aa e Bt aumentaram o índice de velocidade de germinação e o número de brotações nas duas variedades. Na var. RB867515, a velocidade de germinação ainda foi positivamente influenciada pela inoculação mista, sendo o mesmo observado pela inoculação de Gd na var. IACSP95-5000.

  18. Dynamics and controls of heterotrophic prokaryotic production in the western tropical South Pacific Ocean: links with diazotrophic and photosynthetic activity

    Science.gov (United States)

    Van Wambeke, France; Gimenez, Audrey; Duhamel, Solange; Dupouy, Cécile; Lefevre, Dominique; Pujo-Pay, Mireille; Moutin, Thierry

    2018-05-01

    Heterotrophic prokaryotic production (BP) was studied in the western tropical South Pacific (WTSP) using the leucine technique, revealing spatial and temporal variability within the region. Integrated over the euphotic zone, BP ranged from 58 to 120 mg C m-2 d-1 within the Melanesian Archipelago, and from 31 to 50 mg C m-2 d-1 within the western subtropical gyre. The collapse of a bloom was followed during 6 days in the south of Vanuatu using a Lagrangian sampling strategy. During this period, rapid evolution was observed in the three main parameters influencing the metabolic state: BP, primary production (PP) and bacterial growth efficiency. With N2 fixation being one of the most important fluxes fueling new production, we explored relationships between BP, PP and N2 fixation rates over the WTSP. The contribution of N2 fixation rates to bacterial nitrogen demand ranged from 3 to 81 %. BP variability was better explained by the variability of N2 fixation rates than by that of PP in surface waters of the Melanesian Archipelago, which were characterized by N-depleted layers and low DIP turnover times (TDIP 100 h), deeper in the Melanesian Archipelago, or within the entire euphotic zone in the subtropical gyre. The bacterial carbon demand to gross primary production ratio ranged from 0.75 to 3.1. These values are discussed in the framework of various assumptions and conversion factors used to estimate this ratio, including the methodological errors, the daily variability of BP, the bacterial growth efficiency and one bias so far not considered: the ability for Prochlorococcus to assimilate leucine in the dark.

  19. Changes in labile phosphorus forms during maturation of vermicompost enriched with phosphorus-solubilizing and diazotrophic bacteria.

    Science.gov (United States)

    Busato, Jader G; Lima, Lívia S; Aguiar, Natália O; Canellas, Luciano P; Olivares, Fábio L

    2012-04-01

    The aim of this study was to assess the effect of N(2)-fixing and P-solubilizing bacteria during maturation of vermicompost on phosphorus availability. A bacterial suspension containing Burkholderia silvatlantica, Burkholderia spp. and Herbaspirillum seropedicae was applied at the initial stage of vermicomposting. At the end of the incubation time (120days), the nitrogen content had increased by18% compared to uninoculated vermicompost. Water-soluble P was 106% higher in inoculated vermicompost while resin-extractable P increased during the initial vermicomposting stage and was 21% higher at 60days, but was the same in inoculated and uninoculated mature compost. The activity of acid phosphatase was 43% higher in inoculated than uninoculated vermicompost. These data suggest that the introduction of the mixed culture had beneficial effects on vermicompost maturation. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Metaproteomic identification of diazotrophic methanotrophs and their localization in root tissues of field-grown rice plants.

    Science.gov (United States)

    Bao, Zhihua; Okubo, Takashi; Kubota, Kengo; Kasahara, Yasuhiro; Tsurumaru, Hirohito; Anda, Mizue; Ikeda, Seishi; Minamisawa, Kiwamu

    2014-08-01

    In a previous study by our group, CH4 oxidation and N2 fixation were simultaneously activated in the roots of wild-type rice plants in a paddy field with no N input; both processes are likely controlled by a rice gene for microbial symbiosis. The present study examined which microorganisms in rice roots were responsible for CH4 oxidation and N2 fixation under the field conditions. Metaproteomic analysis of root-associated bacteria from field-grown rice (Oryza sativa Nipponbare) revealed that nitrogenase complex-containing nitrogenase reductase (NifH) and the alpha subunit (NifD) and beta subunit (NifK) of dinitrogenase were mainly derived from type II methanotrophic bacteria of the family Methylocystaceae, including Methylosinus spp. Minor nitrogenase proteins such as Methylocella, Bradyrhizobium, Rhodopseudomonas, and Anaeromyxobacter were also detected. Methane monooxygenase proteins (PmoCBA and MmoXYZCBG) were detected in the same bacterial group of the Methylocystaceae. Because these results indicated that Methylocystaceae members mediate both CH4 oxidation and N2 fixation, we examined their localization in rice tissues by using catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). The methanotrophs were localized around the epidermal cells and vascular cylinder in the root tissues of the field-grown rice plants. Our metaproteomics and CARD-FISH results suggest that CH4 oxidation and N2 fixation are performed mainly by type II methanotrophs of the Methylocystaceae, including Methylosinus spp., inhabiting the vascular bundles and epidermal cells of rice roots. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  1. Decoupling of ammonium regulation and ntcA transcription in the diazotrophic marine cyanobacterium Trichodesmium sp. IMS101.

    Science.gov (United States)

    Post, Anton F; Rihtman, Branko; Wang, Qingfeng

    2012-03-01

    Nitrogen (N) physiology in the marine cyanobacterium Trichodesmium IMS101 was studied along with transcript accumulation of the N-regulatory gene ntcA and of two of its target genes: napA (nitrate assimilation) and nifH (N(2) fixation). N(2) fixation was impaired in the presence of nitrite, nitrate and urea. Strain IMS101 was capable of growth on these combined N sources at ammonium. Whereas ecologically relevant N concentrations (2-20 μM) suppressed growth and assimilation, much higher concentrations were required to affect transcript levels. Transcripts of nifH accumulated under nitrogen-fixing conditions; these transcript levels were maintained in the presence of nitrate (100 μM) and ammonium (20 μM). However, nifH transcript levels were below detection at ammonium concentrations >20 μM. napA mRNA was found at low levels in both N(2)-fixing and ammonium-utilizing filaments, and it accumulated in filaments grown with nitrate. The positive effect of nitrate on napA transcription was abolished by ammonium additions of >200 μM. This effect was restored upon addition of the glutamine synthetase inhibitor L-methionin-DL-sulfoximine. Surprisingly, ntcA transcript levels remained high in the presence of ammonium, even at elevated concentrations. These findings indicate that ammonium repression is decoupled from transcriptional activation of ntcA in Trichodesmium IMS101.

  2. The small unicellular diazotrophic symbiont, UCYN-A, is a key player in the marine nitrogen cycle

    DEFF Research Database (Denmark)

    Martínez-Pérez, Clara; Mohr, Wiebke; Löscher, Carolin R

    2016-01-01

    to live in symbioses with specific eukaryotic algae. Single-cell analyses showed that both algae-UCYN-A symbioses actively fixed N2, contributing ∼20% to N2 fixation in the tropical North Atlantic, revealing their significance in this region. These symbioses had growth rates five to ten times higher than...

  3. RNA-seq analyses reveal insights into the function of respiratory nitrate reductase of the diazotroph Herbaspirillum seropedicae.

    Science.gov (United States)

    Bonato, Paloma; Batista, Marcelo B; Camilios-Neto, Doumit; Pankievicz, Vânia C S; Tadra-Sfeir, Michelle Z; Monteiro, Rose Adele; Pedrosa, Fabio O; Souza, Emanuel M; Chubatsu, Leda S; Wassem, Roseli; Rigo, Liu Un

    2016-09-01

    Herbaspirillum seropedicae is a nitrogen-fixing β-proteobacterium that associates with roots of gramineous plants. In silico analyses revealed that H. seropedicae genome has genes encoding a putative respiratory (NAR) and an assimilatory nitrate reductase (NAS). To date, little is known about nitrate metabolism in H. seropedicae, and, as this bacterium cannot respire nitrate, the function of NAR remains unknown. This study aimed to investigate the function of NAR in H. seropedicae and how it metabolizes nitrate in a low aerated-condition. RNA-seq transcriptional profiling in the presence of nitrate allowed us to pinpoint genes important for nitrate metabolism in H. seropedicae, including nitrate transporters and regulatory proteins. Additionally, both RNA-seq data and physiological characterization of a mutant in the catalytic subunit of NAR (narG mutant) showed that NAR is not required for nitrate assimilation but is required for: (i) production of high levels of nitrite, (ii) production of NO and (iii) dissipation of redox power, which in turn lead to an increase in carbon consumption. In addition, wheat plants showed an increase in shoot dry weight only when inoculated with H. seropedicae wild type, but not with the narG mutant, suggesting that NAR is important to H. seropedicae-wheat interaction. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  4. Vibrio plantisponsor sp. nov., a diazotrophic bacterium isolated from a mangrove associated wild rice (Porteresia coarctata Tateoka)

    Digital Repository Service at National Institute of Oceanography (India)

    Rameshkumar, N.; Gomez-Gil, B.; Sproer, C.; Lang, E.; Kumar, N.D.; Krishnamurthi, S.; Nair, S.; Roque, A.

    nifH amplicon was purified using Eppendorf Perfectprep Gel Cleanup Kit (according to the manufacturer’s instructions). The purified nifH amplicon was used directly for DNA sequencing with the dideoxy chain termination method with the Big...Dye terminator kit (Applied Biosystems) using the above mentioned nifH primers, and the reaction products were analysed by capillary electrophoresis on an ABI 310 Genetic Analyzer (Applied Biosystems). For phylogenetic analysis, partial nifH nucleotide...

  5. Novel European free-living, non-diazotrophic Bradyrhizobium isolates from contrasting soils that lack nodulation and nitrogen fixation genes - a genome comparison

    Science.gov (United States)

    Jones, Frances Patricia; Clark, Ian M.; King, Robert; Shaw, Liz J.; Woodward, Martin J.; Hirsch, Penny R.

    2016-05-01

    The slow-growing genus Bradyrhizobium is biologically important in soils, with different representatives found to perform a range of biochemical functions including photosynthesis, induction of root nodules and symbiotic nitrogen fixation and denitrification. Consequently, the role of the genus in soil ecology and biogeochemical transformations is of agricultural and environmental significance. Some isolates of Bradyrhizobium have been shown to be non-symbiotic and do not possess the ability to form nodules. Here we present the genome and gene annotations of two such free-living Bradyrhizobium isolates, named G22 and BF49, from soils with differing long-term management regimes (grassland and bare fallow respectively) in addition to carbon metabolism analysis. These Bradyrhizobium isolates are the first to be isolated and sequenced from European soil and are the first free-living Bradyrhizobium isolates, lacking both nodulation and nitrogen fixation genes, to have their genomes sequenced and assembled from cultured samples. The G22 and BF49 genomes are distinctly different with respect to size and number of genes; the grassland isolate also contains a plasmid. There are also a number of functional differences between these isolates and other published genomes, suggesting that this ubiquitous genus is extremely heterogeneous and has roles within the community not including symbiotic nitrogen fixation.

  6. Ideonella azotifigens sp. nov., an aerobic diazotroph of the Betaproteobacteria isolated from grass rhizosphere soil, and emended description of the genus Ideonella.

    Science.gov (United States)

    Noar, Jesse D; Buckley, Daniel H

    2009-08-01

    Strain 1a22T, a nitrogen-fixing bacterium, was isolated from soil associated with the rhizosphere of a perennial grass growing in a fallow agricultural field in Ithaca, New York, USA. Analysis of the 16S rRNA gene sequence placed the strain in the Rubrivivax-Roseateles-Leptothrix-Azohydromonas-Aquincola-Ideonella branch of the Betaproteobacteria and the closest characterized relative was the type strain of Ideonella dechloratans (97.7% 16S rRNA sequence similarity). Cells of strain 1a22T were Gram-negative, motile, straight rods, which formed polyhydroxybutyrate-like granules and were positive for oxidase and weakly positive for catalase. Cells were chemo-organotrophic, unable to grow by reduction of chlorate or nitrate and grew exclusively through aerobic respiration. Growth with mannitol on N-free solid media caused the strain to produce copious amounts of slime. The G+C content of the genomic DNA was 67.4 mol%. The major cellular fatty acids were C16:1 cis-9 and C16:0 and cells contained significant amounts of the hydroxy fatty acids C10:0 3-OH, C12:0 2-OH and C12:0 3-OH. Based on DNA-DNA hybridization studies, 16S rRNA gene sequence analysis, fatty acid analysis, and morphological and physiological characteristics, strain 1a22T represents a novel species in the genus Ideonella, for which the name Ideonella azotifigens sp. nov. is proposed. The type strain of Ideonella azotifigens is 1a22T (=JCM 15503T=DSM 21438T).

  7. A common transport system for methionine, L-methionine-DL-sulfoximine (MSX), and phosphinothricin (PPT) in the diazotrophic cyanobacterium Nostoc muscorum.

    Science.gov (United States)

    Singh, Arvind Kumar; Syiem, Mayashree B; Singh, Rajkumar S; Adhikari, Samrat; Rai, Amar Nath

    2008-05-01

    We present evidence, for the first time, of the occurrence of a transport system common for amino acid methionine, and methionine/glutamate analogues L-methionine-DL-sulfoximine (MSX) and phosphinothricin (PPT) in cyanobacterium Nostoc muscorum. Methionine, which is toxic to cyanobacterium, enhanced its nitrogenase activity at lower concentrations. The cyanobacterium showed a biphasic pattern of methionine uptake activity that was competitively inhibited by the amino acids alanine, isoleucine, leucine, phenylalanine, proline, valine, glutamine, and asparagine. The methionine/glutamate analogue-resistant N. muscorum strains (MSX-R and PPT-R strains) also showed methionine-resistant phenotype accompanied by a drastic decrease in 35S methionine uptake activity. Treatment of protein extracts from these mutant strains with MSX and PPT reduced biosynthetic glutamine synthetase (GS) activity only in vitro and not in vivo. This finding implicated that MSX- and PPT-R phenotypes may have arisen due to a defect in their MSX and PPT transport activity. The simultaneous decrease in methionine uptake activity and in vitro sensitivity toward MSX and PPT of GS protein in MSX- and PPT-R strains indicated that methionine, MSX, and PPT have a common transport system that is shared by other amino acids as well in N. muscorum. Such information can become useful for isolation of methionine-producing cyanobacterial strains.

  8. Chemical characterization of the gels produced by the diazotrophic bacteria Rhizobium tropici and Mesorhizobium sp; Caracterizacao quimica dos geis produzidos pelas bacterias diazotroficas Rhizobium tropici e Mesorhizobium sp.

    Energy Technology Data Exchange (ETDEWEB)

    Monteiro, Nilson Kobori [Departamento de Engenharia e Tecnologia de Alimentos, Instituto de Biociencias, Letras e Ciencias Exatas, Universidade Estadual Paulista, Sao Jose do Rio Preto - SP (Brazil); Aranda-Selverio, Gabriel; Exposti, Diego Tadeu Degli; Silva, Maria de Lourdes Corradi da [Departamento de Fisica, Quimica e Biologia, Faculdade de Ciencias e Tecnologia, Universidade Estadual Paulista, Presidente Prudente - SP (Brazil); Lemos, Eliana Gertrudes Macedo; Campanharo, Joao Carlos [Departamento de Tecnologia, Faculdade de Ciencias Agrarias e Veterinaria, Universidade Estadual Paulista, Jaboticabal - SP (Brazil); Silveira, Joana Lea Meira [Departamento de Bioquimica e Biologia Molecular, Universidade Federal do Parana, Curitiba - PR (Brazil)

    2012-07-01

    The exopolysaccharides with characteristics of gel produced by Rhizobium tropici (EPSRT) and Mesorhizobium sp (EPSMR) are acidic heteropolysaccharide composed mainly of glucose and galactose in a molar ratio of 4:1 and 5:1 respectively, with traces of mannose ({approx} 1%). Chemical analysis showed the presence of uronic acid, pyruvate and acetyl-substituents in the structures of both polymers. Experiments of gel permeation chromatography and polyacrylamide gel electrophoresis showed that EPSRT and EPSMR are homogeneous molecules with low grade of polydispersity. The EPS were characterized using spectroscopic techniques of FT-IR, {sup 1}H and {sup 13}C-NMR. (author)

  9. OBTAINING AND ACTIVITY OF SILVER NANOPARTICLES BASED ON THE EXOPOLYSACCHARIDE OF DIAZOTROPHIC STRAIN Bradyrhizobium japonicum 36 AND AgNO3

    Directory of Open Access Journals (Sweden)

    B. A. Rasulov

    2014-12-01

    Full Text Available The aim of the research was to obtain silver nanoparticles and to study of its antibacterial activity against pathogenic cultures Escherichia coli ATCC11229 and Staphylococcus aureus ATCC6538. Obtained complex was characterized by UV-Vis spectroscopy, X-ray diffraction (XRD and transmission electron microscopy (TEM. Colloid solutions of the complex absorbed radiation in the visible regions of 420–460 nm, which confirmed the formation of silver nanoparticles. The size of synthesized SNP varied from 6 to 50 nm. There has been confirmed presence in them of atomic silver. Obtained compound possessed antibacterial activity against pathogens, such as Escherichia coli ATCC11229 and Staphylococcus aureus ATCC6538.

  10. Transcriptional profiling of nitrogen fixation and the role of NifA in the diazotrophic endophyte Azoarcus sp. strain BH72.

    Directory of Open Access Journals (Sweden)

    Abhijit Sarkar

    Full Text Available BACKGROUND: The model endophyte Azoarcus sp. strain BH72 is known to contribute fixed nitrogen to its host Kallar grass and also expresses nitrogenase genes endophytically in rice seedlings. Availability of nitrogen is a signal regulating the transcription of nitrogenase genes. Therefore, we analysed global transcription in response to differences in the nitrogen source. METHODOLOGY/PRINCIPAL FINDINGS: A DNA microarray, comprising 70-mer oligonucleotides representing 3989 open reading frames of the genome of strain BH72, was used for transcriptome studies. Transcription profiles of cells grown microaerobically on N2 versus ammonium were compared. Expression of 7.2% of the genes was significantly up-regulated, and 5.8% down-regulated upon N2 fixation, respectively. A parallel genome-wide prediction of σ(54-type promoter elements mapped to the upstream region of 38 sequences of which 36 were modulated under the N2 response. In addition to modulation of genes related to N2 fixation, the expressions of gene clusters that might be related to plant-microbe interaction and of several transcription factors were significantly enhanced. While comparing under N2-fixation conditions the transcriptome of wild type with a nifLA(- insertion mutant, NifA being the essential transcriptional activator for nif genes, 24.5% of the genome was found to be affected in expression. A genome-wide prediction of 29 NifA binding sequences matched to 25 of the target genes whose expression was differential during microarray analysis, some of which were putatively negatively regulated by NifA. For selected genes, differential expression was corroborated by real time RT-PCR studies. CONCLUSION/SIGNIFICANCE: Our data suggest that life under conditions of nitrogen fixation is an important part of the lifestyle of strain BH72 in roots, as a wide range of genes far beyond the nif regulon is modulated. Moreover, the NifA regulon in strain BH72 appears to encompass a wider range of cellular functions beyond the regulation of nif genes.

  11. Towards long-read metagenomics: complete assembly of three novel genomes from bacteria dependent on a diazotrophic cyanobacterium in a freshwater lake co-culture.

    Science.gov (United States)

    Driscoll, Connor B; Otten, Timothy G; Brown, Nathan M; Dreher, Theo W

    2017-01-01

    Here we report three complete bacterial genome assemblies from a PacBio shotgun metagenome of a co-culture from Upper Klamath Lake, OR. Genome annotations and culture conditions indicate these bacteria are dependent on carbon and nitrogen fixation from the cyanobacterium Aphanizomenon flos-aquae, whose genome was assembled to draft-quality . Due to their taxonomic novelty relative to previously sequenced bacteria, we have temporarily designated these bacteria as incertae sedis Hyphomonadaceae strain UKL13-1 (3,501,508 bp and 56.12% GC), incertae sedis Betaproteobacterium strain UKL13-2 (3,387,087 bp and 54.98% GC), and incertae sedis Bacteroidetes strain UKL13-3 (3,236,529 bp and 37.33% GC). Each genome consists of a single circular chromosome with no identified plasmids. When compared with binned Illumina assemblies of the same three genomes, there was ~7% discrepancy in total genome length. Gaps where Illumina assemblies broke were often due to repetitive elements. Within these missing sequences were essential genes and genes associated with a variety of functional categories. Annotated gene content reveals that both Proteobacteria are aerobic anoxygenic phototrophs, with Betaproteobacterium UKL13-2 potentially capable of phototrophic oxidation of sulfur compounds. Both proteobacterial genomes contain transporters suggesting they are scavenging fixed nitrogen from A. flos-aquae in the form of ammonium. Bacteroidetes UKL13-3 has few completely annotated biosynthetic pathways, and has a comparatively higher proportion of unannotated genes. The genomes were detected in only a few other freshwater metagenomes, suggesting that these bacteria are not ubiquitous in freshwater systems. Our results indicate that long-read sequencing is a viable method for sequencing dominant members from low-diversity microbial communities, and should be considered for environmental metagenomics when conditions meet these requirements.

  12. Diazotroph-Bacterial Community Structure of Root Nodules Account for Two-Fold Differences in Plant Growth: Consequences for Global Biogeochemical Cycles

    Science.gov (United States)

    Williams, M. A.

    2016-12-01

    The bacterial communities that inhabit and function as mutualists in the nodules of soybean, a major worldwide crop, are a fundamental determinant of plant growth and global nitrogen and carbon cycles. Unfertilized soybean can derive up to 90% of its nitrogen through bacterial-driven diazotrophy. It was the goal of the research in this study to assess whether different bacterial taxa (e.g. Bradyrhizobia spp.) differ in their soybean growth supportive role, which could then feedback to alter global biogeochemical cycling. Using 16S rRNA and NifH genes, nodule bacterial communities were shown to vary across 9 different cultivars of soybean, and that the variation between cultivars were highly correlated to plant yield (97 to 188 bu/Ha) and nitrogen. The relative abundances of gene sequences associated with the closest taxonomic match (NCBI), indicated that several taxa were (r= 0.76) negatively (e.g. Bradyrhizobium sp Ec3.3) or (r= 0.84) positively (e.g. Bradyrhizobium elkanii WSM 2783) correlated with plant yield. Other non-Rhizobiaceae taxa, such as Rhodopseudomonas spp. were also prevalent and correlated with plant yield. Soybeans and other leguminous crops will become increasingly important part of world food production, soil fertility and global biogeochemical cycles with rising population and food demand. The study demonstrates the importance of plant-microbial feedbacks driving plant growth but also ramifications for global cycling of nitrogen and carbon.

  13. Utilization of Vinegar for Isolation of Cellulose Producing Acetic Acid Bacteria

    International Nuclear Information System (INIS)

    Aydin, Y. Andelib; Aksoy, Nuran Deveci

    2010-01-01

    Wastes of traditionally fermented Turkish vinegar were used in the isolation of cellulose producing acetic acid bacteria. Waste material was pre-enriched in Hestrin-Schramm medium and microorganisms were isolated by plating dilution series on HS agar plates The isolated strains were subjected to elaborate biochemical and physiological tests for identification. Test results were compared to those of reference strains Gluconacetobacter xylinus DSM 46604, Gluconacetobacter hansenii DSM 5602 and Gluconacetobacter liquefaciens DSM 5603. Seventeen strains, out of which only three were found to secrete the exopolysaccharide cellulose. The highest cellulose yield was recorded as 0.263±0.02 g cellulose L -1 for the strain AS14 which resembled Gluconacetobacter hansenii in terms of biochemical tests.

  14. Isolation and characterization of a bacterial celluloseproducing ...

    African Journals Online (AJOL)

    year ripened persimmon vinegar, which belonged to the family of Acetobacteracea based on its morphological and physiological characteristics. The phylogenetic position of the isolated strain was most closely related to Gluconacetobacter ...

  15. Untapped Endophytic Colonization and Plant Growth-Promoting Potential of the Genus Novosphingobium to Optimize Rice Cultivation

    OpenAIRE

    Rangjaroen, Chakrapong; Sungthong, Rungroch; Rerkasem, Benjavan; Teaumroong, Neung; Noisangiam, Rujirek; Lumyong, Saisamorn

    2017-01-01

    With the aim of searching for potent diazotrophic bacteria that are free of public health concerns and optimize rice cultivation, the endophytic colonization and plant growth-promoting activities of some endophytic diazotrophic bacteria isolated from rice were evaluated. Among these bacteria, the emerging diazotrophic strains of the genus Novosphingobium effectively associated with rice plant interiors and consequently promoted the growth of rice, even with the lack of a nitrogen source. Thes...

  16. The Role of the Horizontal Gene Pool and Lateral Gene Transfer in Enhancing Microbial Activities in Marine Sediments

    Science.gov (United States)

    2006-05-10

    nifH encoding plasmids of diazotrophic bacteria isolated from roots of a salt marsh grass. Meeting Abstract, 105th General Meeting of the American...When the method was applied to 100 endogenous plasmids isolated from cultivated marine diazotrophs from salt marsh grass rhizoplane niches remarkably...Beeson, K.E., D.L. Erdner, C.E. Bagwell, C.R. Lovell, and P.A. Sobecky. 2002. Differentiation of plasmids in marine diazotroph assemblages

  17. Polyaromatic hydrocarbon (PAH) degradation potential of a new acid tolerant, diazotrophic P-solubilizing and heavy metal resistant bacterium Cupriavidus sp. MTS-7 isolated from long-term mixed contaminated soil.

    Science.gov (United States)

    Kuppusamy, Saranya; Thavamani, Palanisami; Megharaj, Mallavarapu; Lee, Yong Bok; Naidu, Ravi

    2016-11-01

    An isolate of Cupriavidus (strain MTS-7) was identified from a long-term PAHs and heavy metals mixed contaminated soil with the potential to biodegrade both LMW and HMW PAHs with added unique traits of acid and alkali tolerance, heavy metal tolerance, self-nutrient assimilation by N fixation and P solubilization. This strain completely degraded the model 3 (150 mg L(-1) Phe), 4 (150 mg L(-1) Pyr) and 5 (50 mg L(-1) BaP) ring PAHs in 4, 20 and 30 days, respectively. It could mineralize 90-100% of PAHs (200 mg L(-1) of Phe and Pyr) within 15 days across pH ranging from 5 to 8 and even in the presence of toxic metal contaminations. During biodegradation, the minimum inhibitory concentrations were 5 (Cu(2+)) and 3 (Cd(2+), Pb(2+), Zn(2+)) mg L(-1) of the potentially bioavailable metal ions and over 17 mg L(-1) metal levels was lethal for the microbe. Further, it could fix 217-274 μg mL(-1) of N and solubilize 79-135 μg mL(-1) of P while PAHs degradation. MTS-7 as a superior candidate could be thus used in the enhanced bioaugmentation and/or phytoremediation of long-term mixed contaminated sites. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Selection and evaluation of reference genes for RT-qPCR expression studies on Burkholderia tropica strain Ppe8, a sugarcane-associated diazotrophic bacterium grown with different carbon sources or sugarcane juice.

    Science.gov (United States)

    da Silva, Paula Renata Alves; Vidal, Marcia Soares; de Paula Soares, Cleiton; Polese, Valéria; Simões-Araújo, Jean Luís; Baldani, José Ivo

    2016-11-01

    Among the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness by evaluating the relative expression of genes involved in glycogen and trehalose biosynthesis when strain Ppe8 was grown on different carbon sources and sugarcane juice.

  19. Comparison of Bacterial Cellulose Production among Different Strains and Fermented Media

    Directory of Open Access Journals (Sweden)

    Maryam Jalili Tabaii

    2015-12-01

    Full Text Available The effect of different carbon sources on bacterial cellulose production by Gluconacetobacter xylinus (PTCC 1734 and two newly isolated strains (from vinegar under static culture conditions was studied. The production of bacterial cellulose was examined in modified Hestrin-Shramm medium by replacing D-glucose with other carbon sources. The results showed that the yield and characteristics of bacterial cellulose were influenced by the type of carbon source. Glycerol gave the highest yield in all of the studied strains (6%, 9.7% and 3.8% for S, A2 strain and Gluconacetobacter xylinus (PTCC 1734, respectively. The maximum dry bacterial cellulose weight in the glycerol containing medium is due to A2 strain (1.9 g l-1 in comparison to Gluconacetobacter xylinus as reference strain (0.76 g l-1. Although all of the studied strains were in Gluconacetobacter family, each used different sugars for maximum production after glycerol (mannitol and fructose for two newly isolated strains and glucose for Gluconacetobacter xylinus. The maximum moisture content was observed when sucrose and food-grade sucrose were used as carbon source. Contrary to expectations, while the maximum thickness of bacterial cellulose membrane was attained when glycerol was used, bacterial cellulose from glycerol had less moisture content than the others. The oxidized cellulose showed antibacterial activities, which makes it as a good candidate for food-preservatives.

  20. Molecular Ecology of nifH Genes and Transcripts Along a Chronosequence in Revegetated Areas of the Tengger Desert.

    Science.gov (United States)

    Wang, Jin; Bao, Jing-Ting; Li, Xin-Rong; Liu, Yu-Bing

    2016-01-01

    The colonization and succession of diazotrophs are essential for the development of organic soil layers in desert. We examined the succession of diazotrophs in the well-established revegetated areas representing a chronosequence of 0 year (control), 22 years (restored artificially since 1981), 57 years (restored artificially since 1956), and more than 100 years (restored naturally) to determine the community assembly and active expression of diazotrophs. The pyrosequencing data revealed that Alphaproteobacteria-like diazotrophs predominated in the topsoil of our mobile dune site, while cyanobacterial diazotrophs predominated in the revegetated sites. The cyanobacterial diazotrophs were primarily composed of the heterocystous genera Anabaena, Calothrix, Cylindrospermum, Nodularia, Nostoc, Trichormus, and Mastigocladus. Almost all the nifH sequences belonged to the Cyanobacteria phylum (all the relative abundance values >99.1 %) at transcript level and all the active cyanobacterial diazotrophs distributed in the families Nostocaceae and Rivulariaceae. The most dominant active cyanobacterial genus was Cylindrospermum in all the samples. The rank abundance and community analyses demonstrated that most of the diazotrophic diversity originated from the "rare" species, and all the DNA-based diazotrophic libraries were richer and more diverse than their RNA-based counterparts in the revegetated sites. Significant differences in the diazotrophic community and their active population composition were observed among the four research sites. Samples from the 1981-revegetating site (predominated by cyanobacterial crusts) showed the highest nitrogenase activity, followed by samples from the naturally revegetating site (predominated by lichen crusts), the 1956-revegetating site (predominated by moss crusts), and the mobile dune site (without crusts). Collectively, our data highlight the importance of nitrogen fixation by the primary successional desert topsoil and suggest

  1. Metabolic adaptation, a specialized leaf organ structure and vascular responses to diurnal N2 fixation by nostoc azollae sustain the astonishing productivity of azolla ferns without nitrogen fertilizer

    NARCIS (Netherlands)

    Brouwer, Paul; Bräutigam, Andrea; Buijs, Valerie A.; Tazelaar, Anne O.E.; van der Werf, Adrie; Schlüter, Urte; Reichart, Gert-Jan; Bolger, Anthony; Usadel, Björn; Weber, Andreas P.M.; Schluepmann, Henriette

    2017-01-01

    Sustainable agriculture demands reduced input of man-made nitrogen (N) fertilizer, yet N2 fixation limits the productivity of crops with heterotrophic diazotrophic bacterial symbionts. We investigated floating ferns from the genus Azolla that host phototrophic diazotrophic Nostoc azollae in leaf

  2. Metabolic adaptation, a specialized leaf organ structure and vascular responses to diurnal N

    NARCIS (Netherlands)

    Brouwer, Paul; Bräutigam, Andrea; Buijs, Valerie A.; Tazelaar, Anne O.E.; Werf, van der Adrie; Schlüter, Urte; Reichart, Gert Jan; Bolger, Anthony; Usadel, Björn; Weber, Andreas P.M.; Schluepmann, Henriette

    2017-01-01

    Sustainable agriculture demands reduced input of man-made nitrogen (N) fertilizer, yet N2 fixation limits the productivity of crops with heterotrophic diazotrophic bacterial symbionts. We investigated floating ferns from the genus Azolla that host phototrophic diazotrophic Nostoc azollae in leaf

  3. A Comprehensive Modeling Approach Towards Understanding and Prediction of the Alaskan Coastal System Response to Changes in an Ice-diminished Arctic

    Science.gov (United States)

    2009-09-30

    ammonifying and nitrifying bacteria ; six groups of phytoplankton (diatoms, dinoflagellates, coccolithophores, colonial prymnesiophytes, microflagellates...and diazotrophs ), for which cell counts were made at the species level during 2002, 2003, and 2004 cruises of the Polar Star, Healy, Mirai, Louis St...groups of backscattering coccolithophores and diazotrophs , compared to the other phytoplankton groups. Finally, food web closures is effected in the

  4. Water absorption and maintenance of nanofiber cellulose ...

    African Journals Online (AJOL)

    DR. NJ TONUKARI

    2012-05-17

    May 17, 2012 ... Physiochemical properties of bacterial cellulose producing by Gluconacetobacter rhaeticus TL-2C was ... shape of the mold (Czaja et al., 2006). ... impurity, and then it was freeze-dried and ground to a fine ... Figure 1. Microstructure and chemical structure of bacterial cellulose producing G. rhaeticus TL-2C.

  5. GenBank blastx search result: AK287855 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287855 J065196O16 AB091060.1 AB091060 Gluconacetobacter xylinus acsAB, acsC, acsD genes for cellulose... synthase subunit AB, cellulose synthase subunit C, cellulose synthase subunit D, complete cds. BCT 4e-15 0 ...

  6. GenBank blastx search result: AK242831 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242831 J090067L08 AB091060.1 AB091060 Gluconacetobacter xylinus acsAB, acsC, acsD genes for cellulose... synthase subunit AB, cellulose synthase subunit C, cellulose synthase subunit D, complete cds. BCT 1e-14 1 ...

  7. Symbiosis revisited : Phosphorus and acid buffering stimulate N2 fixation but not Sphagnum growth

    NARCIS (Netherlands)

    Van Den Elzen, Eva; Kox, Martine A R; Harpenslager, Sarah F.; Hensgens, Geert; Fritz, Christian; Jetten, Mike S M; Ettwig, Katharina F.; Lamers, Leon P M

    2017-01-01

    In pristine Sphagnum-dominated peatlands, (di)nitrogen (N2) fixing (diazotrophic) microbial communities associated with Sphagnum mosses contribute substantially to the total nitrogen input, increasing carbon sequestration. The rates of symbiotic nitrogen fixation reported for Sphagnum peatlands,

  8. Nitrogen fixation rates in the eastern Arabian Sea

    Digital Repository Service at National Institute of Oceanography (India)

    Ahmed, A.; Gauns, M.; Kurian, S.; Bardhan, P.; Pratihary, A.K.; Naik, H.; Shenoy, D.M.; Naqvi, S.W.A.

    The Arabian Sea experiences bloom of the diazotroph Trichodesmium during certain times of the year when optimal sea surface temperature and oligotrophic condition favour their growth. We measured nitrogen fixation rates in the euphotic zone during...

  9. Use of 15N-isotope dilution for quantification of nitrogen fixation in Saccharum SPP

    International Nuclear Information System (INIS)

    Singh, M.; Singh, G.B.; Joshi, B.B.

    1994-01-01

    The objectives of present study were to quantify and compare the amount of nitrogen fixed by diazotrophes associated with roots of Saccharum spontaneum, S. barberi and S. sinense using Sclerotachya fusca as a non-fixing control. 5 refs

  10. Genetically modified cyanobacterium Nostoc muscorum ...

    Indian Academy of Sciences (India)

    Madhu

    diazotrophic cultures of the cyanobacterium N. muscorum in quantities of 5 .... growth media and then used for estimation of their characteristics. Each reading is ..... 1224–1232. Christian J H B 1950 The influence of nutrition on the water rela-.

  11. Oxygen and the light-dark cycle of nitrogenase activity in two unicellular cyanobacteria

    NARCIS (Netherlands)

    Compaore, J.; Stal, L.J.

    2010-01-01

    Cyanobacteria capable of fixing dinitrogen exhibit various strategies to protect nitrogenase from inactivation by oxygen. The marine Crocosphaera watsonii WH8501 and the terrestrial Gloeothece sp. PCC6909 are unicellular diazotrophic cyanobacteria that are capable of aerobic nitrogen fixation. These

  12. Nitrogenase genes in non-cyanobacterial plankton: prevalence, diversity, and regulation in marine waters

    DEFF Research Database (Denmark)

    Riemann, Lasse; Farnelid, H.; Steward, G.F.

    2010-01-01

    Marine waters are generally considered to be nitrogen (N) limited and are therefore favourable environments for diazotrophs, i.e. organisms converting atmospheric N2 into ammonium or nitrogen oxides available for growth. In some regions, this import of N supports up to half of the primary...... productivity. Diazotrophic Cyanobacteria appear to be the major contributors to marine N2 fixation in surface waters, whereas the contribution of heterotrophic or chemoautotrophic diazotrophs to this process is usually regarded inconsequential. Culture-independent studies reveal that non......-cyanobacterial diazotrophs are diverse, widely distributed, and actively expressing the nitrogenase gene in marine and estuarine environments. The detection of nifH genes and nifH transcripts, even in N-replete marine waters, suggests that N2 fixation is an ecologically important process throughout the oceans. Because...

  13. Nitrogen-fixing bacteria in Mediterranean seagrass (Posidonia oceanica) roots

    KAUST Repository

    Garcias Bonet, Neus; Arrieta, J M; Duarte, Carlos M.; Marbà , Nú ria

    2016-01-01

    of nitrogen for P. oceanica. The low diversity of nitrogen-fixing bacteria reported here suggests species-specific relationships between diazotrophs and P. oceanica, revealing possible symbiotic interactions that could play a major role in nitrogen acquisition

  14. Occurrence of cyanobacteria - diatom symbiosis in the Bay of Bengal: Implications in biogeochemistry

    Digital Repository Service at National Institute of Oceanography (India)

    Kulkarni, V.V.; Chitari, R.R.; Narale, D.D.; Patil, J.S.; Anil, A.C.

    cyanobacterial endosymbiont of several diatom genera, are the most important diazotrophs accounting for approx. 63% of pelagic N sub(2) -fixation in the world’s oceans. However, information on R. intracellularis is lacking, even though its distribution...

  15. Beijerinckia derxii stimulates the viability of non-N2-fixing bacteria in nitrogen-free media

    Directory of Open Access Journals (Sweden)

    Barbosa Heloiza R.

    2000-01-01

    Full Text Available The interactions between the nitrogen-fixing microorganism Beijerinckia derxii with two non-diazotrophic bacteria, either Escherichia coli or a facultative sulphur-oxidizing chemolitotroph, were studied in mixed cultures. Direct and indirect contact between B. derxii and E. coli were tested. B. derxii increased CFU numbers and/or maintained the viability of the non-diazotrophic bacteria, but neither growth nor nitrogenase activity of the nitrogen-fixing bacterium were affected by either partner.

  16. Corals Form Characteristic Associations with Symbiotic Nitrogen-Fixing Bacteria

    Science.gov (United States)

    Lema, Kimberley A.; Willis, Bette L.

    2012-01-01

    The complex symbiotic relationship between corals and their dinoflagellate partner Symbiodinium is believed to be sustained through close associations with mutualistic bacterial communities, though little is known about coral associations with bacterial groups able to fix nitrogen (diazotrophs). In this study, we investigated the diversity of diazotrophic bacterial communities associated with three common coral species (Acropora millepora, Acropora muricata, and Pocillopora damicormis) from three midshelf locations of the Great Barrier Reef (GBR) by profiling the conserved subunit of the nifH gene, which encodes the dinitrogenase iron protein. Comparisons of diazotrophic community diversity among coral tissue and mucus microenvironments and the surrounding seawater revealed that corals harbor diverse nifH phylotypes that differ between tissue and mucus microhabitats. Coral mucus nifH sequences displayed high heterogeneity, and many bacterial groups overlapped with those found in seawater. Moreover, coral mucus diazotrophs were specific neither to coral species nor to reef location, reflecting the ephemeral nature of coral mucus. In contrast, the dominant diazotrophic bacteria in tissue samples differed among coral species, with differences remaining consistent at all three reefs, indicating that coral-diazotroph associations are species specific. Notably, dominant diazotrophs for all coral species were closely related to the bacterial group rhizobia, which represented 71% of the total sequences retrieved from tissue samples. The species specificity of coral-diazotroph associations further supports the coral holobiont model that bacterial groups associated with corals are conserved. Our results suggest that, as in terrestrial plants, rhizobia have developed a mutualistic relationship with corals and may contribute fixed nitrogen to Symbiodinium. PMID:22344646

  17. Aeração e adição de sais na produção de ácido indol acético por bactérias diazotróficas Aeration and salt effects on indol acetic production by diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Tharwat El-Sayed El-Desouk Radwan

    2005-10-01

    Full Text Available Foi analisada a produção de compostos indólicos por Azospirillum brasilense Cd, A. lipoferum Br 17, Herbaspirillum seropedicae Z 67, H. rubrisubalbicans M4 e a estirpe 34 isolada de arroz, que não se enquadra em nenhuma das espécies de Herbaspirillum já descritas, em relação a diferentes condições de aeração e concentrações de sais. A maior aeração do meio propiciou aumento na produção de compostos indólicos pelas bactérias testadas. Foi verificado aumento desses compostos, em culturas estáticas, em meio sem nitrogênio no caso de Azospirillum, e na presença de N para as estirpes de Herbaspirillum. O aumento da concentração de sais no meio de cultivo inibiu a produção de compostos indólicos, embora tenha sido observado um pequeno aumento quando a concentração de CaCl2 foi de 1 g L-1. O efeito mais deletério da salinidade foi observado com a presença de NaHCO3, seguido de NaCl e Na2SO4. Azospirillum produziu mais compostos indólicos em meio semi-sólido e Herbaspirillum em meio líquido, mas em menor nível.The production of indolic compounds by Azospirillum brasilense Cd, A. lipoferum Br 17, Herbaspirillum seropedicae Z 67, H. rubrisubalbicans M4, and strain 34 isolated from rice, which does not fit into the described Herbaspirillum species, was measured under aeration ratio and salt concentrations. Aeration of the medium increased growth and production of indole compounds by these bacteria. Under static condition, the production was higher both in nitrogen-free medium for Azospirillum, and in amended N medium for the Herbaspirillum strains. Increasing salt concentration into the medium inhibited the production of indole compounds, although a small increase in production was observed, when CaCl2 concentration was raised above 1 g L-1. Deleterious effect of salinity was more pronounced in the presence of NaHCO3, followed by NaCl and Na2SO4. Azospirillum produced more indolic compounds in semi-solid cultures, and Herbaspirillum in liquid medium, but at lower levels.

  18. Effect of neem cake/fertilizers on symbiotic and non-symbiotic N2 fixing bacteria

    International Nuclear Information System (INIS)

    Akhtar, S.; Solangi, A.H.; Gilani, G.; Pirzada, M.H.

    2002-01-01

    Neem cake amendment in soil at 1.3% no adverse effect on the population of four symbiotic Rhizobium species viz., japonicum, R. leguminosarum, R. Phaseoli and R. Fredii and three non-symbiotic free living nitrogen fixers bacteria viz., Pseudomonas diazotrophicus, Klebsiella planticola and Enterobacter cloacae. Neem cake extracted with n-hexane stimulated the growth of Rhizobium species in vitro, whereas Neem cake expeller extracted neither inhibited nor stimulated the growth of Rhizobium species except for R. Fredii, whose was slightly retarded. The fertilizers (urea, NPK and DAP) had no adverse effect on these bacteria even at the dosage ten times higher the recommended dose. (author)

  19. Synthesis of thermoplastic starch-bacterial cellulose nanocomposites via in situ fermentation

    OpenAIRE

    Osorio, Marlon A.; Restrepo, David; Velásquez-Cock, Jorge A.; Zuluaga, Robin O.; Montoya, Ursula; Rojas, Orlando; Gañán, Piedad F.; Marin, Diana; Castro, Cristina I.

    2014-01-01

    In this paper, a nanocomposite based on thermoplastic starch (TPS) reinforced with bacterial cellulose (BC) nanoribbons was synthesized by in situ fermentation and chemical crosslinking. BC nanoribbons were produced by a Colombian native strain of Gluconacetobacter medellinensis; the nanocomposite was plasticized with glycerol and crosslinked with citric acid. The reinforcement percentage in the nanocomposites remained constant throughout the fermentation time because of the TPS absorption ca...

  20. Dissolved Organic Matter Influences N2 Fixation in the New Caledonian Lagoon (Western Tropical South Pacific

    Directory of Open Access Journals (Sweden)

    Mar Benavides

    2018-03-01

    Full Text Available Specialized prokaryotes performing biological dinitrogen (N2 fixation (“diazotrophs” provide an important source of fixed nitrogen in oligotrophic marine ecosystems such as tropical and subtropical oceans. In these waters, cyanobacterial photosynthetic diazotrophs are well known to be abundant and active, yet the role and contribution of non-cyanobacterial diazotrophs are currently unclear. The latter are not photosynthetic (here called “heterotrophic” and hence require external sources of organic matter to sustain N2 fixation. Here we added the photosynthesis inhibitor 3-(3,4-dichlorophenyl-1,1-dimethylurea (DCMU to estimate the N2 fixation potential of heterotrophic diazotrophs as compared to autotrophic ones. Additionally, we explored the influence of dissolved organic matter (DOM on these diazotrophs along a coast to open ocean gradient in the surface waters of a subtropical coral lagoon (New Caledonia. Total N2 fixation (samples not amended with DCMU ranged from 0.66 to 1.32 nmol N L−1 d−1. The addition of DCMU reduced N2 fixation by >90%, suggesting that the contribution of heterotrophic diazotrophs to overall N2 fixation activity was minor in this environment. Higher contribution of heterotrophic diazotrophs occurred in stations closer to the shore and coincided with the decreasing lability of DOM, as shown by various colored DOM and fluorescent DOM (CDOM and FDOM indices. We tested the response of diazotrophs (in terms of nifH gene expression and bulk N2 fixation rates upon the addition of a mix of carbohydrates (“DOC” treatment, amino acids (“DON” treatment, and phosphonates and phosphomonesters (“DOP” treatment. While nifH expression increased significantly in Trichodesmium exposed to the DOC treatment, bulk N2 fixation rates increased significantly only in the DOP treatment. The lack of nifH expression by gammaproteobacteria, in any of the DOM addition treatments applied, questions the contribution of non

  1. Response of free-living nitrogen-fixing microorganisms to land use change in the Amazon rainforest.

    Science.gov (United States)

    Mirza, Babur S; Potisap, Chotima; Nüsslein, Klaus; Bohannan, Brendan J M; Rodrigues, Jorge L M

    2014-01-01

    The Amazon rainforest, the largest equatorial forest in the world, is being cleared for pasture and agricultural use at alarming rates. Tropical deforestation is known to cause alterations in microbial communities at taxonomic and phylogenetic levels, but it is unclear whether microbial functional groups are altered. We asked whether free-living nitrogen-fixing microorganisms (diazotrophs) respond to deforestation in the Amazon rainforest, using analysis of the marker gene nifH. Clone libraries were generated from soil samples collected from a primary forest, a 5-year-old pasture originally converted from primary forest, and a secondary forest established after pasture abandonment. Although diazotroph richness did not significantly change among the three plots, diazotroph community composition was altered with forest-to-pasture conversion, and phylogenetic similarity was higher among pasture communities than among those in forests. There was also 10-fold increase in nifH gene abundance following conversion from primary forest to pasture. Three environmental factors were associated with the observed changes: soil acidity, total N concentration, and C/N ratio. Our results suggest a partial restoration to initial levels of abundance and community structure of diazotrophs following pasture abandonment, with primary and secondary forests sharing similar communities. We postulate that the response of diazotrophs to land use change is a direct consequence of changes in plant communities, particularly the higher N demand of pasture plant communities for supporting aboveground plant growth.

  2. Environmental conditions outweigh geographical contiguity in determining the similarity of nifH-harboring microbial communities in sediments of two disconnected marginal seas

    Directory of Open Access Journals (Sweden)

    Haixia Zhou

    2016-07-01

    Full Text Available Ecological evidence suggests that heterotrophic diazotrophs fueled by organic carbon respiration in sediments play an important role in marine nitrogen fixation. However, fundamental knowledge about the identities, abundance, diversity, biogeography and controlling environmental factors of nitrogen-fixing communities in open ocean sediments is still elusive. Surprisingly, little is known also about nitrogen-fixing communities in sediments of the more research-accessible marginal seas. Here we report on an investigation of the environmental geochemistry and putative diazotrophic microbiota in the sediments of Bohai Sea, an eutrophic marginal sea of the western Pacific Ocean. Diverse and abundant nifH gene sequences were identified and sulfate-reducing bacteria were found to be the dominant putative nitrogen-fixing microbes. Community statistical analyses suggested bottom water temperature, bottom water chlorophyll a content (or the covarying turbidity and sediment porewater Eh (or the covarying pH as the most significant environmental factors controlling the structure and spatial distribution of the putative diazotrophic communities, while sediment Hg content, sulfide content and porewater SiO32--Si content were identified as the key environmental factors correlated positively with the nifH gene abundance in Bohai Sea sediments. Comparative analyses between the Bohai Sea and the northern South China Sea identified a significant composition difference of the putative diazotrophic communities in sediments between the shallow-water (estuarine and nearshore and deep-water (offshore and deep-sea environments, and sediment porewater dissolved oxygen content, water depth and in situ temperature as the key environmental factors tentatively controlling the species composition, community structure and spatial distribution of the marginal sea sediment nifH-harboring microbiota. This confirms the ecophysiological specialization and niche differentiation

  3. Mesozooplankton Graze on Cyanobacteria in the Amazon River Plume and Western Tropical North Atlantic

    Directory of Open Access Journals (Sweden)

    Brandon J. Conroy

    2017-08-01

    Full Text Available Diazotrophic cyanobacteria, those capable of fixing di-nitrogen (N2, are considered one of the major sources of new nitrogen (N in the oligotrophic tropical ocean, but direct incorporation of diazotrophic N into food webs has not been fully examined. In the Amazon River-influenced western tropical North Atlantic (WTNA, diatom diazotroph associations (DDAs and the filamentous colonial diazotrophs Trichodesmium have seasonally high abundances. We sampled epipelagic mesozooplankton in the Amazon River plume and WTNA in May–June 2010 to investigate direct grazing by mesozooplankton on two DDA populations: Richelia associated with Rhizosolenia diatoms (het-1 and Hemiaulus diatoms (het-2, and on Trichodesmium using highly specific qPCR assays targeting nitrogenase genes (nifH. Both DDAs and Trichodesmium occurred in zooplankton gut contents, with higher detection of het-2 predominantly in calanoid copepods (2.33–16.76 nifH copies organism-1. Abundance of Trichodesmium was low (2.21–4.03 nifH copies organism-1, but they were consistently detected at high salinity stations (>35 in calanoid copepods. This suggests direct grazing on DDAs, Trichodesmium filaments and colonies, or consumption as part of sinking aggregates, is common. In parallel with the qPCR approach, a next generation sequencing analysis of 16S rRNA genes identified that cyanobacterial assemblage associated with zooplankton guts was dominated by the non-diazotrophic unicellular phylotypes Synechococcus (56% and Prochlorococcus (26%. However, in two separate calanoid copepod samples, two unicellular diazotrophs Candidatus Atelocyanobacterium thalassa (UCYN-A and Crocosphaera watsonii (UCYN-B were present, respectively, as a small component of cyanobacterial assemblages (<2%. This study represents the first evidence of consumption of DDAs, Trichodesmium, and unicellular cyanobacteria by calanoid copepods in an area of the WTNA known for high carbon export. These diazotroph populations

  4. Incorporation of nitrogen from N2 fixation into amino acids of zooplankton

    DEFF Research Database (Denmark)

    Loick-Wilde, Natalie; Dutz, Jörg; Miltner, Anja

    2012-01-01

    quantified the direct incorporation of 15N tracer from N2-fixing N. spumigena (diazotroph nitrogen) and ammonium-utilizing R. salina into the amino acid nitrogen (AA-N) of zooplankton using complementary gas chromatography– combustion–isotope ratio mass spectrometry, gas chromatography–mass spectrometry...... consistently low in E. affinis when exposed to N. spumigena, suggesting that these animals were reluctant to feed on N. spumigena. Essential isoleucine received most of the diazotroph nitrogen in field zooplankton, while nonessential amino acids received most 15N tracer in E. affinis. N. spumigena was clearly...... an important amino acid nitrogen source for Baltic Sea zooplankton...

  5. Diversity and activity of nitrogen fixing archaea and bacteria associated with micro-environments of wetland rice

    Science.gov (United States)

    Schmidt, Hannes; Woebken, Dagmar

    2017-04-01

    Wetland rice is one of the world's most important crop plants. The cultivation on waterlogged paddy soils is strongly limited by nitrogen (N), which is typically supplied by industrial fertilizers that are not only costly but also exhibit hazardous effects on the environment. It has been reported that "Biological Nitrogen Fixation" through N2-fixing bacteria and archaea (diazotrophs) can alleviate the N-shortage in rice cultivation, thus carrying out an important ecosystem function. However, our understanding of the diversity and in situ N2 fixation activity of diazotrophs in flooded rice fields is still rudimentary. Moreover, knowledge on the impact of biochemical gradients established by root activity (i.e. exudation, radial oxygen loss) on the functioning of N-fixing microorganisms in paddy soil ecosystems is limited. We aimed at studying underlying processes on biologically relevant scales. Greenhouse studies were performed to identify key factors that control rice-diazotroph association and related N2 fixation activities. Paddy soils of different geographical origin were cultivated with two commercially used genotypes of wetland rice. Samples were separated into bulk soil, rhizosphere soil, rhizoplane, and roots at flowering stage of rice plant development. These samples were subjected to functional assays and various molecular biological techniques in order to analyze the associated diazotroph communities. Based on Illumina amplicon sequencing of nifH genes and transcripts, we show that the diversity and potential activity of diazotroph communities varies according to micro-environments. We will comparatively discuss the influence of (a) the soil microbial "seed bank" and (b) plant genotype in shaping the respective microbiomes and selecting for potentially active diazotrophs. Actual N2 fixation activities of soil-genotype combinations and micro-environments will be shown on the basis of incubation assays using 15N2-containing atmospheres. Areas of potential

  6. Cyanobacteria Occurrence and Nitrogen Fixation Rates in the ...

    African Journals Online (AJOL)

    The occurrence and biological nitrogen fixation rates of epiphytic and benthic diazotrophs were studied in seagrass meadows at sites with seaweed farms and at a control site without seaweed farms from two locations, Chwaka Bay and Jambiani, along the east coast of. Zanzibar. Ten species of cyanobacteria were ...

  7. Nitrogenase Activity in Intertidal Sediment Along the Tanzanian ...

    African Journals Online (AJOL)

    significant differences in nitrogenase activity between the upper, mid and lower littoral zones, and between the three sampling locations, i.e. Tanga, Dar es Salaam and Mafia. A nitrogen fixation rate of 38 mmol N m-2y-1 was estimated. We conclude that diazotrophs contribute a substantial amount of nitrogen input to the ...

  8. Temperature induced changes in the heterocyst glycolipid composition of N

    NARCIS (Netherlands)

    Bauersachs, T.; Stal, L.J.; Grego, M.; Schwark, L.; Schwark, L.

    2014-01-01

    We investigated the effect of temperature on the heterocyst glycolipid (HG) composition of the diazotrophic heterocystous cyanobacteria Anabaena sp. strain CCY9613 and Nostoc sp. strain CCY9926 grown at 9, 12, 16, 20 and 24 degrees C. Both strains contained an overall similar composition of

  9. Temperature induced changes in the heterocyst glycolipid composition of N2-fixing heterocystous cyanobacteria

    NARCIS (Netherlands)

    Bauersachs, T.; Stal, L.J.; Grego, M.; Schwark, L.

    2014-01-01

    We investigated the effect of temperature on the heterocyst glycolipid (HG) composition of the diazotrophic heterocystous cyanobacteria Anabaena sp. strain CCY9613 and Nostoc sp. strain CCY9926 grown at 9, 12, 16, 20 and 24 °C. Both strains contained an overall similar composition of heterocyst

  10. Oxygen and temperature in relation to nitrogen fixation in cyanobacteria: In the daily life of cyanobacteria

    NARCIS (Netherlands)

    Compaoré, J.

    2010-01-01

    The results described in this thesis reveal that unicellular diazotrophic cyanobacteria are unable to fix N2 under fully aerobic conditions. This suggests that in their natural environment these organisms must be exposed to O2 concentrations well below air saturation at least during the periods that

  11. Nitrogen cycling in summer active perennial grass systems in South Australia: Non-symbiotic nitrogen fixation

    NARCIS (Netherlands)

    Gupta, V.V.S.R.; Kroker, S.J.; Hicks, M.; Davoren, W.; Descheemaeker, K.K.E.; Llewellyn, R.

    2014-01-01

    Non-symbiotic nitrogen (N2) fixation by diazotrophic bacteria is a potential source for biological N inputs in non-leguminous crops and pastures. Perennial grasses generally add larger quantities of above- and belowground plant residues to soil, and so can support higher levels of soil biological

  12. Microbial quality of soil from the Pampa biome in response to different grazing pressures

    Directory of Open Access Journals (Sweden)

    Rafael S. Vargas

    2015-06-01

    Full Text Available The aim of this study was to evaluate the impact of different grazing pressures on the activity and diversity of soil bacteria. We performed a long-term experiment in Eldorado do Sul, southern Brazil, that assessed three levels of grazing pressure: high pressure (HP, with 4% herbage allowance (HA, moderate pressure (MP, with 12% HA, and low pressure (LP, with 16% HA. Two reference areas were also assessed, one of never-grazed native vegetation (NG and another of regenerated vegetation after two years of grazing (RG. Soil samples were evaluated for microbial biomass and enzymatic (β-glucosidase, arylsulfatase and urease activities. The structure of the bacterial community and the population of diazotrophic bacteria were evaluated by RFLP of the 16S rRNA and nifH genes, respectively. The diversity of diazotrophic bacteria was assessed by partial sequencing of the 16S rDNA gene. The presence of grazing animals increased soil microbial biomass in MP and HP. The structures of the bacterial community and the populations of diazotrophic bacteria were altered by the different grazing managements, with a greater diversity of diazotrophic bacteria in the LP treatment. Based on the characteristics evaluated, the MP treatment was the most appropriate for animal production and conservation of the Pampa biome.

  13. What Is the True Nitrogenase Reaction? A Guided Approach

    Science.gov (United States)

    Ipata, Piero L.; Pesi, Rossana

    2015-01-01

    Only diazotrophic bacteria, called "Rizhobia," living as symbionts in the root nodules of leguminous plants and certain free-living prokaryotic cells can fix atmospheric N[subscript 2]. In these microorganisms, nitrogen fixation is carried out by the nitrogenase protein complex. However, the reduction of nitrogen to ammonia has an…

  14. Biological nitrogen fixation is not a major contributor to the nitrogen demand of a commercially growth South African sugarcane cultivar

    NARCIS (Netherlands)

    Hoefsloot, G.; Termorshuizen, A.J.; Watt, D.A.; Cramer, M.D.

    2005-01-01

    It has previously been reported that endophytic diazotrophic bacteria contribute significantly to the nitrogen budgets of some graminaceous species. In this study the contribution of biological nitrogen fixation to the N-budget of a South African sugarcane cultivar was evaluated using 15N natural

  15. Differential accumulation of nif structural gene mRNA in Azotobacter vinelandii.

    Science.gov (United States)

    Hamilton, Trinity L; Jacobson, Marty; Ludwig, Marcus; Boyd, Eric S; Bryant, Donald A; Dean, Dennis R; Peters, John W

    2011-09-01

    Northern analysis was employed to investigate mRNA produced by mutant strains of Azotobacter vinelandii with defined deletions in the nif structural genes and in the intergenic noncoding regions. The results indicate that intergenic RNA secondary structures effect the differential accumulation of transcripts, supporting the high Fe protein-to-MoFe protein ratio required for optimal diazotrophic growth.

  16. Potential for nitrogen fixation in fungus-growing termite symbiosis

    DEFF Research Database (Denmark)

    Sapountzis, Panagiotis; de Verges, Jane; Rousk, Kathrin

    2016-01-01

    Termites host a gut microbiota of diverse and essential symbionts that enable specialization on dead plant material; an abundant, but nutritionally imbalanced food source. To supplement the severe shortage of dietary nitrogen (N), some termite species make use of diazotrophic bacteria to fix atmo...

  17. KONSISTENSI PRODUKSI NATA DALAM MEDIA FERMENTASI YANG MENGANDUNG HIDROLISAT UBI KAYU

    Directory of Open Access Journals (Sweden)

    Lutfansyah Muchtar

    2017-11-01

    Full Text Available Nata de coco is known as a gelatinous substance, white to creamy-yellow, firm, glossy and not sticky, produced by some bacteria, which forms on the surface of coconut water medium during fermentation. Acetobacter xylinum (current name: Gluconacetobacter xylinus is the common bacteria which synthesized nata de coco. Besides coconut water, there are some alternatives for making nata, such as cassava. Gluconacetobacter sp. was employed in this study to produce nata de coco from cassava hydrolysate medium (U and compared it qualitatively and quantitatively to coconut water medium (K. In U medium, glucose was the main product derived from hydrolysis of cassava. Fermentation was conducted three times with each former fermentations became sources of inocula for the next fermentations, respectively. All data were analysed with IBM SPSS Statistics 23. Fermentation product of nata from U medium were relatively stable in thickness, wet weight, dry weight, yield, water content, and log from total bacteria (quantitative and in colour, texture, transparency, and surface of nata (qualitative. Glucose was rapidly consumed by bacteria then other sugar (fructose and sucrose. Glucose consumption and nata yield was affected by the number of bacteria. However, nata yield was not significantly influenced by glucose consumption. Nata yield was possibly influenced by other factors such as nutrient content in U medium. Growth dynamics of Gluconacetobacter sp. was measured in log cfu mL-1 on Hassid-Barker Agar (HBA medium and relative number of 16S rRNA genes. Bacterial growth during fermentation in U medium were relatively more stable than fermentation on K medium although the yield in K medium gradually increased and were higher than U medium.

  18. Nanomaterials from bacterial cellulose for antimicrobial wound dressing

    Science.gov (United States)

    Liyaskina, E.; Revin, V.; Paramonova, E.; Nazarkina, M.; Pestov, N.; Revina, N.; Kolesnikova, S.

    2017-01-01

    Bacterial nanocellulose (BNC) is widely used in biomedical applications. BNC has attracted increasing attention as a novel wound dressing material, but it has no antimicrobial activity. To get over this problem in the present study the BNC was saturated with antibiotic fusidic acid (FA). The subject of the experiment was BNC, produced by bacteria Gluconacetobacter sucrofermentans B-11267. The resulting biocomposites have high antibiotic activity against Staphylococcus aureus and can be used in medicine as a wound dressing. The structure of BNC was analyzed by atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR).

  19. Nanomaterials from bacterial cellulose for antimicrobial wound dressing

    International Nuclear Information System (INIS)

    Liyaskina, E; Revin, V; Paramonova, E; Nazarkina, M; Pestov, N; Revina, N; Kolesnikova, S

    2017-01-01

    Bacterial nanocellulose (BNC) is widely used in biomedical applications. BNC has attracted increasing attention as a novel wound dressing material, but it has no antimicrobial activity. To get over this problem in the present study the BNC was saturated with antibiotic fusidic acid (FA). The subject of the experiment was BNC, produced by bacteria Gluconacetobacter sucrofermentans B-11267. The resulting biocomposites have high antibiotic activity against Staphylococcus aureus and can be used in medicine as a wound dressing. The structure of BNC was analyzed by atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR). (paper)

  20. Preparation and characterization of biocellulose membranes obtained by biosynthesis of bacteria komagataeibacter rhaeticus

    International Nuclear Information System (INIS)

    Machado, Rachel Temperani A.; Tercjak, Agnieszka; Gutierrez, Juncal; Barud, Hernane S.

    2015-01-01

    This work shows preparation and characterization of bacterial cellulose membranes (CB) produced by Komagataeibacter rhaeticus strain, and strain from ATCC bank, Gluconacetobacter hansenii (ATCC 23769) used as a comparative. Physicochemical assay were done, such as Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD), Scanning Electron Microscopy couplet to Energy-dispersive X-ray spectroscopy (SEM/EDX). SEM images reveled a compact structure formed by cellulose nanofibers. FTIR spectra are shown characteristics bands of cellulosic materials and XRD shows cellulose native structure, type I cellulose. (author)

  1. Desenvolvimento de membranas compósitas à base de nanocelulose para pilhas de combustível

    OpenAIRE

    Martins, Ana Patrícia Carvalho

    2016-01-01

    A presente dissertação tem como objetivo o desenvolvimento de membranas polieletrólitas baseadas em celulose bacteriana (CB) para aplicação em pilhas de combustível de condução protónica, que se espera ser uma solução ambientalmente sustentável. A CB é um material celulósico, com uma estrutura nanofibrilar tridimensional, biossintetizada por algumas estirpes de bactérias não patogénicas como é o caso da Gluconacetobacter sacchari. Devido a algumas das suas propriedades, nomeadamente a sua ele...

  2. Preparation and characterization of biocellulose membranes obtained by biosynthesis of bacteria komagataeibacter rhaeticus; Preparacao e caracterizacao de membranas de biocelulose obtidas pela biossintese da bacteria komagataeibacter rhaeticus

    Energy Technology Data Exchange (ETDEWEB)

    Machado, Rachel Temperani A. [Centro Universitario de Araraquara (QUIMERA), Araraquara, SP (Brazil). Lab. de Quimica Medicinal e Medicina Regenerativa; Trovatti, Eliane; Ribeiro, Sidney J.L. [Universidade Estadual Paulista Julio de Mesquita Filho (UNESP), Araraquara, SP (Brazil). Instituto de Quimica; Berreta, Andresa A. [Apis Flora LTDA, Ribeirao Preto, SP (Brazil); Tercjak, Agnieszka; Gutierrez, Juncal; Barud, Hernane S., E-mail: hernane.barud@gmail.com, E-mail: hsbarud@uniara.com.br [Group ' Materials + Technologies' (GMT), Department of Chemical and Environmental Engineering, Polytechnic School, University of the Basque Country (UPV/EHU), San Sebastian (Spain)

    2015-07-01

    This work shows preparation and characterization of bacterial cellulose membranes (CB) produced by Komagataeibacter rhaeticus strain, and strain from ATCC bank, Gluconacetobacter hansenii (ATCC 23769) used as a comparative. Physicochemical assay were done, such as Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD), Scanning Electron Microscopy couplet to Energy-dispersive X-ray spectroscopy (SEM/EDX). SEM images reveled a compact structure formed by cellulose nanofibers. FTIR spectra are shown characteristics bands of cellulosic materials and XRD shows cellulose native structure, type I cellulose. (author)

  3. Distribution of nitrogen fixation and nitrogenase-like sequences amongst microbial genomes

    Science.gov (United States)

    2012-01-01

    Background The metabolic capacity for nitrogen fixation is known to be present in several prokaryotic species scattered across taxonomic groups. Experimental detection of nitrogen fixation in microbes requires species-specific conditions, making it difficult to obtain a comprehensive census of this trait. The recent and rapid increase in the availability of microbial genome sequences affords novel opportunities to re-examine the occurrence and distribution of nitrogen fixation genes. The current practice for computational prediction of nitrogen fixation is to use the presence of the nifH and/or nifD genes. Results Based on a careful comparison of the repertoire of nitrogen fixation genes in known diazotroph species we propose a new criterion for computational prediction of nitrogen fixation: the presence of a minimum set of six genes coding for structural and biosynthetic components, namely NifHDK and NifENB. Using this criterion, we conducted a comprehensive search in fully sequenced genomes and identified 149 diazotrophic species, including 82 known diazotrophs and 67 species not known to fix nitrogen. The taxonomic distribution of nitrogen fixation in Archaea was limited to the Euryarchaeota phylum; within the Bacteria domain we predict that nitrogen fixation occurs in 13 different phyla. Of these, seven phyla had not hitherto been known to contain species capable of nitrogen fixation. Our analyses also identified protein sequences that are similar to nitrogenase in organisms that do not meet the minimum-gene-set criteria. The existence of nitrogenase-like proteins lacking conserved co-factor ligands in both diazotrophs and non-diazotrophs suggests their potential for performing other, as yet unidentified, metabolic functions. Conclusions Our predictions expand the known phylogenetic diversity of nitrogen fixation, and suggest that this trait may be much more common in nature than it is currently thought. The diverse phylogenetic distribution of nitrogenase

  4. Distribution of nitrogen fixation and nitrogenase-like sequences amongst microbial genomes

    Directory of Open Access Journals (Sweden)

    Dos Santos Patricia C

    2012-05-01

    Full Text Available Abstract Background The metabolic capacity for nitrogen fixation is known to be present in several prokaryotic species scattered across taxonomic groups. Experimental detection of nitrogen fixation in microbes requires species-specific conditions, making it difficult to obtain a comprehensive census of this trait. The recent and rapid increase in the availability of microbial genome sequences affords novel opportunities to re-examine the occurrence and distribution of nitrogen fixation genes. The current practice for computational prediction of nitrogen fixation is to use the presence of the nifH and/or nifD genes. Results Based on a careful comparison of the repertoire of nitrogen fixation genes in known diazotroph species we propose a new criterion for computational prediction of nitrogen fixation: the presence of a minimum set of six genes coding for structural and biosynthetic components, namely NifHDK and NifENB. Using this criterion, we conducted a comprehensive search in fully sequenced genomes and identified 149 diazotrophic species, including 82 known diazotrophs and 67 species not known to fix nitrogen. The taxonomic distribution of nitrogen fixation in Archaea was limited to the Euryarchaeota phylum; within the Bacteria domain we predict that nitrogen fixation occurs in 13 different phyla. Of these, seven phyla had not hitherto been known to contain species capable of nitrogen fixation. Our analyses also identified protein sequences that are similar to nitrogenase in organisms that do not meet the minimum-gene-set criteria. The existence of nitrogenase-like proteins lacking conserved co-factor ligands in both diazotrophs and non-diazotrophs suggests their potential for performing other, as yet unidentified, metabolic functions. Conclusions Our predictions expand the known phylogenetic diversity of nitrogen fixation, and suggest that this trait may be much more common in nature than it is currently thought. The diverse phylogenetic

  5. OCORRÊNCIA E ATIVIDADE DE BACTÉRIAS DIAZOTRÓFICAS EM FORRAGEIRAS CULTIVADAS NA REGIÃO SEMIÁRIDA NO BRASIL

    Directory of Open Access Journals (Sweden)

    MÁRCIA CARNEIRO MONTEIRO DOS SANTOS

    2013-01-01

    Full Text Available Diazotrophic bacteria play important role on economy nitrogen fertilizer in forage glasses and environmental sustainability. With the objective of evaluate the occurrence of diazotrophic bacteria associated associated with native forage at two different times and identify isolates with potential for fixing N2 in the semiarid region. Was conducted in the Soil Laboratory of the Centro de Saúde e Tecnologia Rural in Federal University of Campina Grande, Campus de Patos, Paraíba. A study was carried out at the Soil Laboratory of the Centro de Saúde e Tecnologia Rural of the Federal University of Campina Grande, in Patos, Paraíba. The objective this study was to verify in the native diazotrophic endophytic bacteria associated with forage glasses and to identify isolates in NBF hability, during the rainy and dry seasons in semiarid region. Root samples of three forage grasses [Andropogon gayanus Kunth (Andropogon, Panicum maximum Jacq. cv. Tanzania (Tanzânia and Cenchrus ciliaris L. (Buffel] cultivated in the field experiment was utilized. The bacteria were isolated in NFb culture media free-N specific for Azospirillum spp., the isolates was quantified, phenotypically characterized, purified. From caracterized after, was evaluated for their nitrogenase activity in vitro. The resultas show diazotrophics bacteria growth in the three forage grasses tested. The ocorrency diazotrophics bacteria be influencied by genotipic plant and seasonal variation. A more density of NSDB of the genus Azospirillum associated in the root grasses were obtained in dry season. The NSDB isolates, show a high potencial for new studies on the genetic caracteristcs and avaliation the inoculo potential to (BNF associated on grasses in semiarid region.

  6. Assessing the loading and release of metronidazole from bacterial cellulose film as a pharmaceutical dressing

    Directory of Open Access Journals (Sweden)

    Mohammad Ali Salehi

    2017-08-01

    Full Text Available Background: Bacterial cellulose membrane (BCM produced by Gluconacetobacter xylinus is an advantageous bacterial product and because of its unique properties could be used as an ideal dressing. The aim of this study was to consider the capability of this biomaterial in the release of Metronidazole. In the case of proving this capability, it provides the basis for the production of a dressing containing this type of antibiotic. Materials and Methods: In this study, BCM was initially synthesized by Gluconacetobacter xylinus. The BCM was loaded by Metronidazole. Then the release process was considered in distilled water and buffer phosphate Saline. The ultra violet spectrophotometry was applied for measuring the concentration of the released drug. Results: The chemical structure of bacterial cellulose was confirmed by Fourier Transform Infrared (FT-IR spectroscopy. The release of Metronidazole in distilled water and phosphate buffered Saline was reached to 84.27% and 84.71%, respectively. Due to higher release in phosphate buffered Saline media, it seems that the trend of release in vitro provides efficient results. Conclusion: Results of this study provides the basis for future research on supplying an ideal dressing from this microbial product.

  7. Komagataeibacter rhaeticus as an alternative bacteria for cellulose production.

    Science.gov (United States)

    Machado, Rachel T A; Gutierrez, Junkal; Tercjak, Agnieszka; Trovatti, Eliane; Uahib, Fernanda G M; Moreno, Gabriela de Padua; Nascimento, Andresa P; Berreta, Andresa A; Ribeiro, Sidney J L; Barud, Hernane S

    2016-11-05

    A strain isolated from Kombucha tea was isolated and used as an alternative bacterium for the biosynthesis of bacterial cellulose (BC). In this study, BC generated by this novel bacterium was compared to Gluconacetobacter xylinus biosynthesized BC. Kinetic studies reveal that Komagataeibacter rhaeticus was a viable bacterium to produce BC according to yield, thickness and water holding capacity data. Physicochemical properties of BC membranes were investigated by UV-vis and Fourier transform infrared spectroscopies (FTIR), thermogravimetrical analysis (TGA) and X-ray diffraction (XRD). Additionally, scanning electron microscopy (SEM) and atomic force microscopy (AFM) were also used for morphological characterization. Mechanical properties at nano and macroscale were studied employing PeakForce quantitative nanomechanical property mapping (QNM) and dynamic mechanical analyzer (DMA), respectively. Results confirmed that BC membrane biosynthesized by Komagataeibacter rhaeticus had similar physicochemical, morphological and mechanical properties than BC membrane produced by Gluconacetobacter xylinus and can be widely used for the same applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Effects of Altered Temperature & Precipitation on Soil Bacterial & Microfaunal Communities as Mediated by Biological Soil Crusts

    Energy Technology Data Exchange (ETDEWEB)

    Neher, Deborah A. [University of Vermont

    2004-08-31

    With increased temperatures in our original pot study we observed a decline in lichen/moss crust cover and with that a decline in carbon and nitrogen fixation, and thus a probable decline of C and N input into crusts and soils. Soil bacteria and fauna were affected negatively by increased temperature in both light and dark crusts, and with movement from cool to hot and hot to hotter desert climates. Crust microbial biomass and relative abundance of diazotrophs was reduced greatly after one year, even in pots that were not moved from their original location, although no change in diazotroph community structure was observed. Populations of soil fauna moved from cool to hot deserts were affected more negatively than those moved from hot to hotter deserts.

  9. Study on the improvement of dinitrogen fixation on rice-bacteria associative system

    International Nuclear Information System (INIS)

    Huang Yide; Zhu Hairong; Qiu Yuansheng; Iyama, S.; Fuji, T.; Hirota, Y.

    1991-01-01

    Inoculation of improved associative diazotrophs Klebsiella oxytoca NG 13 or Enterobacter cloacae E26 was performed in association with three varietes of rice (Oryza sativa L.), namely Indica type C544, Japonica type T65 and Harebare. These improved diazotrophs carried a nifA containing plasmid pMC73A derived from Klebsiella pneumoniae M5al. In pot experiments the rice seedlings were planted in non-sterilized soil and grown under flooded conditions in the green hours. By using 15 N dilution technique it has shown that inoculation of these improved bacterial strains (NG13/pMC73A or E26/pMC73A) stimulated the growth rate and increased the nitrogen content of rice plants in comparison with that of Nif - phenotype K.oxytoca NG1389 and E.cloacae E262. The nitrogen-fixing rate was 10-35% Ndfa after 120 days cultivation

  10. BACTÉRIAS DIAZOTRÓFICAS E ADUBAÇÃO NITROGENADA EM CULTIVARES DE ARROZ

    Directory of Open Access Journals (Sweden)

    SALOMÃO LIMA GUIMARÃES

    2010-01-01

    Full Text Available The purpose of this study was to evaluate the effect of diazotrophic bacteria inoculation and nitrogen fertilization on grain yield and nitrogen accumulation in rice plants. We used seeds of rice cultivars IR42 and IAC4440, which were inoculated with peat containing the strains ZAE94 (BR 11417 - Herbaspirillum seropedicae and M130 (BR 11340 - Burkholderia sp.. We used the first 20 cm of an Albaqualf Ecological Series. The experiment was conducted in a completely randomized design with four replications and four plants per pot. We applied three levels of nitrogen (0, 50 and 100 kg N ha--1 and evaluated the parameters of shoot dry weight, nitrogen accumulation in shoots and grains, in addition to grain yield and the quantification of population of diazotrophic plants. Positive effects of inoculation were observed in both cultivars studied in all parameters evaluated, mainly in the accumulation of nitrogen in plants and grains.

  11. The Biology of Heterotrophic N2-fixing Bacteria in Marine and Estuarine Waters

    DEFF Research Database (Denmark)

    Bentzon-Tilia, Mikkel

    Biological nitrogen (N)2 fixation is of paramount importance for marine N cycling and for life in the oceans in general. It represents the sole mechanism by which microorganisms can channel inert atmospheric N2 gas into biomass and hence it may fuel a significant fraction of primary production...... heterotrophic isolates, this thesis aims at addressing these unknowns. It was found that heterotrophic diazotrophs were present and active in environments previously not associated with N2 fixation e.g. suboxic basins of the Baltic Sea and estuarine surface waters. In these environments they contributed...... with significant amounts fixed N2, suggesting that a reevaluation of the significance of N fixation in suboxic waters and estuarine coastal waters is warranted. It was also documented that heterotrophic diazotrophs could be enriched in culture based on their ability to utilize N2 as the sole N source...

  12. Producción, evaluación y caracterización microbiológica de un biofertilizante artesanal para la fijación de nitrógeno en especies vegetales forrajeras.

    OpenAIRE

    Reyes Villegas, María Isabel

    2016-01-01

    The increased production of grasslands in Ecuador has necessitated the use of chemical fertilizers, as in the case of commercial urea, which has caused impacts on agricultural land. As an alternative to this chemical fertilizer , developed , tested and characterized a craft biofertilizer made from roots of alfalfa ( Medicago sativa) , White Clover ( Trifolium repens ) and rye grass ( Lolium perenne), which has diazotrophic microbiological activity of bacterial species such...

  13. Interconnection of nitrogen fixers and iron in the Pacific Ocean: Theory and numerical simulations

    OpenAIRE

    Dutkiewicz, S.; Ward, B. A.; Monteiro, F.; Follows, M. J.

    2012-01-01

    We examine the interplay between iron supply, iron concentrations and phytoplankton communities in the Pacific Ocean. We present a theoretical framework which considers the competition for iron and nitrogen resources between phytoplankton to explain where nitrogen fixing autotrophs (diazotrophs, which require higher iron quotas, and have slower maximum growth) can co-exist with other phytoplankton. The framework also indicates that iron and fixed nitrogen concentrations can be strongly contro...

  14. The cyanobacterium Mastigocladus fulfills the nitrogen demand of a terrestrial hot spring microbial mat.

    Science.gov (United States)

    Estrella Alcamán, María; Fernandez, Camila; Delgado, Antonio; Bergman, Birgitta; Díez, Beatriz

    2015-10-01

    Cyanobacteria from Subsection V (Stigonematales) are important components of microbial mats in non-acidic terrestrial hot springs. Despite their diazotrophic nature (N2 fixers), their impact on the nitrogen cycle in such extreme ecosystems remains unknown. Here, we surveyed the identity and activity of diazotrophic cyanobacteria in the neutral hot spring of Porcelana (Northern Patagonia, Chile) during 2009 and 2011-2013. We used 16S rRNA and the nifH gene to analyze the distribution and diversity of diazotrophic cyanobacteria. Our results demonstrate the dominance of the heterocystous genus Mastigocladus (Stigonematales) along the entire temperature gradient of the hot spring (69-38 °C). In situ nitrogenase activity (acetylene reduction), nitrogen fixation rates (cellular uptake of (15)N2) and nifH transcription levels in the microbial mats showed that nitrogen fixation and nifH mRNA expression were light-dependent. Nitrogen fixation activities were detected at temperatures ranging from 58 °C to 46 °C, with maximum daily rates of 600 nmol C2H4 cm(-2) per day and 94.1 nmol N cm(-2) per day. These activity patterns strongly suggest a heterocystous cyanobacterial origin and reveal a correlation between nitrogenase activity and nifH gene expression during diurnal cycles in thermal microbial mats. N and C fixation in the mats contributed ~3 g N m(-2) per year and 27 g C m(-2) per year, suggesting that these vital demands are fully met by the diazotrophic and photoautotrophic capacities of the cyanobacteria in the Porcelana hot spring.

  15. Elevated CO2 enhances nitrogen fixation and growth in the marine cyanobacterium Trichodesmium

    Czech Academy of Sciences Publication Activity Database

    Levitan, O.; Rosenberg, G.; Šetlík, Ivan; Šetlíková, Eva; Grígel, Juraj; Klepetář, Jiří; Prášil, Ondřej; Berman-Frank, I.

    2007-01-01

    Roč. 13, č. 2 (2007), s. 531-538 ISSN 1354-1013 R&D Projects: GA MŠk 1P05ME824; GA ČR GA206/05/0335; GA MŽP SL/1/6/04 Institutional research plan: CEZ:AV0Z50200510 Keywords : ccm * cyanobacteria * diazotrophs Subject RIV: EE - Microbiology, Virology Impact factor: 4.786, year: 2007

  16. Influence of Lipid Composition in Amplifying or Ameliorating Toxicant Effects on Phytoplankton.

    Science.gov (United States)

    1992-04-30

    431-442. Robertson, B.R., and D.K. Button. 1989. Characterizing aquatic bacteria according to population, cell size, and apparent DNA content by flow...rapid preparation lornahene,. G . 198 1. Formation of hydrocarbons by bacteria of fatty acid esters for gas chromatographic analysis. A4nal. atid algae... diazotrophic cyanobacterium Trichodesmium thiebautii. Limnol Oceanogr 35:1832-1837 Volkman JK, Jeffrey SW, Nichols PD, Rogers GI, Garland CD (1989) Fatty

  17. [Assimilation of biological nitrogen by European beaver].

    Science.gov (United States)

    Vecherskiĭ, M V; Naumova, E I; Kostina, N V; Umarov, M M

    2009-01-01

    Nitrogenase activity, the abundance of diazotrophic bacteria, the structure and functional characteristics of the complex of microorganisms, and the content of nitrogen and carbon were determined in the contents of the gastrointestinal tract of the European beaver. A high nitrogen-fixing activity in the large intestine correlated with an increase in nitrogen content in the chyme upon its transfer over the gastrointestinal tract. It is assumed that microbial nitrogen fixation plays a major role in nitrogen nutrition of the European beaver.

  18. Mesopelagic N2 Fixation Related to Organic Matter Composition in the Solomon and Bismarck Seas (Southwest Pacific.

    Directory of Open Access Journals (Sweden)

    Mar Benavides

    Full Text Available Dinitrogen (N2 fixation was investigated together with organic matter composition in the mesopelagic zone of the Bismarck (Transect 1 and Solomon (Transect 2 Seas (Southwest Pacific. Transparent exopolymer particles (TEP and the presence of compounds sharing molecular formulae with saturated fatty acids and sugars, as well as dissolved organic matter (DOM compounds containing nitrogen (N and phosphorus (P were higher on Transect 1 than on Transect 2, while oxygen concentrations showed an opposite pattern. N2 fixation rates (up to ~1 nmol N L-1 d-1 were higher in Transect 1 than in Transect 2, and correlated positively with TEP, suggesting a dependence of diazotroph activity on organic matter. The scores of the multivariate ordination of DOM molecular formulae and their relative abundance correlated negatively with bacterial abundances and positively with N2 fixation rates, suggesting an active bacterial exploitation of DOM and its use to sustain diazotrophic activity. Sequences of the nifH gene clustered with Alpha-, Beta-, Gamma- and Deltaproteobacteria, and included representatives from Clusters I, III and IV. A third of the clone library included sequences close to the potentially anaerobic Cluster III, suggesting that N2 fixation was partially supported by presumably particle-attached diazotrophs. Quantitative polymerase chain reaction (qPCR primer-probe sets were designed for three phylotypes and showed low abundances, with a phylotype within Cluster III at up to 103 nifH gene copies L-1. These results provide new insights into the ecology of non-cyanobacterial diazotrophs and suggest that organic matter sustains their activity in the mesopelagic ocean.

  19. A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

    Science.gov (United States)

    Ng, Gordon; Tom, Curtis G S; Park, Angela S; Zenad, Lounis; Ludwig, Robert A

    2009-01-01

    Nitrogen (N(2)) fixation also yields hydrogen (H(2)) at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2) as sole N-source) bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase), has nevertheless been presumed responsible for recycling such endogenous hydrogen. As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase) was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase). An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2)-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. Representative of aerobic N(2)-fixing and H(2)-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2) respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2), specifically that produced by N(2) fixation. To benefit human civilization, H(2) has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As such, the reversible, group-4 Ni,Fe-hydrogenases, such

  20. Is the distribution of nitrogen-fixing cyanobacteria in the oceans related to temperature?

    Science.gov (United States)

    Stal, Lucas J

    2009-07-01

    Approximately 50% of the global natural fixation of nitrogen occurs in the oceans supporting a considerable part of the new primary production. Virtually all nitrogen fixation in the ocean occurs in the tropics and subtropics where the surface water temperature is 25°C or higher. It is attributed almost exclusively to cyanobacteria. This is remarkable firstly because diazotrophic cyanobacteria are found in other environments irrespective of temperature and secondly because primary production in temperate and cold oceans is generally limited by nitrogen. Cyanobacteria are oxygenic phototrophic organisms that evolved a variety of strategies protecting nitrogenase from oxygen inactivation. Free-living diazotrophic cyanobacteria in the ocean are of the non-heterocystous type, namely the filamentous Trichodesmium and the unicellular groups A-C. I will argue that warm water is a prerequisite for these diazotrophic organisms because of the low-oxygen solubility and high rates of respiration allowing the organism to maintain anoxic conditions in the nitrogen-fixing cell. Heterocystous cyanobacteria are abundant in freshwater and brackish environments in all climatic zones. The heterocyst cell envelope is a tuneable gas diffusion barrier that optimizes the influx of both oxygen and nitrogen, while maintaining anoxic conditions inside the cell. It is not known why heterocystous cyanobacteria are absent from the temperate and cold oceans and seas.

  1. Microbial dinitrogen fixation in coral holobionts exposed to thermal stress and bleaching.

    Science.gov (United States)

    Cardini, Ulisse; van Hoytema, Nanne; Bednarz, Vanessa N; Rix, Laura; Foster, Rachel A; Al-Rshaidat, Mamoon M D; Wild, Christian

    2016-09-01

    Coral holobionts (i.e., coral-algal-prokaryote symbioses) exhibit dissimilar thermal sensitivities that may determine which coral species will adapt to global warming. Nonetheless, studies simultaneously investigating the effects of warming on all holobiont members are lacking. Here we show that exposure to increased temperature affects key physiological traits of all members (herein: animal host, zooxanthellae and diazotrophs) of both Stylophora pistillata and Acropora hemprichii during and after thermal stress. S. pistillata experienced severe loss of zooxanthellae (i.e., bleaching) with no net photosynthesis at the end of the experiment. Conversely, A. hemprichii was more resilient to thermal stress. Exposure to increased temperature (+ 6°C) resulted in a drastic increase in daylight dinitrogen (N2 ) fixation, particularly in A. hemprichii (threefold compared with controls). After the temperature was reduced again to in situ levels, diazotrophs exhibited a reversed diel pattern of activity, with increased N2 fixation rates recorded only in the dark, particularly in bleached S. pistillata (twofold compared to controls). Concurrently, both animal hosts, but particularly bleached S. pistillata, reduced both organic matter release and heterotrophic feeding on picoplankton. Our findings indicate that physiological plasticity by coral-associated diazotrophs may play an important role in determining the response of coral holobionts to ocean warming. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  2. Climate change and regulation of hepatotoxin production in Cyanobacteria.

    Science.gov (United States)

    Gehringer, Michelle M; Wannicke, Nicola

    2014-04-01

    Harmful, bloom-forming cyanobacteria (CyanoHABs) are occurring with increasing regularity in freshwater and marine ecosystems. The most commonly occurring cyanobacterial toxins are the hepatotoxic microcystin and nodularin. These cyclic hepta- and pentapeptides are synthesised nonribosomally by the gene products of the toxin gene clusters mcy and nda, respectively. Understanding of the regulation of hepatotoxin production is incomplete, although there is strong evidence supporting the roles of iron, light, higher nitrate availability and inorganic carbon in modulating microcystin levels. The majority of these studies have focused on the unicellular freshwater, microcystin-producing strain of Microcystis aeruginosa, with little attention being paid to terrestrial or marine toxin producers. This review intends to investigate the regulation of microcystin and nodularin production in unicellular and filamentous diazotrophic cyanobacteria against the background of changing climate conditions. Special focus is given to diazotrophic filamentous cyanobacteria, for example Nodularia spumigena, capable of regulating their nitrogen levels by actively fixing dinitrogen. By combining data from significant studies, an overall scheme of the regulation of toxin production is presented, focussing specifically on nodularin production in diazotrophs against the background of increasing carbon dioxide concentrations and temperatures envisaged under current climate change models. Furthermore, the risk of sustaining and spreading CyanoHABs in the future ocean is evaluated. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  3. Monitoring the persistence of a genetically modified strain of the Azorhizobium caulinodans in the rhizosphere of Triticum aestivum L.

    Directory of Open Access Journals (Sweden)

    Jan Jacek Slaski

    2013-01-01

    Full Text Available A protocol was developed to monitor persistence and spread of a genetically modified strain of the growth promoting diazotroph Azorhizobium caulinodans used to inoculate field grown wheat. The protocol was used to identify an endogluconase (egl overproducing strain of A. caulinodans isolated from inoculated soils. A detection limit of 2500 cfu g-1 and 108_1010cfu g-1 of soil was determined using the BIOLOG® fingerprinting method and PCR technique, respectively. Since none of the tested samples were positive for the bacteria or the Egl 1 gene construct, it was concluded that A. caulinodans ORS 571 pGV910-C1 did not persist or spread in any of the tested field locations. This result may be due to low soil temperatures and competition of indigenous microorganisms: environmental factors that were not favorable for the diazotroph to thrive in the test locations. The application of different inoculation methodologies, as well as the study of other rhizobial genera for the inoculation of wheat in further experiments, is strongly recommended. Key words: detection, diazotroph, endogluconase, ORS 571, wheat

  4. The damage caused by Callosobruchus maculatus on cowpea grains is dependent on the plant genotype.

    Science.gov (United States)

    Torres, Elida Barros; Nóbrega, Rafaela S A; Fernandes-Júnior, Paulo Ivan; Silva, Luciana Barboza; Dos Santos Carvalho, Gabriel; Marinho, Rita de Cassia Nunes; Pavan, Bruno E

    2016-09-01

    Beans from cowpea cultivars fertilized with mineral N or inoculated with various rhizobium strains may contain different nitrogen concentrations and nitrogen metabolite composition, which affects the beans' defense mechanisms against pests. In this study, the population growth of Callosobruchus maculatus reared on beans from four cowpea cultivars fertilized with different nitrogen sources was evaluated. The factors tested were beans from four cowpea cultivars and seven different nitrogen sources: mineral N fertilization, inoculation with five strains of symbiotic diazotrophic bacteria, and soil nitrogen (absolute control). BRS Tapaihum and BRS Acauã cultivars had lower cumulative emergence and instantaneous rate of population growth of the insects compared with other cultivars, indicating antixenosis resistance against C. maculatus. Inoculation of BRS Acauã cultivar with the diazotrophic bacteria strain BR 3299 resulted in higher mortality of C. maculatus. For BRS Tapaihum cultivar, inoculation with diazotrophic bacteria strains BR3267, BR 3262 and BR 3299, and nitrogen fertilization resulted in higher mortality among C. maculatus. BRS Tapaihum and BRS Acauã cultivars showed the lowest cumulative insect emergence and instantaneous rates of population growth, and the highest insect mortality, mainly when the grains were obtained from plants inoculated with rhizobial strains. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  5. Photoperiod length paces the temporal orchestration of cell cycle and carbon-nitrogen metabolism in Crocosphaera watsonii.

    Science.gov (United States)

    Dron, Anthony; Rabouille, Sophie; Claquin, Pascal; Talec, Amélie; Raimbault, Virginie; Sciandra, Antoine

    2013-12-01

    We analysed the effect of photoperiod length (PPL) (16:8 and 8:16 h of light-dark regime, named long and short PPL, respectively) on the temporal orchestration of the two antagonistic, carbon and nitrogen acquisitions in the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii strain WH8501 growing diazotrophically. Carbon and nitrogen metabolism were monitored at high frequency, and their patterns were compared with the cell cycle progression. The oxygen-sensitive N2 fixation process occurred mainly during the dark period, where photosynthesis cannot take place, inducing a light-dark cycle of cellular C : N ratio. Examination of circadian patterns in the cell cycle revealed that cell division occurred during the midlight period, (8 h and 4 h into the light in the long and short PPL conditions, respectively), thus timely separated from the energy-intensive diazotrophic process. Results consistently show a nearly 5 h time lag between the end of cell division and the onset of N2 fixation. Shorter PPLs affected DNA compaction of C. watsonii cells and also led to a decrease in the cell division rate. Therefore, PPL paces the growth of C. watsonii: a long PPL enhances cell division while a short PPL favours somatic growth (biomass production) with higher carbon and nitrogen cell contents. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  6. Nitrogen-fixing bacteria in Mediterranean seagrass (Posidonia oceanica) roots

    KAUST Repository

    Garcias Bonet, Neus

    2016-03-09

    Biological nitrogen fixation by diazotrophic bacteria in seagrass rhizosphere and leaf epiphytic community is an important source of nitrogen required for plant growth. However, the presence of endophytic diazotrophs remains unclear in seagrass tissues. Here, we assess the presence, diversity and taxonomy of nitrogen-fixing bacteria within surface-sterilized roots of Posidonia oceanica. Moreover, we analyze the nitrogen isotopic signature of seagrass tissues in order to notice atmospheric nitrogen fixation. We detected nitrogen-fixing bacteria by nifH gene amplification in 13 out of the 78 roots sampled, corresponding to 9 locations out of 26 meadows. We detected two different types of bacterial nifH sequences associated with P. oceanica roots, which were closely related to sequences previously isolated from the rhizosphere of a salt marsh cord grass and a putative anaerobe. Nitrogen content of seagrass tissues showed low isotopic signatures in all the sampled meadows, pointing out the atmospheric origin of the assimilated nitrogen by seagrasses. However, this was not related with the presence of endophytic nitrogen fixers, suggesting the nitrogen fixation occurring in rhizosphere and in the epiphytic community could be an important source of nitrogen for P. oceanica. The low diversity of nitrogen-fixing bacteria reported here suggests species-specific relationships between diazotrophs and P. oceanica, revealing possible symbiotic interactions that could play a major role in nitrogen acquisition by seagrasses in oligotrophic environments where they form lush meadows.

  7. Incorporating nitrogen fixing cyanobacteria in the global biogeochemical model HAMOCC

    Science.gov (United States)

    Paulsen, Hanna; Ilyina, Tatiana; Six, Katharina

    2015-04-01

    Nitrogen fixation by marine diazotrophs plays a fundamental role in the oceanic nitrogen and carbon cycle as it provides a major source of 'new' nitrogen to the euphotic zone that supports biological carbon export and sequestration. Since most global biogeochemical models include nitrogen fixation only diagnostically, they are not able to capture its spatial pattern sufficiently. Here we present the incorporation of an explicit, dynamic representation of diazotrophic cyanobacteria and the corresponding nitrogen fixation in the global ocean biogeochemical model HAMOCC (Hamburg Ocean Carbon Cycle model), which is part of the Max Planck Institute for Meteorology Earth system model (MPI-ESM). The parameterization of the diazotrophic growth is thereby based on available knowledge about the cyanobacterium Trichodesmium spp., which is considered as the most significant pelagic nitrogen fixer. Evaluation against observations shows that the model successfully reproduces the main spatial distribution of cyanobacteria and nitrogen fixation, covering large parts of the tropical and subtropical oceans. Besides the role of cyanobacteria in marine biogeochemical cycles, their capacity to form extensive surface blooms induces a number of bio-physical feedback mechanisms in the Earth system. The processes driving these interactions, which are related to the alteration of heat absorption, surface albedo and momentum input by wind, are incorporated in the biogeochemical and physical model of the MPI-ESM in order to investigate their impacts on a global scale. First preliminary results will be shown.

  8. Influence of External Nitrogen on Nitrogenase Enzyme Activity and Auxin Production in Herbaspirillum seropedicae (Z78).

    Science.gov (United States)

    Yin, Tan Tzy; Pin, Ui Li; Ghazali, Amir Hamzah Ahmad

    2015-04-01

    The production of nitrogenase enzyme and auxins by free living diazotrophs has the potential to influence the growth of host plants. In this study, diazotrophs were grown in the presence of various concentrations of nitogen (N) to determine the optimal concentration of N for microbial growth stimulation, promotion of gaseous N (N2) fixation, and phytohormone production. Therefore, we investigate whether different levels of N supplied to Herbaspirillum seropedicae (Z78) have significant effects on nitrogenase activity and auxin production. The highest nitrogenase activity and the lowest auxin production of H. seropedicae (Z78) were both recorded at 0 gL(-1) of NH4Cl. Higher levels of external N caused a significant decrease in the nitrogenase activity and an increased production of auxins. In a subsequent test, two different inoculum sizes of Z78 (10(6) and 10(12) cfu/ml) were used to study the effect of different percentages of acetylene on nitrogenase activity of the inoculum via the acetylene reduction assay (ARA). The results showed that the optimal amount of acetylene required for nitrogenase enzyme activity was 5% for the 10(6) cfu/ml inoculum, whereas the higher inoculum size (10(12) cfu/ml) required at least 10% of acetylene for optimal nitrogenase activity. These findings provide a clearer understanding of the effects of N levels on diazotrophic nitrogenase activity and auxin production, which are important factors influencing plant growth.

  9. Diversity of the Bacterial Microbiome in the Roots of Four Saccharum Species: S. spontaneum, S. robustum, S. barberi, and S. officinarum

    Directory of Open Access Journals (Sweden)

    Meng Dong

    2018-02-01

    Full Text Available Endophytic bacteria are nearly ubiquitously present in the internal tissues of plants, and some endophytes can promote plant growth. In this study, we sampled the roots of four ancestral species of sugarcane (two genotypes per species and two sugarcane cultivars, and used 16S rRNA and nifH gene sequencing to characterize the root endophytic bacterial communities and diazotroph diversity. A total of 7,198 operational taxonomic units (OTUs were detected for the endophytic bacteria community. The endophytic bacterial communities exhibited significantly different α- and β-diversities. From the 202 detected families in the sugarcane roots, a core microbiome containing 13 families was identified. The nifH gene was successfully detected in 9 of 30 samples from the four sugarcane species assayed, and 1,734 OTUs were merged for endophytic diazotrophs. In the tested samples, 43 families of endophytic diazotrophs were detected, and six families showed differences across samples. Among the 20 most abundant detected genera, 10 have been reported to be involved in nitrogen fixation in sugarcane. These findings demonstrate the diversity of the microbial communities in different sugarcane germplasms and shed light on the mechanism of biological nitrogen fixation in sugarcane.

  10. Functional Single-Cell Approach to Probing Nitrogen-Fixing Bacteria in Soil Communities by Resonance Raman Spectroscopy with 15N2 Labeling.

    Science.gov (United States)

    Cui, Li; Yang, Kai; Li, Hong-Zhe; Zhang, Han; Su, Jian-Qiang; Paraskevaidi, Maria; Martin, Francis L; Ren, Bin; Zhu, Yong-Guan

    2018-04-17

    Nitrogen (N) fixation is the conversion of inert nitrogen gas (N 2 ) to bioavailable N essential for all forms of life. N 2 -fixing microorganisms (diazotrophs), which play a key role in global N cycling, remain largely obscure because a large majority are uncultured. Direct probing of active diazotrophs in the environment is still a major challenge. Herein, a novel culture-independent single-cell approach combining resonance Raman (RR) spectroscopy with 15 N 2 stable isotope probing (SIP) was developed to discern N 2 -fixing bacteria in a complex soil community. Strong RR signals of cytochrome c (Cyt c, frequently present in diverse N 2 -fixing bacteria), along with a marked 15 N 2 -induced Cyt c band shift, generated a highly distinguishable biomarker for N 2 fixation. 15 N 2 -induced shift was consistent well with 15 N abundance in cell determined by isotope ratio mass spectroscopy. By applying this biomarker and Raman imaging, N 2 -fixing bacteria in both artificial and complex soil communities were discerned and imaged at the single-cell level. The linear band shift of Cyt c versus 15 N 2 percentage allowed quantification of N 2 fixation extent of diverse soil bacteria. This single-cell approach will advance the exploration of hitherto uncultured diazotrophs in diverse ecosystems.

  11. A new species of Burkholderia isolated from sugarcane roots promotes plant growth

    Science.gov (United States)

    Paungfoo-Lonhienne, Chanyarat; Lonhienne, Thierry G A; Yeoh, Yun Kit; Webb, Richard I; Lakshmanan, Prakash; Chan, Cheong Xin; Lim, Phaik-Eem; Ragan, Mark A; Schmidt, Susanne; Hugenholtz, Philip

    2014-01-01

    Sugarcane is a globally important food, biofuel and biomaterials crop. High nitrogen (N) fertilizer rates aimed at increasing yield often result in environmental damage because of excess and inefficient application. Inoculation with diazotrophic bacteria is an attractive option for reducing N fertilizer needs. However, the efficacy of bacterial inoculants is variable, and their effective formulation remains a knowledge frontier. Here, we take a new approach to investigating diazotrophic bacteria associated with roots using culture-independent microbial community profiling of a commercial sugarcane variety (Q208A) in a field setting. We first identified bacteria that were markedly enriched in the rhizosphere to guide isolation and then tested putative diazotrophs for the ability to colonize axenic sugarcane plantlets (Q208A) and promote growth in suboptimal N supply. One isolate readily colonized roots, fixed N2 and stimulated growth of plantlets, and was classified as a new species, Burkholderia australis sp. nov. Draft genome sequencing of the isolate confirmed the presence of nitrogen fixation. We propose that culture-independent identification and isolation of bacteria that are enriched in rhizosphere and roots, followed by systematic testing and confirming their growth-promoting capacity, is a necessary step towards designing effective microbial inoculants. PMID:24350979

  12. Aphotic N2 fixation along an oligotrophic to ultraoligotrophic transect in the western tropical South Pacific Ocean

    Science.gov (United States)

    Benavides, Mar; Shoemaker, Katyanne M.; Moisander, Pia H.; Niggemann, Jutta; Dittmar, Thorsten; Duhamel, Solange; Grosso, Olivier; Pujo-Pay, Mireille; Hélias-Nunige, Sandra; Fumenia, Alain; Bonnet, Sophie

    2018-05-01

    The western tropical South Pacific (WTSP) Ocean has been recognized as a global hot spot of dinitrogen (N2) fixation. Here, as in other marine environments across the oceans, N2 fixation studies have focused on the sunlit layer. However, studies have confirmed the importance of aphotic N2 fixation activity, although until now only one had been performed in the WTSP. In order to increase our knowledge of aphotic N2 fixation in the WTSP, we measured N2 fixation rates and identified diazotrophic phylotypes in the mesopelagic layer along a transect spanning from New Caledonia to French Polynesia. Because non-cyanobacterial diazotrophs presumably need external dissolved organic matter (DOM) sources for their nutrition, we also identified DOM compounds using Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) with the aim of searching for relationships between the composition of DOM and non-cyanobacterial N2 fixation in the aphotic ocean. N2 fixation rates were low (average 0.63 ± 0.07 nmol N L-1 d-1) but consistently detected across all depths and stations, representing ˜ 6-88 % of photic N2 fixation. N2 fixation rates were not significantly correlated with DOM compounds. The analysis of nifH gene amplicons revealed a wide diversity of non-cyanobacterial diazotrophs, mostly matching clusters 1 and 3. Interestingly, a distinct phylotype from the major nifH subcluster 1G dominated at 650 dbar, coinciding with the oxygenated Subantarctic Mode Water (SAMW). This consistent pattern suggests that the distribution of aphotic diazotroph communities is to some extent controlled by water mass structure. While the data available are still too scarce to elucidate the distribution and controls of mesopelagic non-cyanobacterial diazotrophs in the WTSP, their prevalence in the mesopelagic layer and the consistent detection of active N2 fixation activity at all depths sampled during our study suggest that aphotic N2 fixation may contribute significantly to fixed

  13. Symbiosis revisited: phosphorus and acid buffering stimulate N2 fixation but not Sphagnum growth

    Science.gov (United States)

    van den Elzen, Eva; Kox, Martine A. R.; Harpenslager, Sarah F.; Hensgens, Geert; Fritz, Christian; Jetten, Mike S. M.; Ettwig, Katharina F.; Lamers, Leon P. M.

    2017-03-01

    In pristine Sphagnum-dominated peatlands, (di)nitrogen (N2) fixing (diazotrophic) microbial communities associated with Sphagnum mosses contribute substantially to the total nitrogen input, increasing carbon sequestration. The rates of symbiotic nitrogen fixation reported for Sphagnum peatlands, are, however, highly variable, and experimental work on regulating factors that can mechanistically explain this variation is largely lacking. For two common fen species (Sphagnum palustre and S. squarrosum) from a high nitrogen deposition area (25 kg N ha-1 yr-1), we found that diazotrophic activity (as measured by 15 - 15N2 labeling) was still present at a rate of 40 nmol N gDW-1 h-1. This was surprising, given that nitrogen fixation is a costly process. We tested the effects of phosphorus availability and buffering capacity by bicarbonate-rich water, mimicking a field situation in fens with stronger groundwater or surface water influence, as potential regulators of nitrogen fixation rates and Sphagnum performance. We expected that the addition of phosphorus, being a limiting nutrient, would stimulate both diazotrophic activity and Sphagnum growth. We indeed found that nitrogen fixation rates were doubled. Plant performance, in contrast, did not increase. Raised bicarbonate levels also enhanced nitrogen fixation, but had a strong negative impact on Sphagnum performance. These results explain the higher nitrogen fixation rates reported for minerotrophic and more nutrient-rich peatlands. In addition, nitrogen fixation was found to strongly depend on light, with rates 10 times higher in light conditions suggesting high reliance on phototrophic organisms for carbon. The contrasting effects of phosphorus and bicarbonate on Sphagnum spp. and their diazotrophic communities reveal strong differences in the optimal niche for both partners with respect to conditions and resources. This suggests a trade-off for the symbiosis of nitrogen fixing microorganisms with their Sphagnum

  14. Heterotrophic bacterial production and metabolic balance during the VAHINE mesocosm experiment in the New Caledonia lagoon

    Science.gov (United States)

    Van Wambeke, France; Pfreundt, Ulrike; Barani, Aude; Berthelot, Hugo; Moutin, Thierry; Rodier, Martine; Hess, Wolfgang R.; Bonnet, Sophie

    2016-06-01

    Studies investigating the fate of diazotrophs through the microbial food web are lacking, although N2 fixation can fuel up to 50 % of new production in some oligotrophic oceans. In particular, the role played by heterotrophic prokaryotes in this transfer is largely unknown. In the frame of the VAHINE (VAriability of vertical and tropHIc transfer of diazotroph derived N in the south wEst Pacific) experiment, three replicate large-volume (˜ 50 m3) mesocosms were deployed for 23 days in the new Caledonia lagoon and were intentionally fertilized on day 4 with dissolved inorganic phosphorus (DIP) to stimulate N2 fixation. We specifically examined relationships between heterotrophic bacterial production (BP) and N2 fixation or primary production, determined bacterial growth efficiency and established carbon budgets. BP was statistically higher during the second phase of the experiment (P2: days 15-23), when chlorophyll biomass started to increase compared to the first phase (P1: days 5-14). Phosphatase alkaline activity increased drastically during the second phase of the experiment, showing adaptations of microbial populations after utilization of the added DIP. Notably, among autotrophs, Synechococcus abundances increased during P2, possibly related to its capacity to assimilate leucine and to produce alkaline phosphatase. Bacterial growth efficiency based on the carbon budget (27-43 %), was notably higher than generally cited for oligotrophic environments and discussed in links with the presence of abundant species of bacteria expressing proteorhodopsin. The main fates of gross primary production (particulate + dissolved) were respiration (67 %) and export through sedimentation (17 %). BP was highly correlated with particulate primary production and chlorophyll biomass during both phases of the experiment but was slightly correlated, and only during P2 phase, with N2 fixation rates. Heterotrophic bacterial production was strongly stimulated after mineral N enrichment

  15. Aphotic N2 fixation along an oligotrophic to ultraoligotrophic transect in the western tropical South Pacific Ocean

    Directory of Open Access Journals (Sweden)

    M. Benavides

    2018-05-01

    Full Text Available The western tropical South Pacific (WTSP Ocean has been recognized as a global hot spot of dinitrogen (N2 fixation. Here, as in other marine environments across the oceans, N2 fixation studies have focused on the sunlit layer. However, studies have confirmed the importance of aphotic N2 fixation activity, although until now only one had been performed in the WTSP. In order to increase our knowledge of aphotic N2 fixation in the WTSP, we measured N2 fixation rates and identified diazotrophic phylotypes in the mesopelagic layer along a transect spanning from New Caledonia to French Polynesia. Because non-cyanobacterial diazotrophs presumably need external dissolved organic matter (DOM sources for their nutrition, we also identified DOM compounds using Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS with the aim of searching for relationships between the composition of DOM and non-cyanobacterial N2 fixation in the aphotic ocean. N2 fixation rates were low (average 0.63 ± 0.07 nmol N L−1 d−1 but consistently detected across all depths and stations, representing ∼ 6–88 % of photic N2 fixation. N2 fixation rates were not significantly correlated with DOM compounds. The analysis of nifH gene amplicons revealed a wide diversity of non-cyanobacterial diazotrophs, mostly matching clusters 1 and 3. Interestingly, a distinct phylotype from the major nifH subcluster 1G dominated at 650 dbar, coinciding with the oxygenated Subantarctic Mode Water (SAMW. This consistent pattern suggests that the distribution of aphotic diazotroph communities is to some extent controlled by water mass structure. While the data available are still too scarce to elucidate the distribution and controls of mesopelagic non-cyanobacterial diazotrophs in the WTSP, their prevalence in the mesopelagic layer and the consistent detection of active N2 fixation activity at all depths sampled during our study suggest that aphotic N2

  16. Exploring microbial succession and diversity during solid-state fermentation of Tianjin duliu mature vinegar.

    Science.gov (United States)

    Nie, Zhiqiang; Zheng, Yu; Wang, Min; Han, Yue; Wang, Yuenan; Luo, Jianmei; Niu, Dandan

    2013-11-01

    Tianjin duliu mature vinegar was one of famous Chinese traditional vinegars. The unique flavor and taste of vinegar are mainly generated by the multitudinous microorganisms during fermentation. In this research, the composition and succession of microbial communities in the entire solid-state fermentation were investigated, including starter daqu and acetic acid fermentation (AAF). Molds and yeasts in daqu, including Aspergillus, Saccharomycopsis and Pichia, decreased in AAF. The bacterial compositions increased from four genera in daqu to more than 13 genera in AAF. Principal component analysis showed that Acetobacter, Gluconacetobacter, Lactobacillus and Nostoc were dominant bacteria that were correlated well with AAF process. In the early fermentation period, lactic acid bacteria (LAB) decreased while acetic acid bacteria and Nostoc increased rapidly with the accumulation of total acids. Then, the abundance and diversity of LAB increased (more than 80%), indicating that LAB had important influences on the flavor and taste of vinegar. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Characterization of purified bacterial cellulose focused on its use on paper restoration.

    Science.gov (United States)

    Santos, Sara M; Carbajo, José M; Quintana, Ester; Ibarra, David; Gomez, Nuria; Ladero, Miguel; Eugenio, M Eugenia; Villar, Juan C

    2015-02-13

    Bacterial cellulose (BC) synthesized by Gluconacetobacter sucrofermentans CECT 7291 seems to be a good option for the restoration of degraded paper. In this work BC layers are cultivated and purified by two different methods: an alkaline treatment when the culture media contains ethanol and a thermal treatment if the media is free from ethanol. The main goal of these tests was the characterization of BC layers measured in terms of tear and burst indexes, optical properties, SEM, X-ray diffraction, FTIR, degree of polymerization, static and dynamic contact angles, and mercury intrusion porosimetry. The BC layers were also evaluated in the same terms after an aging treatment. Results showed that BC has got high crystallinity index, low internal porosity, good mechanical properties and high stability over time, especially when purified by the alkaline treatment. These features make BC an adequate candidate for degraded paper reinforcement. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Different temperatures select distinctive acetic acid bacteria species and promotes organic acids production during Kombucha tea fermentation.

    Science.gov (United States)

    De Filippis, Francesca; Troise, Antonio Dario; Vitaglione, Paola; Ercolini, Danilo

    2018-08-01

    Kombucha is a traditional beverage produced by tea fermentation, carried out by a symbiotic consortium of bacteria and yeasts. Acetic Acid Bacteria (AAB) usually dominate the bacterial community of Kombucha, driving the fermentative process. The consumption of this beverage was often associated to beneficial effects for the health, due to its antioxidant and detoxifying properties. We characterized bacterial populations of Kombucha tea fermented at 20 or 30 °C by using culture-dependent and -independent methods and monitored the concentration of gluconic and glucuronic acids, as well as of total polyphenols. We found significant differences in the microbiota at the two temperatures. Moreover, different species of Gluconacetobacter were selected, leading to a differential abundance of gluconic and glucuronic acids. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Symbiosis between microorganisms from kombucha and kefir: Potential significance to the enhancement of kombucha function.

    Science.gov (United States)

    Yang, Zhiwei; Zhou, Feng; Ji, Baoping; Li, Bo; Luo, Yangchao; Yang, Li; Li, Tao

    2010-01-01

    Gluconacetobacter sp. A4 (G. sp. A4), which had strong ability to produce d-saccharic acid 1, 4 lactone (DSL), was the key functional bacteria isolated from the kombucha preserved. This paper investigated the interaction between G. sp. A4 and ten different strains of lactic acid bacteria (LAB) obtained from kefir. The result suggested that the LAB promoted DSL production of G. sp. A4 to different extents, ranging from 4.86% to 86.70%. Symbiosis between G. sp. A4 and LAB was studied. LAB's metabolites, xylitol, and acetic acid, were utilized by G. sp. A4, and it promoted the growth of G. sp. A4 and yield of DSL. Therefore, in developing starter cultures for kombucha fermentation process, a mixed flora of LAB and G. sp. A4 would be the optimal combination.

  20. Optimitzación de la biosíntesis de nanocelulosa bacteriana por fermentación

    OpenAIRE

    García González, Alberto

    2017-01-01

    La cel·lulosa bacteriana és un biopolímer obtingut per fermentació amb diversos microorganismes, dels quals l’espècie més eficient en la producció és la Gluconacetobacter xylinus sucrofermentans. La principal diferencia entre la cel·lulosa bacteriana y la resta de cel·luloses que existeixen és la seva puresa, per això, és un del biopolímer més atractius per les seves possibles aplicacions en diverses àrees com són l’alimentació, la farmàcia i la medicina. A més de la seva puresa, les seves...

  1. Propriedades ópticas de suspensões coloidais e filmes à base de nanocelulose

    OpenAIRE

    Santos, Daniele Bueno dos [UNESP

    2012-01-01

    A celulose bacteriana (CB), produzida na superfície do meio de cultura da bactéria Gluconacetobacter xylinus foi utilizada para a produção de nanocristais de celulose, ou seja, cristalitos de celulose de dimensões nanométricas isolados por meio de hidrólise ácida, que em suspensão aquosa podem se auto-organizar para a formação de fase líquido cristalina nemática quiral. Foi realizada a hidrólise ácida da CB usando ácido sulfúrico 64% (m/m) a uma temperatura de 50 °C, o tempo de hidrólise foi ...

  2. Antisera production to detect indoleacetic acid in cultures of plant-growth promoting bacteria

    International Nuclear Information System (INIS)

    Rojas, Marcia M; Hernandez, Annia; Rives, Narovis; Tejera, Berto; Acebo, Yanelis; Heydrich, Mayra

    2012-01-01

    Rabbit polyclonal antisera against indoleacetic acid (IAA) bound to nitrocellulose membrane were obtained, which exhibited a high titer and specificity. The dot immunobinding technique with colloidal gold was used to detect auxin production by several strains belonging to Gluconacetobacter, Herbaspirillum, Azospirillum, Pseudomonas, Burkholderia and Bacillus genera, using culture supernatants as antigens. Moreover, auxin production was quantified by the Salkowski's method to corroborate the previous results. It was found that that all the studied microorganisms produce IAA and the feasibility of using these antisera to detect the metabolite was confirmed. Taking into account the potentialities of plant growth promoting bacteria as biofertilizers, the use of these antisera for a rapid and easy detection of IAA in bacteria associated with important crops is thus recommended.

  3. [Comparative genomics and evolutionary analysis of CRISPR loci in acetic acid bacteria].

    Science.gov (United States)

    Xia, Kai; Liang, Xin-le; Li, Yu-dong

    2015-12-01

    The clustered regularly interspaced short palindromic repeat (CRISPR) is a widespread adaptive immunity system that exists in most archaea and many bacteria against foreign DNA, such as phages, viruses and plasmids. In general, CRISPR system consists of direct repeat, leader, spacer and CRISPR-associated sequences. Acetic acid bacteria (AAB) play an important role in industrial fermentation of vinegar and bioelectrochemistry. To investigate the polymorphism and evolution pattern of CRISPR loci in acetic acid bacteria, bioinformatic analyses were performed on 48 species from three main genera (Acetobacter, Gluconacetobacter and Gluconobacter) with whole genome sequences available from the NCBI database. The results showed that the CRISPR system existed in 32 species of the 48 strains studied. Most of the CRISPR-Cas system in AAB belonged to type I CRISPR-Cas system (subtype E and C), but type II CRISPR-Cas system which contain cas9 gene was only found in the genus Acetobacter and Gluconacetobacter. The repeat sequences of some CRISPR were highly conserved among species from different genera, and the leader sequences of some CRISPR possessed conservative motif, which was associated with regulated promoters. Moreover, phylogenetic analysis of cas1 demonstrated that they were suitable for classification of species. The conservation of cas1 genes was associated with that of repeat sequences among different strains, suggesting they were subjected to similar functional constraints. Moreover, the number of spacer was positively correlated with the number of prophages and insertion sequences, indicating the acetic acid bacteria were continually invaded by new foreign DNA. The comparative analysis of CRISR loci in acetic acid bacteria provided the basis for investigating the molecular mechanism of different acetic acid tolerance and genome stability in acetic acid bacteria.

  4. CARACTERIZACIÓN DE BACTERIAS DIAZOTRÓFICAS ASIMBIÓTICAS ASOCIADAS AL EUCALIPTO (Eucalyptus sp. EN CODAZZI, CESAR.

    Directory of Open Access Journals (Sweden)

    Dolly Melissa Obando Castellanos

    2010-09-01

    Full Text Available Se evaluó el efecto de las épocas climáticas (lluvia y sequía y del estrato de la muestra (Suelo rizosférico, raíces y hojas sobre la población de los géneros Azotobacter, Beijerinckia, Derxia, Azospirillum, Herbaspirillum, Gluconacetobacter y Burkholderia en el Eucalipto (Eucalyptus sp.. Así mismo, se evalúo su capacidad en la producción de compuestos indólicos como promotores del crecimiento vegetal y su actividad de reducción de acetileno como indicador de la fijación biológica de nitrógeno. Los resultados no registraron diferencias estadísticas significativas en el test de Tukey (P ≤ 0.05 en la población con respecto a la época climática. Con respecto al estrato de muestra, los aislamientos tentativos a Herbaspirillum sp. y Azospirillum sp. presentaron diferencias significativas en suelo rizosférico y raíces. Se obtuvieron 44 aislamientos de los cuales se agruparon por caracterización fenotípica como: 14 presuntivos del género Beijerinckia sp., 12 de Azotobacter sp., 8 de Derxia sp., 4 de Herbarpirillum sp., 5 de Azospirillum sp., 1 de Gluconacetobacter sp. y 1 de Burkholderia sp. Por su alto potencial fueron seleccionados y criopreservados los aislamientos C27, C26, C25 y C45, las cuales presentaron los mejores valores de eficiencia in vitro, superando valores de producción de las cepas de referencia utilizadas (A. chroococcum (AC-01 y A. brasilense (SP7.

  5. Microbiological investigations on the water of a thermal bath at Budapest.

    Science.gov (United States)

    Szuróczki, Sára; Kéki, Zsuzsa; Káli, Szandra; Lippai, Anett; Márialigeti, Károly; Tóth, Erika

    2016-06-01

    Thermal baths are unique aquatic environments combining a wide variety of natural and anthropogenic ecological factors, which also appear in their microbiological state. There is limited information on the microbiology of thermal baths in their complexity, tracking community shifts from the thermal wells to the pools. In the present study, the natural microbial community of well and pool waters in Gellért bath was studied in detail by cultivation-based techniques. To isolate bacteria, 10% R2A and minimal synthetic media (with "bath water") with agar-agar and gellan gum were used after prolonged incubation time; moreover, polyurethane blocks covered with media were also applied. Strains were identified by sequencing their 16S rRNA gene after grouping them by amplified rDNA restriction analysis. From each sample, the dominance of Alphaproteobacteria was characteristic though their diversity differed among samples. Members of Actinobacteria, Firmicutes, Beta- and Gamma-proteobacteria, Deinococcus-Thermus, and Bacteroidetes were also identified. Representatives of Deinococcus-Thermus phylum appeared only in the pool water. The largest groups in the pool water belonged to the Tistrella and Chelatococcus genera. The most dominant member in the well water was a new taxon, its similarity to Hartmannibacter diazotrophicus as closest relative was 93.93%.

  6. Ammonium photo-production by heterocytous cyanobacteria: potentials and constraints.

    Science.gov (United States)

    Grizeau, Dominique; Bui, Lan Anh; Dupré, Catherine; Legrand, Jack

    2016-08-01

    Over the last decades, production of microalgae and cyanobacteria has been developed for several applications, including novel foods, cosmetic ingredients and more recently biofuel. The sustainability of these promising developments can be hindered by some constraints, such as water and nutrient footprints. This review surveys data on N2-fixing cyanobacteria for biomass production and ways to induce and improve the excretion of ammonium within cultures under aerobic conditions. The nitrogenase complex is oxygen sensitive. Nevertheless, nitrogen fixation occurs under oxic conditions due to cyanobacteria-specific characteristics. For instance, in some cyanobacteria, the vegetative cell differentiation in heterocyts provides a well-adapted anaerobic microenvironment for nitrogenase protection. Therefore, cell cultures of oxygenic cyanobacteria have been grown in laboratory and pilot photobioreactors (Dasgupta et al., 2010; Fontes et al., 1987; Moreno et al., 2003; Nayak & Das, 2013). Biomass production under diazotrophic conditions has been shown to be controlled by environmental factors such as light intensity, temperature, aeration rate, and inorganic carbon concentration, also, more specifically, by the concentration of dissolved oxygen in the culture medium. Currently, there is little information regarding the production of extracellular ammonium by heterocytous cyanobacteria. This review compares the available data on maximum ammonium concentrations and analyses the specific rate production in cultures grown as free or immobilized filamentous cyanobacteria. Extracellular production of ammonium could be coupled, as suggested by recent research on non-diazotrophic cyanobacteria, to that of other high value metabolites. There is little information available regarding the possibility for using diazotrophic cyanobacteria as cellular factories may be in regard of the constraints due to nitrogen fixation.

  7. Prone to fix: Resilience of the active nitrogen-fixing rice root microbiome

    Science.gov (United States)

    Hurek, Thomas; Sabale, Mugdha; Sarkar, Abhijit; Pees, Tobias; Reinhold-Hurek, Barbara

    2016-04-01

    Due to water consumption, many lowland rice areas in Asia are undergoing a transition that involves adoption of new management strategies, with crop rotations encompassing a non-flooded crop, including maize. Shifting from flooded to non-flooded cropping is likely to affect microbial nitrogen cycling. For analysis of the root-associated microbiome of rice and maize in response to flooding or nitrogen fertilizer, we combine methods of microbial ecology (Next-Generation sequencing of amplicons), and a reductionist approach with pure cultures of the endophytic diazotroph Azoarus sp.. Field plots of the ICON project (Introducing non-flooded crops in rice-dominated landscapes: Impact on Carbon, nitrogen and water budgets) at the International Rice Research Institute in the Philippines were analyzed. Root-associated activity of nitrogenase gene expression was assessed by quantitative RT-PCR of nifH. For rice, expression levels were surprisingly stable, in response to non-flooded versus flooded conditions, or in response to conventional nitrogen fertilizer applications versus lack of N-fertilizer. In contrast, the active diazotrophic population of maize roots was not resistant to N-fertilization, nifH expression strongly decreased. Concordant changes in the diazotrophic resident or active communities were detected by nifH amplicon sequence analysis, based on bacterial DNA or mRNA, respectively. For high-resolution analyses of the endobiome in gnotobiotic culture, we developed a dual fluorescence reporter system for Azoarcus sp. BH72 which allows to quantify and visualize epi- and endophytic gene expression by concfocal microscopy (CLSM). This allowed us to demonstrate sites of active nitrogen fixation (gene expression) in association with rice roots. We confirmed that at low nitrogen fertilizer levels, endophytic nifH gene expression persisted in rice roots, while it was repressed in maize roots. This supports our observation of remarkable stability of nitrogen fixation

  8. Bacteroidales ectosymbionts of gut flagellates shape the nitrogen-fixing community in dry-wood termites

    Science.gov (United States)

    Desai, Mahesh S; Brune, Andreas

    2012-01-01

    Although it is well documented that the lack of nitrogen in the diet of wood-feeding termites is compensated by the nitrogen-fixing capacity of their gut microbiota, the bacteria responsible for this activity are largely unknown. Here, we analyzed the diversity and expression of nitrogenase genes (homologs of nifH) in four species of dry-wood termites (Kalotermitidae), which thrive on a particularly nitrogen-poor resource. Although each species harbored a highly diverse suite of termite-specific homologs in their microliter-sized hindgut, only a core set related to nifH genes of Treponema and Azoarcus spp., ‘Azobacteroides pseudotrichonymphae', the first member of the Bacteroidales identified as a diazotroph, and termite-gut-specific anfH genes of hitherto unknown origin were preferentially expressed. Transcription patterns corroborated that the populations of active diazotrophs differ fundamentally between termite genera. Capillary-picked suspensions of the flagellates Devescovina arta and Snyderella tabogae revealed that their bacterial ectosymbionts each possess two paralogs of nifH, which apparently have been acquired consecutively during evolution of Bacteroidales, but only one of them (anfH) is actively expressed. Transcription patterns correlated neither with the molybdenum content of the diet nor with intestinal hydrogen concentrations, measured with microsensors. We propose that the nitrogen-fixing community in different dry-wood termites is shaped by the symbionts of their specific flagellate populations. Our findings suggest that the diazotrophic nature of ‘Armantifilum devescovinae' has an important role in the nitrogen metabolism of dry-wood termites and is the driving force of co-evolution with its flagellate host. PMID:22189498

  9. Biogeochemical tracers of the marine cyanobacterium Trichodesmium

    Science.gov (United States)

    Carpenter, Edward J.; Harvey, H. Rodger; Fry, Brian; Capone, Douglas G.

    1997-01-01

    We examined the utility of several biogeochemical tracers for following the fate of the planktonic diazotrophic cyanobacterium Trichodesmium in the sea. The presence of a (CIO) fatty acid previously reported was observed in a culture of Trichodesmium but was not found in natural samples. This cyanobacterium had high concentrations of C 14 and C 16 acids, with lesser amounts of several saturated and unsaturated C 18 fatty acids. This composition was similar to that of other marine cyanobacteria. The major hydrocarbon identified was the C 17n-alkane, which was present in all samples from the five stations examined. Sterols common to algae and copepods were observed in many samples along with hopanoids representative of bacteria, suggesting a varied community structure in colonies collected from different stations. We found no unique taxonomic marker of Trichodesmium among the sterols. Measurements of the σ 15N and σ 13C in Trichodesmium samples from the SW Sargasso and NW Caribbean Seas averaged -0.4960 (range from -0.7 to -0.25960) and -12.9%0 (range from -15.2 to -11.9960), respectively, thus confirming previous observations that this cyanobacterial diazotroph has both the lowest σ 15N and highest σ 13C of any marine phytoplankter observed to date. A culture of Trichodesmium grown under diazotrophic conditions had a σ 15N between -1.3 and -3.6960. Our results support the supposition that the relatively low σ 15N and high σ 13C values observed in suspended and sediment-trapped material from some tropical and subtropical seas result from substantial input of C and N by Trichodesmium.

  10. Effect of inorganic nitrogenous fertilizer on productivity of recently reclaimed saline sodic soils with and without biofertilizer.

    Science.gov (United States)

    Mehdi, S M; Sarfraz, M; Shabbir, G; Abbas, G

    2007-07-15

    Saline sodic soils after reclamation become infertile due to leaching of most of the nutrients along with salts from the rooting medium. Microbes can play a vital role in the productivity improvement of such soils. In this study a saline sodic field having EC, 6.5 dS m(-1), pH, 9.1 and gypsum requirement (GR) 3.5 tons acre(-1) was reclaimed by applying gypsum at the rate of 100% GR. Rice and wheat crops were transplanted/sown for three consecutive years. Inorganic nitrogenous fertilizer was used with and without biofertilizers i.e., Biopower (Azospirillum) for rice and diazotroph inoculums for wheat. Nitrogen was applied at the rate of 0, 75% of recommended dose (RD), RD, 125% of RD and 150% of RD. Recommended dose of P without K was applied to all the plots. Biopower significantly improved Paddy and straw yield of rice over inorganic nitrogenous fertilizer. In case of wheat diazotroph inoculum improved grain and straw yield significantly over inorganic nitrogenous fertilizer. Among N fertilizer rates, RD + 25% additional N fertilizer was found to be the best dose for rice and wheat production in recently reclaimed soils. Nitrogen concentration and its uptake by paddy, grain and straw were also increased by biopower and diazotroph inoculum over inorganic nitrogenous fertilizer. Among N fertilizer rates, RD + 25% additional N fertilizer was found to be the best dose for nitrogen concentration and its uptake by paddy, grain and straw. Total soil N, available P and extractable K were increased while salinity/sodicity parameters were decreased with the passage of time. The productivity of the soil was improved more by biofertilizers over inorganic N fertilizers.

  11. Nitrogenase gene amplicons from global marine surface waters are dominated by genes of non-cyanobacteria.

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    Hanna Farnelid

    Full Text Available Cyanobacteria are thought to be the main N(2-fixing organisms (diazotrophs in marine pelagic waters, but recent molecular analyses indicate that non-cyanobacterial diazotrophs are also present and active. Existing data are, however, restricted geographically and by limited sequencing depths. Our analysis of 79,090 nitrogenase (nifH PCR amplicons encoding 7,468 unique proteins from surface samples (ten DNA samples and two RNA samples collected at ten marine locations world-wide provides the first in-depth survey of a functional bacterial gene and yield insights into the composition and diversity of the nifH gene pool in marine waters. Great divergence in nifH composition was observed between sites. Cyanobacteria-like genes were most frequent among amplicons from the warmest waters, but overall the data set was dominated by nifH sequences most closely related to non-cyanobacteria. Clusters related to Alpha-, Beta-, Gamma-, and Delta-Proteobacteria were most common and showed distinct geographic distributions. Sequences related to anaerobic bacteria (nifH Cluster III were generally rare, but preponderant in cold waters, especially in the Arctic. Although the two transcript samples were dominated by unicellular cyanobacteria, 42% of the identified non-cyanobacterial nifH clusters from the corresponding DNA samples were also detected in cDNA. The study indicates that non-cyanobacteria account for a substantial part of the nifH gene pool in marine surface waters and that these genes are at least occasionally expressed. The contribution of non-cyanobacterial diazotrophs to the global N(2 fixation budget cannot be inferred from sequence data alone, but the prevalence of non-cyanobacterial nifH genes and transcripts suggest that these bacteria are ecologically significant.

  12. Temporal and spatial variability of biological nitrogen fixation off the upwelling system of central Chile (35-38.5°S)

    Science.gov (United States)

    Fernandez, Camila; González, Maria Lorena; Muñoz, Claudia; Molina, Veronica; Farias, Laura

    2015-05-01

    Although N2 fixation could represent a supplementary source of bioavailable nitrogen in coastal upwelling areas and underlying oxygen minimum zones (OMZs), the limited data available prevent assessing its variability and biogeochemical significance. Here we report the most extensive N2 fixation data set gathered to date in the upwelling area off central Chile (36°S). It covers interannual to high frequency time scales in an area of about 82,500 km2 in the eastern South Pacific (ESP). Because heterotrophic N2 fixation may be regulated by DOM availability in the ESP, we conducted experiments at different oxygen conditions and included DOM amendments in order to test diazotrophic activity. Rates in the euphotic zone showed strong temporal variability which resulted in values reaching 0.5 nmol L-1 d-1 in 2006 (average 0.32 ± 0.17 nmol L-1 d-1) and up to 126.8 nmol L-1 d-1 (average 24.75 ± 37.9 nmol L-1 d-1) in 2011. N2 fixation in subsurface suboxic conditions (1.5 ± 1.16 nmol L-1 d-1) also occurred mainly during late summer and autumn while virtually absent in winter. The diversity of diazotrophs was dominated by heterotrophs, with higher richness in surface compared to OMZ waters. Rates in oxygen depleted conditions could exceed values obtained in the euphotic layer, but rates were not dependent on the availability of dissolved organic matter. N2 fixation also showed a positive correlation with total chlorophyll and the C:N ratio of phytoplankton, but not to the P excess compared to N. We conclude that the diazotrophic community responds to the composition of phytoplankton rather than the extent of N deficiency and the availability of bulk DOM in this system.

  13. Microbiome of Trichodesmium Colonies from the North Pacific Subtropical Gyre

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    Mary R. Gradoville

    2017-07-01

    Full Text Available Filamentous diazotrophic Cyanobacteria of the genus Trichodesmium, often found in colonial form, provide an important source of new nitrogen to tropical and subtropical marine ecosystems. Colonies are composed of several clades of Trichodesmium in association with a diverse community of bacterial and eukaryotic epibionts. We used high-throughput 16S rRNA and nifH gene sequencing, carbon (C and dinitrogen (N2 fixation assays, and metagenomics to describe the diversity and functional potential of the microbiome associated with Trichodesmium colonies collected from the North Pacific Subtropical Gyre (NPSG. The 16S rRNA and nifH gene sequences from hand-picked colonies were predominantly (>99% from Trichodesmium Clade I (i.e., T. thiebautii, which is phylogenetically and ecologically distinct from the Clade III IMS101 isolate used in most laboratory studies. The bacterial epibiont communities were dominated by Bacteroidetes, Alphaproteobacteria, and Gammaproteobacteria, including several taxa with a known preference for surface attachment, and were relatively depleted in the unicellular Cyanobacteria and small photoheterotrophic bacteria that dominate NPSG surface waters. Sequencing the nifH gene (encoding a subcomponent of the nitrogenase enzyme identified non-Trichodesmium diazotrophs that clustered predominantly among the Cluster III nifH sequence-types that includes putative anaerobic diazotrophs. Trichodesmium colonies may represent an important habitat for these Cluster III diazotrophs, which were relatively rare in the surrounding seawater. Sequence analyses of nifH gene transcripts revealed several cyanobacterial groups, including heterocystous Richelia, associated with the colonies. Both the 16S rRNA and nifH datasets indicated strong differences between Trichodesmium epibionts and picoplankton in the surrounding seawater, and also between the epibionts inhabiting Trichodesmium puff and tuft colony morphologies. Metagenomic and 16S r

  14. A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

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    Gordon Ng

    Full Text Available BACKGROUND: Nitrogen (N(2 fixation also yields hydrogen (H(2 at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2 as sole N-source bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase, has nevertheless been presumed responsible for recycling such endogenous hydrogen. METHODS AND FINDINGS: As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase. An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. CONCLUSIONS: Representative of aerobic N(2-fixing and H(2-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2 respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2, specifically that produced by N(2 fixation. To benefit human civilization, H(2 has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As

  15. Nutrient uplift in a cyclonic eddy increases diversity, primary productivity and iron demand of microbial communities relative to a western boundary current

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    Martina A. Doblin

    2016-04-01

    Full Text Available The intensification of western boundary currents in the global ocean will potentially influence meso-scale eddy generation, and redistribute microbes and their associated ecological and biogeochemical functions. To understand eddy-induced changes in microbial community composition as well as how they control growth, we targeted the East Australian Current (EAC region to sample microbes in a cyclonic (cold-core eddy (CCE and the adjacent EAC. Phototrophic and diazotrophic microbes were more diverse (2–10 times greater Shannon index in the CCE relative to the EAC, and the cell size distribution in the CCE was dominated (67% by larger micro-plankton $(\\geq 20\\lrm{\\mu }\\mathrm{m}$ ≥ 20 μ m , as opposed to pico- and nano-sized cells in the EAC. Nutrient addition experiments determined that nitrogen was the principal nutrient limiting growth in the EAC, while iron was a secondary limiting nutrient in the CCE. Among the diazotrophic community, heterotrophic NifH gene sequences dominated in the EAC and were attributable to members of the gamma-, beta-, and delta-proteobacteria, while the CCE contained both phototrophic and heterotrophic diazotrophs, including Trichodesmium, UCYN-A and gamma-proteobacteria. Daily sampling of incubation bottles following nutrient amendment captured a cascade of effects at the cellular, population and community level, indicating taxon-specific differences in the speed of response of microbes to nutrient supply. Nitrogen addition to the CCE community increased picoeukaryote chlorophyll a quotas within 24 h, suggesting that nutrient uplift by eddies causes a ‘greening’ effect as well as an increase in phytoplankton biomass. After three days in both the EAC and CCE, diatoms increased in abundance with macronutrient (N, P, Si and iron amendment, whereas haptophytes and phototrophic dinoflagellates declined. Our results indicate that cyclonic eddies increase delivery of nitrogen to the upper ocean to potentially

  16. Covariation of viral parameters with bacterial assemblage richness and diversity in the water column and sediments

    Science.gov (United States)

    Hewson, Ian; Fuhrman, Jed A.

    2007-05-01

    Viruses are hypothesized to maintain diversity in microbial assemblages by regulating the abundance of dominant competitors and thereby allowing less-dominant competitors to persist in assemblages; however, there have been few empirical data sets to support this idea. In this study, we examined the relationship between the ratio of viral abundance to bacterial abundance, viral production, and the relative richness and diversity of bacterial assemblage fingerprints, in samples taken from geographically widespread locations (North Pacific gyre, the Amazon River plume and adjacent North Atlantic gyre, Gulf of Mexico, Southern California Bight and Arafura—Coral Seas) which are oligo- to mesotrophic. Bacterial assemblage richness and diversity as measured by automated rRNA intergenic spacer (ARISA) fingerprinting were significantly and positively correlated with the ratio of virus abundance to bacteria abundance (VBR) and to the rate of virus production only in the oligotrophic North Pacific gyre. ARISA fingerprint richness/diversity were not significantly correlated to viral parameters when assessed across all samples in surface waters, suggesting there is not a singular global quantitative relationship between viral pressure and host diversity within well evolved host/virus systems in different geographic locations in plankton. In sediments off Southern California, viral parameters significantly and negatively correlated with ARISA diversity, suggesting strong viral interactions in this habitat. To examine covariation of viral parameters and the relative abundance and diversity of rarer bacterial taxa (i.e., less-dominant competitor), the richness and diversity of diazotroph communities was measured using terminal restriction fragment length polymorphism (TRFLP) of a portion ( nifH) of the nitrogenase gene. The richness and diversity of diazotrophic communities were significantly and negatively correlated with viral parameters across all locations. Since diazotrophs

  17. Genome Wide Transcriptional Profiling of Herbaspirillum seropedicae SmR1 Grown in the presence of naringenin

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    Michele Zibetti Tadra-Sfeir

    2015-05-01

    Full Text Available Herbaspirillum seropedicae is a diazotrophic bacterium which associates endophytically with economically important gramineae. Flavonoids such as naringenin, have been shown to have an effect on the interaction between H. seropedicae and its host plants. We used a high-throughput sequencing based method (RNA-Seq to access the influence of naringenin on the whole transcriptome profile of H. seropedicae. Three hundred and four genes were downregulated and seventy seven were upregulated by naringenin. Data analysis revealed that genes related to bacterial flagella biosynthesis, chemotaxis and biosynthesis of peptidoglycan were repressed by naringenin. Moreover, genes involved in aromatic metabolism and multidrug transport efllux were actived.

  18. Genome wide transcriptional profiling of Herbaspirillum seropedicae SmR1 grown in the presence of naringenin.

    Science.gov (United States)

    Tadra-Sfeir, Michelle Z; Faoro, Helisson; Camilios-Neto, Doumit; Brusamarello-Santos, Liziane; Balsanelli, Eduardo; Weiss, Vinicius; Baura, Valter A; Wassem, Roseli; Cruz, Leonardo M; De Oliveira Pedrosa, Fábio; Souza, Emanuel M; Monteiro, Rose A

    2015-01-01

    Herbaspirillum seropedicae is a diazotrophic bacterium which associates endophytically with economically important gramineae. Flavonoids such as naringenin have been shown to have an effect on the interaction between H. seropedicae and its host plants. We used a high-throughput sequencing based method (RNA-Seq) to access the influence of naringenin on the whole transcriptome profile of H. seropedicae. Three hundred and four genes were downregulated and seventy seven were upregulated by naringenin. Data analysis revealed that genes related to bacterial flagella biosynthesis, chemotaxis and biosynthesis of peptidoglycan were repressed by naringenin. Moreover, genes involved in aromatic metabolism and multidrug transport efllux were actived.

  19. A two-dimensional proteome reference map of Herbaspirillum seropedicae proteins.

    Science.gov (United States)

    Chaves, Daniela Fojo Seixas; Ferrer, Pércio Pereira; de Souza, Emanuel Maltempi; Gruz, Leonardo Magalhães; Monteiro, Rose Adele; de Oliveira Pedrosa, Fábio

    2007-10-01

    Herbaspirillum seropedicae is an endophytic diazotroph associated with economically important crops such as rice, sugarcane, and wheat. Here, we present a 2-D reference map for H. seropedicae. Using MALDI-TOF-MS we identified 205 spots representing 173 different proteins with a calculated average of 1.18 proteins/gene. Seventeen hypothetical or conserved hypothetical ORFs were shown to code for true gene products. These data will support the genome annotation process and provide a basis on which to undertake comparative proteomic studies.

  20. Carbon and nitrogen stable isotope ratios of pelagic zooplankton elucidate ecohydrographic features in the oligotrophic Red Sea

    KAUST Repository

    Kürten, Benjamin

    2015-11-10

    Although zooplankton occupy key roles in aquatic biogeochemical cycles, little is known about the pelagic food web and trophodynamics of zooplankton in the Red Sea. Natural abundance stable isotope analysis (SIA) of carbon (δ13C) and N (δ15N) is one approach to elucidating pelagic food web structures and diet assimilation Integrating the combined effects of ecological processes and hydrography, ecohydrographic features often translate into geographic patterns in δ13C and δ15N values at the base of food webs. This is due, for example, to divergent 15N abundances in source end-members (deep water sources: high δ15N, diazotrophs: low δ15N). Such patterns in the spatial distributions of stable isotope values were coined isoscapes. Empirical data of atmospheric, oceanographic, and biological processes, which drive the ecohydrographic gradients of the oligotrophic Red Sea, are under-explored and some rather anticipated than proven. Specifically, five processes underpin Red Sea gradients: a) monsoon-related intrusions of nutrient-rich Indian Ocean water; b) basin scale thermohaline circulation; c) mesoscale eddy activity that causes up-welling of deep water nutrients into the upper layer; d) the biological fixation of atmospheric nitrogen (N2) by diazotrophs; and e) the deposition of aerosol-derived N. This study assessed relationships between environmental samples (nutrients, chlorophyll a), oceanographic data (temperature, salinity, current velocity [ADCP]), particulate organic matter (POM), and net-phytoplankton, with the δ13C and δ15N values of zooplankton collected in spring 2012 from 16°28’ to 26°57’N along the central axis of the Red Sea. The δ15N of bulk POM and most zooplankton taxa increased from North (Duba) to South (Farasan). The potential contribution of deep water nutrient-fueled phytoplankton, POM, and diazotrophs varied among sites. Estimates suggested higher diazotroph contributions in the North, a greater contribution of POM in the South

  1. Insights Into the Biogeochemical Cycling of Iron, Nitrate, and Phosphate Across a 5,300 km South Pacific Zonal Section (153°E-150°W)

    Science.gov (United States)

    Ellwood, Michael J.; Bowie, Andrew R.; Baker, Alex; Gault-Ringold, Melanie; Hassler, Christel; Law, Cliff S.; Maher, William A.; Marriner, Andrew; Nodder, Scott; Sander, Sylvia; Stevens, Craig; Townsend, Ashley; van der Merwe, Pier; Woodward, E. Malcolm S.; Wuttig, Kathrin; Boyd, Philip W.

    2018-02-01

    Iron, phosphate, and nitrate are essential nutrients for phytoplankton growth, and hence, their supply into the surface ocean controls oceanic primary production. Here we present a GEOTRACES zonal section (GP13; 30-33°S, 153°E-150°W) extending eastward from Australia to the oligotrophic South Pacific Ocean gyre outlining the concentrations of these key nutrients. Surface dissolved iron concentrations are elevated at >0.4 nmol L-1 near continental Australia (west of 165°E) and decreased eastward to ≤0.2 nmol L-1 (170°W-150°W). The supply of dissolved iron into the upper ocean (nitrate concentrations averaged 5 ± 4 nmol L-1 between 170°W and 150°W, while surface water phosphate concentrations averaged 58 ± 30 nmol L-1. The supply of nitrogen into the upper ocean is primarily from deeper waters (24-1647 μmol m-2 d-1) with atmospheric deposition and nitrogen fixation contributing leading to almost quantitative removal of nitrate. The supply stoichiometry for iron and nitrogen relative to phosphate at and above the DCM declines eastward leading to two biogeographical provinces: one with diazotroph production and the other without diazotroph production.

  2. Efecto del uso del suelo sobre rizobacterias fosfatosolubizadoras y diazotroficas en el distrito de riego del río zulia,norte de santander (colombia

    Directory of Open Access Journals (Sweden)

    Ronal Fernando-Cañon

    2009-07-01

    Full Text Available It was quantified the population of diazotrophic and phosphate solubilizer bacteria with the aim of determine the effect of the use of soils during different periods of intervention with culture of rice in three agro-ecological zones of the District of Irrigation of the Zulia river. The results showed that the management of the culture of rice in the different studied zones significantly influenced the population of microorganisms in all the studied culture media, excepting the semi-solid culture media JMV. The populations of the genera Azospirillum spp., Azotobacter spp., Beijerinckia spp., were influenced by the fertility of the soils in the zones of Buena Esperanza and Restauración compared with Limoncito, where the quantities of organic matter and nutritional escential elements were lower. It were obtained 28 isolations of entophytic, associative, free-living diazotrophic and phosphate solubilizer rhizobacteria according with their macroscopic characteristics in the culture media Batata, JMV, Ashby and nutritive, from the analysis of the population in the zones of Buena Esperanza, Restauración and Limoncito. These isolations were purified and preserved in sterilized saline solution (0.85% NaCl at 4°C in the Laboratory of Microbiology of the Colombian Agricultural Institute, ICA, with the purpose to be used in subsequent studies about their potential as biofertilizers in rice cultured soils of the department.

  3. Beneficial effects of aluminum enrichment on nitrogen-fixing cyanobacteria in the South China Sea.

    Science.gov (United States)

    Liu, Jiaxing; Zhou, Linbin; Ke, Zhixin; Li, Gang; Shi, Rongjun; Tan, Yehui

    2018-04-01

    Few studies focus on the effects of aluminum (Al) on marine nitrogen-fixing cyanobacteria, which play important roles in the ocean nitrogen cycling. To examine the effects of Al on the nitrogen-fixing cyanobacteria, bioassay experiments in the oligotrophic South China Sea (SCS) and culture of Crocosphaera watsonii in the laboratory were conducted. Field data showed that 200 nM Al stimulated the growth and the nitrogenase gene expression of Trichodesmium and unicellular diazotrophic cyanobacterium group A, and the nitrogen fixation rates of the whole community. Laboratory experiments demonstrated that Al stimulated the growth and nitrogen fixation of C. watsonii under phosphorus limited conditions. Both field and laboratory results indicated that Al could stimulate the growth of diazotrophs and nitrogen fixation in oligotrophic oceans such as the SCS, which is likely related to the utilization of phosphorus, implying that Al plays an important role in the ocean nitrogen and carbon cycles by influencing nitrogen fixation. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. N2-fixation and seedling growth promotion of lodgepole pine by endophytic Paenibacillus polymyxa.

    Science.gov (United States)

    Anand, Richa; Grayston, Susan; Chanway, Christopher

    2013-08-01

    We inoculated lodgepole pine (Pinus contorta var. latifolia (Dougl.) Engelm.) with Paenibacillus polymyxa P2b-2R, a diazotrophic bacterium previously isolated from internal stem tissue of a naturally regenerating pine seedling to evaluate biological nitrogen fixation and seedling growth promotion by this microorganism. Seedlings generated from pine seed inoculated with strain P2b-2R were grown for up to 13 months in a N-limited soil mix containing 0.7 mM available N labeled as Ca((15)NO3)2 to facilitate detection of N2-fixation. Strain P2b-2R developed a persistent endophytic population comprising 10(2)-10(6) cfu g(-1) plant tissue inside pine roots, stems, and needles during the experiment. At the end of the growth period, P2b-2R had reduced seedling mortality by 14 % and (15)N foliar N abundance 79 % and doubled foliar N concentration and seedling biomass compared to controls. Our results suggest that N2-fixation by P. polymyxa enhanced growth of pine seedlings and support the hypothesis that plant-associated diazotrophs capable of endophytic colonization can satisfy a significant proportion of the N required by tree seedlings growing under N-limited conditions.

  5. Fungal endophyte Phomopsis liquidambari affects nitrogen transformation processes and related microorganisms in the rice rhizosphere

    Directory of Open Access Journals (Sweden)

    Bo eYang

    2015-09-01

    Full Text Available The endophytic fungus Phomopsis liquidambari performs an important ecosystem service by assisting its host with acquiring soil nitrogen (N, but little is known regarding how this fungus influences soil N nutrient properties and microbial communities. In this study, we investigated the impact of P. liquidambari on N dynamics,the abundance and composition of N cycling genes in rhizosphere soil treated with three levels of N (urea. Ammonia-oxidizing archaea (AOA, ammonia-oxidizing bacteria (AOB and diazotrophs were assayed using quantitative real-time polymerase chain reaction and denaturing gradient gel electrophoresis at four rice growing stages (S0: before planting, S1: tillering stage, S2: grain filling stage, and S3: ripening stage. A significant increase in the available nitrate and ammonium contents was found in the rhizosphere soil of endophyte-infected rice under low N conditions. Moreover, P. liquidambari significantly increased the potential nitrification rates (PNR, affected the abundance and community structure of AOA, AOB and diazotrophs under low N conditions in the S1 and S2 stages. The root exudates were determined due to their important role in rhizosphere interactions. P. liquidambari colonization altered the exudation of organic compounds by rice roots and P. liquidambari increased the concentration of soluble saccharides, total free amino acids and organic acids

  6. Biological nitrogen fixation in the oxygen-minimum region of the eastern tropical North Pacific ocean.

    Science.gov (United States)

    Jayakumar, Amal; Chang, Bonnie X; Widner, Brittany; Bernhardt, Peter; Mulholland, Margaret R; Ward, Bess B

    2017-10-01

    Biological nitrogen fixation (BNF) was investigated above and within the oxygen-depleted waters of the oxygen-minimum zone of the Eastern Tropical North Pacific Ocean. BNF rates were estimated using an isotope tracer method that overcame the uncertainty of the conventional bubble method by directly measuring the tracer enrichment during the incubations. Highest rates of BNF (~4 nM day -1 ) occurred in coastal surface waters and lowest detectable rates (~0.2 nM day -1 ) were found in the anoxic region of offshore stations. BNF was not detectable in most samples from oxygen-depleted waters. The composition of the N 2 -fixing assemblage was investigated by sequencing of nifH genes. The diazotrophic assemblage in surface waters contained mainly Proteobacterial sequences (Cluster I nifH), while both Proteobacterial sequences and sequences with high identities to those of anaerobic microbes characterized as Clusters III and IV type nifH sequences were found in the anoxic waters. Our results indicate modest input of N through BNF in oxygen-depleted zones mainly due to the activity of proteobacterial diazotrophs.

  7. Paving a Path to Understanding Metabolic Responses to Iron Bioavailability: Global Proteomic Analysis of Crocosphaera watsonii

    Science.gov (United States)

    Gauglitz, J.; McIlvin, M. R.; Moran, D. M.; Waterbury, J. B.; Saito, M. A.

    2016-02-01

    Marine diazotrophic cyanobacteria provide a key source of new nitrogen into the oceans and are important contributors to primary production. The geographic distribution of these cyanobacteria is impacted by available iron and phosphorus as well as environmental conditions such as temperature, however available iron concentrations are thought to be particularly critical due to the high demand for iron in cellular processes. Iron bioavailability and microorganismal adaptations to low iron environments may thus play a key role in dictating community structure, however the mechanisms by which cyanobacteria acquire iron and regulate its uptake are not well defined. In this study, the unicellular diazotroph, Crocosphaera watsonii WH8501, was acclimated to a range of bioavailable iron concentrations (from 0.001nM to 8.13nM Fe') using trace metal clean culturing techniques and the proteomes were analyzed by LC/MS-MS. Physiological and proteomic data indicate three distinct phenotypic ranges: iron-replete, iron-limited, and iron-starved. Trends in photosynthetic, carbon fixation and iron storage proteins across the iron gradient indicate that the C. watsonii proteome responds directly to iron availability. Further analysis of relative protein expression, which describes the physiological state of the cell, will lead to insights into how C. watsonii is able to adapt to iron-limited conditions and the resulting biogeochemical implications will be discussed.

  8. Identification of nitrogen-fixing genes and gene clusters from metagenomic library of acid mine drainage.

    Science.gov (United States)

    Dai, Zhimin; Guo, Xue; Yin, Huaqun; Liang, Yili; Cong, Jing; Liu, Xueduan

    2014-01-01

    Biological nitrogen fixation is an essential function of acid mine drainage (AMD) microbial communities. However, most acidophiles in AMD environments are uncultured microorganisms and little is known about the diversity of nitrogen-fixing genes and structure of nif gene cluster in AMD microbial communities. In this study, we used metagenomic sequencing to isolate nif genes in the AMD microbial community from Dexing Copper Mine, China. Meanwhile, a metagenome microarray containing 7,776 large-insertion fosmids was constructed to screen novel nif gene clusters. Metagenomic analyses revealed that 742 sequences were identified as nif genes including structural subunit genes nifH, nifD, nifK and various additional genes. The AMD community is massively dominated by the genus Acidithiobacillus. However, the phylogenetic diversity of nitrogen-fixing microorganisms is much higher than previously thought in the AMD community. Furthermore, a 32.5-kb genomic sequence harboring nif, fix and associated genes was screened by metagenome microarray. Comparative genome analysis indicated that most nif genes in this cluster are most similar to those of Herbaspirillum seropedicae, but the organization of the nif gene cluster had significant differences from H. seropedicae. Sequence analysis and reverse transcription PCR also suggested that distinct transcription units of nif genes exist in this gene cluster. nifQ gene falls into the same transcription unit with fixABCX genes, which have not been reported in other diazotrophs before. All of these results indicated that more novel diazotrophs survive in the AMD community.

  9. Synthesis of silver nanoparticles on the basis of low and high molar mass exopolysaccharides of Bradyrhizobium japonicum 36 and its antimicrobial activity against some pathogens.

    Science.gov (United States)

    Rasulov, Bakhtiyor; Rustamova, Nigora; Yili, Abulimiti; Zhao, Hai-Qing; Aisa, Haji A

    2016-07-01

    Silver nanoparticles (SNPs) were synthesized on the basis of exopolysaccharides (low and high molar mass) of diazotrophic Bradyrhizobium japonicum 36 strain. The synthesis of SNPs was carried out by direct reduction of silver nitrate with ethanol-insoluble (high molar mass, HMW) and ethanol-soluble (low molar mass, LMW) fractions of exopolysaccharides (EPS), produced by diazotrophic strain B. japonicum 36. SNPs were characterized using UV-vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). SNPs synthesized on the basis of LMW EPS absorbed radiation in the visible regions of 420 nm, whereas SNPs based on the HMW EPS have a wavelength maximum at 450 nm because of the strong SPR transition. Moreover, the antibacterial and antifungal activities of the SNPs were examined in vitro against Escherichia coli, Staphylococcus aureus, and Candida albicans. SNPs synthesized on the basis of LMW EPS were active than those synthesized on the basis of HMW EPS. Besides, UV-visible spectroscopic evaluation confirmed that SNPs synthesized on the basis of LMW EPS were far more stable than those obtained on the basis of HMW EPS.

  10. Identification of nitrogen-fixing genes and gene clusters from metagenomic library of acid mine drainage.

    Directory of Open Access Journals (Sweden)

    Zhimin Dai

    Full Text Available Biological nitrogen fixation is an essential function of acid mine drainage (AMD microbial communities. However, most acidophiles in AMD environments are uncultured microorganisms and little is known about the diversity of nitrogen-fixing genes and structure of nif gene cluster in AMD microbial communities. In this study, we used metagenomic sequencing to isolate nif genes in the AMD microbial community from Dexing Copper Mine, China. Meanwhile, a metagenome microarray containing 7,776 large-insertion fosmids was constructed to screen novel nif gene clusters. Metagenomic analyses revealed that 742 sequences were identified as nif genes including structural subunit genes nifH, nifD, nifK and various additional genes. The AMD community is massively dominated by the genus Acidithiobacillus. However, the phylogenetic diversity of nitrogen-fixing microorganisms is much higher than previously thought in the AMD community. Furthermore, a 32.5-kb genomic sequence harboring nif, fix and associated genes was screened by metagenome microarray. Comparative genome analysis indicated that most nif genes in this cluster are most similar to those of Herbaspirillum seropedicae, but the organization of the nif gene cluster had significant differences from H. seropedicae. Sequence analysis and reverse transcription PCR also suggested that distinct transcription units of nif genes exist in this gene cluster. nifQ gene falls into the same transcription unit with fixABCX genes, which have not been reported in other diazotrophs before. All of these results indicated that more novel diazotrophs survive in the AMD community.

  11. Oscillating behavior of carbohydrate granule formation and dinitrogen fixation in the cyanobacterium Cyanothece sp. strain ATCC 51142

    Science.gov (United States)

    Schneegurt, M. A.; Sherman, D. M.; Nayar, S.; Sherman, L. A.; Mitchell, C. A. (Principal Investigator)

    1994-01-01

    It has been shown that some aerobic, unicellular, diazotrophic cyanobacteria temporally separate photosynthetic O2 evolution and oxygen-sensitive N2 fixation. Cyanothece sp. ATCC strain 51142 is an aerobic, unicellular, diazotrophic cyanobacterium that fixes N2 during discrete periods of its cell cycle. When the bacteria are maintained under diurnal light-dark cycles, N2 fixation occurs in the dark. Similar cycling is observed in continuous light, implicating a circadian rhythm. Under N2-fixing conditions, large inclusion granules form between the thylakoid membranes. Maximum granulation, as observed by electron microscopy, occurs before the onset of N2 fixation, and the granules decrease in number during the period of N2 fixation. The granules can be purified from cell homogenates by differential centrifugation. Biochemical analyses of the granules indicate that these structures are primarily carbohydrate, with some protein. Further analyses of the carbohydrate have shown that it is a glucose polymer with some characteristics of glycogen. It is proposed that N2 fixation is driven by energy and reducing power stored in these inclusion granules. Cyanothece sp. strain ATCC 51142 represents an excellent experimental organism for the study of the protective mechanisms of nitrogenase, metabolic events in cyanobacteria under normal and stress conditions, the partitioning of resources between growth and storage, and biological rhythms.

  12. Chasing after Non-cyanobacterial Nitrogen Fixation in Marine Pelagic Environments

    Directory of Open Access Journals (Sweden)

    Pia H. Moisander

    2017-09-01

    Full Text Available Traditionally, cyanobacterial activity in oceanic photic layers was considered responsible for the marine pelagic dinitrogen (N2 fixation. Other potentially N2-fixing bacteria and archaea have also been detected in the pelagic water column, however, the activity and importance of these non-cyanobacterial diazotrophs (NCDs remain poorly constrained. In this perspective we summarize the N2 fixation rates from recently published studies on photic and aphotic layers that have been attributed to NCD activity via parallel molecular measurements, and discuss the status, challenges, and data gaps in estimating non-cyanobacterial N2 fixation NCNF in the ocean. Rates attributed to NCNF have generally been near the detection limit thus far (<1 nmol N L−1 d−1. Yet, if considering the large volume of the dark ocean, even low rates of NCNF could make a significant contribution to the new nitrogen input to the ocean. The synthesis here shows that nifH transcription data for NCDs have been reported in only a few studies where N2 fixation rates were detected in the absence of diazotrophic cyanobacteria. In addition, high apparent diversity and regional variability in the NCDs complicate investigations of these communities. Future studies should focus on further investigating impacts of environmental drivers including oxygen, dissolved organic matter, and dissolved inorganic nitrogen on NCNF. Describing the ecology of NCDs and accurately measuring NCNF rates, are critical for a future evaluation of the contribution of NCNF to the marine nitrogen budget.

  13. Decreasing Fertilizer use by Optimizing Plant-microbe Interactions for Sustainable Supply of Nitrogen for Bioenergy Crops

    Science.gov (United States)

    Schicklberger, M. F.; Huang, J.; Felix, P.; Pettenato, A.; Chakraborty, R.

    2013-12-01

    Nitrogen (N) is an essential component of DNA and proteins and consequently a key element of life. N often is limited in plants, affecting plant growth and productivity. To alleviate this problem, tremendous amounts of N-fertilizer is used, which comes at a high economic price and heavy energy demand. In addition, N-fertilizer also significantly contributes to rising atmospheric greenhouse gas concentrations. Therefore, the addition of fertilizer to overcome N limitation is highly undesirable. To explore reduction in fertilizer use our research focuses on optimizing the interaction between plants and diazotrophic bacteria, which could provide adequate amounts of N to the host-plant. Therefore we investigated the diversity of microbes associated with Tobacco (Nicotiana tabacum) and Switchgrass (Panicum virgatum), considered as potential energy crop for bioenergy production. Several bacterial isolates with representatives from Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, Bacteriodetes and Bacilli were obtained from the roots, leaves, rhizoplane and rhizosphere of these plants. Majority of these isolates grew best with simple sugars and small organic acids. As shown by PCR amplification of nifH, several of these isolates are potential N2-fixing bacteria. We investigated diazotrophs for their response to elevated temperature and salinity (two common climate change induced stresses found on marginal lands), their N2-fixing ability, and their response to root exudates (which drive microbial colonization of the plant). Together this understanding is necessary for the development of eco-friendly, economically sustainable energy crops by decreasing their dependency on fertilizer.

  14. Endophytic Colonization and In Planta Nitrogen Fixation by a Herbaspirillum sp. Isolated from Wild Rice Species

    Science.gov (United States)

    Elbeltagy, Adel; Nishioka, Kiyo; Sato, Tadashi; Suzuki, Hisa; Ye, Bin; Hamada, Toru; Isawa, Tsuyoshi; Mitsui, Hisayuki; Minamisawa, Kiwamu

    2001-01-01

    Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium. Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum. Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus. To examine whether Herbaspirillum sp. strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria. Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds. Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds. Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp. strain B501 colonized mainly intercellular spaces in the leaves of wild rice. Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties. Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O. officinalis, was detected by the acetylene reduction and 15N2 gas incorporation assays. Therefore, we conclude that Herbaspirillum sp. strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O. officinalis. PMID:11679357

  15. Transcriptional Profiling of Nitrogen Fixation in Azotobacter vinelandii▿†

    Science.gov (United States)

    Hamilton, Trinity L.; Ludwig, Marcus; Dixon, Ray; Boyd, Eric S.; Dos Santos, Patricia C.; Setubal, João C.; Bryant, Donald A.; Dean, Dennis R.; Peters, John W.

    2011-01-01

    Most biological nitrogen (N2) fixation results from the activity of a molybdenum-dependent nitrogenase, a complex iron-sulfur enzyme found associated with a diversity of bacteria and some methanogenic archaea. Azotobacter vinelandii, an obligate aerobe, fixes nitrogen via the oxygen-sensitive Mo nitrogenase but is also able to fix nitrogen through the activities of genetically distinct alternative forms of nitrogenase designated the Vnf and Anf systems when Mo is limiting. The Vnf system appears to replace Mo with V, and the Anf system is thought to contain Fe as the only transition metal within the respective active site metallocofactors. Prior genetic analyses suggest that a number of nif-encoded components are involved in the Vnf and Anf systems. Genome-wide transcription profiling of A. vinelandiicultured under nitrogen-fixing conditions under various metal amendments (e.g., Mo or V) revealed the discrete complement of genes associated with each nitrogenase system and the extent of cross talk between the systems. In addition, changes in transcript levels of genes not directly involved in N2fixation provided insight into the integration of central metabolic processes and the oxygen-sensitive process of N2fixation in this obligate aerobe. The results underscored significant differences between Mo-dependent and Mo-independent diazotrophic growth that highlight the significant advantages of diazotrophic growth in the presence of Mo. PMID:21724999

  16. Heterotrophic N2-fixation contributes to nitrogen economy of a common wetland sedge, Schoenoplectus californicus.

    Science.gov (United States)

    Rejmánková, Eliška; Sirová, Dagmara; Castle, Stephanie T; Bárta, Jiří; Carpenter, Heather

    2018-01-01

    A survey of the ecological variability within 52 populations of Schoenoplectus californicus (C.A. Mey.) Soják across its distributional range revealed that it is commonly found in nitrogen (N) limited areas, but rarely in phosphorus limited soils. We explored the hypothesis that S. californicus supplements its nitrogen demand by bacterial N2-fixation processes associated with its roots and rhizomes. We estimated N2-fixation of diazotrophs associated with plant rhizomes and roots from several locations throughout the species' range and conducted an experiment growing plants in zero, low, and high N additions. Nitrogenase activity in rhizomes and roots was measured using the acetylene reduction assay. The presence of diazotrophs was verified by the detection of the nifH gene. Nitrogenase activity was restricted to rhizomes and roots and it was two orders of magnitude higher in the latter plant organs (81 and 2032 nmol C2H4 g DW-1 d-1, respectively). Correspondingly, 40x more nifH gene copies were found on roots compared to rhizomes. The proportion of the nifH gene copies in total bacterial DNA was positively correlated with the nitrogenase activity. In the experiment, the contribution of fixed N to the plant N content ranged from 13.8% to 32.5% among clones from different locations. These are relatively high values for a non-cultivated plant and justify future research on the link between N-fixing bacteria and S. californicus production.

  17. Nitrogen fixation in Red Sea seagrass meadows

    KAUST Repository

    Abdallah, Malak

    2017-05-01

    Seagrasses are key coastal ecosystems, providing many ecosystem services. Seagrasses increase biodiversity as they provide habitat for a large set of organisms. In addition, their structure provides hiding places to avoid predation. Seagrasses can grow in shallow marine coastal areas, but several factors regulate their growth and distribution. Seagrasses can uptake different kinds of organic and inorganic nutrients through their leaves and roots. Nitrogen and phosphorous are the most important nutrients for seagrass growth. Biological nitrogen fixation is the conversion of atmospheric nitrogen into ammonia by diazotrophic bacteria. This process provides a significant source of nitrogen for seagrass growth. The nitrogen fixation is controlled by the nif genes which are found in diazotrophs. The main goal of the project is to measure nitrogen fixation rates on seagrass sediments, in order to compare among various seagrass species from the Red Sea. Moreover, we will compare the fixing rates of the Vegetated areas with the bare sediments. This project will help to ascertain the role of nitrogen fixing bacteria in the development of seagrass meadows.

  18. Identification of Nitrogen-Fixing Genes and Gene Clusters from Metagenomic Library of Acid Mine Drainage

    Science.gov (United States)

    Yin, Huaqun; Liang, Yili; Cong, Jing; Liu, Xueduan

    2014-01-01

    Biological nitrogen fixation is an essential function of acid mine drainage (AMD) microbial communities. However, most acidophiles in AMD environments are uncultured microorganisms and little is known about the diversity of nitrogen-fixing genes and structure of nif gene cluster in AMD microbial communities. In this study, we used metagenomic sequencing to isolate nif genes in the AMD microbial community from Dexing Copper Mine, China. Meanwhile, a metagenome microarray containing 7,776 large-insertion fosmids was constructed to screen novel nif gene clusters. Metagenomic analyses revealed that 742 sequences were identified as nif genes including structural subunit genes nifH, nifD, nifK and various additional genes. The AMD community is massively dominated by the genus Acidithiobacillus. However, the phylogenetic diversity of nitrogen-fixing microorganisms is much higher than previously thought in the AMD community. Furthermore, a 32.5-kb genomic sequence harboring nif, fix and associated genes was screened by metagenome microarray. Comparative genome analysis indicated that most nif genes in this cluster are most similar to those of Herbaspirillum seropedicae, but the organization of the nif gene cluster had significant differences from H. seropedicae. Sequence analysis and reverse transcription PCR also suggested that distinct transcription units of nif genes exist in this gene cluster. nifQ gene falls into the same transcription unit with fixABCX genes, which have not been reported in other diazotrophs before. All of these results indicated that more novel diazotrophs survive in the AMD community. PMID:24498417

  19. Herbaspirillum seropedicae signal transduction protein PII is structurally similar to the enteric GlnK.

    Science.gov (United States)

    Machado Benelli, Elaine; Buck, Martin; Polikarpov, Igor; Maltempi de Souza, Emanuel; Cruz, Leonardo M; Pedrosa, Fábio O

    2002-07-01

    PII-like proteins are signal transduction proteins found in bacteria, archaea and eukaryotes. They mediate a variety of cellular responses. A second PII-like protein, called GlnK, has been found in several organisms. In the diazotroph Herbaspirillum seropedicae, PII protein is involved in sensing nitrogen levels and controlling nitrogen fixation genes. In this work, the crystal structure of the unliganded H. seropedicae PII was solved by X-ray diffraction. H. seropedicae PII has a Gly residue, Gly108 preceding Pro109 and the main-chain forms a beta turn. The glycine at position 108 allows a bend in the C-terminal main-chain, thereby modifying the surface of the cleft between monomers and potentially changing function. The structure suggests that the C-terminal region of PII proteins may be involved in specificity of function, and nonenteric diazotrophs are found to have the C-terminal consensus XGXDAX(107-112). We are also proposing binding sites for ATP and 2-oxoglutarate based on the structural alignment of PII with PII-ATP/GlnK-ATP, 5-carboxymethyl-2-hydroxymuconate isomerase and 4-oxalocrotonate tautomerase bound to the inhibitor 2-oxo-3-pentynoate.

  20. Five decades of N2 fixation research in the North Atlantic Ocean

    Directory of Open Access Journals (Sweden)

    Mar eBenavides

    2015-06-01

    Full Text Available Dinitrogen (N2 fixation (the reduction of atmospheric N2 to ammonium by specialized prokaryotic microbes, represents an important input of fixed nitrogen and contributes significantly to primary productivity in the oceans. Marine N2 fixation was discovered in the North Atlantic Ocean (NA in the 1960s. Ever since, the NA has been subject to numerous studies that have looked into the diversity and abundance of N2-fixing microbes (diazotrophs, the spatial and temporal variability of N2 fixation rates, and the range of physical and chemical variables that control them. The NA provides 10-25% of the globally fixed N2, ranking as the third basin with the largest N2 fixation inputs in the world’s oceans. This basin suffers a chronic depletion in phosphorus availability, more aeolian dust deposition than any other basin in the world’s oceans, and significant nutrient inputs from important rivers like the Amazon and the Congo. These characteristics make it unique in comparison with other oceanic basins. After five decades of intensive research, here we present a comprehensive review of our current understanding of diazotrophic activity in the NA from both a geochemical and biological perspective. We discuss the advantages and disadvantages of current methods, future perspectives, and questions which remain to be answered.

  1. Linkage of the Nit1C gene cluster to bacterial cyanide assimilation as a nitrogen source.

    Science.gov (United States)

    Jones, Lauren B; Ghosh, Pallab; Lee, Jung-Hyun; Chou, Chia-Ni; Kunz, Daniel A

    2018-05-21

    A genetic linkage between a conserved gene cluster (Nit1C) and the ability of bacteria to utilize cyanide as the sole nitrogen source was demonstrated for nine different bacterial species. These included three strains whose cyanide nutritional ability has formerly been documented (Pseudomonas fluorescens Pf11764, Pseudomonas putida BCN3 and Klebsiella pneumoniae BCN33), and six not previously known to have this ability [Burkholderia (Paraburkholderia) xenovorans LB400, Paraburkholderia phymatum STM815, Paraburkholderia phytofirmans PsJN, Cupriavidus (Ralstonia) eutropha H16, Gluconoacetobacter diazotrophicus PA1 5 and Methylobacterium extorquens AM1]. For all bacteria, growth on or exposure to cyanide led to the induction of the canonical nitrilase (NitC) linked to the gene cluster, and in the case of Pf11764 in particular, transcript levels of cluster genes (nitBCDEFGH) were raised, and a nitC knock-out mutant failed to grow. Further studies demonstrated that the highly conserved nitB gene product was also significantly elevated. Collectively, these findings provide strong evidence for a genetic linkage between Nit1C and bacterial growth on cyanide, supporting use of the term cyanotrophy in describing what may represent a new nutritional paradigm in microbiology. A broader search of Nit1C genes in presently available genomes revealed its presence in 270 different bacteria, all contained within the domain Bacteria, including Gram-positive Firmicutes and Actinobacteria, and Gram-negative Proteobacteria and Cyanobacteria. Absence of the cluster in the Archaea is congruent with events that may have led to the inception of Nit1C occurring coincidentally with the first appearance of cyanogenic species on Earth, dating back 400-500 million years.

  2. Perturbation of gut bacteria induces a coordinated cellular immune response in the purple sea urchin larva

    Science.gov (United States)

    CH Ho, Eric; Buckley, Katherine M; Schrankel, Catherine S; Schuh, Nicholas W; Hibino, Taku; Solek, Cynthia M; Bae, Koeun; Wang, Guizhi; Rast, Jonathan P

    2016-01-01

    The purple sea urchin (Strongylocentrotus purpuratus) genome sequence contains a complex repertoire of genes encoding innate immune recognition proteins and homologs of important vertebrate immune regulatory factors. To characterize how this immune system is deployed within an experimentally tractable, intact animal, we investigate the immune capability of the larval stage. Sea urchin embryos and larvae are morphologically simple and transparent, providing an organism-wide model to view immune response at cellular resolution. Here we present evidence for immune function in five mesenchymal cell types based on morphology, behavior and gene expression. Two cell types are phagocytic; the others interact at sites of microbial detection or injury. We characterize immune-associated gene markers for three cell types, including a perforin-like molecule, a scavenger receptor, a complement-like thioester-containing protein and the echinoderm-specific immune response factor 185/333. We elicit larval immune responses by (1) bacterial injection into the blastocoel and (2) seawater exposure to the marine bacterium Vibrio diazotrophicus to perturb immune state in the gut. Exposure at the epithelium induces a strong response in which pigment cells (one type of immune cell) migrate from the ectoderm to interact with the gut epithelium. Bacteria that accumulate in the gut later invade the blastocoel, where they are cleared by phagocytic and granular immune cells. The complexity of this coordinated, dynamic inflammatory program within the simple larval morphology provides a system in which to characterize processes that direct both aspects of the echinoderm-specific immune response as well as those that are shared with other deuterostomes, including vertebrates. PMID:27192936

  3. Study on the Use of Microbial Cellulose as a Biocarrier for 1,3-Dihydroxy-2-Propanone and Its Potential Application in Industry

    Directory of Open Access Journals (Sweden)

    Lidia Stasiak-Różańska

    2018-04-01

    Full Text Available Can microbial cellulose (MC be used as a bio-carrier for 1,3-dihydroxy-2-propanone (DHA? The aim of this study was to examine the possibility of using MC as a biomaterial for DHA transferring into the stratum corneum and inducing changes in skin color. The MC patches were obtained from Gluconacetobacter xylinus strain and incubated in solutions with various concentrations of DHA (g·L−1: 20; 50; 80; 110 at 22 °C for 24 h. Afterwards; the patches were applied onto the skin for 15, 30, or 60 min. Skin color changes were assessed visually compared to a control patches without DHA. The intensity of skin color was increasing with the increase of DHA concentration and time of patches application. Application of MC patches with DHA (50 g·L−1 for 30 min ensured the color which was considered the closest to the desired natural tan effect. MC patches containing DHA can be biocarriers enabling DHA transport into the stratum corneum and causing skin color changes. Study results indicate a new possibility for industrial applications of MC; e.g., as a biocarrier in masking the symptoms of vitiligo or production of self-tanning agents in the form of masks.

  4. Cost-effective production of bacterial cellulose using acidic food industry by-products.

    Science.gov (United States)

    Revin, Victor; Liyaskina, Elena; Nazarkina, Maria; Bogatyreva, Alena; Shchankin, Mikhail

    2018-03-13

    To reduce the cost of obtaining bacterial cellulose, acidic by-products of the alcohol and dairy industries were used without any pretreatment or addition of other nitrogen sources. Studies have shown that the greatest accumulation of bacterial cellulose (6.19g/L) occurs on wheat thin stillage for 3 days of cultivation under dynamic conditions, which is almost 3 times higher than on standard Hestrin and Schramm medium (2.14g/L). The use of whey as a nutrient medium makes it possible to obtain 5.45g/L bacterial cellulose under similar conditions of cultivation. It is established that the pH of the medium during the growth of Gluconacetobacter sucrofermentans B-11267 depends on the feedstock used and its initial value. By culturing the bacterium on thin stillage and whey, there is a decrease in the acidity of the waste. It is shown that the infrared spectra of bacterial cellulose obtained in a variety of environments have a similar character, but we found differences in the micromorphology and crystallinity of the resulting biopolymer. Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  5. Monitoring the microbial community during solid-state acetic acid fermentation of Zhenjiang aromatic vinegar.

    Science.gov (United States)

    Xu, Wei; Huang, Zhiyong; Zhang, Xiaojun; Li, Qi; Lu, Zhenming; Shi, Jinsong; Xu, Zhenghong; Ma, Yanhe

    2011-09-01

    Zhenjiang aromatic vinegar is one of the most famous Chinese traditional vinegars. In this study, change of the microbial community during its fermentation process was investigated. DGGE results showed that microbial community was comparatively stable, and the diversity has a disciplinary series of changes during the fermentation process. It was suggested that domestication of microbes and unique cycle-inoculation style used in the fermentation of Zhenjiang aromatic vinegar were responsible for comparatively stable of the microbial community. Furthermore, two clone libraries were constructed. The results showed that bacteria presented in the fermentation belonged to genus Lactobacillus, Acetobacter, Gluconacetobacter, Staphylococcus, Enterobacter, Pseudomonas, Flavobacterium and Sinorhizobium, while the fungi were genus Saccharomyces. DGGE combined with clone library analysis was an effective and credible technique for analyzing the microbial community during the fermentation process of Zhenjiang aromatic vinegar. Real-time PCR results suggested that the biomass showed a "system microbes self-domestication" process in the first 5 days, then reached a higher level at the 7th day before gradually decreasing until the fermentation ended at the 20th day. This is the first report to study the changes of microbial community during fermentation process of Chinese traditional solid-state fermentation of vinegar. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. Strain typing of acetic acid bacteria responsible for vinegar production by the submerged elaboration method.

    Science.gov (United States)

    Fernández-Pérez, Rocío; Torres, Carmen; Sanz, Susana; Ruiz-Larrea, Fernanda

    2010-12-01

    Strain typing of 103 acetic acid bacteria isolates from vinegars elaborated by the submerged method from ciders, wines and spirit ethanol, was carried on in this study. Two different molecular methods were utilised: pulsed field gel electrophoresis (PFGE) of total DNA digests with a number of restriction enzymes, and enterobacterial repetitive intergenic consensus (ERIC) - PCR analysis. The comparative study of both methods showed that restriction fragment PFGE of SpeI digests of total DNA was a suitable method for strain typing and for determining which strains were present in vinegar fermentations. Results showed that strains of the species Gluconacetobacter europaeus were the most frequent leader strains of fermentations by the submerged method in the studied vinegars, and among them strain R1 was the predominant one. Results showed as well that mixed populations (at least two different strains) occurred in vinegars from cider and wine, whereas unique strains were found in spirit vinegars, which offered the most stressing conditions for bacterial growth. Copyright © 2010 Elsevier Ltd. All rights reserved.

  7. Proteome analysis of Acetobacter pasteurianus during acetic acid fermentation.

    Science.gov (United States)

    Andrés-Barrao, Cristina; Saad, Maged M; Chappuis, Marie-Louise; Boffa, Mauro; Perret, Xavier; Ortega Pérez, Ruben; Barja, François

    2012-03-16

    Acetic acid bacteria (AAB) are Gram-negative, strictly aerobic microorganisms that show a unique resistance to ethanol (EtOH) and acetic acid (AcH). Members of the Acetobacter and Gluconacetobacter genera are capable of transforming EtOH into AcH via the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) enzymes and are used for the industrial production of vinegar. Several mechanisms have been proposed to explain how AAB resist high concentrations of AcH, such as the assimilation of acetate through the tricarboxylic acid (TCA) cycle, the export of acetate by various transporters and modifications of the outer membrane. However, except for a few acetate-specific proteins, little is known about the global proteome responses to AcH. In this study, we used 2D-DIGE to compare the proteome of Acetobacter pasteurianus LMG 1262(T) when growing in glucose or ethanol and in the presence of acetic acid. Interesting protein spots were selected using the ANOVA p-value of 0.05 as threshold and 1.5-fold as the minimal level of differential expression, and a total of 53 proteins were successfully identified. Additionally, the size of AAB was reduced by approximately 30% in length as a consequence of the acidity. A modification in the membrane polysaccharides was also revealed by PATAg specific staining. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Acetic acid bacteria: A group of bacteria with versatile biotechnological applications.

    Science.gov (United States)

    Saichana, Natsaran; Matsushita, Kazunobu; Adachi, Osao; Frébort, Ivo; Frebortova, Jitka

    2015-11-01

    Acetic acid bacteria are gram-negative obligate aerobic bacteria assigned to the family Acetobacteraceae of Alphaproteobacteria. They are members of the genera Acetobacter, Gluconobacter, Gluconacetobacter, Acidomonas, Asaia, Kozakia, Swaminathania, Saccharibacter, Neoasaia, Granulibacter, Tanticharoenia, Ameyamaea, Neokomagataea, and Komagataeibacter. Many strains of Acetobacter and Komagataeibacter have been known to possess high acetic acid fermentation ability as well as the acetic acid and ethanol resistance, which are considered to be useful features for industrial production of acetic acid and vinegar, the commercial product. On the other hand, Gluconobacter strains have the ability to perform oxidative fermentation of various sugars, sugar alcohols, and sugar acids leading to the formation of several valuable products. Thermotolerant strains of acetic acid bacteria were isolated in order to serve as the new strains of choice for industrial fermentations, in which the cooling costs for maintaining optimum growth and production temperature in the fermentation vessels could be significantly reduced. Genetic modifications by adaptation and genetic engineering were also applied to improve their properties, such as productivity and heat resistance. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose

    Directory of Open Access Journals (Sweden)

    Lin eChen

    2015-11-01

    Full Text Available The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled 7 times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors.

  10. Enriched glucose and dextrin mannitol-based media modulates fibroblast behavior on bacterial cellulose membranes

    International Nuclear Information System (INIS)

    Stumpf, Taisa R.; Pértile, Renata A.N.; Rambo, Carlos R.; Porto, Luismar M.

    2013-01-01

    Bacterial cellulose (BC) produced by Gluconacetobacter hansenii is a suitable biopolymer for biomedical applications. In order to modulate the properties of BC and expand its use as substrate for tissue engineering mainly in the form of biomembranes, glucose or dextrin were added into a BC fermentation mannitol-based medium (BCGl and BCDe, respectively) under static culture conditions. SEM images showed effects on fiber density and porosity on both sides of the BC membranes. Both enriched media decreased the BET surface area, water holding capacity, and rehydration rate. Fourier transform infrared (attenuated total reflectance mode) spectroscopy (FTIR-ATR) analysis revealed no change in the chemical structure of BC. L929 fibroblast cells were seeded on all BC-based membranes and evaluated in aspects of cell adhesion, proliferation and morphology. BCG1 membranes showed the highest biological performance and hold promise for the use in tissue engineering applications. - Highlights: • Glucose and dextrin were used to modify culture media for BC production. • Microarchitecture of BC was different depending on the enriching agent. • Fibroblasts adhered on the surface of BC modified microarchitectures. • Fibroblasts adhered on glucose modified BC exhibited healthy cell morphology

  11. In vitro chondrogenesis with lysozyme susceptible bacterial cellulose as a scaffold.

    Science.gov (United States)

    Yadav, Vikas; Sun, Lin; Panilaitis, Bruce; Kaplan, David L

    2015-12-01

    A current focus of tissue engineering is the use of adult human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes for cartilage repair. Several natural and synthetic polymers (including cellulose) have been explored as a biomaterial scaffold for cartilage tissue engineering. While bacterial cellulose (BC) has been used in tissue engineering, its lack of degradability in vivo and high crystallinity restricts widespread applications in the field. Recently we reported the formation of a novel bacterial cellulose that is lysozyme-susceptible and -degradable in vivo from metabolically engineered Gluconacetobacter xylinus. Here we report the use of this modified bacterial cellulose (MBC) for cartilage tissue engineering using hMSCs. MBC's glucosaminoglycan-like chemistry, combined with in vivo degradability, suggested opportunities to exploit this novel polymer in cartilage tissue engineering. We have observed that, like BC, MBC scaffolds support cell attachment and proliferation. Chondrogenesis of hMSCs in the MBC scaffolds was demonstrated by real-time RT-PCR analysis for cartilage-specific extracellular matrix (ECM) markers (collagen type II, aggrecan and SOX9) as well as histological and immunohistochemical evaluations of cartilage-specific ECM markers. Further, the attachment, proliferation, and differentiation of hMSCs in MBC showed unique characteristics. For example, after 4 weeks of cultivation, the spatial cell arrangement and collagen type-II and ACAN distribution resembled those in native articular cartilage tissue, suggesting promise for these novel in vivo degradable scaffolds for chondrogenesis. Copyright © 2013 John Wiley & Sons, Ltd.

  12. Fabrication of antimicrobial bacterial cellulose-Ag/AgCl nanocomposite using bacteria as versatile biofactory

    Energy Technology Data Exchange (ETDEWEB)

    Liu Chuang [Tianjin University, Key Laboratory for Green Technology, School of Chemical Engineering and Technology (China); Yang Dong; Wang Yuangui [Tianjin University, Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of Ministry of Education, School of Chemical Engineering and Technology (China); Shi Jiafu; Jiang Zhongyi, E-mail: zhyjiang@tju.edu.cn [Tianjin University, Key Laboratory for Green Technology, School of Chemical Engineering and Technology (China)

    2012-08-15

    In nature, a number of nanocomposites are formed through biomineralization-relevant processes under mild conditions. In the present study, a total 'biologic' route to fabricate nanocomposite is reported. Non-pathogenic bacteria, Gluconacetobacter xylinum, was utilized as a versatile biofactory, which produced biopolymer bacterial cellulose (BC) and induced the formation of Ag/AgCl nanoparticles, yielding BC-Ag/AgCl nanocomposite. Scanning electron microscopy revealed that nanoparticles with average size of 17.4 nm were randomly embedded into the BC network; transmission electron microscopy and X-ray diffraction confirmed that the nanoparticles were mixtures of face-centered cubic silver and silver chloride nanoparticles. Moreover, the content of silver in the BC nanocomposite is around 0.05 wt%, determined by atomic absorption spectrometry and X-ray photoelectron spectroscopy analysis. The entire process of nanocomposite fabrication was conducted at ambient environment without utilizing toxic agents or producing hazardous products, which is not only environmentally friendly but also with less chances to generate harmful products to human bodies as biomedical materials. The resultant nanocomposite displayed the desirable activity in inhibiting bacterial growth of both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli microorganisms on agar plate and in liquid culture, indicating the potential of the material as antimicrobial wound dressing materials. This work demonstrated the feasibility of using microorganism to fabricate nanocomposite, especially for biomedical materials.

  13. Bacterial cellulose production from cotton-based waste textiles: enzymatic saccharification enhanced by ionic liquid pretreatment.

    Science.gov (United States)

    Hong, Feng; Guo, Xiang; Zhang, Shuo; Han, Shi-fen; Yang, Guang; Jönsson, Leif J

    2012-01-01

    Cotton-based waste textiles were explored as alternative feedstock for production of bacterial cellulose (BC) by Gluconacetobacter xylinus. The cellulosic fabrics were treated with the ionic liquid (IL) 1-allyl-3-methylimidazolium chloride ([AMIM]Cl). [AMIM]Cl caused 25% inactivation of cellulase activity at a concentration as low as of 0.02 g/mL and decreased BC production during fermentation when present in concentrations higher than 0.0005 g/mL. Therefore, removal of residual IL by washing with hot water was highly beneficial to enzymatic saccharification as well as BC production. IL-treated fabrics exhibited a 5-7-fold higher enzymatic hydrolysis rate and gave a seven times larger yield of fermentable sugars than untreated fabrics. BC from cotton cloth hydrolysate was obtained at an yield of 10.8 g/L which was 83% higher than that from the culture grown on glucose-based medium. The BC from G. xylinus grown on IL-treated fabric hydrolysate had a 79% higher tensile strength than BC from glucose-based culture medium which suggests that waste cotton pretreated with [AMIM]Cl has potential to serve as a high-quality carbon source for BC production. Copyright © 2011 Elsevier Ltd. All rights reserved.

  14. Unraveling microbial ecology of industrial-scale Kombucha fermentations by metabarcoding and culture-based methods.

    Science.gov (United States)

    Coton, Monika; Pawtowski, Audrey; Taminiau, Bernard; Burgaud, Gaëtan; Deniel, Franck; Coulloumme-Labarthe, Laurent; Fall, Abdoulaye; Daube, Georges; Coton, Emmanuel

    2017-05-01

    Kombucha, historically an Asian tea-based fermented drink, has recently become trendy in Western countries. Producers claim it bears health-enhancing properties that may come from the tea or metabolites produced by its microbiome. Despite its long history of production, microbial richness and dynamics have not been fully unraveled, especially at an industrial scale. Moreover, the impact of tea type (green or black) on microbial ecology was not studied. Here, we compared microbial communities from industrial-scale black and green tea fermentations, still traditionally carried out by a microbial biofilm, using culture-dependent and metabarcoding approaches. Dominant bacterial species belonged to Acetobacteraceae and to a lesser extent Lactobacteriaceae, while the main identified yeasts corresponded to Dekkera, Hanseniaspora and Zygosaccharomyces during all fermentations. Species richness decreased over the 8-day fermentation. Among acetic acid bacteria, Gluconacetobacter europaeus, Gluconobacter oxydans, G. saccharivorans and Acetobacter peroxydans emerged as dominant species. The main lactic acid bacteria, Oenococcus oeni, was strongly associated with green tea fermentations. Tea type did not influence yeast community, with Dekkera bruxellensis, D. anomala, Zygosaccharomyces bailii and Hanseniaspora valbyensis as most dominant. This study unraveled a distinctive core microbial community which is essential for fermentation control and could lead to Kombucha quality standardization. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Microbially influenced corrosion communities associated with fuel-grade ethanol environments.

    Science.gov (United States)

    Williamson, Charles H D; Jain, Luke A; Mishra, Brajendra; Olson, David L; Spear, John R

    2015-08-01

    Microbially influenced corrosion (MIC) is a costly problem that impacts hydrocarbon production and processing equipment, water distribution systems, ships, railcars, and other types of metallic infrastructure. In particular, MIC is known to cause considerable damage to hydrocarbon fuel infrastructure including production, transportation, and storage systems, often times with catastrophic environmental contamination results. As the production and use of alternative fuels such as fuel-grade ethanol (FGE) increase, it is important to consider MIC of engineered materials exposed to these "newer fuels" as they enter existing infrastructure. Reports of suspected MIC in systems handling FGE and water prompted an investigation of the microbial diversity associated with these environments. Small subunit ribosomal RNA gene pyrosequencing surveys indicate that acetic-acid-producing bacteria (Acetobacter spp. and Gluconacetobacter spp.) are prevalent in environments exposed to FGE and water. Other microbes previously implicated in corrosion, such as sulfate-reducing bacteria and methanogens, were also identified. In addition, acetic-acid-producing microbes and sulfate-reducing microbes were cultivated from sampled environments containing FGE and water. Results indicate that complex microbial communities form in these FGE environments and could cause significant MIC-related damage that may be difficult to control. How to better manage these microbial communities will be a defining aspect of improving mitigation of global infrastructure corrosion.

  16. Diversity of the microbiota involved in wine and organic apple cider submerged vinegar production as revealed by DHPLC analysis and next-generation sequencing.

    Science.gov (United States)

    Trček, Janja; Mahnič, Aleksander; Rupnik, Maja

    2016-04-16

    Unfiltered vinegar samples collected from three oxidation cycles of the submerged industrial production of each, red wine and organic apple cider vinegars, were sampled in a Slovene vinegar producing company. The samples were systematically collected from the beginning to the end of an oxidation cycle and used for culture-independent microbial analyses carried out by denaturing high pressure liquid chromatography (DHPLC) and Illumina MiSeq sequencing of 16S rRNA gene variable regions. Both approaches showed a very homogeneous bacterial structure during wine vinegar production but more heterogeneous during organic apple cider vinegar production. In all wine vinegar samples Komagataeibacter oboediens (formerly Gluconacetobacter oboediens) was a predominating species. In apple cider vinegar the acetic acid and lactic acid bacteria were two major groups of bacteria. The acetic acid bacterial consortium was composed of Acetobacter and Komagataeibacter with the Komagataeibacter genus outcompeting the Acetobacter in all apple cider vinegar samples at the end of oxidation cycle. Among the lactic acid bacterial consortium two dominating genera were identified, Lactobacillus and Oenococcus, with Oenococcus prevailing with increasing concentration of acetic acid in vinegars. Unexpectedly, a minor genus of the acetic acid bacterial consortium in organic apple cider vinegar was Gluconobacter, suggesting a possible development of the Gluconobacter population with a tolerance against ethanol and acetic acid. Among the accompanying bacteria of the wine vinegar, the genus Rhodococcus was detected, but it decreased substantially by the end of oxidation cycles. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Fabrication of antimicrobial bacterial cellulose–Ag/AgCl nanocomposite using bacteria as versatile biofactory

    International Nuclear Information System (INIS)

    Liu Chuang; Yang Dong; Wang Yuangui; Shi Jiafu; Jiang Zhongyi

    2012-01-01

    In nature, a number of nanocomposites are formed through biomineralization-relevant processes under mild conditions. In the present study, a total “biologic” route to fabricate nanocomposite is reported. Non-pathogenic bacteria, Gluconacetobacter xylinum, was utilized as a versatile biofactory, which produced biopolymer bacterial cellulose (BC) and induced the formation of Ag/AgCl nanoparticles, yielding BC–Ag/AgCl nanocomposite. Scanning electron microscopy revealed that nanoparticles with average size of 17.4 nm were randomly embedded into the BC network; transmission electron microscopy and X-ray diffraction confirmed that the nanoparticles were mixtures of face-centered cubic silver and silver chloride nanoparticles. Moreover, the content of silver in the BC nanocomposite is around 0.05 wt%, determined by atomic absorption spectrometry and X-ray photoelectron spectroscopy analysis. The entire process of nanocomposite fabrication was conducted at ambient environment without utilizing toxic agents or producing hazardous products, which is not only environmentally friendly but also with less chances to generate harmful products to human bodies as biomedical materials. The resultant nanocomposite displayed the desirable activity in inhibiting bacterial growth of both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli microorganisms on agar plate and in liquid culture, indicating the potential of the material as antimicrobial wound dressing materials. This work demonstrated the feasibility of using microorganism to fabricate nanocomposite, especially for biomedical materials.

  18. Enriched glucose and dextrin mannitol-based media modulates fibroblast behavior on bacterial cellulose membranes

    Energy Technology Data Exchange (ETDEWEB)

    Stumpf, Taisa R.; Pértile, Renata A.N. [Integrated Technologies Laboratory, Department of Chemical and Food Engineering (Brazil); Rambo, Carlos R., E-mail: rambo@intelab.ufsc.br [Department of Electrical Engineering, Federal University of Santa Catarina, Florianópolis 88040-900 (Brazil); Porto, Luismar M. [Integrated Technologies Laboratory, Department of Chemical and Food Engineering (Brazil)

    2013-12-01

    Bacterial cellulose (BC) produced by Gluconacetobacter hansenii is a suitable biopolymer for biomedical applications. In order to modulate the properties of BC and expand its use as substrate for tissue engineering mainly in the form of biomembranes, glucose or dextrin were added into a BC fermentation mannitol-based medium (BCGl and BCDe, respectively) under static culture conditions. SEM images showed effects on fiber density and porosity on both sides of the BC membranes. Both enriched media decreased the BET surface area, water holding capacity, and rehydration rate. Fourier transform infrared (attenuated total reflectance mode) spectroscopy (FTIR-ATR) analysis revealed no change in the chemical structure of BC. L929 fibroblast cells were seeded on all BC-based membranes and evaluated in aspects of cell adhesion, proliferation and morphology. BCG1 membranes showed the highest biological performance and hold promise for the use in tissue engineering applications. - Highlights: • Glucose and dextrin were used to modify culture media for BC production. • Microarchitecture of BC was different depending on the enriching agent. • Fibroblasts adhered on the surface of BC modified microarchitectures. • Fibroblasts adhered on glucose modified BC exhibited healthy cell morphology.

  19. Bacterial cellulose membrane as flexible substrate for organic light emitting devices

    International Nuclear Information System (INIS)

    Legnani, C.; Vilani, C.; Calil, V.L.; Barud, H.S.; Quirino, W.G.; Achete, C.A.; Ribeiro, S.J.L.; Cremona, M.

    2008-01-01

    Bacterial cellulose (BC) membranes produced by gram-negative, acetic acid bacteria (Gluconacetobacter xylinus), were used as flexible substrates for the fabrication of Organic Light Emitting Diodes (OLED). In order to achieve the necessary conductive properties indium tin oxide (ITO) thin films were deposited onto the membrane at room temperature using radio frequency (r.f.) magnetron sputtering with an r.f. power of 30 W, at pressure of 8 mPa in Ar atmosphere without any subsequent thermal treatment. Visible light transmittance of about 40% was observed. Resistivity, mobility and carrier concentration of deposited ITO films were 4.90 x 10 -4 Ohm cm, 8.08 cm 2 /V-s and - 1.5 x 10 21 cm -3 , respectively, comparable with commercial ITO substrates. In order to demonstrate the feasibility of devices based on BC membranes three OLEDs with different substrates were produced: a reference one with commercial ITO on glass, a second one with a SiO 2 thin film interlayer between the BC membrane and the ITO layer and a third one just with ITO deposited directly on the BC membrane. The observed OLED luminance ratio was: 1; 0.5; 0.25 respectively, with 2400 cd/m 2 as the value for the reference OLED. These preliminary results show clearly that the functionalized biopolymer, biodegradable, biocompatible bacterial cellulose membranes can be successfully used as substrate in flexible organic optoelectronic devices

  20. In vitro evaluation of osteoblastic cells on bacterial cellulose modified with multi-walled carbon nanotubes as scaffold for bone regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Gutiérrez-Hernández, José Manuel [Coordination for Innovation and Application of Science and Technology, Autonomous University San Luis Potosi, 78000 San Luis Potosi (Mexico); Department of Wood, Cellulose and Paper Research, University Guadalajara, 45110 Guadalajara (Mexico); Escobar-García, Diana María [Laboratory of Basic Sciences, Faculty of Dentistry, Autonomous University San Luis Potosi, 78000 San Luis Potosi (Mexico); Escalante, Alfredo [Department of Wood, Cellulose and Paper Research, University Guadalajara, 45110 Guadalajara (Mexico); Flores, Hector [Laboratory of Basic Sciences, Faculty of Dentistry, Autonomous University San Luis Potosi, 78000 San Luis Potosi (Mexico); González, Francisco Javier [Coordination for Innovation and Application of Science and Technology, Autonomous University San Luis Potosi, 78000 San Luis Potosi (Mexico); Gatenholm, Paul [Chalmers University of Technology, Department of Chemistry and Chemical Engineering, Biopolymer Technology, SE-412 96 Göteborg (Sweden); Toriz, Guillermo, E-mail: gtoriz@dmcyp.cucei.udg.mx [Department of Wood, Cellulose and Paper Research, University Guadalajara, 45110 Guadalajara (Mexico); Chalmers University of Technology, Department of Chemistry and Chemical Engineering, Biopolymer Technology, SE-412 96 Göteborg (Sweden)

    2017-06-01

    In this paper we explore the use of native bacterial cellulose (BC) in combination with functionalized multi-walled carbon nanotubes (MWNTs) as an original biomaterial, suitable three-dimensional (3D) scaffold for osteoblastic cell culture. Functionalized MWNTs were mixed with native BC (secreted by Gluconacetobacter xylinus) with the aim of reinforcing the mechanical properties of BC. The results indicate that BC-MWNTs scaffolds support osteoblast viability, adhesion and proliferation at higher levels as compared to traditional culture substrates. Chemically functionalized MWNTs are also an excellent material to be used as scaffold because these did not affect cell viability and showed an enhanced osteoblast adhesion. These results suggest the potential for this combination of biomaterials, i.e. BC and carbon nanomaterials, as scaffolds for bone regeneration. - Highlights: • Functionalization of multiwalled carbon nanotubes with carboxyl groups for reduces their toxicity against osteoblastic cells. • Use of native bacterial cellulose with functionalized multi-walled carbon nanotubes as scaffolds for tissue engineering. • Bacterial cellulose with multi-walled carbon nanotubes as scaffolds give an excellent option to be used in bone regeneration.

  1. Performance of improved bacterial cellulose application in the production of functional paper.

    Science.gov (United States)

    Basta, A H; El-Saied, H

    2009-12-01

    The purpose of this work was to study the feasibility of producing economic flame retardant bacterial cellulose (BC) and evaluating its behaviour in paper production. This type of BC was prepared by Gluconacetobacter subsp. xylinus and substituting the glucose in the cultivation medium by glucose phosphate as a carbon source; as well as using corn steep liquor as a nitrogen source. The investigated processing technique did not dispose any toxic chemicals that pollute the surroundings or cause unacceptable effluents, making the process environmentally safe. The fire retardant behaviour of the investigated BC has been studied by non-isothermal thermogravimetric analysis (TGA & DTGA). The activation energy of each degradation stage and the order of degradation were estimated using the Coats-Redfern equation and the least square method. Strength, optical properties, and thermogravimetric analysis of BC-phosphate added paper sheets were also tested. The study confirmed that the use of glucose phosphate along with glucose was significant in the high yield production of phosphate containing bacterial cellulose (PCBC1); more so than the use of glucose phosphate alone (PCBC2). Incorporating 5% of the PCBC with wood pulp during paper sheet formation was found to significantly improve kaolin retention, strength, and fire resistance properties as compared to paper sheets produced from incorporating bacterial cellulose (BC). This modified BC is a valuable product for the preparation of specialized paper, in addition to its function as a fillers aid.

  2. Biosynthesis of highly porous bacterial cellulose nanofibers

    Science.gov (United States)

    Hosseini, Hadi; Kokabi, Mehrdad; Mousavi, Seyyed Mohammad

    2018-01-01

    Bacterial cellulose nanofibers (BCNFs) as a sustainable and biodegradable polymer has drawn tremendous research attention in tissue engineering, bacterial sensors and drug delivery due to its extraordinary properties such as high purity, high crystallinity, high water absorption capacity and excellent mechanical strength in the wet state. This awesome properties, is attributed to BCNFs structure, therefore its characterization is important. In this work, the bacterial strain, Gluconacetobacter xylinus (PTCC 1734, obtained from Iranian Research Organization for Science and Technology (IROST)), was used to produce BCNFs hydrogel using bacterial fermentation under static condition at 29 °C for 10 days in the incubator. Then, the biosynthesized BCNFs wet gel, were dried at ambient temperature and pressure and characterized using Brunauer-Emmett-Teller (BET) and Field emission scanning electron microscopy (FE-SEM) analysis. FESEM image displayed highly interconnected and porous structure composed of web-like continuous, nanofibers with an average diameter of 48.5±2.1 nm. BET result analysis depicted BCNFs dried at ambient conditions had IV isotherm type, according to the IUPAC classification, indicating that BCNFs dried at ambient condition is essentially mesoporous. On the other hand, BET results depicted, mesoporous structure is around 85%. In addition, Specific surface area (SBET) obtained 81.45 m2/g. These results are in accordance with the FESEM observation.

  3. Sequence-based analysis of the bacterial and fungal compositions of multiple kombucha (tea fungus) samples.

    Science.gov (United States)

    Marsh, Alan J; O'Sullivan, Orla; Hill, Colin; Ross, R Paul; Cotter, Paul D

    2014-04-01

    Kombucha is a sweetened tea beverage that, as a consequence of fermentation, contains ethanol, carbon dioxide, a high concentration of acid (gluconic, acetic and lactic) as well as a number of other metabolites and is thought to contain a number of health-promoting components. The sucrose-tea solution is fermented by a symbiosis of bacteria and yeast embedded within a cellulosic pellicle, which forms a floating mat in the tea, and generates a new layer with each successful fermentation. The specific identity of the microbial populations present has been the focus of attention but, to date, the majority of studies have relied on culture-based analyses. To gain a more comprehensive insight into the kombucha microbiota we have carried out the first culture-independent, high-throughput sequencing analysis of the bacterial and fungal populations of 5 distinct pellicles as well as the resultant fermented kombucha at two time points. Following the analysis it was established that the major bacterial genus present was Gluconacetobacter, present at >85% in most samples, with only trace populations of Acetobacter detected (kombucha, also being revealed. The yeast populations were found to be dominated by Zygosaccharomyces at >95% in the fermented beverage, with a greater fungal diversity present in the cellulosic pellicle, including numerous species not identified in kombucha previously. Ultimately, this study represents the most accurate description of the microbiology of kombucha to date. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Novel nitrogen-fixing Acetobacter nitrogenifigens sp. nov., isolated from Kombucha tea.

    Science.gov (United States)

    Dutta, Debasree; Gachhui, Ratan

    2006-08-01

    The four nitrogen-fixing bacteria so far described in the family Acetobacteraceae belong to the genera Gluconacetobacter and Acetobacter. Nitrogen-fixing bacterial strain RG1(T) was isolated from Kombucha tea and, based on the phylogenetic analysis of 16S rRNA gene sequence which is supported by a high bootstrap value, was found to belong to the genus Acetobacter. Strain RG1(T) differed from Acetobacter aceti, the nearest member with a 16S rRNA gene sequence similarity of 98.2 %, and type strains of other Acetobacter species with regard to several characteristics of growth features in culture media, growth in nitrogen-free medium, production of gamma-pyrone from glucose and dihydroxyacetone from glycerol. Strain RG1(T) utilized maltose, glycerol, sorbitol, fructose, galactose, arabinose and ethanol, but not methanol as a carbon source. These results, along with electrophoretic mobility patterns of nine metabolic enzymes, suggest that strain RG1(T) represents a novel nitrogen-fixing species. The ubiquinone present was Q-9 and DNA G+C content was 64.1 mol%. Strain RG1(T) exhibited a low value of 2-24 % DNA-DNA relatedness to the type strains of related acetobacters, which placed it as a separate taxon. On the basis of this data, the name Acetobacter nitrogenifigens sp. nov. is proposed, with the type strain RG1(T) (=MTCC 6912(T)=LMG 23498(T)).

  5. In vitro evaluation of osteoblastic cells on bacterial cellulose modified with multi-walled carbon nanotubes as scaffold for bone regeneration

    International Nuclear Information System (INIS)

    Gutiérrez-Hernández, José Manuel; Escobar-García, Diana María; Escalante, Alfredo; Flores, Hector; González, Francisco Javier; Gatenholm, Paul; Toriz, Guillermo

    2017-01-01

    In this paper we explore the use of native bacterial cellulose (BC) in combination with functionalized multi-walled carbon nanotubes (MWNTs) as an original biomaterial, suitable three-dimensional (3D) scaffold for osteoblastic cell culture. Functionalized MWNTs were mixed with native BC (secreted by Gluconacetobacter xylinus) with the aim of reinforcing the mechanical properties of BC. The results indicate that BC-MWNTs scaffolds support osteoblast viability, adhesion and proliferation at higher levels as compared to traditional culture substrates. Chemically functionalized MWNTs are also an excellent material to be used as scaffold because these did not affect cell viability and showed an enhanced osteoblast adhesion. These results suggest the potential for this combination of biomaterials, i.e. BC and carbon nanomaterials, as scaffolds for bone regeneration. - Highlights: • Functionalization of multiwalled carbon nanotubes with carboxyl groups for reduces their toxicity against osteoblastic cells. • Use of native bacterial cellulose with functionalized multi-walled carbon nanotubes as scaffolds for tissue engineering. • Bacterial cellulose with multi-walled carbon nanotubes as scaffolds give an excellent option to be used in bone regeneration.

  6. Production of bacterial cellulose using different carbon sources and culture media.

    Science.gov (United States)

    Mohammadkazemi, Faranak; Azin, Mehrdad; Ashori, Alireza

    2015-03-06

    In this work, the effects of carbon sources and culture media on the production and structural properties of bacterial cellulose (BC) have been studied. BC nanofibers were synthesized using Gluconacetobacter xylinus strain PTCC 1734. Media used were Hestrin-Schramm (H), Yamanaka (Y), and Zhou (Z). Five different carbon sources, namely date syrup, glucose, mannitol, sucrose, and food-grade sucrose were used in these media. All the produced BC pellicles were characterized in terms of dry weight production, biomass yield, thermal stability, crystallinity and morphology by thermogravimetric analysis (TGA), x-ray diffraction (XRD), and field emission scanning electron microscopy (FE-SEM). The obtained results showed that mannitol lead to the highest yield, followed by sucrose. The highest production efficiency of mannitol might be due to the nitrogen source, which plays an important role. The maximum improvement on the thermal stability of the composites was achieved when mannitol was used in H medium. In addition, the crystallinity was higher in BC formed in H medium compared to other media. FE-SEM micrographs illustrated that the BC pellicles, synthesized in the culture media H and Z, were stable, unlike those in medium Y that were unstable. The micrographs of BC produced in media containing mannitol and sucrose provided evidence of the strong interfacial adhesion between the BC fibers without noticeable aggregates. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Improvement production of bacterial cellulose by semi-continuous process in molasses medium.

    Science.gov (United States)

    Cakar, Fatih; Ozer, Işılay; Aytekin, A Özhan; Sahin, Fikrettin

    2014-06-15

    Bacterial cellulose (BC) has unique properties such as structural, functional, physical and chemical. The mass production of BC for industrial application has recently become attractive to produce more economical and high productive cellulose. In this study, to improve the productivity of bacterial cellulose (BC), BC production by Gluconacetobacter xylinus FC01 was investigated in molasses medium with static semi-continuous operation mode. Cell dry weight, polysaccharide, sugar and cellulose concentrations were monitored and cellulose was characterized by Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM). The highest cellulose yield (1.637 g/L) was obtained in SCP50-7d, which molasses of 1/2 ratio for 7 days by static semi-continuous operation mode. The results show that BC can be highly produced by G. xylinus in molasses with static semi-continuous process than batch process. We claimed that low-cost medium with semi-continuous operation mode in static culture is a good candidate for industrial scale BC productions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Molecular assessment of microbiota structure and dynamics along mixed olive oil and winery wastewaters biotreatment.

    Science.gov (United States)

    Eusébio, Ana; Tacão, Marta; Chaves, Sandra; Tenreiro, Rogério; Almeida-Vara, Elsa

    2011-07-01

    The major parcel of the degradation occurring along wastewater biotreatments is performed either by the native microbiota or by added microbial inocula. The main aim of this study was to apply two fingerprinting methods, temperature gradient gel electrophoresis (TGGE) and length heterogeneity-PCR (LH-PCR) analysis of 16S rRNA gene fragments, in order to assess the microbiota structure and dynamics during mixed olive oil and winery wastewaters aerobic biotreatment performed in a jet-loop reactor (JLR). Sequence homology analysis showed the presence of bacterial genera Gluconacetobacter, Klebsiella, Lactobacillus, Novosphingobium, Pseudomonas, Prevotella, Ralstonia, Sphingobium and Sphingomonas affiliated with five main phylogenetic groups: alpha-, beta- and gamma-Proteobacteria, Firmicutes and Bacteroidetes. LH-PCR analysis distinguished eight predominant DNA fragments correlated with the samples showing highest performance (COD removal rates of 67 up to 75%). Cluster analysis of both TGGE and LH-PCR fingerprinting profiles established five main clusters, with similarity coefficients higher than 79% (TGGE) and 62% (LH-PCR), and related with hydraulic retention time, indicating that this was the main factor responsible for the shifts in the microbiota structure. Canonical correspondence analysis revealed that changes observed on temperature and O(2) level were also responsible for shifts in microbiota composition. Community level metabolic profile analysis was used to test metabolic activities in samples. Integrated data revealed that the microbiota structure corresponds to bacterial groups with high degradative potential and good suitability for this type of effluents biotreatments.

  9. Evaluation of bacterial nanocellulose-based uniform wound dressing for large area skin transplantation

    Energy Technology Data Exchange (ETDEWEB)

    Fu, Lina [Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074 (China); National Engineering Research Center for Nano-Medicine, Huazhong University of Science and Technology, Wuhan 430074 (China); Zhou, Ping [Institute of Organ Transplant of Tongji Hospital, Huazhong University of Science and Technology, Wuhan (China); Zhang, Shengmin [Advanced Biomaterials and Tissue Engineering Center, Huazhong University of Science and Technology, Wuhan (China); Yang, Guang, E-mail: yang_sunny@yahoo.com [Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074 (China); National Engineering Research Center for Nano-Medicine, Huazhong University of Science and Technology, Wuhan 430074 (China)

    2013-07-01

    Bacterial nanocellulose (BNC) was biosynthesized by Gluconacetobacter xylinus. The surface area, physicochemical structure and morphology of the materials were characterized. Here provides a method for an efficient production of uniform BNC, which is beneficial for the fast characterization and evaluation of BNC. In vitro cytotoxicity of the materials was evaluated by the proliferation, the adhesion, the viability and the morphology of NIH/3T3 cells. Low cytotoxicity of the BNC was observed, and micrographs demonstrate a good proliferation and adhesion of NIH/3T3 cells on BNC. Large area full-thickness skin defects were made on the back of C57BL/6 mice in animal surgery. The wounds were transplanted with BNC films and the results compared to those in a control group. The rehabilitation of the wound surfaces and the pathological sections of mice were investigated and are discussed. Histological examinations demonstrated faster and better healing effect and lower inflammatory response in the BNC group than those in the control group. Preliminary results on wound dressings from BNC show a curative effect promoting the healing of epithelial tissue. BNC is a promising natural polymer with medical applications in wound dressings. - Highlights: • BNC is expected to be a promising material in wound healing and skin transplantation. • We studied surface area, physicochemical structures and morphology of uniform BNC. • Cyto-evaluation results of BNC-based wound dressing show a good biocompatibility. • Large area skin transplantation experiments suggest a good performance of healing.

  10. Evaluation of bacterial nanocellulose-based uniform wound dressing for large area skin transplantation

    International Nuclear Information System (INIS)

    Fu, Lina; Zhou, Ping; Zhang, Shengmin; Yang, Guang

    2013-01-01

    Bacterial nanocellulose (BNC) was biosynthesized by Gluconacetobacter xylinus. The surface area, physicochemical structure and morphology of the materials were characterized. Here provides a method for an efficient production of uniform BNC, which is beneficial for the fast characterization and evaluation of BNC. In vitro cytotoxicity of the materials was evaluated by the proliferation, the adhesion, the viability and the morphology of NIH/3T3 cells. Low cytotoxicity of the BNC was observed, and micrographs demonstrate a good proliferation and adhesion of NIH/3T3 cells on BNC. Large area full-thickness skin defects were made on the back of C57BL/6 mice in animal surgery. The wounds were transplanted with BNC films and the results compared to those in a control group. The rehabilitation of the wound surfaces and the pathological sections of mice were investigated and are discussed. Histological examinations demonstrated faster and better healing effect and lower inflammatory response in the BNC group than those in the control group. Preliminary results on wound dressings from BNC show a curative effect promoting the healing of epithelial tissue. BNC is a promising natural polymer with medical applications in wound dressings. - Highlights: • BNC is expected to be a promising material in wound healing and skin transplantation. • We studied surface area, physicochemical structures and morphology of uniform BNC. • Cyto-evaluation results of BNC-based wound dressing show a good biocompatibility. • Large area skin transplantation experiments suggest a good performance of healing

  11. Description of a novel approach to engineer cartilage with porous bacterial nanocellulose for reconstruction of a human auricle.

    Science.gov (United States)

    Feldmann, Eva-Maria; Sundberg, J F; Bobbili, B; Schwarz, S; Gatenholm, P; Rotter, N

    2013-11-01

    In this study, we investigated the effects of human primary chondrocytes, derived from routine septorhino- and otoplasties on a novel nondegradable biomaterial. This biomaterial, porous bacterial nanocellulose, is produced by Gluconacetobacter xylinus. Porosity is generated by paraffin beads embedded during the fermentation process. Human primary chondrocytes were able to adhere to bacterial nanocellulose and produce cartilaginous matrix proteins such as aggrecan (after 14 days) and collagen type II (after 21 days) in the presence of differentiation medium. Cells were located within the pores and in a dense cell layer covering the surface of the biomaterial. Cells were able to re-differentiate, as cell shape and extra cellular matrix gene expression showed a chondrogenic phenotype in three-dimensional bacterial nanocellulose culture. Collagen type I and versican expression decreased during three-dimensional culture. Variations in pore sizes of 150-300 µm and 300-500 µm did not influence cartilaginous extra cellular matrix synthesis. Varying seeding densities from 9.95 × 10(2) to 1.99 × 10(3) cells/mm(2) and 3.98 × 10(3) cells/mm(2) did not result in differences in quality of extra cellular matrix neo-synthesis. Our results demonstrated that both nasal and auricular chondrocytes are equally suitable to synthesize new extra cellular matrix on bacterial nanocellulose. Therefore, we propose both cell sources in combination with bacterial nanocellulose as promising candidates for the special needs of auricular reconstruction.

  12. In situ biosynthesis of bacterial nanocellulose-CaCO3 hybrid bionanocomposite: One-step process.

    Science.gov (United States)

    Mohammadkazemi, Faranak; Faria, Marisa; Cordeiro, Nereida

    2016-08-01

    In this work, a simple and green route to the synthesis of the bacterial nanocellulose-calcium carbonate (BNC/CaCO3) hybrid bionanocomposites using one-step in situ biosynthesis was studied. The CaCO3 was incorporated in the bacterial nanocellulose structure during the cellulose biosynthesis by Gluconacetobacter xylinus PTCC 1734 bacteria. Hestrin-Schramm (HS) and Zhou (Z) culture media were used to the hybrid bionanocomposites production and the effect of ethanol addition was investigated. Attenuated total reflection Fourier transform infrared spectroscopy, field emission scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, inverse gas chromatography and thermogravimetric analysis were used to characterize the samples. The experimental results demonstrated that the ethanol and culture medium play an important role in the BNC/CaCO3 hybrid bionanocomposites production, structure and properties. The BNC/CaCO3 biosynthesized in Z culture medium revealed higher O/C ratio and amphoteric surface character, which justify the highest CaCO3 content incorporation. The CaCO3 was incorporated into the cellulosic matrix decreasing the bacterial nanocellulose crystallinity. This work reveals the high potential of in situ biosynthesis of BNC/CaCO3 hybrid bionanocomposites and opens a new way to the high value-added applications of bacterial nanocellulose. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Understanding the Role of Physical Properties of Cellulose on Its Hydrolyzability by Cellulases

    Science.gov (United States)

    O'Dell, Patrick Jonathan

    Cellulose has long been explored as a potential feedstock for biofuel, however the recalcitrance of cellulose makes its conversion into biofuel much more challenging and economically unfavorable compared to well-established processes for converting starch or sugar feedstocks into biofuel. Enzymes capable of hydrolyzing cellulose into soluble sugars, glucose and cellobiose, have been found to work processively along cellulose microfibrils starting from reducing end groups. For this study, cellulose was produced and purified in-house from Gluconacetobacter xylinum cultures, and characterized by quantifying functional groups (aldehyde, ketone, and carboxyl groups) to determine the extent of oxidation of cellulose due to the processing steps. The main goal of this study was to look at the impacts of ultrasonication on cellulose's structure and the enzymatic hydrolyzability of cellulose. A completely randomized experimental design was used to test the effect of ultrasonication time and amplitude (intensity) on changes in cellulose fibril length, degree of polymerization, and rates and extents of hydrolysis. Results indicated that sonication time does significantly impact both the fibril length and average degree of polymerization of cellulose. The impact of ultrasonication on the hydrolyzability of cellulose by commercial cellulase and beta-glucosidase preparations could not be effectively resolved due to high variability in the experimental results. These studies serve as a basis for future studies understanding the role of cellulose microstructure in the mechanism of cellulase hydrolysis of cellulose.

  14. Formation of Highly Twisted Ribbons in a Carboxymethylcellulase Gene-Disrupted Strain of a Cellulose-Producing Bacterium

    Science.gov (United States)

    Sugano, Yasushi; Shoda, Makoto; Sakakibara, Hitoshi; Oiwa, Kazuhiro; Tuzi, Satoru; Imai, Tomoya; Sugiyama, Junji; Takeuchi, Miyuki; Yamauchi, Daisuke

    2013-01-01

    Cellulases are enzymes that normally digest cellulose; however, some are known to play essential roles in cellulose biosynthesis. Although some endogenous cellulases of plants and cellulose-producing bacteria are reportedly involved in cellulose production, their functions in cellulose production are unknown. In this study, we demonstrated that disruption of the cellulase (carboxymethylcellulase) gene causes irregular packing of de novo-synthesized fibrils in Gluconacetobacter xylinus, a cellulose-producing bacterium. Cellulose production was remarkably reduced and small amounts of particulate material were accumulated in the culture of a cmcax-disrupted G. xylinus strain (F2-2). The particulate material was shown to contain cellulose by both solid-state 13C nuclear magnetic resonance analysis and Fourier transform infrared spectroscopy analysis. Electron microscopy revealed that the cellulose fibrils produced by the F2-2 cells were highly twisted compared with those produced by control cells. This hypertwisting of the fibrils may reduce cellulose synthesis in the F2-2 strains. PMID:23243308

  15. Diversity of acetic acid bacteria present in healthy grapes from the Canary Islands.

    Science.gov (United States)

    Valera, Maria José; Laich, Federico; González, Sara S; Torija, Maria Jesús; Mateo, Estibaliz; Mas, Albert

    2011-11-15

    The identification of acetic acid bacteria (AAB) from sound grapes from the Canary Islands is reported in the present study. No direct recovery of bacteria was possible in the most commonly used medium, so microvinifications were performed on grapes from Tenerife, La Palma and Lanzarote islands. Up to 396 AAB were isolated from those microvinifications and identified by 16S rRNA gene sequencing and phylogenetic analysis. With this method, Acetobacter pasteurianus, Acetobacter tropicalis, Gluconobacter japonicus and Gluconacetobacter saccharivorans were identified. However, no discrimination between the closely related species Acetobacter malorum and Acetobacter cerevisiae was possible. As previously described, 16S-23S rRNA gene internal transcribed spacer (ITS) region phylogenetic analysis was required to classify isolates as one of those species. These two species were the most frequently occurring, accounting for more than 60% of the isolates. For typing the AAB isolates, both the Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR and (GTG)5-PCR techniques gave similar resolution. A total of 60 profiles were identified. Thirteen of these profiles were found in more than one vineyard, and only one profile was found on two different islands (Tenerife and La Palma). Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Rapid identification of acetic acid bacteria using MALDI-TOF mass spectrometry fingerprinting.

    Science.gov (United States)

    Andrés-Barrao, Cristina; Benagli, Cinzia; Chappuis, Malou; Ortega Pérez, Ruben; Tonolla, Mauro; Barja, François

    2013-03-01

    Acetic acid bacteria (AAB) are widespread microorganisms characterized by their ability to transform alcohols and sugar-alcohols into their corresponding organic acids. The suitability of matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS) for the identification of cultured AAB involved in the industrial production of vinegar was evaluated on 64 reference strains from the genera Acetobacter, Gluconacetobacter and Gluconobacter. Analysis of MS spectra obtained from single colonies of these strains confirmed their basic classification based on comparative 16S rRNA gene sequence analysis. MALDI-TOF analyses of isolates from vinegar cross-checked by comparative sequence analysis of 16S rRNA gene fragments allowed AAB to be identified, and it was possible to differentiate them from mixed cultures and non-AAB. The results showed that MALDI-TOF MS analysis was a rapid and reliable method for the clustering and identification of AAB species. Copyright © 2012 Elsevier GmbH. All rights reserved.

  17. Metabolic profiling of two maize (Zea mays L.) inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense

    Science.gov (United States)

    Brusamarello-Santos, Liziane Cristina; Gilard, Françoise; Brulé, Lenaïg; Quilleré, Isabelle; Gourion, Benjamin; Ratet, Pascal; Maltempi de Souza, Emanuel; Lea, Peter J.; Hirel, Bertrand

    2017-01-01

    Maize roots can be colonized by free-living atmospheric nitrogen (N2)-fixing bacteria (diazotrophs). However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2), already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+). The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts. Moreover, a number

  18. Reconstruction of the biogeochemistry and ecology of photoautotrophs based on the nitrogen and carbon isotopic compositions of vanadyl porphyrins from Miocene siliceous sediments

    Directory of Open Access Journals (Sweden)

    Y. Kashiyama

    2008-05-01

    Full Text Available We determined both the nitrogen and carbon isotopic compositions of various vanadyl alkylporphyrins isolated from siliceous marine sediments of the Onnagawa Formation (middle Miocene, northeastern Japan to investigate the biogeochemistry and ecology of photoautotrophs living in the paleo-ocean. The distinctive isotopic signals support the interpretations of previous works that the origin of 17-nor-deoxophylloerythroetioporphyrin (DPEP is chlorophylls-c1-3, whereas 8-nor-DPEP may have originated from chlorophylls-a2 or b2 or bacteriochlorophyll-a. Although DPEP and cycloheptanoDPEP are presumably derived from common precursory pigments, their isotopic compositions differed in the present study, suggesting that the latter represents a specific population within the photoautotrophic community. The average δ15N value for the entire photoautotrophic community is estimated to be –2 to +1‰ from the δ15N values of DPEP (–6.9 to –3.6‰; n=7, considering that the empirical isotopic relationships that the tetrapyrrole nuclei of chloropigments are depleted in 15N by ~4.8‰ and enriched in 13C by ~1.8‰ relative to the whole cells. This finding suggests that nitrogen utilized in the primary production was supplied mainly through N2-fixation by diazotrophic cyanobacteria. Based on the δ13C values of DPEP (–17.9 to –15.6‰; n=7, we estimated isotopic fractionation associated with photosynthetic carbon fixation to be 8–14‰. This range suggests the importance of β-carboxylation and/or active transport of the carbon substrate, indicating in turn the substantial contribution of diazotrophic cyanobacteria to primary production. Based on the δ15N values of 17-nor-DPEP (–7.4 to –2.4‰ n=7, the δ15N range of chlorophylls-c-producing algae was estimated to be –3

  19. Metabolic profiling of two maize (Zea mays L. inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense.

    Directory of Open Access Journals (Sweden)

    Liziane Cristina Brusamarello-Santos

    Full Text Available Maize roots can be colonized by free-living atmospheric nitrogen (N2-fixing bacteria (diazotrophs. However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2, already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+. The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts

  20. Metabolic profiling of two maize (Zea mays L.) inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense.

    Science.gov (United States)

    Brusamarello-Santos, Liziane Cristina; Gilard, Françoise; Brulé, Lenaïg; Quilleré, Isabelle; Gourion, Benjamin; Ratet, Pascal; Maltempi de Souza, Emanuel; Lea, Peter J; Hirel, Bertrand

    2017-01-01

    Maize roots can be colonized by free-living atmospheric nitrogen (N2)-fixing bacteria (diazotrophs). However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2), already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+). The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts. Moreover, a number

  1. Draft genome sequence of Bradyrhizobium manausense strain BR 3351T, an effective symbiont isolated from Amazon rainforest.

    Science.gov (United States)

    Simões-Araújo, Jean Luiz; Rumjanek, Norma Gouvêa; Xavier, Gustavo Ribeiro; Zilli, Jerri Édson

    The strain BR 3351 T (Bradyrhizobium manausense) was obtained from nodules of cowpea (Vigna unguiculata L. Walp) growing in soil collected from Amazon rainforest. Furthermore, it was observed that the strain has high capacity to fix nitrogen symbiotically in symbioses with cowpea. We report here the draft genome sequence of strain BR 3351 T . The information presented will be important for comparative analysis of nodulation and nitrogen fixation for diazotrophic bacteria. A draft genome with 9,145,311bp and 62.9% of GC content was assembled in 127 scaffolds using 100bp pair-end Illumina MiSeq system. The RAST annotation identified 8603 coding sequences, 51 RNAs genes, classified in 504 subsystems. Published by Elsevier Editora Ltda.

  2. First Evidence of Palytoxin and 42-Hydroxy-palytoxin in the Marine Cyanobacterium Trichodesmium

    Directory of Open Access Journals (Sweden)

    Dominique Laurent

    2011-03-01

    Full Text Available Marine pelagic diazotrophic cyanobacteria of the genus Trichodesmium (Oscillatoriales are widespread throughout the tropics and subtropics, and are particularly common in the waters of New Caledonia. Blooms of Trichodesmium are suspected to be a potential source of toxins in the ciguatera food chain and were previously reported to contain several types of paralyzing toxins. The toxicity of water-soluble extracts of Trichodesmium spp. were analyzed by mouse bioassay and Neuroblastoma assay and their toxic compounds characterized using liquid chromatography coupled with tandem mass spectrometry techniques. Here, we report the first identification of palytoxin and one of its derivatives, 42-hydroxy-palytoxin, in field samples of Trichodesmium collected in the New Caledonian lagoon. The possible role played by Trichodesmium blooms in the development of clupeotoxism, this human intoxication following the ingestion of plankton-eating fish and classically associated with Ostreopsis blooms, is also discussed.

  3. Improved Alkane Production in Nitrogen-Fixing and Halotolerant Cyanobacteria via Abiotic Stresses and Genetic Manipulation of Alkane Synthetic Genes.

    Science.gov (United States)

    Kageyama, Hakuto; Waditee-Sirisattha, Rungaroon; Sirisattha, Sophon; Tanaka, Yoshito; Mahakhant, Aparat; Takabe, Teruhiro

    2015-07-01

    Cyanobacteria possess the unique capacity to produce alkane. In this study, effects of nitrogen deficiency and salt stress on biosynthesis of alkanes were investigated in three kinds of cyanobacteria. Intracellular alkane accumulation was increased in nitrogen-fixing cyanobacterium Anabaena sp. PCC7120, but decreased in non-diazotrophic cyanobacterium Synechococcus elongatus PCC7942 and constant in a halotolerant cyanobacterium Aphanothece halophytica under nitrogen-deficient condition. We also found that salt stress increased alkane accumulation in Anabaena sp. PCC7120 and A. halophytica. The expression levels of two alkane synthetic genes were not upregulated significantly under nitrogen deficiency or salt stress in Anabaena sp. PCC7120. The transformant Anabaena sp. PCC7120 cells with additional alkane synthetic gene set from A. halophytica increased intracellular alkane accumulation level compared to control cells. These results provide a prospect to improve bioproduction of alkanes in nitrogen-fixing halotolerant cyanobacteria via abiotic stresses and genetic engineering.

  4. Enhanced production of biomass, pigments and antioxidant capacity of a nutritionally important cyanobacterium Nostochopsis lobatus.

    Science.gov (United States)

    Pandey, Usha; Pandey, J

    2008-07-01

    A diazotrophic cyanobacterium Nostochopsis lobatus was evaluated for enhanced production of biomass, pigments and antioxidant capacity. N. lobatus showed potentially high antioxidant capacity (46.12 microM AEAC) with significant improvement under immobilized cell cultures (87.05 microM AEAC). When a mixture of P and Fe was supplemented, biomass, pigments, nutritive value and antioxidant capacity increased substantially at pH 7.8. When considered separately, P appeared to be a better supplement than Fe for the production of biomass, chlorophyll and carotenoids. However, for phycocyanin, phycoerythrin, nutritive value and antioxidant capacity, Fe appeared more effective than P. Our study indicates N. lobatus to be a promising bioresource for enhanced production of nutritionally rich biomass, pigments and antioxidants. The study also suggests that P and Fe are potentially effective supplements for scale-up production for commercial application.

  5. Moorea producens gen. nov., sp. nov. and Moorea bouillonii comb. nov., tropical marine cyanobacteria rich in bioactive secondary metabolites.

    Science.gov (United States)

    Engene, Niclas; Rottacker, Erin C; Kaštovský, Jan; Byrum, Tara; Choi, Hyukjae; Ellisman, Mark H; Komárek, Jiří; Gerwick, William H

    2012-05-01

    The filamentous cyanobacterial genus Moorea gen. nov., described here under the provisions of the International Code of Botanical Nomenclature, is a cosmopolitan pan-tropical group abundant in the marine benthos. Members of the genus Moorea are photosynthetic (containing phycocyanin, phycoerythrin, allophycocyanin and chlorophyll a), but non-diazotrophic (lack heterocysts and nitrogenase reductase genes). The cells (discoid and 25-80 µm wide) are arranged in long filaments (algae blooms and, due to morphological resemblance to the genus Lyngbya, this group has often been incorrectly cited in the literature. We here describe two species of the genus Moorea: Moorea producens sp. nov. (type species of the genus) with 3L(T) as the nomenclature type, and Moorea bouillonii comb. nov. with PNG5-198(R) as the nomenclature type.

  6. System analysis of salt and osmotic stress induced proteins in Nostoc muscorum and Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    Vipin Kaithwas

    2017-06-01

    Full Text Available In this study the proteome response of the two diazotrophic organism’s viz. Nostoc muscorum and Bradyrhizobium japonicum exposed to salt (NaCl and osmotic (sucrose stresses was compared. Out of the total over expressed proteins; we have selected only three over expressed proteins viz. GroEL chaperonin, nitrogenase Mo-Fe protein and argininosuccinate synthase for further analysis, and then we analyzed the amino acid frequencies of all the three over expressed proteins. That led to the conclusion that amino acids e.g. alanine, glycine and valine that were energetically cheaper to produce were showing higher frequencies. This study would help in tracing the phylogenetic relationship between protein families.

  7. Sucrose in Cyanobacteria: From a Salt-Response Molecule to Play a Key Role in Nitrogen Fixation

    Directory of Open Access Journals (Sweden)

    María A. Kolman

    2015-01-01

    Full Text Available In the biosphere, sucrose is mainly synthesized in oxygenic photosynthetic organisms, such as cyanobacteria, green algae and land plants, as part of the carbon dioxide assimilation pathway. Even though its central position in the functional biology of plants is well documented, much less is known about the role of sucrose in cyanobacteria. In those prokaryotes, sucrose accumulation has been associated with salt acclimation, and considered as a compatible solute in low-salt tolerant strains. In the last years, functional characterizations of sucrose metabolizing enzymes, metabolic control analysis, cellular localization of gene expressions, and reverse genetic experiments have revealed that sucrose metabolism is crucial in the diazotrophic growth of heterocystic strains, and besides, that it can be connected to glycogen synthesis. This article briefly summarizes the current state of knowledge of sucrose physiological functions in modern cyanobacteria and how they might have evolved taking into account the phylogenetic analyses of sucrose enzymes.

  8. Energy balance of maize production in Brazil: the energetic constraints of a net positive outcome

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Luis Henrique de Barros; Alves, Bruno Jose Rodrigues; Urquiaga, Segundo

    2008-07-01

    Among the factors used to analyze and to establish the sustainability of a whole agricultural production system, the energy balance is one of the most powerful and robust. The maize production in Brazil is surely the reflex of an energy intensive system that demands many field operations and heavy fertilizer applications, notably nitrogen in urea form. This work presents an energy balance of this major crop adjusted to the Brazilian conditions of cultivation. The input components were grouped based on their energy contents, and the possible improvements in the agricultural practices that could improve energy balance and net energy withdrawn from the farming were considered. The replacement of N synthetic fertilizer by biological nitrogen fixation, whether the process is directly carried out by endophytic diazotroph bacteria or by means of a N{sub 2}- fixing legume culture planted before the main crop as a green-manure is also discussed. (author)

  9. Effects of UV-B and heavy metals on nitrogen and phosphorus metabolism in three cyanobacteria.

    Science.gov (United States)

    Yadav, Shivam; Prajapati, Rajesh; Atri, Neelam

    2016-01-01

    Cyanobacteria sp. (diazotrophic and planktonic) hold a major position in ecosystem, former one due to their intrinsic capability of N2-fixation and later because of mineralization of organic matter. Unfortunately, their exposure to variety of abiotic stresses is unavoidable. Comparative analysis of interactive effect of UV-B and heavy metals (Cd/Zn) on nitrogen and phosphorus metabolism of three cyanobacteria (Anabaena, Microcystis, Nostoc) revealed additive inhibition (χ(2) significant p cyanobacteria suggests UV-B-induced structural change(s) in the enzyme/carriers. Metals seem to compete for the binding sites of the enzymes and carriers; as noticed for Anabaena and Microcystis showing change in Km while no change in the Km value of Nostoc suggests non-competitive nutrient uptake. Higher accumulation and more adverse effect on Na(+) and K(+) efflux proposes Cd as more toxic compared to Zn. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Estimation of nitrogen fixation in Saccharum spp. by 15N dilution method

    International Nuclear Information System (INIS)

    Singh, Mohan

    1994-01-01

    The amount of nitrogen fixed by bacteria associated with the roots of Saccharum spontaneum, S. sinense, and S. barberi has been estimated by 15 N-isotope dilution method using Sclerotachya fusca as a non-fixing control. S. spontaneum produced highest shoot dry weight among the species tested but maximum nitrogen was accumulated by S. barberi. Highest dilution in the 15 N-enrichment was observed in S. spontaneum followed by S. sinense and S. barberi in comparison to the control plant of Sclerotchya fusca. S. spontaneum derived 60 per cent followed by S. sinense 54 per cent and S. barberi 35 per cent of their total nitrogen requirement through fixation of nitrogen by diazotrophic bacteria associated with their roots. (author). 11 refs., 2 tabs

  11. Free-living dinitrogen-fixing bacteria isolated from petroleum refinery oily sludge

    Energy Technology Data Exchange (ETDEWEB)

    Laguerre, G.; Bossand, B.; Bardin, R.

    1987-07-01

    Dinitrogen-fixing activity (acetylene reduction and /sup 15/N/sub 2/ fixation) was found in an oily sludge originating from a petroleum refinery. Two representative dinitrogen-fixing bacterial strains were isolated from this oily waste. Their nitrogenase activity was effective when they were cultivated on sterilized sludge or simple carbon substrates (organic acid salts, sugars). Using the classical methods, these strains could not be unambiguously related to other diazotrophic taxa. The landfarming process is widely used for oily sludge disposal; this study shows that oily sludges are more than a simple carbon input into the soil but that they must also be considered as real sources of dinitrogen-fixing and probably degradative microorganisms.

  12. Global analysis of gene expression dynamics within the marine microbial community during the VAHINE mesocosm experiment in the southwest Pacific

    Science.gov (United States)

    Pfreundt, Ulrike; Spungin, Dina; Bonnet, Sophie; Berman-Frank, Ilana; Hess, Wolfgang R.

    2016-07-01

    Microbial gene expression was followed for 23 days within a mesocosm (M1) isolating 50 m3 of seawater and in the surrounding waters in the Nouméa lagoon, New Caledonia, in the southwest Pacific as part of the VAriability of vertical and tropHIc transfer of diazotroph derived N in the south wEst Pacific (VAHINE) experiment. The aim of VAHINE was to examine the fate of diazotroph-derived nitrogen (DDN) in a low-nutrient, low-chlorophyll ecosystem. On day 4 of the experiment, the mesocosm was fertilized with phosphate. In the lagoon, gene expression was dominated by the cyanobacterium Synechococcus, closely followed by Alphaproteobacteria. In contrast, drastic changes in the microbial community composition and transcriptional activity were triggered within the mesocosm within the first 4 days, with transcription bursts from different heterotrophic bacteria in rapid succession. The microbial composition and activity of the surrounding lagoon ecosystem appeared more stable, although following similar temporal trends as in M1. We detected significant gene expression from Chromerida in M1, as well as the Nouméa lagoon, suggesting these photoautotrophic alveolates were present in substantial numbers in the open water. Other groups contributing substantially to the metatranscriptome were affiliated with marine Euryarchaeota Candidatus Thalassoarchaea (inside and outside) and Myoviridae bacteriophages likely infecting Synechococcus, specifically inside M1. High transcript abundances for ammonium transporters and glutamine synthetase in many different taxa (e.g., Pelagibacteraceae, Synechococcus, Prochlorococcus, and Rhodobacteraceae) was consistent with the known preference of most bacteria for this nitrogen source. In contrast, Alteromonadaceae highly expressed urease genes; Rhodobacteraceae and Prochlorococcus showed some urease expression, too. Nitrate reductase transcripts were detected on day 10 very prominently in Synechococcus and in Halomonadaceae. Alkaline

  13. Cellulose decomposition and associated nitrogen fixation by mixed cultures of Cellulomonas gelida and Azospirillum species or Bacillus macerans

    Energy Technology Data Exchange (ETDEWEB)

    Halsall, D.M.; Gibson, A.H.

    1985-10-01

    Mixed cultures of Cellulomonas gelida plus Azospirillum lipoferum or Azospirillum brasilense and C. gelida plus Bacillus macerans were shown to degrade cellulose and straw and to utilize the energy-yielding products to fix atmospheric nitrogen. This cooperative process was followed over 30 days in sand-based cultures in which the breakdown of 20% of the cellulose and 28 to 30% of the straw resulted in the fixation of 12 to 14.6 mg of N per g of cellulose and 17 to 19 mg of N per g of straw consumed. Cellulomonas species have certain advantages over aerobic cellulose-degrading fungi in being able to degrade cellulose at oxygen concentrations as low as 1% O/sub 2/ (vol/vol) which would allow a close association between cellulose-degrading and microaerobic diazotrophic microorganisms. Cultures inoculated with initially different proportions of A. brasilense and C. gelida all reached a stable ratio of approximately 1 Azospirillum/3 Cellulomonas cells.

  14. Nitrogen-fixing and uricolytic bacteria associated with the gut of Dendroctonus rhizophagus and Dendroctonus valens (Curculionidae: Scolytinae).

    Science.gov (United States)

    Morales-Jiménez, Jesús; Vera-Ponce de León, Arturo; García-Domínguez, Aidé; Martínez-Romero, Esperanza; Zúñiga, Gerardo; Hernández-Rodríguez, César

    2013-07-01

    The bark beetles of the genus Dendroctonus feed on phloem that is a nitrogen-limited source. Nitrogen fixation and nitrogen recycling may compensate or alleviate such a limitation, and beetle-associated bacteria capable of such processes were identified. Raoultella terrigena, a diazotrophic bacteria present in the gut of Dendroctonus rhizophagus and D. valens, exhibited high acetylene reduction activity in vitro with different carbon sources, and its nifH and nifD genes were sequenced. Bacteria able to recycle uric acid were Pseudomonas fluorescens DVL3A that used it as carbon and nitrogen source, Serratia proteomaculans 2A CDF and Rahnella aquatilis 6-DR that used uric acid as sole nitrogen source. Also, this is the first report about the uric acid content in whole eggs, larvae, and adults (male and female) samples of the red turpentine beetle (Dendroctonus valens). Our results suggest that the gut bacteria of these bark beetles could contribute to insect N balance.

  15. Free-living dinitrogen-fixing bacteria isolated from petroleum refinery oily sludge

    International Nuclear Information System (INIS)

    Laguerre, G.; Bossand, B.; Bardin, R.

    1987-01-01

    Dinitrogen-fixing activity (acetylene reduction and 15 N 2 fixation) was found in an oily sludge originating from a petroleum refinery. Two representative dinitrogen-fixing bacterial strains were isolated from this oily waste. Their nitrogenase activity was effective when they were cultivated on sterilized sludge or simple carbon substrates (organic acid salts, sugars). Using the classical methods, these strains could not be unambiguously related to other diazotrophic taxa. The landfarming process is widely used for oily sludge disposal; this study shows that oily sludges are more than a simple carbon input into the soil but that they must also be considered as real sources of dinitrogen-fixing and probably degradative microorganisms

  16. Draft genome sequence of Bradyrhizobium manausense strain BR 3351T, an effective symbiont isolated from Amazon rainforest

    Directory of Open Access Journals (Sweden)

    Jean Luiz Simões-Araújo

    Full Text Available ABSTRACT The strain BR 3351T (Bradyrhizobium manausense was obtained from nodules of cowpea (Vigna unguiculata L. Walp growing in soil collected from Amazon rainforest. Furthermore, it was observed that the strain has high capacity to fix nitrogen symbiotically in symbioses with cowpea. We report here the draft genome sequence of strain BR 3351T. The information presented will be important for comparative analysis of nodulation and nitrogen fixation for diazotrophic bacteria. A draft genome with 9,145,311 bp and 62.9% of GC content was assembled in 127 scaffolds using 100 bp pair-end Illumina MiSeq system. The RAST annotation identified 8603 coding sequences, 51 RNAs genes, classified in 504 subsystems.

  17. Inter-domain cross-talk controls the NifA protein activity of Herbaspirillum seropedicae.

    Science.gov (United States)

    Monteiro, R A; de Souza, E M; Wassem, R; Yates, M G; Pedrosa, F O; Chubatsu, L S

    2001-11-09

    Herbaspirillum seropedicae is an endophytic diazotroph, which colonizes sugar cane, wheat, rice and maize. The activity of NifA, a transcriptional activator of nif genes in H. seropedicae, is controlled by ammonium ions through a mechanism involving its N-terminal domain. Here we show that this domain interacts specifically in vitro with the N-truncated NifA protein, as revealed by protection against proteolysis, and this interaction caused an inhibitory effect on both the ATPase and DNA-binding activities of the N-truncated NifA protein. We suggest that the N-terminal domain inhibits NifA-dependent transcriptional activation by an inter-domain cross-talk between the catalytic domain of the NifA protein and its regulatory N-terminal domain in response to fixed nitrogen.

  18. Fnr is involved in oxygen control of Herbaspirillum seropedicae N-truncated NifA protein activity in Escherichia coli.

    Science.gov (United States)

    Monteiro, Rose A; de Souza, Emanuel M; Yates, M Geoffrey; Pedrosa, Fabio O; Chubatsu, Leda S

    2003-03-01

    Herbaspirillum seropedicae is an endophytic diazotroph belonging to the beta-subclass of the class Proteobacteria, which colonizes many members of the Gramineae. The activity of the NifA protein, a transcriptional activator of nif genes in H. seropedicae, is controlled by ammonium ions through its N-terminal domain and by oxygen through mechanisms that are not well understood. Here we report that the NifA protein of H. seropedicae is inactive and more susceptible to degradation in an fnr Escherichia coli background. Both ef