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Sample records for endophyte gluconacetobacter diazotrophicus

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

    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

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

    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.

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

    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.

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

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

    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

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

    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

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

    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.

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

    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...

  9. 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

    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

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

    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. Natural Endophytic Occurrence of Acetobacter diazotrophicus in Pineapple Plants.

    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.

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

    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

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

    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.

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

    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.

  15. 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.

    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.

  16. 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

    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

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

    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.

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

    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.

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

    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. Endophytic colonization of plant roots by nitrogen-fixing bacteria

    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)

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

    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.

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

    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.

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

    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)

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

    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. 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

    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.

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

    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.

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

    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 ...

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

    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% ...

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

    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.

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

    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

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

    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)).

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

    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).

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

    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...

  14. Endophytes as sources of antibiotics.

    Martinez-Klimova, Elena; Rodríguez-Peña, Karol; Sánchez, Sergio

    2017-06-15

    Until a viable alternative can be accessible, the emergence of resistance to antimicrobials requires the constant development of new antibiotics. Recent scientific efforts have been aimed at the bioprospecting of microorganisms' secondary metabolites, with special emphasis on the search for antimicrobial natural products derived from endophytes. Endophytes are microorganisms that inhabit the internal tissues of plants without causing apparent harm to the plant. The present review article compiles recent (2006-2016) literature to provide an update on endophyte research aimed at finding metabolites with antibiotic activities. We have included exclusively information on endophytes that produce metabolites capable of inhibiting the growth of bacterial, fungal and protozoan pathogens of humans, animals and plants. Where available, the identified metabolites have been listed. In this review, we have also compiled a list of the bacterial and fungal phyla that have been isolated as endophytes as well as the plant families from which the endophytes were isolated. The majority of endophytes that produce antibiotic metabolites belong to either phylum Ascomycota (kingdom Fungi) or to phylum Actinobacteria (superkingdom Bacteria). Endophytes that produce antibiotic metabolites were predominant, but certainly not exclusively, from the plant families Fabaceae, Lamiaceae, Asteraceae and Araceae, suggesting that endophytes that produce antimicrobial metabolites are not restricted to a reduced number of plant families. The locations where plants (and inhabiting endophytes) were collected from, according to the literature, have been mapped, showing that endophytes that produce bioactive compounds have been collected globally. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. 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

    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

  16. Beauveria bassiana as an endophyte

    McKinnon, Aimee C.; Saari, Susanna Talvikki; Moran-Diez, Maria E.

    2017-01-01

    In the last decade there has been increased focus on the potential of endophytic Beauveria bassiana for the biocontrol of insect herbivores. Generally, detection of endophytes is acknowledged to be problematic and recovery method-dependent. Herein, we critically analyse the methodology reported...... for the detection of B. bassiana as endophytes following experimental inoculation. In light of the methodology, we further review the effects of endophytic B. bassiana on insect herbivores. Our review indicated the need for stringent protocols for surface sterilisation including thorough experimental controls....... For molecular detection protocols by PCR, residual DNA from surface inocula must also be considered. The biocontrol potential of B. bassiana endophytes appears promising although both negative and neutral effects on insect herbivores were reported and there remains ambiguity with respect to the location...

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

    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% ...

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

    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% ...

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

    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.

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

    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.

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

    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.

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

    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.

  3. Fungal Endophytes: Beyond Herbivore Management

    Bamisope S. Bamisile

    2018-03-01

    Full Text Available The incorporation of entomopathogenic fungi as biocontrol agents into Integrated Pest Management (IPM programs without doubt, has been highly effective. The ability of these fungal pathogens such as Beauveria bassiana and Metarhizium anisopliae to exist as endophytes in plants and protect their colonized host plants against the primary herbivore pests has widely been reported. Aside this sole role of pest management that has been traditionally ascribed to fungal endophytes, recent findings provided evidence of other possible functions as plant yield promoter, soil nutrient distributor, abiotic stress and drought tolerance enhancer in plants. However, reports on these additional important effects of fungal endophytes on the colonized plants remain scanty. In this review, we discussed the various beneficial effects of endophytic fungi on the host plants and their primary herbivore pests; as well as some negative effects that are relatively unknown. We also highlighted the prospects of our findings in further increasing the acceptance of fungal endophytes as an integral part of pest management programs for optimized crop production.

  4. Fungal endophytes: modifiers of plant disease.

    Busby, Posy E; Ridout, Mary; Newcombe, George

    2016-04-01

    Many recent studies have demonstrated that non-pathogenic fungi within plant microbiomes, i.e., endophytes ("endo" = within, "phyte" = plant), can significantly modify the expression of host plant disease. The rapid pace of advancement in endophyte ecology warrants a pause to synthesize our understanding of endophyte disease modification and to discuss future research directions. We reviewed recent literature on fungal endophyte disease modification, and here report on several emergent themes: (1) Fungal endophyte effects on plant disease span the full spectrum from pathogen antagonism to pathogen facilitation, with pathogen antagonism most commonly reported. (2) Agricultural plant pathosystems are the focus of research on endophyte disease modification. (3) A taxonomically diverse group of fungal endophytes can influence plant disease severity. And (4) Fungal endophyte effects on plant disease severity are context-dependent. Our review highlights the importance of fungal endophytes for plant disease across a broad range of plant pathosystems, yet simultaneously reveals that complexity within plant microbiomes presents a significant challenge to disentangling the biotic environmental factors affecting plant disease severity. Manipulative studies integrating eco-evolutionary approaches with emerging molecular tools will be poised to elucidate the functional importance of endophytes in natural plant pathosystems that are fundamental to biodiversity and conservation.

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

    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

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

    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.

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

    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.

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

    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.

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

    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

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

    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.

  11. Laparoscopic partial nephrectomy for endophytic hilar tumors

    Di Pierro, G B; Tartaglia, N; Aresu, L

    2014-01-01

    To analyze feasibility and outcomes of laparoscopic partial nephrectomy (LPN) for endophytic hilar tumors in low-intermediate (ASA I-II) risk patients.......To analyze feasibility and outcomes of laparoscopic partial nephrectomy (LPN) for endophytic hilar tumors in low-intermediate (ASA I-II) risk patients....

  12. Bacterial Endophyte Colonization and Distribution within Plants

    Shyam L. Kandel

    2017-11-01

    Full Text Available The plant endosphere contains a diverse group of microbial communities. There is general consensus that these microbial communities make significant contributions to plant health. Both recently adopted genomic approaches and classical microbiology techniques continue to develop the science of plant-microbe interactions. Endophytes are microbial symbionts residing within the plant for the majority of their life cycle without any detrimental impact to the host plant. The use of these natural symbionts offers an opportunity to maximize crop productivity while reducing the environmental impacts of agriculture. Endophytes promote plant growth through nitrogen fixation, phytohormone production, nutrient acquisition, and by conferring tolerance to abiotic and biotic stresses. Colonization by endophytes is crucial for providing these benefits to the host plant. Endophytic colonization refers to the entry, growth and multiplication of endophyte populations within the host plant. Lately, plant microbiome research has gained considerable attention but the mechanism allowing plants to recruit endophytes is largely unknown. This review summarizes currently available knowledge about endophytic colonization by bacteria in various plant species, and specifically discusses the colonization of maize plants by Populus endophytes.

  13. The isolation and characterization of endophytic microorganisms ...

    Fungi were identified by distinguishing between reproductive structures using a microculture technique. While observing diaphanized root fragments, we found arbuscular mycorrhizal fungi (AMF) and dark septate endophytic (DSE) fungi in the fine and coarse roots of H. marrubioides. The endophytic CR was more ...

  14. Isolation and characterization of beneficial indigenous endophytic ...

    Plant-associated bacteria that live inside plant tissues without causing any damage to plants are defined as endophytic bacteria. The present study was carried out to analyze the phenotypic and genotypic diversity of endophytic bacteria associated with Amaranthus hybridus, Solanum lycopersicum and Cucurbita maxima.

  15. A endophytic fungus, Ramichloridium cerophilum, promotes growth ...

    Tuoyo Aghomotsegin

    2016-06-22

    Jun 22, 2016 ... A fungal endophyte, Ramichloridium cerophilum, was identified as a Class 2 endophytes species ... The mycorrhizal symbiosis between plants and fungi is common and .... growing fungal colony and placed into a sterile plastic pot and .... bacteria associated with the roots of Chinese cabbage (Brassica.

  16. Fungal endophytes: diversity and functional roles

    Rodriguez, R.J.; White, J.F.; Arnold, A.E.; Redman, R.S.

    2009-01-01

    All plants in natural ecosystems appear to be symbiotic with fungal endophytes. This highly diverse group of fungi can have profound impacts on plant communities through increasing fitness by conferring abiotic and biotic stress tolerance, increasing biomass and decreasing water consumption, or decreasing fitness by altering resource allocation. Despite more than 100 yr of research resulting in thousands of journal articles, the ecological significance of these fungi remains poorly characterized. Historically, two endophytic groups (clavicipitaceous (C) and nonclavicipitaceous (NC)) have been discriminated based on phylogeny and life history traits. Here, we show that NC-endophytes represent three distinct functional groups based on host colonization and transmission, in planta biodiversity and fitness benefits conferred to hosts. Using this framework, we contrast the life histories, interactions with hosts and potential roles in plant ecophysiology of C- and NC-endophytes, and highlight several key questions for future work in endophyte biology.

  17. Fungal endophytes for sustainable crop production.

    Lugtenberg, Ben J J; Caradus, John R; Johnson, Linda J

    2016-12-01

    This minireview highlights the importance of endophytic fungi for sustainable agriculture and horticulture production. Fungal endophytes play a key role in habitat adaptation of plants resulting in improved plant performance and plant protection against biotic and abiotic stresses. They encode a vast variety of novel secondary metabolites including volatile organic compounds. In addition to protecting plants against pathogens and pests, selected fungal endophytes have been used to remove animal toxicities associated with fungal endophytes in temperate grasses, to create corn and rice plants that are tolerant to a range of biotic and abiotic stresses, and for improved management of post-harvest control. We argue that practices used in plant breeding, seed treatments and agriculture, often caused by poor knowledge of the importance of fungal endophytes, are among the reasons for the loss of fungal endophyte diversity in domesticated plants and also accounts for the reduced effectiveness of some endophyte strains to confer plant benefits. We provide recommendations on how to mitigate against these negative impacts in modern agriculture. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

    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

  19. Exploitation of endophytes for sustainable agricultural intensification.

    Le Cocq, Kate; Gurr, Sarah J; Hirsch, Penny R; Mauchline, Tim H

    2017-04-01

    Intensive agriculture, which depends on unsustainable levels of agrochemical inputs, is environmentally harmful, and the expansion of these practices to meet future needs is not economically feasible. Other options should be considered to meet the global food security challenge. The plant microbiome has been linked to improved plant productivity and, in this microreview, we consider the endosphere - a subdivision of the plant microbiome. We suggest a new definition of microbial endophyte status, the need for synergy between fungal and bacterial endophyte research efforts, as well as potential strategies for endophyte application to agricultural systems. © 2016 THE AUTHORS. MOLECULAR PLANT PATHOLOGY PUBLISHED BY BRITISH SOCIETY FOR PLANT PATHOLOGY AND JOHN WILEY & SONS LTD.

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

    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.

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

    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.

  2. Bioactive endophytes warrant intensified exploration and conservation.

    Smith, Stephen A; Tank, David C; Boulanger, Lori-Ann; Bascom-Slack, Carol A; Eisenman, Kaury; Kingery, David; Babbs, Beatrice; Fenn, Kathleen; Greene, Joshua S; Hann, Bradley D; Keehner, Jocelyn; Kelley-Swift, Elizabeth G; Kembaiyan, Vivek; Lee, Sun Jin; Li, Puyao; Light, David Y; Lin, Emily H; Ma, Cong; Moore, Emily; Schorn, Michelle A; Vekhter, Daniel; Nunez, Percy V; Strobel, Gary A; Donoghue, Michael J; Strobel, Scott A

    2008-08-25

    A key argument in favor of conserving biodiversity is that as yet undiscovered biodiversity will yield products of great use to humans. However, the link between undiscovered biodiversity and useful products is largely conjectural. Here we provide direct evidence from bioassays of endophytes isolated from tropical plants and bioinformatic analyses that novel biology will indeed yield novel chemistry of potential value. We isolated and cultured 135 endophytic fungi and bacteria from plants collected in Peru. nrDNAs were compared to samples deposited in GenBank to ascertain the genetic novelty of cultured specimens. Ten endophytes were found to be as much as 15-30% different than any sequence in GenBank. Phylogenetic trees, using the most similar sequences in GenBank, were constructed for each endophyte to measure phylogenetic distance. Assays were also conducted on each cultured endophyte to record bioactivity, of which 65 were found to be bioactive. The novelty of our contribution is that we have combined bioinformatic analyses that document the diversity found in environmental samples with culturing and bioassays. These results highlight the hidden hyperdiversity of endophytic fungi and the urgent need to explore and conserve hidden microbial diversity. This study also showcases how undergraduate students can obtain data of great scientific significance.

  3. Bioactive endophytes warrant intensified exploration and conservation.

    Stephen A Smith

    2008-08-01

    Full Text Available A key argument in favor of conserving biodiversity is that as yet undiscovered biodiversity will yield products of great use to humans. However, the link between undiscovered biodiversity and useful products is largely conjectural. Here we provide direct evidence from bioassays of endophytes isolated from tropical plants and bioinformatic analyses that novel biology will indeed yield novel chemistry of potential value.We isolated and cultured 135 endophytic fungi and bacteria from plants collected in Peru. nrDNAs were compared to samples deposited in GenBank to ascertain the genetic novelty of cultured specimens. Ten endophytes were found to be as much as 15-30% different than any sequence in GenBank. Phylogenetic trees, using the most similar sequences in GenBank, were constructed for each endophyte to measure phylogenetic distance. Assays were also conducted on each cultured endophyte to record bioactivity, of which 65 were found to be bioactive.The novelty of our contribution is that we have combined bioinformatic analyses that document the diversity found in environmental samples with culturing and bioassays. These results highlight the hidden hyperdiversity of endophytic fungi and the urgent need to explore and conserve hidden microbial diversity. This study also showcases how undergraduate students can obtain data of great scientific significance.

  4. Genetic compatibility determines endophyte-grass combinations.

    Kari Saikkonen

    Full Text Available Even highly mutually beneficial microbial-plant interactions, such as mycorrhizal- and rhizobial-plant exchanges, involve selfishness, cheating and power-struggles between the partners, which depending on prevailing selective pressures, lead to a continuum of interactions from antagonistic to mutualistic. Using manipulated grass-endophyte combinations in a five year common garden experiment, we show that grass genotypes and genetic mismatches constrain genetic combinations between the vertically (via host seeds transmitted endophytes and the out-crossing host, thereby reducing infections in established grass populations. Infections were lost in both grass tillers and seedlings in F(1 and F(2 generations, respectively. Experimental plants were collected as seeds from two different environments, i.e., meadows and nearby riverbanks. Endophyte-related benefits to the host included an increased number of inflorescences, but only in meadow plants and not until the last growing season of the experiment. Our results illustrate the importance of genetic host specificity and trans-generational maternal effects on the genetic structure of a host population, which act as destabilizing forces in endophyte-grass symbioses. We propose that (1 genetic mismatches may act as a buffering mechanism against highly competitive endophyte-grass genotype combinations threatening the biodiversity of grassland communities and (2 these mismatches should be acknowledged, particularly in breeding programmes aimed at harnessing systemic and heritable endophytes to improve the agriculturally valuable characteristics of cultivars.

  5. Diversity of endophytic fungi in Glycine max.

    Fernandes, Elio Gomes; Pereira, Olinto Liparini; da Silva, Cynthia Cânedo; Bento, Claudia Braga Pereira; de Queiroz, Marisa Vieira

    2015-12-01

    Endophytic fungi are microorganisms that live within plant tissues without causing disease during part of their life cycle. With the isolation and identification of these fungi, new species are being discovered, and ecological relationships with their hosts have also been studied. In Glycine max, limited studies have investigated the isolation and distribution of endophytic fungi throughout leaves and roots. The distribution of these fungi in various plant organs differs in diversity and abundance, even when analyzed using molecular techniques that can evaluate fungal communities in different parts of the plants, such as denaturing gradient gel electrophoresis (DGGE). Our results show there is greater species richness of culturable endophytic filamentous fungi in the leaves G. max as compared to roots. Additionally, the leaves had high values for diversity indices, i.e. Simpsons, Shannon and Equitability. Conversely, dominance index was higher in roots as compared to leaves. The fungi Ampelomyces sp., Cladosporium cladosporioides, Colletotrichum gloeosporioides, Diaporthe helianthi, Guignardia mangiferae and Phoma sp. were more frequently isolated from the leaves, whereas the fungi Fusarium oxysporum, Fusarium solani and Fusarium sp. were prevalent in the roots. However, by evaluating the two communities by DGGE, we concluded that the species richness was higher in the roots than in the leaves. UPGMA analysis showed consistent clustering of isolates; however, the fungus Leptospora rubella, which belongs to the order Dothideales, was grouped among species of the order Pleosporales. The presence of endophytic Fusarium species in G. max roots is unsurprising, since Fusarium spp. isolates have been previously described as endophyte in other reports. However, it remains to be determined whether the G. max Fusarium endophytes are latent pathogens or non-pathogenic forms that benefit the plant. This study provides a broader knowledge of the distribution of the fungal

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

    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)

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

    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.

  8. Rock-degrading endophytic bacteria in cacti

    M. Esther Puente; Ching Y. Li; Yoav Bashan

    2009-01-01

    A plant-bacterium association of the cardon cactus (Pachycereus pringlei) and endophytic bacteria promotes establishment of seedlings and growth on igneous rocks without soil. These bacteria weather several rock types and minerals, unbind significant amounts of useful minerals for plants from the rocks, fix in vitro N2. produce...

  9. Potential biosurfactant producing endophytic and epiphytic fungi ...

    Potential biosurfactant producing endophytic and epiphytic fungi, isolated from macrophytes in the Negro River in Manaus, Amazonas, Brazil. ... Solms and Cyperus ligularis L., macrophytes collected from oil-contaminated waters, were studied to assess their potential for producing biosurfactants; the most promising ones ...

  10. Sebacinales Everywhere: Previously Overlooked Ubiquitous Fungal Endophytes

    Weiss, M.; Sýkorová, Zuzana; Garnica, S.; Riess, K.; Martos, F.; Krause, C.; Oberwinkler, F.; Bauer, R.; Redecker, D.

    2011-01-01

    Roč. 6, č. 2 (2011), s. 1-7 E-ISSN 1932-6203 Institutional research plan: CEZ:AV0Z60050516 Keywords : Sebacinales * endophytes * mycorrhiza Subject RIV: EF - Botanics Impact factor: 4.092, year: 2011

  11. The secret world of endophytes in perspective

    This work in Fungal Ecology is focused on the group of plant symbionts that have been termed collectively ‘microbial endophytes’. Broadly, microbial endophytes are commonly considered to be any of a diverse group of bacteria, cyanobacteria, or fungi that colonize internal tissues of plants. After ...

  12. Phyllosticta capitalensis, a widespread endophyte of plants

    Wikee, S.; Lombard, L.; Crous, P.W.; Nakashima, C.; Motohashi, K.; Chukeatirote, E.; Alias, S.A.; McKenzie, E.H.C.; Hyde, K.D.

    2013-01-01

    Phyllosticta capitalensis is an endophyte and weak plant pathogen with a worldwide distribution presently known from 70 plant families. This study isolated P. capitalensis from different host plants in northern Thailand, and determined their different life modes. Thirty strains of P. capitalensis

  13. Botrallin from the endophytic fungus Hyalodendriella sp ...

    use

    2011-12-12

    Dec 12, 2011 ... Bioassay-guided fractionation of the crude methanol extract of the mycelia from the endophytic fungus. Hyalodendriella sp. Ponipodef12, associated with the hybrid 'Neva' of Populus deltoides Marsh × P. nigra L., led to the isolation of one compound coded as P12-1 which was identified as botrallin (1,7-.

  14. Genomic DNA extraction and barcoding of endophytic fungi.

    Diaz, Patricia L; Hennell, James R; Sucher, Nikolaus J

    2012-01-01

    Endophytes live inter- and/or intracellularly inside healthy aboveground tissues of plants without causing disease. Endophytic fungi are found in virtually every vascular plant species examined. The origins of this symbiotic relationship between endophytes go back to the emergence of vascular plants. Endophytic fungi receive nutrition and protection from their hosts while the plants benefit from the production of fungal secondary metabolites, which enhance the host plants' resistance to herbivores, pathogens, and various abiotic stresses. Endophytic fungi have attracted increased interest as potential sources of secondary metabolites with agricultural, industrial, and medicinal use. This chapter provides detailed protocols for isolation of genomic DNA from fungal endophytes and its use in polymerase chain reaction-based amplification of the internal transcribed spacer region between the conserved flanking regions of the small and large subunit of ribosomal RNA for barcoding purposes.

  15. Mechanisms Involved in Nematode Control by Endophytic Fungi.

    Schouten, Alexander

    2016-08-04

    Colonization of plants by particular endophytic fungi can provide plants with improved defenses toward nematodes. Evidently, such endophytes can be important in developing more sustainable agricultural practices. The mechanisms playing a role in this quantitative antagonism are poorly understood but most likely multifactorial. This knowledge gap obstructs the progress regarding the development of endophytes or endophyte-derived constituents into biocontrol agents. In part, this may be caused by the fact that endophytic fungi form a rather heterogeneous group. By combining the knowledge of the currently characterized antagonistic endophytic fungi and their effects on nematode behavior and biology with the knowledge of microbial competition and induced plant defenses, the various mechanisms by which this nematode antagonism operates or may operate are discussed. Now that new technologies are becoming available and more accessible, the currently unresolved mechanisms can be studied in greater detail than ever before.

  16. Endophytic fungi reduce leaf-cutting ant damage to seedlings

    Bittleston, L. S.; Brockmann, F.; Wcislo, W.; Van Bael, S. A.

    2011-01-01

    Our study examines how the mutualism between Atta colombica leaf-cutting ants and their cultivated fungus is influenced by the presence of diverse foliar endophytic fungi (endophytes) at high densities in tropical leaf tissues. We conducted laboratory choice trials in which ant colonies chose between Cordia alliodora seedlings with high (Ehigh) or low (Elow) densities of endophytes. The Ehigh seedlings contained 5.5 times higher endophyte content and a greater diversity of fungal morphospecies than the Elow treatment, and endophyte content was not correlated with leaf toughness or thickness. Leaf-cutting ants cut over 2.5 times the leaf area from Elow relative to Ehigh seedlings and had a tendency to recruit more ants to Elow plants. Our findings suggest that leaf-cutting ants may incur costs from cutting and processing leaves with high endophyte loads, which could impact Neotropical forests by causing variable damage rates within plant communities. PMID:20610420

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

    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

  18. Beech cupules share endophytic fungi with leaves and twigs

    Tateno, Osamu; Hirose, Dai; Osono, Takashi; Takeda, Hiroshi

    2015-01-01

    Endophytic mycobiota on leaves, twigs and cupules of Fagus crenata were investigated using a culture-dependent method over a growing season to test the hypothesis that endophytic fungi of cupule (a woody phyllome) share some components of the endophytic fungal assemblages with both leaves and twigs. A total of 14 fungal taxa were isolated, and the most frequent taxon was Phomopsis sp., followed by Xylaria sp., Ascochyta fagi and Geniculosporium sp. The compositions of fungal assemblages of le...

  19. Consumption of Endophyte Infected Fescue During Gestation in Beef Cows

    Oliver, Katherine Rene

    2016-01-01

    Tall fescue is a widely grown, cool season grass prevalent in the eastern United States that is known for its resistance to abiotic and biotic stresses. A main reason for tall fescue's resistance to these stresses is attributed to the presence of a fungal endophyte. Unfortunately, this endophyte also adversely affects cattle production. Cows consuming the ergot alkaloids produced by these endophytes can exhibit decreased feed intake, growth performance, organ vasoconstriction, and increased...

  20. Antifungal activity and molecular identification of endophytic fungi ...

    Antifungal activity and molecular identification of endophytic fungi from the angiosperm Rhodomyrtus tomentosa. Juthatip Jeenkeawpieam, Souwalak Phongpaichit, Vatcharin Rukachaisirikul, Jariya Sakayaroj ...

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

    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.

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

    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

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

    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

  4. Fungal endophytes of sorghum in Burkina Faso

    Zida, E P; Thio, I G; Néya, B J

    2014-01-01

    A survey was conducted to assess the natural occurrence and distribution of fungal endophytes in sorghum in relation to plant performance in two distinct agro-ecological zones in Burkina Faso. Sorghum farm-saved seeds were sown in 48 farmers’ fields in Sahelian and North Sudanian zones to produce...... sorghum plants. In each field, leaf samples were collected from five well-developed (performing) and five less-developed (non-performing) plants at 3-5 leaf stage, while at plant maturity leaf, stem and root samples were collected from the same plants and fungal endophytes were isolated. A total of 39...... fungal species belonging to 25 genera were isolated. The most represented genera included Fusarium, Leptosphaeria, Curvularia, Nigrospora and Penicillium. The genera Fusarium and Penicillium occurred significantly higher in performing plants as compared to non-performing plants while the genera...

  5. Search for endophytic diazotrophs in barley seeds

    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.

  6. Bacterial endophytes enhance competition by invasive plants.

    Rout, Marnie E; Chrzanowski, Thomas H; Westlie, Tara K; DeLuca, Thomas H; Callaway, Ragan M; Holben, William E

    2013-09-01

    Invasive plants can alter soil microbial communities and profoundly alter ecosystem processes. In the invasive grass Sorghum halepense, these disruptions are consequences of rhizome-associated bacterial endophytes. We describe the effects of N2-fixing bacterial strains from S. halepense (Rout and Chrzanowski, 2009) on plant growth and show that bacteria interact with the plant to alter soil nutrient cycles, enabling persistence of the invasive. • We assessed fluxes in soil nutrients for ∼4 yr across a site invaded by S. halepense. We assayed the N2-fixing bacteria in vitro for phosphate solubilization, iron chelation, and production of the plant-growth hormone indole-3-acetic acid (IAA). We assessed the plant's ability to recruit bacterial partners from substrates and vertically transmit endophytes to seeds and used an antibiotic approach to inhibit bacterial activity in planta and assess microbial contributions to plant growth. • We found persistent alterations to eight biogeochemical cycles (including nitrogen, phosphorus, and iron) in soils invaded by S. halepense. In this context, three bacterial isolates solubilized phosphate, and all produced iron siderophores and IAA in vitro. In growth chamber experiments, bacteria were transmitted vertically, and molecular analysis of bacterial community fingerprints from rhizomes indicated that endophytes are also horizontally recruited. Inhibiting bacterial activity with antibiotics resulted in significant declines in plant growth rate and biomass, with pronounced rhizome reductions. • This work suggests a major role of endophytes on growth and resource allocation of an invasive plant. Indeed, bacterial isolate physiology is correlated with invader effects on biogeochemical cycles of nitrogen, phosphate, and iron.

  7. Interactions among endophytic bacteria and fungi: effects and ...

    Madhu

    The colonization of plants by putative endophytes has been visualized by using laser scanning confocal microscope (Coombs and Franco 2003). Endophytes promote the growth of plants in various ways, for example through secretion of plant growth regulators;. e.g. indole-acetic acid (Lee et al 2004), via phosphate-.

  8. Endophytic Fungi as Novel Resources of natural Therapeutics

    Maheshwari Rajamanikyam

    2017-08-01

    Full Text Available ABSTRACT Fungal endophytes constitute a major part of the unexplored fungal diversity. Endophytic fungi (EF are an important source for novel, potential and active metabolites. Plant-endophyte interaction and endophyte -endophyte interactions study provide insights into mutualism and metabolite production by fungi. Bioactive compounds produced by endophytes main function are helping the host plants to resist external biotic and abiotic stress, which benefit the host survival in return. These organisms mainly consist of members of the Ascomycota, Basidiomycota, Zygomycota and Oomycota. Recently, the genome sequencing technology has emerged as one of the most efficient tools that can provide whole information of a genome in a small period of time. Endophytes are fertile ground for drug discovery. EFare considered as the hidden members of the microbial world and represent an underutilized resource for new therapeutics and compounds. Endophytes are rich source of natural products displaying broad spectrum of biological activities like anticancer, antibacterial, antiviral, immunomodulatory, antidiabetic, antioxidant, anti-arthritis and anti-inflammatory.

  9. Plant growth promoting potential of endophytic bacteria isolated ...

    Endophytic microorganisms are able to promote plant growth through various mechanisms, such as production of plant hormones and antimicrobial substances, as well as to provide the soil with nutrients, for instance, inorganic phosphate. This study aimed to evaluate the potential of endophytic bacteria isolated from ...

  10. Induction of abiotic stress tolerance in plants by endophytic microbes.

    Lata, R; Chowdhury, S; Gond, S K; White, J F

    2018-04-01

    Endophytes are micro-organisms including bacteria and fungi that survive within healthy plant tissues and promote plant growth under stress. This review focuses on the potential of endophytic microbes that induce abiotic stress tolerance in plants. How endophytes promote plant growth under stressful conditions, like drought and heat, high salinity and poor nutrient availability will be discussed. The molecular mechanisms for increasing stress tolerance in plants by endophytes include induction of plant stress genes as well as biomolecules like reactive oxygen species scavengers. This review may help in the development of biotechnological applications of endophytic microbes in plant growth promotion and crop improvement under abiotic stress conditions. Increasing human populations demand more crop yield for food security while crop production is adversely affected by abiotic stresses like drought, salinity and high temperature. Development of stress tolerance in plants is a strategy to cope with the negative effects of adverse environmental conditions. Endophytes are well recognized for plant growth promotion and production of natural compounds. The property of endophytes to induce stress tolerance in plants can be applied to increase crop yields. With this review, we intend to promote application of endophytes in biotechnology and genetic engineering for the development of stress-tolerant plants. © 2018 The Society for Applied Microbiology.

  11. Antagonistic bioactivity of an endophytic bacterium isolated from ...

    Antagonistic bioactivity of an endophytic bacterium isolated from Epimedium brevicornu Maxim. R He, G Wang, X Liu, C Zhang, F Lin. Abstract. Endophytic bacteria are one of the most potential biological control agents in plant disease protection. The aim of this work was to evaluate the antimicrobial activities of a strain of ...

  12. Screening mycotoxins for quorum inhibition in a biocontrol bacterial endophyte

    Bacterial endophytes are used as biocontrol organisms for plant pathogens such as the maize endophyte Fusarium verticillioides and its production of fumonisin mycotoxins. However, such applications are not always predictable and efficient. Bacteria communicate via cell-dependent signals, which are r...

  13. Mechanisms Involved in Nematode Control by Endophytic Fungi

    Schouten, Sander

    2016-01-01

    Colonization of plants by particular endophytic fungi can provide plants with improved defenses toward nematodes. Evidently, such endophytes can be important in developing more sustainable agricultural practices. The mechanisms playing a role in this quantitative antagonism are poorly understood

  14. Endophytic bacteria with potential for bioremediation of petroleum ...

    Endophytic microorganisms live inside plants and show no apparent damage for the host. They often assist in plants' survival and facilitate their growth, or they can metabolize organic contaminants. This study aimed to isolate and identify the endophytic bacteria of plants present in impacted areas, as well as to test their ...

  15. Enhanced resistance to Spodoptera litura in endophyte infected cauliflower plants.

    Thakur, Abhinay; Kaur, Sanehdeep; Kaur, Amarjeet; Singh, Varinder

    2013-04-01

    Endophytic fungi, which live within host plant tissues without causing any visible symptom of disease, are important mediators of plant-herbivore interactions. These endophytes enhance resistance of host plant against insect herbivores mainly by productions of various alkaloid based defensive compounds in the plant tissue or through alterations of plant nutritional quality. Two endophytic fungi, i.e., Nigrospora sp. and Cladosporium sp., were isolated from Tinospora cordifolia (Thunb.) Miers, a traditional indian medicinal plant. Cauliflower (Brassica oleracea L.) plants were inoculated with these two endophytic fungi. The effect of endophyte infected and uninfected cauliflower plants were measured on the survival and development of Spodoptera litura (Fab.), a polyphagous pest. Endophyte infected cauliflower plants showed resistance to S. litura in the form of significant increase in larval and pupal mortality in both the fungi. Inhibitory effects of endophytic fungi also were observed on adult emergence, longevity, reproductive potential, as well as hatchability of eggs. Thus, it is concluded that antibiosis to S. litura could be imparted by artificial inoculation of endophytes and this could be used to develop alternative ecologically safe control strategies.

  16. Endophytes: exploitation as a tool in plant protection

    Devanushi Dutta

    2014-10-01

    Full Text Available Endophytes are symptomless fungal or bacterial microorganisms found in almost all living plant species reported so far. They are the plant-associated microbes that form symbiotic association with their host plants by colonizing the internal tissues, which has made them valuable for agriculture as a tool in improving crop performance. Many fungal endophytes produce secondary metabolites such as auxin, gibberellin etc that helps in growth and development of the host plant. Some of these compounds are antibiotics having antifungal, antibacterial and insecticidal properties, which strongly inhibit the growth of other microorganisms, including plant pathogens. This article reviews the endophyte isolated from different plants, mode of endophytic infection and benefits derived by the host plant as a result of endophytism.

  17. Rethinking production of Taxol® (paclitaxel) using endophyte biotechnology.

    Kusari, Souvik; Singh, Satpal; Jayabaskaran, Chelliah

    2014-06-01

    Taxol® (generic name paclitaxel) represents one of the most clinically valuable natural products known to mankind in the recent past. More than two decades have elapsed since the notable discovery of the first Taxol®-producing endophytic fungus, which was followed by a plethora of reports on other endophytes possessing similar biosynthetic potential. However, industrial-scale Taxol® production using fungal endophytes, although seemingly promising, has not seen the light of the day. In this opinion article, we embark on the current state of knowledge on Taxol® biosynthesis focusing on the chemical ecology of its producers, and ask whether it is actually possible to produce Taxol® using endophyte biotechnology. The key problems that have prevented the exploitation of potent endophytic fungi by industrial bioprocesses for sustained production of Taxol® are discussed. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Endophytic actinobacteria of medicinal plants: diversity and bioactivity.

    Golinska, Patrycja; Wypij, Magdalena; Agarkar, Gauravi; Rathod, Dnyaneshwar; Dahm, Hanna; Rai, Mahendra

    2015-08-01

    Endophytes are the microorganisms that exist inside the plant tissues without having any negative impact on the host plant. Medicinal plants constitute the huge diversity of endophytic actinobacteria of economical importance. These microbes have huge potential to synthesis of numerous novel compounds that can be exploited in pharmaceutical, agricultural and other industries. It is of prime importance to focus the present research on practical utilization of this microbial group in order to find out the solutions to the problems related to health, environment and agriculture. An extensive characterization of diverse population of endophytic actinobacteria associated with medicinal plants can provide a greater insight into the plant-endophyte interactions and evolution of mutualism. In the present review, we have discussed the diversity of endophytic actinobacteria of from medicinal plants their multiple bioactivities.

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

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

    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

  1. Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria.

    Beltran-Garcia, Miguel J; White, James F; Prado, Fernanda M; Prieto, Katia R; Yamaguchi, Lydia F; Torres, Monica S; Kato, Massuo J; Medeiros, Marisa H G; Di Mascio, Paolo

    2014-11-06

    Plants form symbiotic associations with endophytic bacteria within tissues of leaves, stems, and roots. It is unclear whether or how plants obtain nitrogen from these endophytic bacteria. Here we present evidence showing nitrogen flow from endophytic bacteria to plants in a process that appears to involve oxidative degradation of bacteria. In our experiments we employed Agave tequilana and its seed-transmitted endophyte Bacillus tequilensis to elucidate organic nitrogen transfer from (15)N-labeled bacteria to plants. Bacillus tequilensis cells grown in a minimal medium with (15)NH4Cl as the nitrogen source were watered onto plants growing in sand. We traced incorporation of (15)N into tryptophan, deoxynucleosides and pheophytin derived from chlorophyll a. Probes for hydrogen peroxide show its presence during degradation of bacteria in plant tissues, supporting involvement of reactive oxygen in the degradation process. In another experiment to assess nitrogen absorbed as a result of endophytic colonization of plants we demonstrated that endophytic bacteria potentially transfer more nitrogen to plants and stimulate greater biomass in plants than heat-killed bacteria that do not colonize plants but instead degrade in the soil. Findings presented here support the hypothesis that some plants under nutrient limitation may degrade and obtain nitrogen from endophytic microbes.

  2. Endophytic Fungal Diversity in Medicinal Plants of Western Ghats, India

    Monnanda Somaiah Nalini

    2014-01-01

    Full Text Available Endophytes constitute an important component of microbial diversity, and in the present investigation, seven plant species with rich ethnobotanical uses representing six families were analyzed for the presence of endophytic fungi from their natural habitats during monsoon (May/June and winter (November/December seasons of 2007. Fungal endophytes were isolated from healthy plant parts such as stem, root, rhizome, and inflorescence employing standard isolation methods. One thousand five hundred and twenty-nine fungal isolates were obtained from 5200 fragments. Stem fragments harbored more endophytes (80.37% than roots (19.22%. 31 fungal taxa comprised of coelomycetes (65%, hyphomycetes (32%, and ascomycetes (3%. Fusarium, Acremonium, Colletotrichum, Chaetomium, Myrothecium, Phomopsis, and Pestalotiopsis spp. were commonly isolated. Diversity indices differed significantly between the seasons (P<0.001. Species richness was greater for monsoon isolations than winter. Host specificity was observed for few fungal endophytes. UPGMA cluster analysis grouped the endophytes into distinct clusters on the basis of genetic distance. This study is the first report on the diversity and host-specificity of endophytic fungal taxa were from the semi evergreen forest type in Talacauvery subcluster of Western Ghats.

  3. [Isolation and physiological characteristics of endophytic actinobacteria from medicinal plants].

    Du, Huijing; Su, Jing; Yu, Liyan; Zhang, Yuqin

    2013-01-04

    To isolate, incubate and characterize cultivable endophytic antinobacteria from medicinal plants, and analyze the diversity of the endophytic antinobacteria, then explore the novel microbial resources. Ten media were used to isolate endophytic antinobacteria from 37 fresh medicinal plant tissue samples. The optimal cultivation conditions for endophytic antinobacteria were determined by comparison. Based on the morphology of the colonies and cells of the new isolates, we chose 174 isolates to analyze their 16S rRNA gene sequences and the diversity of the medicinal plant endophytic antinobacteria. The physiological characteristics of 27 representative strains were studied using Biolog GEN III MicroPlates, API 50CH and API ZYM kits. In total 940 endophytics affiliated to 47 genera of 30 families were isolated, among which more than 600 actinobacteria belonged to 34 genera and 7 unknown taxa. Good growth of the endophytic antinobacteria on PYG (peptone-yeast-glycerol) medium with pH 7.2 at 28-32 degrees C was observed. Physiological characteristics differences of these isolates related to their phylogenetic relationships. Greater differences were shown among the strains from the same host plants than those from differ,ent plants grown in the same area. There are great diverse endophytic actinobacteria inside the medicinal plants. No direct relationship of the endophytic actinobacteria from medicinal plants with the host plants in the sole carbon source utilization, fermentation of carbon sources to produce acid and the enzyme activities was found, while it seemed that the physiological characteristics of the isolates related to the geographical distribution of their host.

  4. Sesquiterpenes from the Endophyte Glomerella cingulata.

    Liu, Yunbao; Li, Yong; Liu, Zhen; Li, Li; Qu, Jing; Ma, Shuanggang; Chen, Ridao; Dai, Jungui; Yu, Shishan

    2017-10-27

    From the cultured endophytic fungus Glomerella cingulata isolated from a toxic plant, Gelsemium elegans, one new phenanthrene (1), four new sesquiterpenes (2-5), and three known sesquiterpenes (6-8) were isolated. Their structures were elucidated using spectroscopic methods. Based on the ECD calculations, the absolute configurations of the new compounds were determined. Compounds 2, 4, and 5 inhibited lipopolysaccharide (LPS)-induced NO production in BV2 cells by 50.6, 36.1, and 29.4%, respectively, at 1 μM (positive control curcumin, IC 50 = 4.0 μM).

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

    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

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

    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.

  7. Biodegradation of Polyester Polyurethane by Endophytic Fungi▿

    Russell, Jonathan R.; Huang, Jeffrey; Anand, Pria; Kucera, Kaury; Sandoval, Amanda G.; Dantzler, Kathleen W.; Hickman, DaShawn; Jee, Justin; Kimovec, Farrah M.; Koppstein, David; Marks, Daniel H.; Mittermiller, Paul A.; Núñez, Salvador Joel; Santiago, Marina; Townes, Maria A.; Vishnevetsky, Michael; Williams, Neely E.; Vargas, Mario Percy Núñez; Boulanger, Lori-Ann; Bascom-Slack, Carol; Strobel, Scott A.

    2011-01-01

    Bioremediation is an important approach to waste reduction that relies on biological processes to break down a variety of pollutants. This is made possible by the vast metabolic diversity of the microbial world. To explore this diversity for the breakdown of plastic, we screened several dozen endophytic fungi for their ability to degrade the synthetic polymer polyester polyurethane (PUR). Several organisms demonstrated the ability to efficiently degrade PUR in both solid and liquid suspensions. Particularly robust activity was observed among several isolates in the genus Pestalotiopsis, although it was not a universal feature of this genus. Two Pestalotiopsis microspora isolates were uniquely able to grow on PUR as the sole carbon source under both aerobic and anaerobic conditions. Molecular characterization of this activity suggests that a serine hydrolase is responsible for degradation of PUR. The broad distribution of activity observed and the unprecedented case of anaerobic growth using PUR as the sole carbon source suggest that endophytes are a promising source of biodiversity from which to screen for metabolic properties useful for bioremediation. PMID:21764951

  8. Antimicrobial chemical constituents from endophytic fungus Phomasp.

    Hidayat Hussain; Siegfried Draeger; Barbara Schulz; Karsten Krohn; Ines Kock; Ahmed Al-Harrasi; Ahmed Al-Rawahi; Ghulam Abbas; Ivan R Green; Afzal Shah; Amin Badshah; Muhammad Saleem

    2014-01-01

    Objective:To evaluate the antimicrobial potential of different extracts of the endophytic fungus Phomasp. and the tentative identification of their active constituents.Methods:The extract and compounds were screened for antimicrobial activity using theAgarWellDiffusionMethod. Four compounds were purified using column chromatography and their structures were assigned using1H and13CNMR spectra,DEPT,2DCOSY,HMQC andHMBC experiments.Results:The ethyl acetate fraction ofPhomasp. showed good antifungal, antibacterial, and algicidal properties.One new dihydrofuran derivative, named phomafuranol(1), together with three known compounds, phomalacton(2),(3R)-5-hydroxymellein(3) and emodin(4) were isolated from the ethyl acetate fraction ofPhomasp.Preliminary studies indicated that phomalacton(2) displayed strong antibacterial, good antifungal and antialgal activities.Similarly(3R)-5-hydroxymellein (3) and emodin(4) showed good antifungal, antibacterial and algicidal properties.Conclusions:Antimicrobial activities of the ethyl acetate fraction of the endophytic fungusPhomasp. and isolated compounds clearly demonstrate thatPhomasp. and its active compounds represent a great potential for the food, cosmetic and pharmaceutical industries.

  9. Endophytes in commercial micropropagation - friend or foe?

    Rödel, Philipp

    2016-07-01

    Full Text Available Medicinal and aromatic plants are superorganisms like all plant species- naturally colonized by bacteria, fungi and protists. Micropropagated plants are facing different challenges under in vitro and ex vitro conditions: Mixotrophic growth under low light conditions on artificial nutrient media, poor gas exchange in small vessels, abiotic stress, bad rooting, transplanting stress, low survival rate during acclimatization in greenhouse. The use of endophytes in micropropagation can improve plant growth, yield, and health and induce tolerance to abiotic and biotic stress. A tool for the use of competent endophytes in micropropagation under in vitro and ex vitro conditions is “biotization” of plantlets with useful bacterial and fungal inocula. Fungal inocula which are used commercially are e.g. arbuscular mycorrhizal fungi in form of spores and extraradical mycelium on different carrier materials like expanded clay, vermiculite, sand or peat. Furthermore representatives of the root fungal genus Trichoderma are applied as spores formulated in powder. Plantgrowth promoting rhizobacteria of the important genera Bacillus, Pseudomonas, Azospirillum and Azotobacter in form of lyophilised endospores/bacterial cells in powder or liquid formulation are also available on the market.

  10. Banana Musa tissue culture plants enhanced by endophytic fungi

    Mo

    Merging biotechnology with biological control: Banana Musa tissue culture plants enhanced by endophytic .... While working in the laminar flow cabinet, sterile filter papers were placed in ..... University of Bonn, Bonn, Germany. Niere, B., 2001.

  11. Characterization of indole acetic acid endophyte producers in ...

    Valued Acer Customer

    2015-02-18

    Feb 18, 2015 ... the number and identity of the isolated endophyte phytobacteria in L. gibba plants .... mL was taken from each sample and placed on plates containing ... databases using Basic Logical Alignment Search Tool analysis ...

  12. Antioxidants in mangrove plants and endophytic fungal associations

    Ravindran, C.; Naveenan, T.; Varatharajan, G.R.; Rajasabapathy, R.; Meena, R.M.

    different mangrove species and the predominant endophytic fungus Aspergillus flavus were analyzed using various in vitro assay systems (such as iron chelating capacity, reducing power, and hydroxyl radicals/ hydrogen peroxide/l-diphenyl-2-picrylhydrazyl...

  13. 7 CFR 201.58d - Fungal endophyte test.

    2010-01-01

    ... staining solution. Slightly crush the seeds. Use caution to prevent carryover hyphae of fungal endophyte... compound microscope at 100-400x magnification, scoring a seed as positive if any identifiable hyphae are...

  14. Endophytic bacteria with plant growth promoting and biocontrol abilities

    Malfanova, Natalia V.

    2013-01-01

    Since global food insecurity is one of the major problems faced by humanity, there is a necessity to increase plant productivity. For this, biofungicides and biofertilizers present an ecologically friendly alternative to their chemical counterparts. Among these bioinoculants, endophytic bacteria

  15. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity

    Puji Astuti; Sudarsono Sudarsono; Khoirun Nisak; Giri Wisnu Nugroho

    2014-01-01

    Purpose: Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Methods: Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatograp...

  16. Bacterial endophytes of perennial crops for management of plant disease

    Melnick, Rachel L.; Bailey, B.A.; Backman, Paul A.

    2013-01-01

    Metadata only record Bacterial endophytes, microorganisms which inhabit the internal tissues of plants, can suppress disease and are often used as a biological control in annual crops. Less research, however, has been applied to the use of bacterial endophytes to prevent disease in perennial crops, which presents a more complex challenge. However, exploration of their potential as a biological control in perennial crops has been limited. This chapter assembles current knowledge on the subj...

  17. Differential methods of localisation of fungal endophytes in the seagrasses

    S. Raja

    2016-07-01

    Full Text Available Sections of three seagrass species (Halophila ovalis, Cymodocea serrulata and Halodule pinifolia were assessed for endophytes based on differential staining using light and fluorescence microscopy method. Acridine orange and aniline blue detected endophytic fungi in 20% and 10% of the segments, respectively, whereas lactophenol cotton blue was more sensitive to detect the fungal hyphae in 70% of the segments. Hyphae were the principal fungal structures generally observed under the cuticle, within the epidermal cells, mesophyll (Parenchyma cells and occasionally within the vascular tissue that varied in type, size and location within the leaf tissue. Present study also recorded the sporulation for the first time from the seagrass endophytes. Successfully amplified products of the ITS region of endophytic fungal DNA, directly from seagrass tissue and also from culture-dependent fungal DNA clearly depicted the presence of endophytic fungi in H. ovalis with two banding patterns (903 and 1381 bp confirming the presence of two dominant fungal genera. The fingerprinting of endophytic fungal community within the seagrass tissue was assessed using denaturing gradient gel electrophoresis (DGGE that derived with multiple bands that clarified the presence of more than one taxon within the seagrass tissue.

  18. Antimicrobial fungal endophytes from the botanical medicine goldenseal (Hydrastis canadensis).

    Egan, Joseph M; Kaur, Amninder; Raja, Huzefa A; Kellogg, Joshua J; Oberlies, Nicholas H; Cech, Nadja B

    2016-09-01

    The potential of fungal endophytes to alter or contribute to plant chemistry and biology has been the topic of a great deal of recent interest. For plants that are used medicinally, it has been proposed that endophytes might play an important role in biological activity. With this study, we sought to identify antimicrobial fungal endophytes from the medicinal plant goldenseal ( Hydrastis canadensis L., Ranunculaceae), a plant used in traditional medicine to treat infection. A total of 23 fungal cultures were obtained from surface-sterilized samples of H. canadensis roots, leaves and seeds. Eleven secondary metabolites were isolated from these fungal endophytes, five of which had reported antimicrobial activity. Hydrastis canadensis plant material was then analyzed for the presence of fungal metabolites using liquid chromatography coupled to high resolving power mass spectrometry. The antimicrobial compound alternariol monomethyl ether was detected both as a metabolite of the fungal endophyte Alternaria spp. isolated from H. canadensis seeds, and as a component of an extract from the H. canadensis seed material. Notably, fungi of the Alternaria genus were isolated from three separate accessions of H. canadensis plant material collected in a time period spanning 5 years. The concentration of alternariol monomethyl ether (991 mg/kg in dry seed material) was in a similar range to that previously reported for metabolites of ecologically important fungal endophytes. The seed extracts themselves, however, did not possess antimicrobial activity.

  19. Seasonal variation of bacterial endophytes in urban trees

    Shu Yi eShen

    2015-05-01

    Full Text Available Bacterial endophytes, non-pathogenic bacteria residing within plants, contribute to the growth and development of plants and their ability to adapt to adverse conditions. In order to fully exploit the capabilities of these bacteria, it is necessary to understand the extent to which endophytic communities vary between species and over time. The endophytes of Acer negundo, Ulmus pumila and Ulmus parvifolia were sampled over three seasons and analyzed using culture dependent and independent methods (culture on two media, terminal restriction fragment length polymorphism, and tagged pyrosequencing of 16S ribosomal amplicons. The majority of culturable endophytes isolated were Actinobacteria, and all the samples harbored Bacillus, Curtobacterium, Frigoribacterium, Methylobacterium, Paenibacilllus and Sphingomonas species. Regardless of culture medium used, only the culturable communities obtained in the winter for A. negundo could be distinguished from those of Ulmus spp.. In contrast, the nonculturable communities were dominated by Proteobacteria and Actinobacteria, particularly Erwinia, Ralstonia and Sanguibacter spp.. The presence and abundance of various bacterial classes and phyla changed with the changing seasons. Multivariate analysis on the culture independent data revealed significant community differences between the endophytic communities of A. negundo and Ulmus spp., but overall season was the main determinant of endophytic community structure. This study suggests investigations of the studies ofendophytic populations of urban trees should expect to find significant seasonal and species-specific community differences and sampling should proceed accordingly.

  20. Molecular dynamics in germinating, endophyte-colonized quinoa seeds

    2017-01-01

    Aims The pseudo-cereal quinoa has an outstanding nutritional value. Seed germination is unusually fast, and plant tolerance to salt stress exceptionally high. Seemingly all seeds harbor bacterial endophytes. This work examines mitogen-activated protein kinase (MAPK) activities during early development. It evaluates possible contribution of endophytes to rapid germination and plant robustness. Methods MAPK activities were monitored in water- and NaCl-imbibed seeds over a 4-h-period using an immunoblot-based approach. Cellulolytic and pectinolytic abilities of bacteria were assessed biochemically, and cellular movement, biofilm, elicitor and antimicrobial compound synthesis genes sequenced. GyrA-based, cultivation-independent studies provided first insight into endophyte diversity. Results Quinoa seeds and seedlings exhibit remarkably complex and dynamic MAPK activity profiles. Depending on seed origin, variances exist in MAPK patterns and probably also in endophyte assemblages. Mucilage-degrading activities enable endophytes to colonize seed surfaces of a non-host species, chia, without apparent adverse effects. Conclusions Owing to their motility, cell wall-loosening and elicitor-generating abilities, quinoa endophytes have the potential to drive cell expansion, move across cell walls, generate damage-associated molecular patterns and activate MAPKs in their host. Bacteria may thus facilitate rapid germination and confer a primed state directly upon seed rehydration. Transfer into non-native crops appears both desirable and feasible. PMID:29416180

  1. Phytohormones in plant-endophyte interactions: investigating the role of these compounds in the recruitment of tomato root fungal endophytes

    Manzotti, Andrea; Jørgensen, Hans Jørgen Lyngs; Collinge, David B.

    in this interaction, but little is known about the specific way by which they influence the recruitment and the colonization of the host tissues. The aim of the current project is to go deeper into the role of these signalling compounds in plant-endophyte interactions. The isolation of endophytic fungi from tomato......-colonization frequency appears to be influenced by the presence/absence of specific phytohormones. In order to obtain a deeper understanding of the role of these compounds in the plant-endophyte interaction, the selected isolates are currently being screened using confocal microscopy and qPCR in order to find candidates...... whose colonization rate is critically affected by the phytohormones of interest. A transcriptomic analysis of tomato plants inoculated with the isolates selected from the screening will provide further clues as to which physiological mechanisms, associated with endophyte recruitment, are influenced...

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

    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 ...

  3. Culturable bacterial endophytes isolated from Mangrove tree (Rhizophora apiculata Blume) enhance seedling growth in Rice

    Deivanai, Subramanian; Bindusara, Amitraghata Santhanam; Prabhakaran, Guruswamy; Bhore, Subhash Janardhan

    2014-01-01

    Background: Endophytic bacteria do have several potential applications in medicine and in other various sectors of biotechnology including agriculture. Bacterial endophytes need to be explored for their potential applications in agricultural biotechnology. One of the potential applications of bacterial endophytes in agricultural is to enhance the growth of the agricultural crops. Hence, this study was undertaken to explore the plant growth promoting potential application of bacterial endophyt...

  4. Bioactivity of fungal endophytes as a function of endophyte taxonomy and the taxonomy and distribution of their host plants.

    Sarah J Higginbotham

    Full Text Available Fungal endophytes--fungi that grow within plant tissues without causing immediate signs of disease--are abundant and diverse producers of bioactive secondary metabolites. Endophytes associated with leaves of tropical plants are an especially exciting and relatively untapped source of novel compounds. However, one major challenge in drug discovery lies in developing strategies to efficiently recover highly bioactive strains. As part of a 15-year drug discovery project, foliar endophytes were isolated from 3198 plant samples (51 orders, 105 families and at least 232 genera of angiosperms and ferns collected in nine geographically distinct regions of Panama. Extracts from culture supernatants of >2700 isolates were tested for bioactivity (in vitro percent inhibition of growth, % IG against a human breast cancer cell line (MCF-7 and the causative agents of malaria, leishmaniasis, and Chagas' disease. Overall, 32.7% of endophyte isolates were highly active in at least one bioassay, including representatives of diverse fungal lineages, host lineages, and collection sites. Up to 17% of isolates tested per assay were highly active. Most bioactive strains were active in only one assay. Fungal lineages differed in the incidence and degree of bioactivity, as did fungi from particular plant taxa, and greater bioactivity was observed in endophytes isolated from plants in cloud forests vs. lowland forests. Our results suggest that using host taxonomy and forest type to tailor plant collections, and selecting endophytes from specific orders or families for cultivation, will markedly increase the efficiency and efficacy of discovering bioactive metabolites for particular pharmaceutical targets.

  5. Resources and testing of endophyte-infected germplasm in national grass repository collections

    A. D. Wilson

    1996-01-01

    Clavicipitaceous endophytes have been known to exist in grasses since the discovery of an endophyte in seeds of damel (Lolium temulentum L.) by Vogl in 1898 (26). The oldest known specimens of damel with endophytic mycelium were seeds retrieved from a pharoah's tomb in an Egyptian pyramid dating back to 3400 B.C. (16). Subsequent work by...

  6. Diversity and Biological Activities of Endophytic Fungi Associated with Micropropagated Medicinal Plant Echinacea purpurea (L.) Moench

    2012-08-01

    1105 Diversity and Biological Activities of Endophytic Fungi Associated with Micropropagated Medicinal Plant Echinacea purpurea (L.) Moench Luiz H...fungal community and micropropagated clones of E. purpurea was re-established after acclimatization to soil and the endophytic fungi produced compounds...Diversity and Biological Activities of Endophytic Fungi Associated with Micropropagated Medicinal Plant Echinacea purpurea (L.) Moench 5a. CONTRACT

  7. Properties of bacterial endophytes and their proposed role in plant growth

    Hardoim, P.R.; Overbeek, van L.S.; Elsas, van J.D.

    2008-01-01

    Bacterial endophytes live inside plants for at least part of their life cycle. Studies of the interaction of endophytes with their host plants and their function within their hosts are important to address the ecological relevance of endophytes. The modulation of ethylene levels in plants by

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

    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.

  9. Fungi with multifunctional lifestyles: endophytic insect pathogenic fungi.

    Barelli, Larissa; Moonjely, Soumya; Behie, Scott W; Bidochka, Michael J

    2016-04-01

    This review examines the symbiotic, evolutionary, proteomic and genetic basis for a group of fungi that occupy a specialized niche as insect pathogens as well as endophytes. We focus primarily on species in the genera Metarhizium and Beauveria, traditionally recognized as insect pathogenic fungi but are also found as plant symbionts. Phylogenetic evidence suggests that these fungi are more closely related to grass endophytes and diverged from that lineage ca. 100 MYA. We explore how the dual life cycles of these fungi as insect pathogens and endophytes are coupled. We discuss the evolution of insect pathogenesis while maintaining an endophytic lifestyle and provide examples of genes that may be involved in the transition toward insect pathogenicity. That is, some genes for insect pathogenesis may have been co-opted from genes involved in endophytic colonization. Other genes may be multifunctional and serve in both lifestyle capacities. We suggest that their evolution as insect pathogens allowed them to effectively barter a specialized nitrogen source (i.e. insects) with host plants for photosynthate. These ubiquitous fungi may play an important role as plant growth promoters and have a potential reservoir of secondary metabolites.

  10. Antimicrobial activities of endophytic fungi isolated from Ophiopogon japonicus (Liliaceae).

    Liang, Hanqiao; Xing, Yongmei; Chen, Juan; Zhang, Dawei; Guo, Shunxing; Wang, Chunlan

    2012-11-28

    Drug resistance in bacteria has become a global concern and the search for new antibacterial agents is urgent and ongoing. Endophytes provide an abundant reservoir of bioactive metabolites for medicinal exploitation, and an increasing number of novel compounds are being isolated from endophytic fungi. Ophiopogon japonicus, containing compounds with antibacterial activity, is a traditional Chinese medicinal plant used for eliminating phlegm, relieving coughs, latent heat in the lungs, and alleviating diabetes mellitus. We investigated the antimicrobial activities of 30 strains of O. japonicus. Fungal endophytes were isolated from roots and stems of O. japonicus collected from Chongqing City, southwestern China. Mycelial extracts (MC) and fermentation broth (FB) were tested for antimicrobial activity using peptide deformylase (PDF) inhibition fluorescence assays and MTT cell proliferation assays. A total of 30 endophytic strains were isolated from O. japonicus; 22 from roots and eight from stems. 53.33% of the mycelial extracts (MC) and 33.33% of the fermentation broths (FB) displayed potent inhibition of PDF. 80% of MC and 33.33% of FB significantly inhibited Staphylococcus aureus. 70% of MC and 36.67% of FB showed strong activities against Cryptococcus neoformans. None showed influence on Escherichia coli. The secondary metabolites of endophytic fungi from O. japonicus are potential antimicrobial agents.

  11. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity.

    Astuti, Puji; Sudarsono, Sudarsono; Nisak, Khoirun; Nugroho, Giri Wisnu

    2014-12-01

    Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatography (TLC) - bioautography was performed to localize the bioactive components within the extract. TLC visualization detection reagents were used to preliminary analyze phytochemical groups of the bioactive compounds. Three endophytic fungi were obtained, two of them showed promising potential. Agar diffusion method showed that endophytic fungi CAL-2 exhibited antimicrobial activity against P. aeruginosa, B. subtilis, S. aureus and S. thypi, whilst CAS-1 inhibited the growth of B. subtilis. TLC bioautography of ethyl acetate extract of CAL-2 revealed at least three bands exhibited antimicrobial activity and at least two bands showed inhibition of B. subtilis growth. Preliminary analysis of the crude extracts suggests that bioactive compounds within CAL-2 extract are terpenoids, phenolics and phenyl propanoid compounds whilst the antimicrobial agents within CAS-1 extract are terpenoids, propylpropanoids, alkaloids or heterocyclic nitrogen compounds. These data suggest the potential of endophytic fungi of C. amboinicus as source for antimicrobial agents.

  12. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity

    Puji Astuti

    2014-12-01

    Full Text Available Purpose: Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Methods: Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatography (TLC - bioautography was performed to localize the bioactive components within the extract. TLC visualization detection reagents were used to preliminary analyze phytochemical groups of the bioactive compounds. Results: Three endophytic fungi were obtained, two of them showed promising potential. Agar diffusion method showed that endophytic fungi CAL-2 exhibited antimicrobial activity against P. aeruginosa, B. subtilis, S. aureus and S. thypi, whilst CAS-1 inhibited the growth of B. subtilis. TLC bioautography of ethyl acetate extract of CAL-2 revealed at least three bands exhibited antimicrobial activity and at least two bands showed inhibition of B. subtilis growth. Preliminary analysis of the crude extracts suggests that bioactive compounds within CAL-2 extract are terpenoids, phenolics and phenyl propanoid compounds whilst the antimicrobial agents within CAS-1 extract are terpenoids, propylpropanoids, alkaloids or heterocyclic nitrogen compounds. Conclusion: These data suggest the potential of endophytic fungi of C. amboinicus as source for antimicrobial agents.

  13. Exploitation of endophytic fungus as a potential source of biofuel

    Nawed Anjum

    2016-06-01

    Full Text Available Biofuel demand is unquestionable in order to reduce greenhouse gaseous emission which can lead to climatic changes and global warming effect. Finding sufficient supply of clean energy for the upcoming is one of the society’s most daunting challenges and is directly linked with global stability, economic prosperity and quality of life. Endophytic microbes reside in the healthy part of the plant without causing any symptoms of disease. It is well known that the endophytic microbes produces wide variety of bioactive compound having, antibacterial, antifungal, antiviral, antitumor, antioxidant, antiinflammatory, immunosuppressive drugs, and volatile organic compounds having similarity with conventional diesel fuel. Now the endophytic fungi, have also been known to possess a suitable lipid matrix at high concentrations and volatile organic compounds having similarity with conventional diesel fuel that make them promising sources for next generation biofuels. This would be more efficient and having lesser number of biosynthetic steps in production, can be brought to immediate use in the existing internal combustion engines without taking about any major modification in automobile design. The present article therefore aims to review the current status of research in the field of alternative source of energy emphasizing endophytic fungi as a source of biofuel precursor, in order to encourage and generate interest among research groups across India and the world for initiating and undertaking more enthusiastic and intensive research activity on endophytic fungi from the Indian subcontinent having the potential to make fuel-related hydrocarbons.

  14. Endohyphal bacterium enhances production of indole-3-acetic acid by a foliar fungal endophyte.

    Michele T Hoffman

    Full Text Available Numerous plant pathogens, rhizosphere symbionts, and endophytic bacteria and yeasts produce the important phytohormone indole-3-acetic acid (IAA, often with profound effects on host plants. However, to date IAA production has not been documented among foliar endophytes -- the diverse guild of primarily filamentous Ascomycota that live within healthy, above-ground tissues of all plant species studied thus far. Recently bacteria that live within hyphae of endophytes (endohyphal bacteria have been detected, but their effects have not been studied previously. Here we show not only that IAA is produced in vitro by a foliar endophyte (here identified as Pestalotiopsis aff. neglecta, Xylariales, but that IAA production is enhanced significantly when the endophyte hosts an endohyphal bacterium (here identified as Luteibacter sp., Xanthomonadales. Both the endophyte and the endophyte/bacterium complex appear to rely on an L-tryptophan dependent pathway for IAA synthesis. The bacterium can be isolated from the fungus when the symbiotic complex is cultivated at 36°C. In pure culture the bacterium does not produce IAA. Culture filtrate from the endophyte-bacterium complex significantly enhances growth of tomato in vitro relative to controls and to filtrate from the endophyte alone. Together these results speak to a facultative symbiosis between an endophyte and endohyphal bacterium that strongly influences IAA production, providing a new framework in which to explore endophyte-plant interactions.

  15. Arbuscular mycorrhizal fungus inoculation reduces the drought-resistance advantage of endophyte-infected versus endophyte-free Leymus chinensis.

    Liu, Hui; Chen, Wei; Wu, Man; Wu, Rihan; Zhou, Yong; Gao, Yubao; Ren, Anzhi

    2017-11-01

    Grasses can be infected simultaneously by endophytic fungi and arbuscular mycorrhizal (AM) fungi. In this study, we tested the hypothesis that endophyte-associated drought resistance of a native grass was affected by an AM fungus. In a greenhouse experiment, we compared the performance of endophyte-infected (EI) and endophyte-free (EF) Leymus chinensis, a dominant species native to the Inner Mongolia steppe, under altered water and AM fungus availability. The results showed that endophyte infection significantly increased drought resistance of the host grass, but the beneficial effects were reduced by AM fungus inoculation. In the mycorrhizal-non-inoculated (MF) treatment, EI plants accumulated significantly more biomass, had greater proline and total phenolic concentration, and lower malondialdehyde concentration than EF plants. In the mycorrhizal-inoculation (MI) treatment, however, no significant difference occurred in either growth or physiological characters measured between EI and EF plants. AM fungus inoculation enhanced drought resistance of EF plants but had no significant effect on drought resistance of EI plants, thus AM fungus inoculation reduced the difference between EI and EF plants. Our findings highlight the importance of interactions among multiple microorganisms for plant performance under drought stress.

  16. Secondary metabolites from the endophytic fungus Talaromyces pinophilus.

    Vinale, F; Nicoletti, R; Lacatena, F; Marra, R; Sacco, A; Lombardi, N; d'Errico, G; Digilio, M C; Lorito, M; Woo, S L

    2017-08-01

    Endophytic fungi have a great influence on plant health and growth, and are an important source of bioactive natural compounds. Organic extracts obtained from the culture filtrate of an endophytic strain of Talaromyces pinophilus isolated from strawberry tree (Arbutus unedo) were studied. Metabolomic analysis revealed the presence of three bioactive metabolites, the siderophore ferrirubin, the platelet-aggregation inhibitor herquline B and the antibiotic 3-O-methylfunicone. The latter was the major metabolite produced by this strain and displayed toxic effects against the pea aphid Acyrthosiphon pisum (Homoptera Aphidiidae). This toxicity represents an additional indication that the widespread endophytic occurrence of T. pinophilus may be related to a possible role in defensive mutualism. Moreover, the toxic activity on aphids could promote further study on 3-O-methylfunicone, or its derivatives, as an alternative to synthetic chemicals in agriculture.

  17. Antagonistic Bioactivity of Endophytic Actinomycetes Isolated from Medicinal Plants

    M. Gangwar

    2011-10-01

    Full Text Available Endophytic actinomycetes are promising biocontrol agents for use in agriculture and have been isolated from various plant species. In the present study, 40 endophytic actinomycetes were isolated from roots, stems and leaves of three medicinal plants viz. Aloe vera, Mentha arvensis and Ocimum sanctum. The identification revealed that the majority of the isolates were Streptomyces spp. and the rest were identified as Saccharopolyspora spp., Micromonospora spp. and Actinopolyspora spp. The dual tests revealed that nine endophytic actinomycete isolates displayed a wide spectrum activity against nine fungal phytopathogens. Out of 8 isolates, 90% inhibited the growth of at least one or more phytopathogenic fungi and Saccharopolyspora 0-9 (Out of 8 isolates, 90% inhibited the growth of at least one or more phytopathogenic fungi and Saccharopolyspora 0-9 exhibited antagonistic activity against Aspergillus niger, Aspergillus flavus, Alternaria brassicicola, Botrytis cinerea, Penicillium digitatum, Fusarium oxysporum, Penicillium pinophilum, Phytophthora dresclea and Colletotrichum falcatum.

  18. Endophytes: a treasure house of bioactive compounds of medicinal importance

    Sushanto Gouda

    2016-09-01

    Full Text Available Endophytes are an endosymbiotic group of microorganisms that colonize in plants and microbes that can be readily isolated from any microbial or plant growth medium. They act as reservoirs of novel bioactive secondary metabolites, such as alkaloids, phenolic acids, quinones, steroids, saponins, tannins, and terpenoids that serve as a potential candidate for antimicrobial, anti-insect, anticancer and many more properties. While plant sources are being extensively explored for new chemical entities for therapeutic purposes, endophytic microbes also constitute an important source for drug discovery. This review aims to comprehend the contribution and uses of endophytes as an impending source of drugs against various forms of diseases and other possible medicinal use.

  19. Fungal endophytes characterization from four species of Diplazium Swartz

    Affina-Eliya, A. A.; Noraini, T.; Nazlina, I.; Ruzi, A. R.

    2014-09-01

    Four species on genus Diplazium namely Diplazium tomentosum, D. sorzogonense, D. asperum and D. accedens of Peninsular Malaysia were studied for presence of fungal endophyte. The objective of this study is to characterize fungal endophytes in the rhizome of four Diplazium species. The rhizome was surface sterilized and incubated to isolate fungal endophytes. Characterization of the colonies was performed by macroscopic morphological, microscopic identification, types of hyphae and mycelium, and spore structure. For isolation that produces spores, the structure of conidiophores and conidia were identified. From this study, four fungal have been isolated and determined as Aspergillus sp. (isolates AE 1), Aspergillus fumigatus (isolates AE 2), Aspergillus versicolor (isolates AE 3) and Verticillium sp. (isolates AE 4). The fungal isolates from this study were classified from the same family Moniliaceae.

  20. Isolation and antifungal screening of endophytic fungi from Erigeron canadensis

    Xuelian Bai

    2017-07-01

    Full Text Available Sixteen fungal strains isolated from the Erigeron canadensis, one of traditional Chinese medicines used to treat the pathogenic infection and dysentery, were evaluated for their antifungal activities against one human pathogen Candida albicans, and two phytopathogens, Colletotrichum fructicola and Rhizoctonia cerealis. The bioassay results indicated that the ethyl acetate extract of the fermentation broth of these fungal endophytes had stronger antimicrobial activities. Among these endophytic strains, the ethyl acetate extracts of strains NPR003 and NPR005 showed the strongest inhibitory effects and has potential application in the discovery of new antifungal agents. This was the first report on the isolation of endophytic fungi from E. canadensis and evaluation of their antifungal activities.

  1. Identification of genetic components involved in Lotus-endophyte interactions

    Zgadzaj, Rafal Lukasz

    of growth hormones or nitrogen fixation. However, the genes involved in plant-endophyte interactions and bacterial accomodation within plant tissues are not known. In order to shed some light on such processes, an approach “one host-one endophyte” was chosen. The focus on a single plant species and a single......Endophytes are microorganisms capable of colonising plant tissues without inducing host defense responses. They have a large impact on plants, since they can modulate plant responses to pathogens, herbivores and environmental stress. They can also induce plant growth promotion through synthesis...... bacterial strain aimed at obtaining a reliable and easy to handle system for plant-microsymbiont interaction research. Two different methods were tested for their usefulness in identification of genetic components involved in plant-endophyte interactions. The first method was based on measuring growth...

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

    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.

  3. [Screening endophytic bacteria against plant-parasitic nematodes].

    Peng, Shuang; Yan, Shuzhen; Chen, Shuanglin

    2011-03-01

    Plant-parasite nematode is one of the most important pathogens in plant. Our objective is to screen endophytic bacteria against plant-parasitic nematodes from plant. Endophytic bacteria were isolated and screened by testing their metabolite against Bursaphelenchus xylophilus in vitro. Those strains inhibiting B. xylophilus were selected to culture in liquid medium and fermentation conditions were optimized by orthogonal test. The stability of the antinematode substances was evaluated by various. In addition, four strains were identified by 16SrDNA sequence analysis. In total 13 strains of endophytic bacteria secreting antinematode metabolite were isolated from 6 species of plant. The supernatant of the fermentation broth of these endophytic bacteria gave 100% mortality of nematodes after treated as the follows: 1 ml each was mixed with 0.2 ml of the suspension of nematodes (2000 nematodes/ml) then incubated at 250C for 24 h, some of which could led to leakage or dissolution of nematodes. Among them, four strains, BCM2, SZ5, CCM7 and DP1, showed stronger activity than others. The supernatants diluted three times also gave not less than 95% mortality after 24 h treatment, and those from DP1 and SZ5 even gave 100% mortality. The fermentation conditions of the four strains were optimized and the antinematode activity grew up four times after optimization. The antinematode substances of these strains were found stable when treated with protease or heating or stored at 4 degrees C after 100 days, while instable when treated with acid or alkali. DP1 and CCM7 were identified to be Bacillus subtilis, while SZ5 and BCM2 to be Bacillus cereus. Endophytic bacteria secreting antinematode metabolite were found in economic crops. The metabolite of some strains showed strong and stable antinematode activity. Our results indicate the real potential of biocontrol by endophytic bacteria.

  4. Maize Endophytic Bacterial Diversity as Affected by Soil Cultivation History.

    Correa-Galeote, David; Bedmar, Eulogio J; Arone, Gregorio J

    2018-01-01

    The bacterial endophytic communities residing within roots of maize ( Zea mays L.) plants cultivated by a sustainable management in soils from the Quechua maize belt (Peruvian Andes) were examined using tags pyrosequencing spanning the V4 and V5 hypervariable regions of the 16S rRNA. Across four replicate libraries, two corresponding to sequences of endophytic bacteria from long time maize-cultivated soils and the other two obtained from fallow soils, 793 bacterial sequences were found that grouped into 188 bacterial operational taxonomic units (OTUs, 97% genetic similarity). The numbers of OTUs in the libraries from the maize-cultivated soils were significantly higher than those found in the libraries from fallow soils. A mean of 30 genera were found in the fallow soil libraries and 47 were in those from the maize-cultivated soils. Both alpha and beta diversity indexes showed clear differences between bacterial endophytic populations from plants with different soil cultivation history and that the soils cultivated for long time requires a higher diversity of endophytes. The number of sequences corresponding to main genera Sphingomonas, Herbaspirillum, Bradyrhizobium and Methylophilus in the maize-cultivated libraries were statistically more abundant than those from the fallow soils. Sequences of genera Dyella and Sreptococcus were significantly more abundant in the libraries from the fallow soils. Relative abundance of genera Burkholderia, candidatus Glomeribacter, Staphylococcus, Variovorax, Bacillus and Chitinophaga were similar among libraries. A canonical correspondence analysis of the relative abundance of the main genera showed that the four libraries distributed in two clearly separated groups. Our results suggest that cultivation history is an important driver of endophytic colonization of maize and that after a long time of cultivation of the soil the maize plants need to increase the richness of the bacterial endophytes communities.

  5. Biodegradation of Trichloroethylene by an Endophyte of Hybrid Poplar

    Kang, Jun Won; Khan, Zareen

    2012-01-01

    We isolated and characterized a novel endophyte from hybrid poplar. This unique endophyte, identified as Enterobacter sp. strain PDN3, showed high tolerance to trichloroethylene (TCE). Without the addition of inducers, such as toluene or phenol, PDN3 rapidly reduced TCE levels in medium from 72.4 μM to 30.1 μM in 24 h with a concurrent release of 127 μM chloride ion, and nearly 80% of TCE (55.3 μM) was dechlorinated by PDN3 in 5 days with 166 μM chloride ion production, suggesting TCE degradation. PMID:22367087

  6. The role of the Oregon State University Endophyte Service Laboratory in diagnosing clinical cases of endophyte toxicoses.

    Craig, A Morrie; Blythe, Linda L; Duringer, Jennifer M

    2014-07-30

    The Oregon State University Colleges of Veterinary Medicine and Agricultural Sciences instituted the Endophyte Service Laboratory to aid in diagnosing toxicity problems associated with cool-season grasses in livestock. The endophyte (Neotyphodium coenophalum) present in tall fescue (Festuca arundinacea) produces ergopeptine alkaloids, of which ergovaline is the molecule used to determine exposure and toxicity thresholds for the vasoconstrictive conditions "fescue foot" and "summer slump". Another vasoconstrictive syndrome, "ergotism," is caused by a parasitic fungus, Claviceps purpurea, and its primary toxin, ergotamine. "Ryegrass staggers" is a neurological condition that affects livestock consuming endophyte (Neotyphodium lolii)-infected perennial ryegrass (Lolium perenne) with high levels of lolitrem B. HPLC-fluorescent analytical methods for these mycotoxins are described and were used to determine threshold levels of toxicity for ergovaline and lolitrem B in cattle, sheep, horses, and camels. In addition, six clinical cases in cattle are presented to illustrate diagnosis of these three diseases.

  7. [Diversity and community structure of endophytic fungi from Taxus chinensis var. mairei].

    2014-07-01

    A total of 628 endophytic fungi were isolated from 480 tissue segments of needles and branches of Taxus chinensis var. mairei. According to morphological characteristics and ITS sequences, they represented 43 taxa in 28 genera, of which 10 Hyphomycetes, 20 Coelomycetes, 12 Ascomycetes and 1 unknown fungus. Phomopsis mali was confirmed as the dominant species. In accordance with relative frequency, Alternaria alternata, Aureobasidium pullulans, Colletotrichum boninense, C. gloeosporioides, Epicoccum nigrum , Fungal sp., Fusarium lateritium, Glomerella cingulata, Magnaporthales sp. , Nigrospora oryzae, Pestalotiopsis maculiformans, P. microspora, Peyronellaea glomerata and Xylaria sp. 1 were more common in T. chinensis var. mairei. T. chinensis var. mairei were severely infected by endophytic fungi. Endophytic fungi were found in 81 percent of plant tissues with a high diversity. Distribution ranges of endophytic fungi were influenced by tissue properties. The colonization rate, richness, diversity of endophytic fungi in needles were obviously lower than in branches, and kinds of endophytic fungi between branches were more similar than those in needles, thus endophytic fungi had tissue preference. In addition, tissue age influenced the community structure of endophytic fungi. The elder branch tissues were, the higher colonization rate, richness, diversity of endophytic fungi were. Systematic studying the diversity and community structure of endophytic fungi in T. chinensis var. mairei and clarifying their distribution regularity in plant tissues would offer basic data and scientific basis for their development and utilization. Discussing the presence of fungal pathogens in healthy plant tissues would be of positive significance for source protection of T. chinensis var. mairei.

  8. Endophytic bacteria: prospects and applications for the phytoremediation of organic pollutants.

    Afzal, Muhammad; Khan, Qaiser M; Sessitsch, Angela

    2014-12-01

    Recently, there has been an increased effort to enhance the efficacy of phytoremediation of contaminated environments by exploiting plant-microbe interactions. The combined use of plants and endophytic bacteria is an emerging approach for the clean-up of soil and water polluted with organic compounds. In plant-endophyte partnerships, plants provide the habitat as well as nutrients to their associated endophytic bacteria. In response, endophytic bacteria with appropriate degradation pathways and metabolic activities enhance degradation of organic pollutants, and diminish phytotoxicity and evapotranspiration of organic pollutants. Moreover, endophytic bacteria possessing plant growth-promoting activities enhance the plant's adaptation and growth in soil and water contaminated with organic pollutants. Overall, the application of endophytic bacteria gives new insights into novel protocols to improve phytoremediation efficiency. However, successful application of plant-endophyte partnerships for the clean-up of an environment contaminated with organic compounds depends on the abundance and activity of the degrading endophyte in different plant compartments. Although many endophytic bacteria have the potential to degrade organic pollutants and improve plant growth, their contribution to enhance phytoremediation efficiency is still underestimated. A better knowledge of plant-endophyte interactions could be utilized to increase the remediation of polluted soil environments and to protect the foodstuff by decreasing agrochemical residues in food crops. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Geographic Variation in Festuca rubra L. Ploidy Levels and Systemic Fungal Endophyte Frequencies.

    Serdar Dirihan

    Full Text Available Polyploidy and symbiotic Epichloë fungal endophytes are common and heritable characteristics that can facilitate environmental range expansion in grasses. Here we examined geographic patterns of polyploidy and the frequency of fungal endophyte colonized plants in 29 Festuca rubra L. populations from eight geographic sites across latitudes from Spain to northernmost Finland and Greenland. Ploidy seemed to be positively and negatively correlated with latitude and productivity, respectively. However, the correlations were nonlinear; 84% of the plants were hexaploids (2n = 6x = 42, and the positive correlation between ploidy level and latitude is the result of only four populations skewing the data. In the southernmost end of the gradient 86% of the plants were tetraploids (2n = 4x = 28, whereas in the northernmost end of the gradient one population had only octoploid plants (2n = 8x = 56. Endophytes were detected in 22 out of the 29 populations. Endophyte frequencies varied among geographic sites, and populations and habitats within geographic sites irrespective of ploidy, latitude or productivity. The highest overall endophyte frequencies were found in the southernmost end of the gradient, Spain, where 69% of plants harbored endophytes. In northern Finland, endophytes were detected in 30% of grasses but endophyte frequencies varied among populations from 0% to 75%, being higher in meadows compared to riverbanks. The endophytes were detected in 36%, 30% and 27% of the plants in Faroe Islands, Iceland and Switzerland, respectively. Practically all examined plants collected from southern Finland and Greenland were endophyte-free, whereas in other geographic sites endophyte frequencies were highly variable among populations. Common to all populations with high endophyte frequencies is heavy vertebrate grazing. We propose that the detected endophyte frequencies and ploidy levels mirror past distribution history of F. rubra after the last glaciation

  10. Assessment of endophytic fungi cultural filtrate on soybean seed ...

    Soybean seeds have high amount of isoflavones but its germination is often confronted with a variety of environmental problems resulting in low germination rate and growth. To overcome this in eco-friendly manner, we investigated the influence of cultural filtrate (CF) of gibberellins-producing endophytic fungi on soybean ...

  11. Inoculation, colonization and distribution of fungal endophytes in ...

    Mo

    and for a part or whole of their life cycle live symptomlessly within the plant. ... inoculated in tissue culture banana plants, must occur at high frequencies in the plant and be able to persist in ... For instance, the influence of fungal endophytes.

  12. Cytosporones O, P and Q from an endophytic Cytospora sp

    Abreu, L.M.; Phipps, Richard Kerry; Pfenning, L.H.

    2010-01-01

    Cytosporones O, P and Q, together with the known compounds cytosporones B, C, D, E and dothiorelones A, 13, C. and H were isolated from the ascomycete fungus Cytospora sp. during a chemotaxonomic study Of fungal endophytes belonging to the related genera Cytospora and Phomopsis from Brazil. The s...

  13. Volatile metabolites profiling of a Chinese mangrove endophytic ...

    Pestalotiopsis JCM2A4, an endophytic fungus originally isolated from leaves of the Chinese mangrove plant Rhizophora mucronata, produces a mixture of volatile metabolites. As determined by gas chromatography and gas chromatography/mass spectrometry (GC/GC-MS), 18 compounds representing all of the hexane ...

  14. Endophytic Phytoaugmentation: Treating Wastewater and Runoff Through Augmented Phytoremediation

    Redfern, Lauren K.

    2016-01-01

    Abstract Limited options exist for efficiently and effectively treating water runoff from agricultural fields and landfills. Traditional treatments include excavation, transport to landfills, incineration, stabilization, and vitrification. In general, treatment options relying on biological methods such as bioremediation have the ability to be applied in situ and offer a sustainable remedial option with a lower environmental impact and reduced long-term operating expenses. These methods are generally considered ecologically friendly, particularly when compared to traditional physicochemical cleanup options. Phytoremediation, which relies on plants to take up and/or transform the contaminant of interest, is another alternative treatment method which has been developed. However, phytoremediation is not widely used, largely due to its low treatment efficiency. Endophytic phytoaugmentation is a variation on phytoremediation that relies on augmenting the phytoremediating plants with exogenous strains to stimulate associated plant-microbe interactions to facilitate and improve remediation efficiency. In this review, we offer a summary of the current knowledge as well as developments in endophytic phytoaugmentation and present some potential future applications for this technology. There has been a limited number of published endophytic phytoaugmentation case studies and much remains to be done to transition lab-scale results to field applications. Future research needs include large-scale endophytic phytoaugmentation experiments as well as the development of more exhaustive tools for monitoring plant-microbe-pollutant interactions. PMID:27158249

  15. Identification of a taxol-producing endophytic fungus EFY-36

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... Morphological and molecular methods were used to identify the statues of an isolate, EFY-36, a taxol- ... of the spores. The analysis of endophytic fungus. 18S ribosome RNA sequence used PCR cloning technology. DNA was extracted by the CTAB method. ... of the fungal mycelium (magnification: 400 ×).

  16. Effect of endophytic Fusarium oxysporum on paralysis and mortality ...

    Three bioassays were conducted to investigate the antagonistic effect of secondary metabolites produced by 5 endophytic Fusarium oxysporum isolates from banana (Musa spp.) plants in Kenya, against Pratylenchus goodeyi. Percentage paralyses were recorded 3, 6 and 24 h after exposure to culture filtrates. Percentage ...

  17. Differential endophytic colonization of sorghum plant by eight ...

    Virulence of the conidia before and after endophytic growth phases were assessed using Galleria mellonella larvae mortality bioassay in-vitro. All the strains of the fungi colonised the sorghum plant. The strains of I. farinosa and B. bassiana were detected in the roots, the stem and the leaves while M. anisopliae was ...

  18. antibacterial activity of endophytic fungi isolated from conifers needles

    Ravnikar, Matjaž

    2015-03-11

    Mar 11, 2015 ... taxonomically place fungi producing ones to determined active metabolites. Seventy three strains of endophytic fungi were isolated ... great number of diverse bioactive compounds (Devaraju and Satish, 2010), which have been ... closed with a glass stopper. The extraction solvents utilized were methanol ...

  19. Antagonistic bioactivity of endophytic strains isolated from Salvia ...

    The antibiotic-producing potential of endophytic populations from medical plant of Salvia miltiorrhiza was examined. A total of 63 isolates was screened against five fungal and three bacterial species for the production of antimicrobial compounds. It showed that more isolates was antagonistic to fungi than to bacteria.

  20. Identification of lead-resistant endophytic bacteria isolated from rice

    Perez-Cordero, Alexander; Barraza-Roman, Zafiro; Martinez-Pacheco, Dalila

    2015-01-01

    Resistance of endophytic bacteria in vitro was evaluated at different lead concentrations. The tissue samples of commercial rice varieties at tillering stage were collected during the first half of 2013, in Monteria, Cordoba, Colombia. Each tissue was subjected to surface cleaning. Endophytic bacteria were isolated in agar R_2A medium. The population density (CFU/g tissue) was determined from each tissue by direct counting of R_2A medium surface. Morphotypes were classified by shape, color, size and appearance. A total of 168 morphotypes were isolated from root, tillers and leaf of different commercial varieties of rice. The lead resistance test is performed in vitro, The lead resistance test was performed in vitro, by the suspensions of endophytic bacteria in log phase and inoculation in minimal medium with five concentrations of lead as Pb (NO_3)_2. The experiment was incubated at 32 degrees celsius and agitated at 150 rpm for five days. The measure of turbidimetry at 600 nm was conduced every hour afterstarting the test. Endophytic bacteria showed the ability to grow at concentrations of 100% of Pb as Pb (NO_3) _2. The presence of Burkholderia cepacia and Pseudomonas putida, which showed resistance to differents lead concentration was confirmed as result of the identification with kit API20E. (author) [es

  1. Identification of lead- resistant endophytic bacteria isolated from rice.

    Alexander Pérez-Cordero

    2015-06-01

    Full Text Available   The objective of this study was to evaluate in vitro the endophytic bacteria resistance to different lead concentrations. The sampling was undertaken in the first half of 2013, when tissue samples of commercial varieties of rice at tillering stage were collected in Montería, Cordoba, Colombia. Each tissue was subjected to surface cleaning. Endophytic bacteria in agar R2A medium were isolated. Population density (CFU/g tissue was determined from each tissue, by direct counting of R2A medium surface. morphotypes were classified by shape, color, size, and appearance. A total of 168 morphotypes were isolated from root, tillers, and leaf of different commercial varieties of rice. The lead resistance test was performed in vitro, to do that, suspensions of endophytic bacteria in log phase were prepared and inoculated in minimal medium with five concentrations of lead as Pb(NO32. The experiment was incubated at 32 °C and agitated at 150 rpm, for five days. Every hour afterstarting the test, turbidimetry measuring at 600 nm was conducted. Results showed the ability of endophytic bacteria to grow at concentrations of 100% of Pb as Pb(NO32. The results of the identification with kit API20E confirmed the presence of Burkholderia cepacia and Pseudomonas putida, which showed resistance to different lead concentrations.

  2. Endophytic microorganisms--promising applications in bioremediation of greenhouse gases.

    Stępniewska, Z; Kuźniar, A

    2013-11-01

    Bioremediation is a technique that uses microbial metabolism to remove pollutants. Various techniques and strategies of bioremediation (e.g., phytoremediation enhanced by endophytic microorganisms, rhizoremediation) can mainly be used to remove hazardous waste from the biosphere. During the last decade, this specific technique has emerged as a potential cleanup tool only for metal pollutants. This situation has changed recently as a possibility has appeared for bioremediation of other pollutants, for instance, volatile organic compounds, crude oils, and radionuclides. The mechanisms of bioremediation depend on the mobility, solubility, degradability, and bioavailability of contaminants. Biodegradation of pollutions is associated with microbial growth and metabolism, i.e., factors that have an impact on the process. Moreover, these factors have a great influence on degradation. As a result, recognition of natural microbial processes is indispensable for understanding the mechanisms of effective bioremediation. In this review, we have emphasized the occurrence of endophytic microorganisms and colonization of plants by endophytes. In addition, the role of enhanced bioremediation by endophytic bacteria and especially of phytoremediation is presented.

  3. Does hybridization of endophytic symbionts in a native grass increase fitness in resource-limited environments?

    Faeth, Stanley H.; Oberhofer, Martina; Saari, Susanna Talvikki

    2017-01-01

    to their grass hosts, especially in stressful environments. We tested the hybrid fitness hypothesis (HFH) that hybrid endophytes enhance fitness in stressful environments relative to non-hybrid endophytes. In a long-term field experiment, we monitored growth and reproduction of hybrid-infected (H+), non......-hybrid infected (NH+), naturally endophyte free (E-) plants and those plants from which the endophyte had been experimentally removed (H- and NH-) in resource-rich and resource-poor environments. Infection by both endophyte species enhanced growth and reproduction. H+ plants outperformed NH+ plants in terms...... of growth by the end of the experiment, supporting HFH. However, H+ plants only outperformed NH+ plants in the resource-rich treatment, contrary to HFH. Plant genotypes associated with each endophyte species had strong effects on growth and reproduction. Our results provide some support the HFH hypothesis...

  4. Biodegradation of Mixed PAHs by PAH-Degrading Endophytic Bacteria

    Xuezhu Zhu

    2016-08-01

    Full Text Available Endophytic bacteria can promote plant growth, induce plant defence mechanisms, and increase plant resistance to organic contaminants. The aims of the present study were to isolate highly PAH-degrading endophytic bacteria from plants growing at PAH-contaminated sites and to evaluate the capabilities of these bacteria to degrade polycyclic aromatic hydrocarbons (PAHs in vitro, which will be beneficial for re-colonizing target plants and reducing plant PAH residues through the inoculation of plants with endophytic bacteria. Two endophytic bacterial strains P1 (Stenotrophomonas sp. and P3 (Pseudomonas sp., which degraded more than 90% of phenanthrene (PHE within 7 days, were isolated from Conyza canadensis and Trifolium pretense L., respectively. Both strains could use naphthalene (NAP, PHE, fluorene (FLR, pyrene (PYR, and benzo(apyrene (B(aP as the sole sources of carbon and energy. Moreover, these bacteria reduced the contamination of mixed PAHs at high levels after inoculation for 7 days; strain P1 degraded 98.0% NAP, 83.1% FLR, 87.8% PHE, 14.4% PYR, and 1.6% B(aP, and strain P3 degraded 95.3% NAP, 87.9% FLR, 90.4% PHE, 6.9% PYR, and negligible B(aP. Notably, the biodegradation of PAHs could be promoted through additional carbon and nitrogen nutrients; therein, beef extract was suggested as the optimal co-substrate for the degradation of PAHs by these two strains (99.1% PHE was degraded within 7 days. Compared with strain P1, strain P3 has more potential for the use in the removal of PAHs from plant tissues. These results provide a novel perspective in the reduction of plant PAH residues in PAH-contaminated sites through inoculating plants with highly PAH-degrading endophytic bacteria.

  5. Seed-vectored endophytic bacteria modulate development of rice seedlings.

    Verma, S K; Kingsley, K; Irizarry, I; Bergen, M; Kharwar, R N; White, J F

    2017-06-01

    The aim of the present study was to evaluate the effects of the removal of indigenous bacteria from rice seeds on seedling growth and development. Here we report the presence of three indigenous endophytic bacteria in rice seeds that play important roles in modulating seedling development (shoot and root lengths, and formation of root hairs and secondary roots) and defence against pathogens. Seed-associated bacteria were removed using surface sterilization with NaOCl (bleach) followed by antibiotic treatment. When bacteria were absent, growth of seedlings in terms of root hair development and overall seedling size was less than that of seedlings that contained bacteria. Reactive oxygen staining of seedlings showed that endophytic bacteria became intracellular in root parenchyma cells and root hairs. Roots containing endophytic bacteria were seen to stain densely for reactive oxygen, while roots free of bacteria stained lightly for reactive oxygen. Bacteria were isolated and identified as Enterobacter asburiae (VWB1), Pantoea dispersa (VWB2) and Pseudomonas putida (VWB3) by 16S rDNA sequencing. Bacteria were found to produce indole acetic acid (auxins), inhibited the pathogen Fusarium oxysporum and solubilized phosphate. Reinoculation of bacteria onto seedlings derived from surface-disinfected rice and Bermuda grass seeds significantly restored seedling growth and development. Rice seeds harbour indigenous bacterial endophytes that greatly influence seedling growth and development, including root and shoot lengths, root hair formation and disease susceptibility of rice seedlings. This study shows that seeds of rice naturally harbour bacterial endophytes that play key roles in modulation of seedling development. © 2017 The Society for Applied Microbiology.

  6. Culturable endophytic bacterial communities associated with field-grown soybean.

    de Almeida Lopes, K B; Carpentieri-Pipolo, V; Oro, T H; Stefani Pagliosa, E; Degrassi, G

    2016-03-01

    Assess the diversity of the culturable endophytic bacterial population associated with transgenic and nontransgenic soybean grown in field trial sites in Brazil and characterize them phenotypically and genotypically focusing on characteristics related to plant growth promotion. Endophytic bacteria were isolated from roots, stems and leaves of soybean cultivars (nontransgenic (C) and glyphosate-resistant (GR) transgenic soybean), including the isogenic BRS133 and BRS245RR. Significant differences were observed in bacterial densities in relation to genotype and tissue from which the isolates were obtained. The highest number of bacteria was observed in roots and in GR soybean. Based on characteristics related to plant growth promotion, 54 strains were identified by partial 16S rRNA sequence analysis, with most of the isolates belonging to the species Enterobacter ludwigii and Variovorax paradoxus. Among the isolates, 44·4% were able to either produce indoleacetic acid (IAA) or solubilize phosphates, and 9·2% (all from GR soybean) presented both plant growth-promoting activities. The results from this study indicate that the abundance of endophytic bacterial communities of soybean differs between cultivars and in general it was higher in the transgenic cultivars than in nontransgenic cultivars. BRS 245 RR exhibited no significant difference in abundance compared to nontransgenic BRS133. This suggests that the impact of the management used in the GR soybean fields was comparable with the impacts of some enviromental factors. However, the bacterial endophytes associated to GR and nontransgenic soybean were different. The soybean-associated bacteria showing characteristics related to plant growth promotion were identified as belonging to the species Pantoea agglomerans and Variovorax paradoxus. Our study demonstrated differences concerning compostion of culturable endophytic bacterial population in nontransgenic and transgenic soybean. © 2016 The Society for Applied

  7. Leaf endophyte load influences fungal garden development in leaf-cutting ants

    Van Bael Sunshine A

    2012-11-01

    Full Text Available Abstract Background Previous work has shown that leaf-cutting ants prefer to cut leaf material with relatively low fungal endophyte content. This preference suggests that fungal endophytes exact a cost on the ants or on the development of their colonies. We hypothesized that endophytes may play a role in their host plants’ defense against leaf-cutting ants. To measure the long-term cost to the ant colony of fungal endophytes in their forage material, we conducted a 20-week laboratory experiment to measure fungal garden development for colonies that foraged on leaves with low or high endophyte content. Results Colony mass and the fungal garden dry mass did not differ significantly between the low and high endophyte feeding treatments. There was, however, a marginally significant trend toward greater mass of fungal garden per ant worker in the low relative to the high endophyte treatment. This trend was driven by differences in the fungal garden mass per worker from the earliest samples, when leaf-cutting ants had been foraging on low or high endophyte leaf material for only 2 weeks. At two weeks of foraging, the mean fungal garden mass per worker was 77% greater for colonies foraging on leaves with low relative to high endophyte loads. Conclusions Our data suggest that the cost of endophyte presence in ant forage material may be greatest to fungal colony development in its earliest stages, when there are few workers available to forage and to clean leaf material. This coincides with a period of high mortality for incipient colonies in the field. We discuss how the endophyte-leaf-cutter ant interaction may parallel constitutive defenses in plants, whereby endophytes reduce the rate of colony development when its risk of mortality is greatest.

  8. Diversity and host range of foliar fungal endophytes: are tropical leaves biodiversity hotspots?

    Arnold, A Elizabeth; Lutzoni, F

    2007-03-01

    Fungal endophytes are found in asymptomatic photosynthetic tissues of all major lineages of land plants. The ubiquity of these cryptic symbionts is clear, but the scale of their diversity, host range, and geographic distributions are unknown. To explore the putative hyperdiversity of tropical leaf endophytes, we compared endophyte communities along a broad latitudinal gradient from the Canadian arctic to the lowland tropical forest of central Panama. Here, we use molecular sequence data from 1403 endophyte strains to show that endophytes increase in incidence, diversity, and host breadth from arctic to tropical sites. Endophyte communities from higher latitudes are characterized by relatively few species from many different classes of Ascomycota, whereas tropical endophyte assemblages are dominated by a small number of classes with a very large number of endophytic species. The most easily cultivated endophytes from tropical plants have wide host ranges, but communities are dominated by a large number of rare species whose host range is unclear. Even when only the most easily cultured species are considered, leaves of tropical trees represent hotspots of fungal species diversity, containing numerous species not yet recovered from other biomes. The challenge remains to recover and identify those elusive and rarely cultured taxa with narrower host ranges, and to elucidate the ecological roles of these little-known symbionts in tropical forests.

  9. Host associations and beta diversity of fungal endophyte communities in New Guinea rainforest trees.

    Vincent, J B; Weiblen, G D; May, G

    2016-02-01

    Processes shaping the distribution of foliar fungal endophyte species remain poorly understood. Despite increasing evidence that these cryptic fungal symbionts of plants mediate interactions with pathogens and herbivores, there remain basic questions regarding the extent to which dispersal limitation and host specificity might shape fungal endophyte community composition in rainforests. To assess the relative importance of spatial pattern and host specificity, we isolated fungi from a sample of mapped trees in lowland Papua New Guinea. Sequences of the internal transcribed spacer (ITS) region were obtained for 2079 fungal endophytes from three sites and clustered into molecular operational taxonomic units (MOTUs) at 95% similarity. Multivariate analyses suggest that host affinity plays a significant role in structuring endophyte community composition whereas there was no evidence of endophyte spatial pattern at the scale of tens to hundreds of metres. Differences in endophyte communities between sampled trees were weakly correlated with variation in foliar traits but not with tree species relatedness. The dominance of relatively few generalist endophytes and the presence of a large number of rare MOTUs was a consistent observation at three sites separated by hundreds of kilometres and regional turnover was low. Host specificity appears to play a relatively weak but more important role than dispersal limitation in shaping the distribution of fungal endophyte communities in New Guinea forests. Our results suggest that in the absence of strong ecological gradients and host turnover, beta diversity of endophyte communities could be low in large areas of contiguous forest. © 2015 John Wiley & Sons Ltd.

  10. The Hidden World within Plants: Ecological and Evolutionary Considerations for Defining Functioning of Microbial Endophytes

    van Overbeek, Leonard S.; Berg, Gabriele; Pirttilä, Anna Maria; Compant, Stéphane; Campisano, Andrea; Döring, Matthias; Sessitsch, Angela

    2015-01-01

    SUMMARY All plants are inhabited internally by diverse microbial communities comprising bacterial, archaeal, fungal, and protistic taxa. These microorganisms showing endophytic lifestyles play crucial roles in plant development, growth, fitness, and diversification. The increasing awareness of and information on endophytes provide insight into the complexity of the plant microbiome. The nature of plant-endophyte interactions ranges from mutualism to pathogenicity. This depends on a set of abiotic and biotic factors, including the genotypes of plants and microbes, environmental conditions, and the dynamic network of interactions within the plant biome. In this review, we address the concept of endophytism, considering the latest insights into evolution, plant ecosystem functioning, and multipartite interactions. PMID:26136581

  11. Biodiversity, Phylogeny, and Antifungal Functions of Endophytic Fungi Associated with Zanthoxylum bungeanum

    Peiqin Li

    2016-09-01

    Full Text Available This study investigated the biodiversity, phylogeny, and antifungal activity of endophytic fungi isolated from Zanthoxylum bungeanum. A total of 940 isolates obtained were grouped into 93 morphotypes, 43 species, and 23 genera, which were authenticated by molecular identification based on rDNA internal transcribed spacer (ITS sequence analysis. A high diversity of endophytic fungi from Z. bungeanum are observed with high species richness S (43, Margalef index D′ (6.1351, Shannon–Wiener index H′ (3.2743, Simpson diversity index Ds (0.9476, PIE index (0.9486, and evenness Pielou index J (0.8705 but a low dominant index λ (0.0524. Significant tissue specificity of the endophytic fungi was observed in Z. bungeanum, and the highest species richness and diversity indexes were obtained in the stem. Phylogenetic analyses of the 93 endophytic isolates were carried out by the neighbor-joining (NJ method to demonstrate their evolutionary processes. Antifungal activities of endophytic fungi were assayed and eight endophytic isolates showed strong and long-lasting inhibition against host pathogenic fungi Fusarium sambucinum and Pseudocercospora zanthoxyli. Here, for the first time, we systematically demonstrate the biodiversity, phylogeny, and antifungal activity of endophytic fungi associated with Z. bungeanum and reveal the value of sampling different tissues of a given plant to obtain the greatest endophyte species diversity, which might offer a framework for further investigation and utilization of endophytic fungi as aunique source of interesting and useful bioactive compounds.

  12. Evaluating Susceptibility to Commercial Fungicide of Endophytic Fungi Isolated from Roses (Rosa hybrida

    Ingrid Carolina Corredor Perilla

    2007-01-01

    Full Text Available Fungal endophytes have shown their potential as biocontrol agents; however, their application in commercial fields remains limited. Continuously applying fungicides to crops (specifically to roses may have harmful effects on endophyte growth. Endophytic fungi were isolated from R. hybrida and their susceptibility to fungicides regularly used for controlling important pathogens was analysed. This was performed in vitro, mixing several fungicide concentrations with standard medium for fungal endophytes; growth inhibition was then measured. The susceptibility of Botrytis cinerea (3015 strain, one of the most important pathogens affecting roses in Colombia, was also assessed using the same protocols. Active ingredients, such as boscalid, captan, iprodione and pyrimethanyl, showed susceptibility ranging from not sensitive (³73.75% to regularly sensitive (³48.75% - <61.25% for 45.45% of the fungal endophytes assessed. Endophytic fungi were highly susceptible to fungicides such as pyrimethanyl, carboxin plus thiram, fludioxonyl plus ciprodinyl and prochloraz. B. cinerea (3015 strain presented high susceptibility (<23.75% to fungicides such as pyrimethanyl, carboxin and thiram, fludioxonil and ciprodinyl, prochloraz. Although B. cinerea showed the greatest growth in controls, the endophytic fungi being assessed grew better in different media with fungicides. The results revealed some of these fungal endophytes’ potential for integrated pest management (IPM in roses in Colombia (3002, 3003, 3004, 3005 and 3006 strains, taking into account correct application time, application frequency and both fungal endophyte and fungicide dosage which may greatly limit fungal endophyte growth.

  13. Exploring the potential of symbiotic fungal endophytes in cereal disease suppression

    O'Hanlon, Karen; Knorr, Kamilla; Jørgensen, Lise Nistrup

    2012-01-01

    , and environmental and health concerns surrounding the use of chemical treatments. There is currently a demand for new disease control strategies, and one such strategy involves the use of symbiotic fungal endophytes as biological control agents against fungal pathogens in cereals. Despite the fact that biological...... control by symbiotic fungal endophytes has been documented, particularly with respect to clavicipitaceous endophytes in C3 cool-season grasses, this area remains relatively underexplored in cereals. We highlight for the first time the potential in using symbiotic fungal endophytes to control foliar cereal...

  14. Community structure of endophytic fungi of four mangrove species in Southern China

    Jia-Long Li

    2016-10-01

    Full Text Available Mangrove forests play an important role in subtropical and tropical coastal ecosystems. Endophytic fungi are widely distributed in various ecosystems and have great contribution to global biodiversity. In order to better understand the effects of mangrove species and tissue types on endophytic fungal community, we investigated cultivable endophytic fungi in leaves and twigs of four mangroves Aegiceras corniculatum, Avicennia marina, Bruguiera gymnorrhiza, and Kandelia candel in Guangxi, China. The four tree species had similar overall colonisation rates of endophytic fungi (24–33%. The colonisation rates of endophytic fungi were higher in twigs (30–58% than in leaves (6–25% in the four plant species. A total of 36 endophytic fungal taxa were identified based on morphological characteristics and molecular data, including 35 Ascomycota and 1 Basidiomycota, dominated by Phomopsis, Phyllosticta, Xylaria, Leptosphaerulina, and Pestalotiopsis. The diversity of endophytic fungi was higher in twigs than in leaves in the four plant species. Some endophytic fungi showed host and tissue preference. The endophytic fungal community composition was different among four mangrove species and between leaf and twig tissues.

  15. EVALUATION OF THE DEVELOPMENT OF MAIZE PLANTS (Zea mays L.) AFTER COLONIZATION BY ENDOPHYTE FUNGUS Fusarium verticillioides

    Gomes, Ulisses de Deus; Orlandelli, Ravely Casarotti; Santos, Mariana Sanches; Polonio, Julio Cesar; Pamphile, João Alencar; Rubin Filho, Celso João

    2013-01-01

    Endophyte fungi inhabit the inside of plants without causing any damage. Benefits from endophyte-plant interactivities include vegetal growth and the plant´s defense against insects and other pathogens. Some endophytes, however, may act as latent pathogens which cause physiological changes and disease symptoms in the host. Current analysis evaluates the development of maize plants colonizer (treatment) and non-colonized (control) with the frequently found endophyte Fusarium verticillioides an...

  16. Detrimental and Neutral Effects of a Wild Grass-Fungal Endophyte Symbiotum on Insect Preference and Performance

    Clement, Stephen L.; Hu, Jinguo; Stewart, Alan V.; Wang, Bingrui; Elberson, Leslie R.

    2011-01-01

    Seed-borne Epichloë/Neotyphodium Glenn, Bacon, Hanlin (Ascomycota: Hypocreales: Clavicipitaceae) fungal endophytes in temperate grasses can provide protection against insect attack with the degree of host resistance related to the grass—endophyte symbiotum and the insect species involved in an interaction. Few experimental studies with wild grass—endophyte symbiota, compared to endophyte-infected agricultural grasses, have tested for anti-insect benefits, let alone for resistance against more...

  17. Bacterial Seed Endophytes of Domesticated Cucurbits Antagonize Fungal and Oomycete Pathogens Including Powdery Mildew

    Khalaf, Eman M.; Raizada, Manish N.

    2018-01-01

    The cucurbit vegetables, including cucumbers, melons and pumpkins, have been cultivated for thousands of years without fungicides. However, their seed germination stage is prone to be infected by soil-borne fungal and oomycete pathogens. Endophytes are symbionts that reside inside plant tissues including seeds. Seed endophytes are founders of the juvenile plant microbiome and can promote host defense at seed germination and later stages. We previously isolated 169 bacterial endophytes associated with seeds of diverse cultivated cucurbits. We hypothesized that these endophytes can antagonize major fungal and oomycete pathogens. Here we tested the endophytes for in vitro antagonism (dual culture assays) against important soil-borne pathogens (Rhizoctonia solani, Fusarium graminearum, Phytophthora capsici, Pythium aphanideratum). The endophytes were also assayed in planta (leaf disk and detached leaf bioassays) for antagonism against a foliar pathogen of global importance, Podosphaera fuliginea, the causative agent of cucurbit powdery mildew. The endophytes were further tested in vitro for secretion of volatile organic compounds (VOCs) known to induce plant defense. Extracellular ribonuclease activity was also tested, as a subset of pathogenesis-related (PR) proteins of plant hosts implicated in suppression of fungal pathogens, displays ribonuclease activity. An unexpected majority of the endophytes (70%, 118/169) exhibited antagonism to the five phytopathogens, of which 68% (50/73) of in vitro antagonists belong to the genera Bacillus and Paenibacillus. All Lactococcus and Pantoea endophytes exhibited anti-oomycete activity. However, amongst the most effective inoculants against Podosphaera fuliginea were Pediococcus and Pantoea endophytes. Interestingly, 67% (113/169) of endophytes emitted host defense inducing VOCs (acetoin/diacetyl) and 62% (104/169) secreted extracellular ribonucleases in vitro, respectively. These results show that seeds of cultivated cucurbits

  18. Bacterial Seed Endophytes of Domesticated Cucurbits Antagonize Fungal and Oomycete Pathogens Including Powdery Mildew

    Eman M. Khalaf

    2018-02-01

    Full Text Available The cucurbit vegetables, including cucumbers, melons and pumpkins, have been cultivated for thousands of years without fungicides. However, their seed germination stage is prone to be infected by soil-borne fungal and oomycete pathogens. Endophytes are symbionts that reside inside plant tissues including seeds. Seed endophytes are founders of the juvenile plant microbiome and can promote host defense at seed germination and later stages. We previously isolated 169 bacterial endophytes associated with seeds of diverse cultivated cucurbits. We hypothesized that these endophytes can antagonize major fungal and oomycete pathogens. Here we tested the endophytes for in vitro antagonism (dual culture assays against important soil-borne pathogens (Rhizoctonia solani, Fusarium graminearum, Phytophthora capsici, Pythium aphanideratum. The endophytes were also assayed in planta (leaf disk and detached leaf bioassays for antagonism against a foliar pathogen of global importance, Podosphaera fuliginea, the causative agent of cucurbit powdery mildew. The endophytes were further tested in vitro for secretion of volatile organic compounds (VOCs known to induce plant defense. Extracellular ribonuclease activity was also tested, as a subset of pathogenesis-related (PR proteins of plant hosts implicated in suppression of fungal pathogens, displays ribonuclease activity. An unexpected majority of the endophytes (70%, 118/169 exhibited antagonism to the five phytopathogens, of which 68% (50/73 of in vitro antagonists belong to the genera Bacillus and Paenibacillus. All Lactococcus and Pantoea endophytes exhibited anti-oomycete activity. However, amongst the most effective inoculants against Podosphaera fuliginea were Pediococcus and Pantoea endophytes. Interestingly, 67% (113/169 of endophytes emitted host defense inducing VOCs (acetoin/diacetyl and 62% (104/169 secreted extracellular ribonucleases in vitro, respectively. These results show that seeds of cultivated

  19. Bacterial Seed Endophytes of Domesticated Cucurbits Antagonize Fungal and Oomycete Pathogens Including Powdery Mildew

    Eman M. Khalaf

    2018-02-01

    Full Text Available The cucurbit vegetables, including cucumbers, melons and pumpkins, have been cultivated for thousands of years without fungicides. However, their seed germination stage is prone to be infected by soil-borne fungal and oomycete pathogens. Endophytes are symbionts that reside inside plant tissues including seeds. Seed endophytes are founders of the juvenile plant microbiome and can promote host defense at seed germination and later stages. We previously isolated 169 bacterial endophytes associated with seeds of diverse cultivated cucurbits. We hypothesized that these endophytes can antagonize major fungal and oomycete pathogens. Here we tested the endophytes for in vitro antagonism (dual culture assays against important soil-borne pathogens (Rhizoctonia solani, Fusarium graminearum, Phytophthora capsici, Pythium aphanidermatum. The endophytes were also assayed in planta (leaf disk and detached leaf bioassays for antagonism against a foliar pathogen of global importance, Podosphaera fuliginea, the causative agent of cucurbit powdery mildew. The endophytes were further tested in vitro for secretion of volatile organic compounds (VOCs known to induce plant defense. Extracellular ribonuclease activity was also tested, as a subset of pathogenesis-related (PR proteins of plant hosts implicated in suppression of fungal pathogens, displays ribonuclease activity. An unexpected majority of the endophytes (70%, 118/169 exhibited antagonism to the five phytopathogens, of which 68% (50/73 of in vitro antagonists belong to the genera Bacillus and Paenibacillus. All Lactococcus and Pantoea endophytes exhibited anti-oomycete activity. However, amongst the most effective inoculants against Podosphaera fuliginea were Pediococcus and Pantoea endophytes. Interestingly, 67% (113/169 of endophytes emitted host defense inducing VOCs (acetoin/diacetyl and 62% (104/169 secreted extracellular ribonucleases in vitro, respectively. These results show that seeds of cultivated

  20. Endophytic fungal communities of Polygonum acuminatum and Aeschynomene fluminensis are influenced by soil mercury contamination.

    Pietro-Souza, William; Mello, Ivani Souza; Vendruscullo, Suzana Junges; Silva, Gilvan Ferreira da; Cunha, Cátia Nunes da; White, James Francis; Soares, Marcos Antônio

    2017-01-01

    The endophytic fungal communities of Polygonum acuminatum and Aeschynomene fluminensis were examined with respect to soil mercury (Hg) contamination. Plants were collected in places with and without Hg+2 for isolation and identification of their endophytic root fungi. We evaluated frequency of colonization, number of isolates and richness, indices of diversity and similarity, functional traits (hydrolytic enzymes, siderophores, indoleacetic acid, antibiosis and metal tolerance) and growth promotion of Aeschynomene fluminensis inoculated with endophytic fungi on soil with mercury. The frequency of colonization, structure and community function, as well as the abundant distribution of taxa of endophytic fungi were influenced by mercury contamination, with higher endophytic fungi in hosts in soil with mercury. The presence or absence of mercury in the soil changes the profile of the functional characteristics of the endophytic fungal community. On the other hand, tolerance of lineages to multiple metals is not associated with contamination. A. fluminensis depends on its endophytic fungi, since plants free of endophytic fungi grew less than expected due to mercury toxicity. In contrast plants containing certain endophytic fungi showed good growth in soil containing mercury, even exceeding growth of plants cultivated in soil without mercury. The data obtained confirm the hypothesis that soil contamination by mercury alters community structure of root endophytic fungi in terms of composition, abundance and species richness. The inoculation of A. fluminensis with certain strains of stress tolerant endophytic fungi contribute to colonization and establishment of the host and may be used in processes that aim to improve phytoremediation of soils with toxic concentrations of mercury.

  1. Isolation of peat swamp forest foliar endophyte fungi as biofertilizer

    Safinah Surya Hakim

    2017-01-01

    Full Text Available Peatland restoration activity is facing many obstacles, particularly in planting techniques and poor nutrient in peat soil. Naturally, endophytic fungi are abundant and have great potential as biofertilizer. This research investigates the potential endophytic fungi isolated from leaves of peat swamp tree species for biofertilizer. Research activities include: exploration, in vitro test to examine the phosphate solubilization and identification. Result showed that there were 360 leave segments collected from 4 sampling locations. The colonization percentage of 222 isolates ranged from 52.17% - 60.17%. Fifty seven morphospecies were selected from 222 isolates. Twelve isolates demonstrated ability to produce clear zones and ten isolates were selected for identification. It is concluded that twelve isolated demonstrated potential ability to produce clear zone and Penicillum citrinum isolate P3.10 was identified as an isolate that show the highest potential ability as a biofertilizer

  2. Root bacterial endophytes alter plant phenotype, but not physiology

    Henning, Jeremiah A.; Weston, David J.; Pelletier, Dale A.

    2016-01-01

    (root:shoot, biomass production, root and leaf growth rates) and physiological traits (chlorophyll content, net photosynthesis, net photosynthesis at saturating light-Asat, and saturating CO2-Amax). Overall, we found that bacterial root endophyte infection increased root growth rate up to 184% and leaf...... growth rate up to 137% relative to non-inoculated control plants, evidence that plants respond to bacteria by modifying morphology. However, endophyte inoculation had no influence on total plant biomass and photosynthetic traits (net photosynthesis, chlorophyll content). In sum, bacterial inoculation did......Plant traits, such as root and leaf area, influence how plants interact with their environment and the diverse microbiota living within plants can influence plant morphology and physiology. Here, we explored how three bacterial strains isolated from the Populus root microbiome, influenced plant...

  3. Endophytic fungi as models for the stereoselective biotransformation of thioridazine.

    Borges, Keyller Bastos; Borges, Warley De Souza; Pupo, Mônica Tallarico; Bonato, Pierina Sueli

    2007-12-01

    The stereoselective kinetic biotransformation of thioridazine, a phenothiazine neuroleptic drug, by endophytic fungi was investigated. In general, the sulfur of lateral chain (position 2) or the sulfur of phenothiazinic ring (position 5) were oxidated yielding the major human metabolites thioridazine-2-sulfoxide and thioridazine-5-sulfoxide. The quantity of metabolites biosynthesized varied among the 12 endophytic fungi evaluated. However, mono-2-sulfoxidation occurred in higher ratio and frequency. Among the 12 fungi evaluated, 4 of them deserve prominence for presenting an evidenced stereoselective biotransformation: Phomopsis sp. (TD2), Glomerella cingulata (VA1), Diaporthe phaseolorum (VR4), and Aspergillus fumigatus (VR12). Both enantiomers of thioridazine were consumed by the fungi; however, the 2-sulfoxidation yielded preferentially the R configuration at the sulfur atom.

  4. Cytotoxic and antibacterial naphthoquinones from an endophytic fungus, Cladosporium sp.

    Md. Imdadul Huque Khan

    Full Text Available Objective: Endophytes have the potential to synthesize various bioactive secondary metabolites. The aim of the study was to find new cytotoxic and antibacterial metabolites from endophytic fungus, Cladosporium sp. isolated from the leaves of Rauwolfia serpentina (L. Benth. ex Kurz. (Fam: Apocyanaceae. Materials and methods: The endophytic fungus was grown on potato dextrose agar medium and extracted using ethyl acetate. Secondary metabolites were isolated by chromatographic separation and re-crystallization, and structures were confirmed by 1H NMR, 13C NMR and mass spectroscopic data. The cytotoxicity was determined by WST-1 assay and brine shrimp lethality bioassay, while antibacterial activity was assessed by disc diffusion method. Results: Two naphthoquinones, namely anhydrofusarubin (1 and methyl ether of fusarubin (2, were isolated from Cladosporium sp. The isolated compounds 1 and 2, by WST-1 assay against human leukemia cells (K-562 showed potential cytotoxicity with IC50 values of 3.97 μg/mL and 3.58 μg/mL, respectively. Initial screening of crude ethyl acetate extract and column fractions F-8 and F-10 exhibited noticeable cytotoxicity to brine shimp nauplii with LC50 values of 42.8, 1.2 and 2.1 μg/mL, respectively. Moreover, the isolated compound 2 (40 μg/disc showed prominent activities against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Bacillus megaterium with an average zone of inhibition of 27 mm, 25 mm, 24 mm and 22 mm, respectively and the activities were compared with kanamycin (30 μg/disc. Conclusion: Our findings indicate that anhydrofusarubin (1 and methyl ether of fusarubin (2 might be useful lead compounds to develop potential cytotoxic and antimicrobial drugs. Keywords: Endophytic fungi, Cladosporium species, Fusarubin, Cytoxicity, Antibacterial activity

  5. Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth

    Castro, Renata Assis; Dourado, Manuella Nóbrega; Almeida, Jaqueline Raquel de; Lacava, Paulo Teixeira; Nave, André; Melo, Itamar Soares de; Azevedo, João Lucio de; Quecine, Maria Carolina

    2018-01-01

    ABSTRACT Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for th...

  6. Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth

    Castro, Renata Assis; Dourado, Manuella Nóbrega; Almeida, Jaqueline Raquel de; Lacava, Paulo Teixeira; Nave, André; Melo, Itamar Soares de; Azevedo, João Lucio de; Quecine, Maria Carolina

    2017-01-01

    Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their abili...

  7. A new dimeric anthraquinone from endophytic Talaromyces sp. YE3016.

    Xie, Xiao-Song; Fang, Xiao-Wei; Huang, Rong; Zhang, Shou-Peng; Wei, Hong-Xia; Wu, Shao-Hua

    2016-08-01

    A new unsymmetrical dimeric anthraquinone, 3-demethyl-3-(2-hydroxypropyl)-skyrin (1) was isolated from the solid-state fermentation extract of an endophytic fungal strain Talaromyces sp. YE 3016, together with five known compounds, skyrin (2), oxyskyrin (3), emodin (4), 1,3,6-trihydroxy-8-methyl-anthraquinone (5) and ergosterol (6). The structure of the new compound was elucidated on the basis of spectroscopic analysis. Compounds 1-3 exhibited moderate cytotoxic activities against MCF-7 cell line.

  8. Phenology of host Chondrus ocellatus with filamentous green endophyte infection

    Choi, Hang Gil; Kim, Changsong; Kim, Young Sik; Lee, Soon Jeong; Park, Myoung Ae; Nam, Ki Wan

    2015-09-01

    Monthly variations in gametophyte and tetrasporophyte biomass of Chondrus ocellatus Holmes, a commercial carragenophyte alga, were examined at wave-exposed and sheltered shore stations of Jungdori, Wando, Korea from September, 2013 to August, 2014. The frequency of infection of the fronds with a green filamentous endophyte was investigated and the endophyte was identified using tufA analysis. Biomass of C. ocellatus was significantly greater at the exposed shore (331.84 g wet wt. m-2) than at the sheltered shore (181 g wet wt. m-2); the average biomass was 259 g wet wt. m-2. Gametophyte biomass of C. ocellatus accounted for 64.25% of the total biomass (259 g wet wt. m-2); tetrasporophyte biomass was 93.05 g wet wt. m-2 (35.93%). Biomass was minimal in winter and maximal in summer at both stations and similar patterns were found for gametophyte and tetrasporophyte biomass. Frond lengths and weights of C. ocellatus were slightly greater at the exposed shore than at the sheltered shore. Fronds of C. ocellatus were infected by a green endophytic species, which grew in between the cortical and medullar tissue and was identified as Ulvella ramosa by tufA analysis. We conclude that the optimal harvesting period of the C. ocellatus field population in terms of biomass might be autumn, after the rapid growth period. Additional in-depth research on the endophytes, such as infection mechanism and frequency, should be performed in order to maintain and manage the field populations of C. ocellatus.

  9. Isolation and characterization of bacterial endophytes of Curcuma longa L.

    Kumar, Ajay; Singh, Ritu; Yadav, Akhilesh; Giri, D. D.; Singh, P. K.; Pandey, Kapil D.

    2016-01-01

    Fourteen endophytic bacterial isolates were isolated from the rhizome of Curcuma longa L. were characterized on the basis of morphology, biochemical characteristics and 16S rRNA gene sequence analysis. The isolates were identified to six strains namely Bacillus cereus (ECL1), Bacillus thuringiensis (ECL2), Bacillus sp. (ECL3), Bacillus pumilis (ECL4), Pseudomonas putida (ECL5), and Clavibacter michiganensis (ECL6). All the strains produced IAA and solubilized phosphate and only two strains pr...

  10. Sunflower growth according to seed inoculation with endophytic bacteria

    Juliana Fernandes dos Santos

    2014-06-01

    Full Text Available The sunflower crop has a great importance worldwide, due to the oil of excellent quality extracted from its seeds and in natura grains that are consumed in various ways. However, drought is one of the main environmental factors that limit its yield. An experiment was carried out under controlled greenhouse conditions, in a completely randomized experimental design, in order to determine the effect of endophytic bacteria inoculation (Bacillus sp. and Enterobacter cloacae on the growth and contents of nutrients and organic solutes, in sunflower leaves and roots under water deficit. Plant height, stem diameter, fresh and dry biomass of shoot and roots, as well as contents of N, P, K, soluble carbohydrates, free proline, free amino acids and soluble proteins, were determined at 35 days after the plant emergence. The water deficit reduced plant growth regardless inoculation. However, under optimum conditions of soil moisture, the combination of both endophytic bacteria increased the sunflower growth. The water deficit also increased the N and K contents in leaves, as well as the organic solutes content in shoots, especially in inoculated plants. These results suggest that the inoculation of endophytic bacteria may increase the capacity of drought stressed plants to perform the osmotic adjustment through a higher accumulation of organic solutes, when compared to plants not inoculated.

  11. Mangrove endophyte promotes reforestation tree (Acacia polyphylla growth

    Renata Assis Castro

    Full Text Available ABSTRACT Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast. Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10 had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48. We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth.

  12. OPTIMIZATION OF ENNIATINS PRODUCTION BY AN ENDOPHYTIC STRAIN FUSARIUM DIMERUM

    Eva Buchtová

    2014-10-01

    Full Text Available The goal of this study was to find suitable composition of cultivation media for enniatin production by isolated endophytic strain Fusarium dimerum. In order to find optimal cultivation media, mono- and di- saccharides, complex nitrogen sources and L-amino acids directed biosynthesis of enniatins were tested. Submerged cultivation experiments were carried out in cultivation flasks. Most promising medium for enniatin accumulation contained fructose, malt extract and peptone for bacteriology. Finally, quite expensive carbon source fructose was replaced by more available syrups. Optimization resulted in 4-times elevated enniatin biosynthesis by metabolites production microorganism. Moreover, this is the strain obtained from Magnolia soulangeana, which has similar metabolites spectrum as the isolated Fusarium dimerum. Comparison of these results with published ones revealed that this endophyte is a potential strain for enniatins biosynthesis in submerged cultivation in which the maximum accumulation 1.27 g.L-1 of enniatin in culture medium was reached in a short period (96 h. The results proved that the endophytic strain F. dimerum may potentially be applied for efficient production of bioactive enniatins.

  13. Isolation and characterization of bacterial endophytes of Curcuma longa L.

    Kumar, Ajay; Singh, Ritu; Yadav, Akhilesh; Giri, D D; Singh, P K; Pandey, Kapil D

    2016-06-01

    Fourteen endophytic bacterial isolates were isolated from the rhizome of Curcuma longa L. were characterized on the basis of morphology, biochemical characteristics and 16S rRNA gene sequence analysis. The isolates were identified to six strains namely Bacillus cereus (ECL1), Bacillus thuringiensis (ECL2), Bacillus sp. (ECL3), Bacillus pumilis (ECL4), Pseudomonas putida (ECL5), and Clavibacter michiganensis (ECL6). All the strains produced IAA and solubilized phosphate and only two strains produced siderophore (ECL3 and ECL5) during plant growth promoting trait analysis. All the endophytic strains utilized glucose, sucrose and yeast extract as a carbon source where as glycine, alanine, cystine and glutamine as nitrogen source. The strains were mostly sensitive to antibiotic chloramphenicol followed by erythromycin while resistant to polymixin B. The endophytic strains effectively inhibit the growth of Escherichia coli, Klebsiella pneumoniae and some of the fungal strain like Fusarium solani and Alterneria alternata. The strain ECL2 and ECL4 tolerated maximum 8 % of NaCl concentration where as strains ECL5 and ECL6 6 % in salinity tolerance.

  14. Research advance on stable mechanism of endophytic fungi to red wine colour during the aging

    Nan, Lijun; Li, Yashan; Cui, Changwei; Ning, Na; Huang, Jing; Xu, Chengdong; Tao, Fang; Zhang, Jinyong

    2018-04-01

    Based on the fact that persistent mutation of vinous color was not conducive to the stabilization of vinous quality during the aging, research advance on the stable mechanism of endophytic fungi to colour of red wine during the aging, including investigative status and developmental dynamic at home and abroad, endophytes and pigment of materials in wine, including effect of endophyte on vinaceous color, and even the application of tracer method in wine was summarized, respectively. The relationship between diversity of community the endophytic fungi and the main pigment material in wine was existent objectively, also included the response mechanism on colony dynamic of endophytic fungi to the various pigment as well as substance related to color, which laid the foundation for exploring the relationships between endophytic fungi and wine color, and the variational mechanism of the color under endophytic fungi during the aging period of wine. Color as an important reference index of wine quality influenced not only the sensory evaluation of consumer, but also the quality of wine because of the self-aggregation or combination of phenolic composition with other substances under the endophytic fungi during the storage. Only steady wine in the color could guarantee the security of quality.

  15. A fungal endophyte helps plants to tolerate root herbivory through changes in gibberellin and jasmonate signaling

    Rebeca Cosme, M.P.

    2016-01-01

    Plant–microbe mutualisms can improve plant defense, but the impact of root endophytes on below-ground herbivore interactions remains unknown. We investigated the effects of the root endophyte Piriformospora indica on interactions between rice (Oryza sativa) plants and its root herbivore rice water

  16. Fungi and bacteria boost resistance to pests and diseases : endophytes a useful addition to pest control

    Messelink, G.

    2017-01-01

    More and more research is revealing that endophytes – microorganisms that live in the plant without harming it – can significantly boost a plant’s resistance to pests. These findings prompted researchers to investigate the potential of endophytes in pest control in greenhouse horticulture.

  17. The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants

    Azevedo, João Lúcio; Araújo, Welington Luiz; Lacava, Paulo Teixeira

    2016-01-01

    Abstract The bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC) and has been associated with important losses in commercial orchards of all sweet orange [Citrus sinensis (L.)] cultivars. The development of this disease depends on the environmental conditions, including the endophytic microbial community associated with the host plant. Previous studies have shown that X. fastidiosa interacts with the endophytic community in xylem vessels as well as in the insect vector, resulting in a lower bacterial population and reduced CVC symptoms. The citrus endophytic bacterium Methylobacterium mesophilicum can trigger X. fastidiosa response in vitro, which results in reduced growth and induction of genes associated with energy production, stress, transport, and motility, indicating that X. fastidiosa has an adaptive response to M. mesophilicum. Although this response may result in reduced CVC symptoms, the colonization rate of the endophytic bacteria should be considered in studies that intend to use this endophyte to suppress CVC disease. Symbiotic control is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace pathogens. Candidate endophytes for symbiotic control of CVC must occupy the xylem of host plants and attach to the precibarium of sharpshooter insects to access the pathogen. In the present review, we focus on interactions between endophytic bacteria from sweet orange plants and X. fastidiosa, especially those that may be candidates for control of CVC. PMID:27727362

  18. An Acremonium endophyte of Lolium perenne associated with hyperthermia of cattle in Pacific County, Washington

    A. D. Wilson; C.C. Gay; S.C. and Fransen

    1992-01-01

    Clavicipitaceous endophytes are well known for causing maladies of livestock. Recent studies of a new syndrome causing hyperthermia of cattle in Pacific County, Washington, prompted surveys of endophytes in pasture grasses of seven affected paddocks. Cattle removed from affected pastures and fed alfalfa became normothermic within 3 days, suggesting a pyrogenic factor...

  19. Endophytic bacteria in cacti seeds can improve the development of cactus seedlings.

    M. Esther Puente; Ching Y. Li; Yoav Bashan

    2009-01-01

    A plant-bacterium association between the giant cardon cactus Pachycereus pringlei and endophytic bacteria help seedlings establish and grow on barren rock, This cactus, together with other desert plants, is responsible for weathering ancient lava flows in the Baja California Peninsula of Mexico.When cardon seeds are inoculated with endophytic...

  20. The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants

    João Lúcio Azevedo

    Full Text Available Abstract The bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC and has been associated with important losses in commercial orchards of all sweet orange [Citrus sinensis (L.] cultivars. The development of this disease depends on the environmental conditions, including the endophytic microbial community associated with the host plant. Previous studies have shown that X. fastidiosa interacts with the endophytic community in xylem vessels as well as in the insect vector, resulting in a lower bacterial population and reduced CVC symptoms. The citrus endophytic bacterium Methylobacterium mesophilicum can trigger X. fastidiosa response in vitro, which results in reduced growth and induction of genes associated with energy production, stress, transport, and motility, indicating that X. fastidiosa has an adaptive response to M. mesophilicum. Although this response may result in reduced CVC symptoms, the colonization rate of the endophytic bacteria should be considered in studies that intend to use this endophyte to suppress CVC disease. Symbiotic control is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace pathogens. Candidate endophytes for symbiotic control of CVC must occupy the xylem of host plants and attach to the precibarium of sharpshooter insects to access the pathogen. In the present review, we focus on interactions between endophytic bacteria from sweet orange plants and X. fastidiosa, especially those that may be candidates for control of CVC.

  1. Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense.

    Sundram, Shamala; Meon, Sariah; Seman, Idris Abu; Othman, Radziah

    2011-08-01

    Endophytic bacteria (Pseudomonas aeruginosa UPMP3 and Burkholderia cepacia UMPB3), isolated from within roots of oil palm (Elaeis guineensis Jacq.) were tested for their presymbiotic effects on two arbuscular mcorrhizal fungi, Glomus intraradices UT126 and Glomus clarum BR152B). These endophytic bacteria were also tested for antagonistic effects on Ganoderma boninense PER 71, a white wood rot fungal pathogen that causes a serious disease in oil palm. Spore germination and hyphal length of each arbuscular mycorrhizal fungal (AMF) pairing with endophytic bacteria was found to be significantly higher than spores plated in the absence of bacteria. Scanning electron microscopy (SEM) showed that the endophytic bacteria were scattered, resting or embedded on the surface hyaline layer or on the degraded walls of AMF spores, possibly feeding on the outer hyaline spore wall. The antagonistic effect of the endophytic bacteria was expressed as severe morphological abnormalities in the hyphal structures of G. boninense PER 71. The effects of the endophytic bacteria on G. boninense PER 71 hyphal structures were observed clearly under SEM. Severe inter-twisting, distortion, lysis and shriveling of the hyphal structures were observed. This study found that the effect of endophytic bacteria on G. intraradices UT126 and G. clarum BR152B resembled that of a mycorrhiza helper bacteria (MHB) association because the association significantly promoted AMF spore germination and hyphal length. However, the endophytic bacteria were extremely damaging to G. boninense PER 71.

  2. Distribution and dispersal of Xylaria endophytes in two tree species in Puerto Rico

    P. Bayman; D. J. Lodge; P. Angulo-Sandoval; Z. Baez-Ortiz

    1998-01-01

    Xylaria species are common endophytes in tropical plants. It is not known, however, whether transmission of Xylaria occurs horizontally or vertically, whether individual Xylaria strains have wide host ranges or are host-specific, or how they are dispersed. We compared frequency of Xylaria endophytes in leaves and seeds of two tree species in Puerto Rico, Casuarina...

  3. Restructuring of endophytic bacterial communities in grapevine yellows-diseased and recovered Vitis vinifera L. plants.

    Bulgari, Daniela; Casati, Paola; Crepaldi, Paola; Daffonchio, Daniele; Quaglino, Fabio; Brusetti, Lorenzo; Bianco, Piero Attilio

    2011-07-01

    Length heterogeneity-PCR assays, combined with statistical analyses, highlighted that the endophytic bacterial community associated with healthy grapevines was characterized by a greater diversity than that present in diseased and recovered plants. The findings suggest that phytoplasmas can restructure the bacterial community by selecting endophytic strains that could elicit a plant defense response.

  4. Vasoconstriction in horses caused by endophyte-infected tall fescue seed is detected with Doppler ultrasonography

    The hypotheses that endophyte (Neotyphodium coenophialum)-infected tall fescue (TF) seed causes vasoconstriction in horses in vivo and that ground seed would cause more pronounced vasoconstriction than whole seed were tested. Ten horses each received 1 of 3 treatments: endophyte-free ground (E–G; n ...

  5. Molecular detection of TasA gene in endophytic Bacillus species ...

    hope&shola

    2012-03-20

    Mar 20, 2012 ... formation in Bacillus species was detected in the endophytic bacteria by polymerase chain reaction. (PCR) amplification. In ten endophytic ... confer a competitive advantage to the spore from the onset of sporulation and later, ... possessing TasA gene (Chen et al., 2007; Gioia et al.,. 2007; Kunst et al., 1997; ...

  6. A New Eudesmane Sesquiterpene from Nigrospora oryzae, an Endophytic Fungus of Aquilaria sinensis

    Dongli Li

    2014-07-01

    Full Text Available A new eudesmane-type sesquiterpene, 11 -hydroxy capitulatin B (1 , along with a known related sesquiterpene, capitulatin B (2, was isolated from the endophytic fungus Nigrospora oryzae A8 from Aquilaria sinensis, the only plant resource for agarwood production in China. This research demonstrates that the endophytic fungi from A. sinensis might play a role in the formation of agarwood.

  7. What Is There in Seeds? Vertically Transmitted Endophytic Resources for Sustainable Improvement in Plant Growth

    Raheem Shahzad

    2018-01-01

    Full Text Available Phytobeneficial microbes, particularly endophytes, such as fungi and bacteria, are concomitant partners of plants throughout its developmental stages, including seed germination, root and stem growth, and fruiting. Endophytic microbes have been identified in plants that grow in a wide array of habitats; however, seed-borne endophytic microbes have not been fully explored yet. Seed-borne endophytes are of great interest because of their vertical transmission; their potential to produce various phytohormones, enzymes, antimicrobial compounds, and other secondary metabolites; and improve plant biomass and yield under biotic and abiotic stresses. This review addresses the current knowledge on endophytes, their ability to produce metabolites, and their influence on plant growth and stress mitigation.

  8. Frequency of endophytic fungi isolated from Dendrobium crumenatum (Pigeon orchid and antimicrobial activity

    WIBOWO MANGUNWARDOYO

    2012-01-01

    Full Text Available Mangunwardoyo W, Suciatmih, Gandjar I. 2012. Frequency of endophytic fungi isolated from Dendrobium crumenatum (Pigeon orchid and antimicrobial activity. Biodiversitas 13: 34-39. The aims of this research was to isolate and study the frequency of endophytic fungi from roots, bulbous, stems, and leaves of Dendrobium crumenatum Sw. (pigeon orchid collected from Tanah Baru housing area, Bogor Botanical Garden, and Herbarium Bogoriense; and to assess for antimicrobial activity against Candida albicans ATCC 2091, Candida tropicalis LIPIMC 203, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6633 and Staphylococcus aureus ATCC 25923. Twelve species of endophytic fungi were identified from 60 samples obtained from D. crumenatum. Guignardia endophyllicola (anamorph: Phyllosticta capitalensis were the dominant endophytic fungi. Screening of the anti-microorganism activity of the endophytic fungi revealed that Fusarium nivale inhibited C albicans and C. tropicalis. All specimens did not inhibit B. subtilis, E. coli, and S. aureus.

  9. Diversity of fungal endophytes in non-native Phragmites australis in the Great Lakes

    Clay, Keith; Shearin, Zachery; Bourke, Kimberly; Bickford, Wesley A.; Kowalski, Kurt P.

    2016-01-01

    Plant–microbial interactions may play a key role in plant invasions. One common microbial interaction takes place between plants and fungal endophytes when fungi asymptomatically colonize host plant tissues. The objectives of this study were to isolate and sequence fungal endophytes colonizing non-native Phragmites australis in the Great Lakes region to evaluate variation in endophyte community composition among three host tissue types and three geographical regions. We collected entire ramets from multiple clones and populations, surface sterilized plant tissues, and plated replicate tissue samples from leaves, stems, and rhizomes on corn meal agar plates to culture and isolate fungal endophytes. Isolates were then subjected to Sanger sequencing of the ITS region of the nuclear ribosomal DNA. Sequences were compared to fungal databases to define operational taxonomic units (OTUs) that were analyzed statistically for community composition. In total, we obtained 173 endophyte isolates corresponding to 55 OTUs, 39 of which were isolated only a single time. The most common OTU corresponded most closely to Sarocladium strictum and comprised 25 % of all fungal isolates. More OTUs were found in stem tissues, but endophyte diversity was greatest in rhizome tissues. PERMANOVA analyses indicated significant differences in endophyte communities among tissue types, geographical regions, and the interaction between those factors, but no differences among individual ramets were detected. The functional role of the isolated endophytes is not yet known, but one genus isolated here (Stagonospora) has been reported to enhance Phragmites growth. Understanding the diversity and functions of Phragmites endophytes may provide targets for control measures based on disrupting host plant/endophyte interactions.

  10. Bioprospecting of South African Plants as a Unique Resource for Bioactive Endophytic Microbes

    Muna Ali Abdalla

    2018-05-01

    Full Text Available South Africa has a long history and strong belief in traditional herbal medicines. Using ethnobotanical knowledge as a lead, a large number of South African medicinal plants have been discovered to possess a wide spectrum of pharmacological properties. In this review, bioprospecting of endophytes is highlighted by following the advantages of the ethnomedicinal approach together with identifying unique medicinal plants where biological activity may be due to endophytes. This review focuses on the current status of South African medicinal plants to motivate the research community to harness the benefits of ethnobotanical knowledge to investigate the presence of endophytic microbes from the most potent South African medicinal plants. The potential chemical diversity and subsequent putative medicinal value of endophytes is deserving of further research. A timely and comprehensive review of literature on recently isolated endophytes and their metabolites was conducted. Worldwide literature from the last 2 years demonstrating the importance of ethnobotanical knowledge as a useful approach to discover endophytic microbes was documented. Information was obtained from scientific databases such as Pubmed, Scopus, Scirus, Google Scholar, Dictionary of Natural Products, Chemical Abstracts Services, official websites, and scientific databases on ethnomedicines. Primary sources such as books, reports, dissertations, and thesises were accessed where available. Recently published information on isolated endophytes with promising bioactivity and their bioactive natural products worldwide (2015-2017 was summarized. The potential value of South African medicinal plants as sources of endophytes is discussed. The insights provided through this study indicate that medicinal plants in South Africa are highly under-investigated sources of potentially useful endophytic microbes. New approaches may be used by medicinal plant scientists for further exploration of natural

  11. Bacterial endophytes from wild maize suppress Fusarium graminearum in modern maize and inhibit mycotoxin accumulation

    Walaa Kamel Mousa

    2015-10-01

    Full Text Available Wild maize (teosinte has been reported to be less susceptible to pests than their modern maize (corn relatives. Endophytes, defined as microbes that inhabit plants without causing disease, are known for their ability to antagonize plant pests and pathogens. We hypothesized that the wild relatives of modern maize may host endophytes that combat pathogens. Fusarium graminearum is the fungus that causes Gibberella Ear Rot (GER in modern maize and produces the mycotoxin, deoxynivalenol (DON. In this study, 215 bacterial endophytes, previously isolated from diverse maize genotypes including wild teosintes, traditional landraces and modern varieties, were tested for their ability to antagonize F. graminearum in vitro. Candidate endophytes were then tested for their ability to suppress GER in modern maize in independent greenhouse trials. The results revealed that three candidate endophytes derived from wild teosintes were most potent in suppressing F. graminearum in vitro and GER in a modern maize hybrid. These wild teosinte endophytes could suppress a broad spectrum of fungal pathogens of modern crops in vitro. The teosinte endophytes also suppressed DON mycotoxin during storage to below acceptable safety threshold levels. A fourth, less robust anti-fungal strain was isolated from a modern maize hybrid. Three of the anti-fungal endophytes were predicted to be Paenibacillus polymyxa, along with one strain of Citrobacter. Microscopy studies suggested a fungicidal mode of action by all four strains. Molecular and biochemical studies showed that the P. polymyxa strains produced the previously characterized anti-Fusarium compound, fusaricidin. Our results suggest that the wild relatives of modern crops may serve as a valuable reservoir for endophytes in the ongoing fight against serious threats to modern agriculture. We discuss the possible impact of crop evolution and domestication on endophytes in the context of plant defense.

  12. An endophytic fungus isolated from finger millet (Eleucine coracona produces anti-fungal natural products

    Walaa Kamel Mousa

    2015-10-01

    Full Text Available Finger millet is an ancient African cereal crop, domesticated 7000 years ago in Ethiopia, reaching India at 3000 BC. Finger millet is reported to be resistant to various fungal pathogens including Fusarium sp. We hypothesized that finger millet may host beneficial endophytes (plant-colonizing microbes that contribute to the antifungal activity. Here we report the first isolation of endophyte(s from finger millet. Five distinct fungal species were isolated from roots and predicted taxonomically based on 18S rDNA sequencing. Extracts from three putative endophytes inhibited growth of F. graminearum and three other pathogenic Fusarium species. The most potent anti-Fusarium strain (WF4, predicted to be a Phoma sp. was confirmed to behave as an endophyte using pathogenicity and confocal microscopy experiments. Bioassay-guided fractionation of the WF4 extract identified four anti-fungal compounds, viridicatol, tenuazonic acid, alternariol and alternariol monomethyl ether. All the purified compounds caused dramatic breakage of F. graminearum hyphae in vitro. These compounds have not previously been reported to have anti-Fusarium activity. None of the compounds, except for tenuazonic acid, have previously been reported to be produced by Phoma. We conclude that the ancient, disease-tolerant crop, finger millet, is a novel source of endophytic anti-fungal natural products. This paper suggests the value of the crops grown by subsistence farmers as sources of endophytes and their natural products. Application of these natural chemicals to solve real world problems will require further validation.

  13. An endophytic fungus isolated from finger millet (Eleusine coracana) produces anti-fungal natural products.

    Mousa, Walaa K; Schwan, Adrian; Davidson, Jeffrey; Strange, Philip; Liu, Huaizhi; Zhou, Ting; Auzanneau, France-Isabelle; Raizada, Manish N

    2015-01-01

    Finger millet is an ancient African cereal crop, domesticated 7000 years ago in Ethiopia, reaching India at 3000 BC. Finger millet is reported to be resistant to various fungal pathogens including Fusarium sp. We hypothesized that finger millet may host beneficial endophytes (plant-colonizing microbes) that contribute to the antifungal activity. Here we report the first isolation of endophyte(s) from finger millet. Five distinct fungal species were isolated from roots and predicted taxonomically based on 18S rDNA sequencing. Extracts from three putative endophytes inhibited growth of F. graminearum and three other pathogenic Fusarium species. The most potent anti-Fusarium strain (WF4, predicted to be a Phoma sp.) was confirmed to behave as an endophyte using pathogenicity and confocal microscopy experiments. Bioassay-guided fractionation of the WF4 extract identified four anti-fungal compounds, viridicatol, tenuazonic acid, alternariol, and alternariol monomethyl ether. All the purified compounds caused dramatic breakage of F. graminearum hyphae in vitro. These compounds have not previously been reported to have anti-Fusarium activity. None of the compounds, except for tenuazonic acid, have previously been reported to be produced by Phoma. We conclude that the ancient, disease-tolerant crop, finger millet, is a novel source of endophytic anti-fungal natural products. This paper suggests the value of the crops grown by subsistence farmers as sources of endophytes and their natural products. Application of these natural chemicals to solve real world problems will require further validation.

  14. A Friendly Relationship between Endophytic Fungi and Medicinal Plants: A Systematic Review

    Jia, Min; Chen, Ling; Xin, Hai-Liang; Zheng, Cheng-Jian; Rahman, Khalid; Han, Ting; Qin, Lu-Ping

    2016-01-01

    Endophytic fungi or endophytes exist widely inside the healthy tissues of living plants, and are important components of plant micro-ecosystems. Over the long period of evolution, some co-existing endophytes and their host plants have established a special relationship with one and another, which can significantly influence the formation of metabolic products in plants, then affect quality and quantity of crude drugs derived from medicinal plants. This paper will focus on the increasing knowledge of relationships between endophytic fungi and medicinal plants through reviewing of published research data obtained from the last 30 years. The analytical results indicate that the distribution and population structure of endophytes can be considerably affected by factors, such as the genetic background, age, and environmental conditions of their hosts. On the other hand, the endophytic fungi can also confer profound impacts on their host plants by enhancing their growth, increasing their fitness, strengthening their tolerances to abiotic and biotic stresses, and promoting their accumulation of secondary metabolites. All the changes are very important for the production of bioactive components in their hosts. Hence, it is essential to understand such relationships between endophytic fungi and their host medicinal plants. Such knowledge can be well exploited and applied for the production of better and more drugs from medicinal plants. PMID:27375610

  15. Isolation and identification of resveratrol-producing endophytes from wine grape Cabernet Sauvignon.

    Liu, Ya; Nan, Lijun; Liu, Junchao; Yan, Haiyan; Zhang, Dianpeng; Han, Xinnian

    2016-01-01

    Obtain endophyte strains with effective resveratrol production from superior grapevine variety Cabernet Sauvignon in Xinjiang and determine related taxonomic position of the strain. Seventy-three strains of endophytes, including 23 strains of bacteria, 14 ones of actinomycetes, 24 fungus and 12 yeasts, were isolated, respectively. The distribution law of endophytes was spring (30.14 %) = summer (30.14 %) < autumn (39.73 %) in different seasons, while the fruit (12.33 %) < leaf (20.55 %) < stem (32.88 %) < root (34.25 %) in different tissues and organs. From the 36 strains of endophytic fungi isolated, seven strains producing polyphenols were screened by ferric chloride-potassium ferricyanide color reaction. C2J6, stable genetic properties producing highly 1.48 mg L(-1) of resveratrol, was identified as Aspergillus niger by 26S rDNA-ITS sequence analysis after thin-layer chromatography sieve analysis, ultra violet wavelength scanning and high performance liquid chromatography, respectively. There were the certain number and kinds of endophytes in the various tissues of Cabernet Sauvignon, which, to a certain extent, reflected the biological diversity of plant endophytes. The fact that the fungus C2J6 producing resveratrol in grape was acquired attested the special ability of the endophytes to produce the same or similar bioactive substances as the host plants.

  16. Salicaceae Endophytes Modulate Stomatal Behavior and Increase Water Use Efficiency in Rice

    Hyungmin Rho

    2018-03-01

    Full Text Available Bacterial and yeast endophytes isolated from the Salicaceae family have been shown to promote growth and alleviate stress in plants from different taxa. To determine the physiological pathways through which endophytes affect plant water relations, we investigated leaf water potential, whole-plant water use, and stomatal responses of rice plants to Salicaceae endophyte inoculation under CO2 enrichment and water deficit. Daytime stomatal conductance and stomatal density were lower in inoculated plants compared to controls. Leaf ABA concentrations increased with endophyte inoculation. As a result, transpirational water use decreased significantly with endophyte inoculation while biomass did not change or slightly increased. This response led to a significant increase in cumulative water use efficiency at harvest. Different endophyte strains produced the same results in host plant water relations and stomatal responses. These stomatal responses were also observed under elevated CO2 conditions, and the increase in water use efficiency was more pronounced under water deficit conditions. The effect on water use efficiency was positively correlated with daily light integrals across different experiments. Our results provide insights on the physiological mechanisms of plant-endophyte interactions involving plant water relations and stomatal functions.

  17. Linking Bacterial Endophytic Communities to Essential Oils: Clues from Lavandula angustifolia Mill

    Emiliani, Giovanni; Maida, Isabel; Perrin, Elena; Chiellini, Carolina; Fondi, Marco; Gallo, Eugenia; Gori, Luigi; Maggini, Valentina; Vannacci, Alfredo; Biffi, Sauro; Firenzuoli, Fabio; Fani, Renato

    2014-01-01

    Endophytic bacteria play a crucial role in plant life and are also drawing much attention for their capacity to produce bioactive compounds of relevant biotechnological interest. Here we present the characterisation of the cultivable endophytic bacteria of Lavandula angustifolia Mill.—a species used since antiquity for its therapeutic properties—since the production of bioactive metabolites from medical plants may reside also in the activity of bacterial endophytes through their direct production, PGPR activity on host, and/or elicitation of plant metabolism. Lavender tissues are inhabited by a tissue specific endophytic community dominated by Proteobacteria, highlighting also their difference from the rhizosphere environment where Actinobacteria and Firmicutes are also found. Leaves' endophytic community resulted as the most diverse from the other ecological niches. Overall, the findings reported here suggest: (i) the existence of different entry points for the endophytic community, (ii) its differentiation on the basis of the ecological niche variability, and (iii) a two-step colonization process for roots endophytes. Lastly, many isolates showed a strong inhibition potential against human pathogens and the molecular characterization demonstrated also the presence of not previously described isolates that may constitute a reservoir of bioactive compounds relevant in the field of pathogen control, phytoremediation, and human health. PMID:24971151

  18. Bacterial endophyte communities of three agricultural important grass species differ in their response towards management regimes

    Wemheuer, Franziska; Kaiser, Kristin; Karlovsky, Petr; Daniel, Rolf; Vidal, Stefan; Wemheuer, Bernd

    2017-01-01

    Endophytic bacteria are critical for plant growth and health. However, compositional and functional responses of bacterial endophyte communities towards agricultural practices are still poorly understood. Hence, we analyzed the influence of fertilizer application and mowing frequency on bacterial endophytes in three agriculturally important grass species. For this purpose, we examined bacterial endophytic communities in aerial plant parts of Dactylis glomerata L., Festuca rubra L., and Lolium perenne L. by pyrotag sequencing of bacterial 16S rRNA genes over two consecutive years. Although management regimes influenced endophyte communities, observed responses were grass species-specific. This might be attributed to several bacteria specifically associated with a single grass species. We further predicted functional profiles from obtained 16S rRNA data. These profiles revealed that predicted abundances of genes involved in plant growth promotion or nitrogen metabolism differed between grass species and between management regimes. Moreover, structural and functional community patterns showed no correlation to each other indicating that plant species-specific selection of endophytes is driven by functional rather than phylogenetic traits. The unique combination of 16S rRNA data and functional profiles provided a holistic picture of compositional and functional responses of bacterial endophytes in agricultural relevant grass species towards management practices.

  19. Assessment of functional traits in the assemblage of endophytic fungi of anacardium othonianum rizzini

    Faria, P. S.; Senabio, J. A.; Soares, M. A.

    2016-01-01

    Plants maintain symbiotic relationships with microorganisms as a strategy to withstand adversities. From this exchange, organisms receive photoassimilates and provide benefits to the plant. Anacardium othonianum Rizzini, locally known as caju-de-arvore-do-cerrado (tree cashew of the cerrado), is a tree species of the family Anacardiaceae nativeto the Midwest region of Brazil. The objective of this study was to characterize the culturable endophytic fungal community, its functional traits and its association with the roots of A. othonianum. The roots of A. othonianum were fragmented (1 cm) and inoculated in medium for the isolation of endophytic microorganisms. The molecular identification of the isolates was performed through the partial sequencing of the internal transcribed spacer (ITS). The endophytic isolates were tested for the synthesis of indole acetic acid (IAA) and phosphate solubilization through the colorimetric method. The root fragments were cleared, stained and examined under a microscope. Structures characteristic of endomycorrhizal and endophytic microorganisms were found on the slides analyzed. A total of 67 fungal strains were isolated and identified in 12 species: Fusarium oxysporum, Bionectria ochroleuca, Periconia macrospinosa, Phomopsis lagerstroemiae, Penicillium kloeckeri, Eupenicillium shearii, Phomopsis asparagi, Penicillium pinophilum, Agaricomycetes sp., Diaporthe sp., Cladosporium cladosporioide sand Paecilomyces lilacinus. All the genera found have been reported in the literature as endophytic species. It can be concluded that A. othonianum maintains associations with endomycorrhizal and endophytic fungi. Twelve endophytic strains were isolated from A. othonianum Rizzini, seven of which have potential for phosphate solubilization and IAA synthesis. (author)

  20. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters.

    Martínez-Rodríguez, Julia del C; De la Mora-Amutio, Marcela; Plascencia-Correa, Luis A; Audelo-Regalado, Esmeralda; Guardado, Francisco R; Hernández-Sánchez, Elías; Peña-Ramírez, Yuri J; Escalante, Adelfo; Beltrán-García, Miguel J; Ogura, Tetsuya

    2014-01-01

    Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost.

  1. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters

    Julia del C. Martínez-Rodríguez

    2014-12-01

    Full Text Available Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI. Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost.

  2. A Legume Genetic Framework Controls Infection of Nodules by Symbiotic and Endophytic Bacteria

    Zgadzaj, Rafal; James, Euan K.; Kelly, Simon; Kawaharada, Yasuyuki; de Jonge, Nadieh; Jensen, Dorthe B.; Madsen, Lene H.; Radutoiu, Simona

    2015-01-01

    Legumes have an intrinsic capacity to accommodate both symbiotic and endophytic bacteria within root nodules. For the symbionts, a complex genetic mechanism that allows mutual recognition and plant infection has emerged from genetic studies under axenic conditions. In contrast, little is known about the mechanisms controlling the endophytic infection. Here we investigate the contribution of both the host and the symbiotic microbe to endophyte infection and development of mixed colonised nodules in Lotus japonicus. We found that infection threads initiated by Mesorhizobium loti, the natural symbiont of Lotus, can selectively guide endophytic bacteria towards nodule primordia, where competent strains multiply and colonise the nodule together with the nitrogen-fixing symbiotic partner. Further co-inoculation studies with the competent coloniser, Rhizobium mesosinicum strain KAW12, show that endophytic nodule infection depends on functional and efficient M. loti-driven Nod factor signalling. KAW12 exopolysaccharide (EPS) enabled endophyte nodule infection whilst compatible M. loti EPS restricted it. Analysis of plant mutants that control different stages of the symbiotic infection showed that both symbiont and endophyte accommodation within nodules is under host genetic control. This demonstrates that when legume plants are exposed to complex communities they selectively regulate access and accommodation of bacteria occupying this specialized environmental niche, the root nodule. PMID:26042417

  3. Fungal Endophytes as a Metabolic Fine-Tuning Regulator for Wine Grape.

    Ming-Zhi Yang

    Full Text Available Endophytes proved to exert multiple effects on host plants, including growth promotion, stress resistance. However, whether endophytes have a role in metabolites shaping of grape has not been fully understood. Eight endophytic fungal strains which originally isolated from grapevines were re-inoculated to field-grown grapevines in this study, and their effects on both leaves and berries of grapevines at maturity stage were assessed, with special focused on secondary metabolites and antioxidant activities. High-density inoculation of all these endophytic fungal strains modified the physio-chemical status of grapevine to different degrees. Fungal inoculations promoted the content of reducing sugar (RS, total flavonoids (TF, total phenols (TPh, trans-resveratrol (Res and activities of phenylalanine ammonia-lyase (PAL, in both leaves and berries of grapevine. Inoculation of endophytic fungal strains, CXB-11 (Nigrospora sp. and CXC-13 (Fusarium sp. conferred greater promotion effects in grape metabolic re-shaping, compared to other used fungal strains. Additionally, inoculation of different strains of fungal endophytes led to establish different metabolites patterns of wine grape. The work implies the possibility of using endophytic fungi as fine-tuning regulator to shape the quality and character of wine grape.

  4. Antifungal and proteolytic activities of endophytic fungi isolated from Piper hispidum Sw

    Ravely Casarotti Orlandelli

    2015-06-01

    Full Text Available Endophytes are being considered for use in biological control, and the enzymes they secrete might facilitate their initial colonization of internal plant tissues and direct interactions with microbial pathogens. Microbial proteases are also biotechnologically important products employed in bioremediation processes, cosmetics, and the pharmaceutical, photographic and food industries. In the present study, we evaluated antagonism and competitive interactions between 98 fungal endophytes and Alternaria alternata, Colletotrichum sp., Phyllosticta citricarpa and Moniliophthora perniciosa. We also examined the proteolytic activities of endophytes grown in liquid medium and conducted cup plate assays. The results showed that certain strains in the assemblage of P. hispidum endophytes are important sources of antifungal properties, primarily Lasiodiplodia theobromae JF766989, which reduced phytopathogen growth by approximately 54 to 65%. We detected 28 endophytes producing enzymatic halos of up to 16.40 mm in diameter. The results obtained in the present study highlight the proteolytic activity of the endophytes Phoma herbarum JF766995 and Schizophyllum commune JF766994, which presented the highest enzymatic halo diameters under at least one culture condition tested. The increased activities of certain isolates in the presence of rice or soy flour as a substrate (with halos up to 17.67 mm in diameter suggests that these endophytes have the potential to produce enzymes using agricultural wastes.

  5. Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth.

    Castro, Renata Assis; Dourado, Manuella Nóbrega; Almeida, Jaqueline Raquel de; Lacava, Paulo Teixeira; Nave, André; Melo, Itamar Soares de; Azevedo, João Lucio de; Quecine, Maria Carolina

    Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000km 2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  6. Comparative analysis of prodigiosin isolated from endophyte Serratia marcescens.

    Khanam, B; Chandra, R

    2018-03-01

    Extraction of pigments from endophytes is an uphill task. Up till now, there are no efficient methods available to extract the maximum amount of prodigiosin from Serratia marcescens. This is one of the important endophytes of Beta vulgaris L. The present work was carried out for the comparative study of six different extraction methods such as homogenization, ultrasonication, freezing and thawing, heat treatment, organic solvents and inorganic acids to evaluate the efficiency of prodigiosin yield. Our results demonstrated that highest extraction was observed in ultrasonication (98·1 ± 1·7%) while the lowest extraction by freezing and thawing (31·8 ± 3·8%) methods. However, thin layer chromatography, high-performance liquid chromatography and Fourier transform infrared data suggest that bioactive pigment in the extract was prodigiosin. To the best of our knowledge, this is the first comprehensive study of extraction methods and identification and purification of prodigiosin from cell biomass of Ser. marcescens isolated from Beta vulgaris L. The prodigiosin family is a potent drug with anticancer, antimalarial, antibacterial, antifungal, antiproliferative and immunosuppressive activities. Moreover, it has immense potential in pharmaceutical, food and textile industries. For the industrial perspective, it is essential to achieve purified, high yield and cost-effective extraction of prodigiosin. To the best of our knowledge, this is the first comprehensive study on prodigiosin extraction and also the first report on endophyte Serratia marcescens isolated from Beta vulgaris L. The significance of our results is to extract high amount and good quality prodigiosin for commercial application. © 2017 The Society for Applied Microbiology.

  7. Antimicrobial activity of Ulva reticulata and its endophytes

    Dhanya, K. I.; Swati, V. I.; Vanka, Kanth Swaroop; Osborne, W. J.

    2016-04-01

    Seaweeds are known to exhibit various antimicrobial properties, since it harbours an enormous range of indigenous bioactive compounds. The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes, thereby exploiting the properties in resisting bacterial diseases. The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate, for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Bacillus subtilis. It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli. Further, Gas chromatography coupled with Mass spectroscopy (GC-MS) revealed the existence of phenol, 3, 5-bis (1, 1-dimethylethyl) in the crude hexane extract which is well-known to possess antibacterial activity. The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas stutzeri by phenotypic and genotypic methods. The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens. The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS. The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts. Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.

  8. Antimicrobials from the marine algal endophyte Penicillium sp.

    Flewelling, Andrew J; Johnson, John A; Gray, Christopher A

    2013-03-01

    An endophytic fungus identified as Penicillium sp. was isolated from the brown alga Fucus spiralis collected from the Shetland Islands, United Kingdom. Bioassay-guided fractionation of an extract of the fungus led to the isolation of cladosporin, epiepoformin, phyllostine, and patulin, all of which showed antimicrobial activity against either Staphylococcus aureus or Pseudomonas aeruginosa. Cladosporin has not previously been identified from a fungus of the genus Penicillium, and, despite being biosynthetically related, epiepoformin, phyllostine and patulin have not been previously reported from one source.

  9. Enantioselective biotransformation of propranolol to the active metabolite 4-hydroxypropranolol by endophytic fungi

    Keyller Bastos Borges

    2011-01-01

    Full Text Available The enantioselective biotransformation of propranolol (Prop by the endophytic fungi Phomopsis sp., Glomerella cingulata, Penicillium crustosum, Chaetomium globosum and Aspergillus fumigatus was investigated by studying the kinetics of the aromatic hydroxylation reaction with the formation of 4-hydroxypropranolol (4-OH-Prop. Both Prop enantiomers were consumed by the fungi in the biotransformation process, but the 4-hydroxylation reaction yielded preferentially (--(S-4-OH-Prop. The quantity of metabolites biosynthesized varied slightly among the evaluated endophytic fungi. These results show that all investigated endophytic fungi could be used as biosynthetic tools in biotransformation processes to obtain the enantiomers of 4-OH-Prop.

  10. Characterization of Five Fungal Endophytes Producing Cajaninstilbene Acid Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.

    Gao, Yuan; Zhao, Jin Tong; Zu, Yuan Gang; Fu, Yu Jie; Wang, Wei; Luo, Meng; Efferth, Thomas

    2011-01-01

    Five fungal endophytes (K4, K5, K6, K9, K14) producing Cajaninstilbene acid (CSA, 3-hydroxy-4-prenyl-5-methoxystilbene-2-carboxylic acid) were isolated from the roots of pigeon pea [Cajanus cajan (L.) Millsp.]. CSA is responsible for the prominent pharmacological activities in pigeon pea. The amount of CSA in culture solution varied among the five fungal endophytes. K4 produced the highest levels of CSA (1037.13 µg/L) among the endophytes tested after incubation for five days. Both morphologi...

  11. Endophytic colonization of tomato plants by the biological control agent Clonostachys rosea

    Høyer, Anna Kaja; Jørgensen, Hans Jørgen Lyngs; Amby, Daniel Buchvaldt

    Fungal endophytes live naturally inside plants without causing symptoms. On the contrary, they can promote plant growth and increase tolerance to abiotic and biotic stress. These beneficial effects have increased the agricultural interest for exploitation of fungal isolates with an endophytic life...... controls seed- and soil-borne diseases and can furthermore promote plant growth. However, it is not known whether IK726 can colonize plants internally and therefore, the objective of the present study was to examine the possibility of an endophytic life-style of IK726 in tomato. Tomato seeds were sown...

  12. Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp.

    Ratnaweera, Pamoda B; de Silva, E Dilip; Williams, David E; Andersen, Raymond J

    2015-07-10

    Opuntia dillenii is an invasive plant well established in the harsh South-Eastern arid zone of Sri Lanka. Evidence suggests it is likely that the endophytic fungal populations of O. dillenii assist the host in overcoming biotic and abiotic stress by producing biologically active metabolites. With this in mind there is potential to discover novel natural products with useful biological activities from this hitherto poorly investigated source. Consequently, an investigation of the antimicrobial activities of the endophytes of O. dillenii, that occupies a unique ecological niche, may well provide useful leads in the discovery of new pharmaceuticals. Endophytic fungi were isolated from the surface sterilized cladodes and flowers of O. dillenii using several nutrient media and the antimicrobial activities were evaluated against three Gram-positive and two Gram-negative bacteria and Candida albicans. The two most bioactive fungi were identified by colony morphology and DNA sequencing. The secondary metabolite of the endophyte Fusarium sp. exhibiting the best activity was isolated via bioassay guided chromatography. The chemical structure was elucidated from the ESIMS and NMR spectroscopic data obtained for the active metabolite. The minimum inhibitory concentrations (MICs) of the active compound were determined. Eight endophytic fungi were isolated from O. dillenii and all except one showed antibacterial activities against at least one of the test bacteria. All extracts were inactive against C. albicans. The most bioactive fungus was identified as Fusarium sp. and the second most active as Aspergillus niger. The structure of the major antibacterial compound of the Fusarium sp. was shown to be the tetramic acid derivative, equisetin. The MIC's for equisetin were 8 μg mL(-1) against Bacillus subtilis, 16 μg mL(-1) against Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus (MRSA). O. dillenii, harbors several endophytic fungi capable of producing

  13. Bioactive Metabolites from Pathogenic and Endophytic Fungi of Forest Trees.

    Masi, Marco; Maddau, Lucia; Linaldeddu, Benedetto Teodoro; Scanu, Bruno; Evidente, Antonio; Cimmino, Alessio

    2018-01-01

    Fungi play an important role in terrestrial ecosystems interacting positively or negatively with plants. These interactions are complex and the outcomes are different depending on the fungal lifestyles, saprotrophic, mutualistic or pathogenic. Furthermore, fungi are well known for producing secondary metabolites, originating from different biosynthetic pathways, which possess biological properties of considerable biotechnological interest. Among the terrestrial ecosystems, temperate forests represent an enormous reservoir of fungal diversity. This review will highlight the goldmine of secondary metabolites produced by pathogenic and endophytic fungi of forest trees with focus on their biological activities. A structured search of bibliographic databases for peer-reviewed research literature was undertaken using a research discovery application providing access to a large and authoritative source of references. The papers selected were examined and the main results were reported and discussed. Two hundred forthy-one papers were included in the review, outlined a large number of secondary metabolites produced by pathogenic and endophiltic fungi and their biological activities, including phytotoxic, antifungal, antioomycetes, antibacterial, brine shrimp lethality, mosquito biting deterrence and larvicidal, cytotoxic, antiproliferative and many other bioactivities. The findings of this review confirm the importance of secondary metabolites produced by pathogenic and endophytic fungi from forest plants growing in temperate regions as an excellent prospects to discover compounds with new bioactivities and mode of actions. In addition, the potential of some metabolites as a source of new drugs and biopesticides is underlined. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Molecular characterization of endophytic fungi associated with the roots of Chenopodium quinoa inhabiting the Atacama Desert, Chile.

    González-Teuber, M; Vilo, C; Bascuñán-Godoy, L

    2017-03-01

    Plant roots can be highly colonized by fungal endophytes. This seems to be of particular importance for the survival of plants inhabiting stressful habitats. This study focused on the Identification of the fungal endophytic community associated with the roots of quinoa plants ( Chenopodium quinoa ) growing near the salt lakes of the Atacama Desert, Chile. One hundred endophytic fungi were isolated from healthy quinoa roots, and the internal transcribed spacer (ITS) region was sequenced for phylogenetic and taxonomic analysis. The isolates were classified into eleven genera and 21 distinct operational taxonomic units (OTUs). Despite a relatively high diversity of root endophytic fungi associated with quinoa plants, the fungal community was dominated by only the Ascomycota phyla. In addition, the most abundant genera were Penicillium , Phoma and Fusarium , which are common endophytes reported in plant roots. This study shows that roots of C . quinoa harbor a diverse group of endophytic fungi. Potential roles of these fungi in plant host tolerance to stressful conditions are discussed.

  15. Effects of natural hybrid and non-hybrid Epichloë endophytes on the response of Hordelymus europaeus to drought stress.

    Oberhofer, Martina; Güsewell, Sabine; Leuchtmann, Adrian

    2014-01-01

    Interspecific hybrid endophytes of the genus Epichloë (Ascomycota, Clavicipitaceae) are prevalent in wild grass populations, possibly because of their larger gene variation, resulting in increased fitness benefits for host plants; however, the reasons are not yet known. We tested hypotheses regarding niche expansion mediated by hybrid endophytes, population-dependent interactions and local co-adaptation in the woodland grass Hordelymus europaeus, which naturally hosts both hybrid and non-hybrid endophyte taxa. Seedlings derived from seeds of four grass populations made endophyte free were re-inoculated with hybrid or non-hybrid endophyte strains, or left endophyte free. Plants were grown in the glasshouse with or without drought treatment. Endophyte infection increased plant biomass and tiller production by 10-15% in both treatments. Endophyte types had similar effects on growth, but opposite effects on reproduction: non-hybrid endophytes increased seed production, whereas hybrid endophytes reduced or prevented it completely. The results are consistent with the observation that non-hybrid endophytes in H. europaeus prevail at dry sites, but cannot explain the prevalence of hybrid endophytes. Thus, our results do not support the hypothesis of niche expansion of hybrid-infected plants. Moreover, plants inoculated with native relative to foreign endophytes yielded higher infections, but both showed similar growth and survival, suggesting weak co-adaptation. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

  16. Heavy metal resistant endophytic fungi isolated from Nypa fruticans in Kuching Wetland National Park

    Choo, Jenny; Sabri, Nuraini Binti Mohd; Tan, Daniel; Mujahid, Aazani; Müller, Moritz

    2015-06-01

    Heavy metal pollution is an environmental issue globally and the aim of this study was to isolate endophytic fungi from mangrove wetlands of Sarawak to assess and test their ability to grow in the presence of various heavy metals (copper (Cu), zinc (Zn), lead (Pb), and chromium (Cr)). Samples of Nypa fruticans were collected from Kuching Wetland National Park (KWNP) for subsequent endophyte isolation. Ninety-three (93) isolates were obtained and assessed and the most resistant isolates (growing at concentrations up to 1000 ppm) were identified using fungal primers ITS 1 and ITS 4. All of the endophytic fungi were identified to be closely related to Pestalotiopsis sp. and this is to our knowledge the first study reporting the ability of Pestalotiopsis sp. to grow at high concentrations of copper, lead, zinc and chromium. Our results highlight the potential of using endophytic fungi for the treatment of heavy metal pollution, for example as biosorbents.

  17. Isolation and characterization of antibacterial compound from a mangrove-endophytic fungus, Penicillium chrysogenum MTCC 5108

    PrabhaDevi; Rodrigues, C.; Naik, C.G.; DeSouza, L.

    Microorganisms, especially endophytic fungi that reside in the tissue of living mangrove plants, seem to play a major role in meeting the general demand for new biologically active substances. During the course of screening for biologically active...

  18. Ecology and Genomic Insights into Plant-Pathogenic and Plant-Nonpathogenic Endophytes.

    Brader, Günter; Compant, Stéphane; Vescio, Kathryn; Mitter, Birgit; Trognitz, Friederike; Ma, Li-Jun; Sessitsch, Angela

    2017-08-04

    Plants are colonized on their surfaces and in the rhizosphere and phyllosphere by a multitude of different microorganisms and are inhabited internally by endophytes. Most endophytes act as commensals without any known effect on their plant host, but multiple bacteria and fungi establish a mutualistic relationship with plants, and some act as pathogens. The outcome of these plant-microbe interactions depends on biotic and abiotic environmental factors and on the genotype of the host and the interacting microorganism. In addition, endophytic microbiota and the manifold interactions between members, including pathogens, have a profound influence on the function of the system plant and the development of pathobiomes. In this review, we elaborate on the differences and similarities between nonpathogenic and pathogenic endophytes in terms of host plant response, colonization strategy, and genome content. We furthermore discuss environmental effects and biotic interactions within plant microbiota that influence pathogenesis and the pathobiome.

  19. The Hidden World within Plants: Ecological and Evolutionary Considerations for Defining Functioning of Microbial Endophytes.

    Hardoim, Pablo R; van Overbeek, Leonard S; Berg, Gabriele; Pirttilä, Anna Maria; Compant, Stéphane; Campisano, Andrea; Döring, Matthias; Sessitsch, Angela

    2015-09-01

    All plants are inhabited internally by diverse microbial communities comprising bacterial, archaeal, fungal, and protistic taxa. These microorganisms showing endophytic lifestyles play crucial roles in plant development, growth, fitness, and diversification. The increasing awareness of and information on endophytes provide insight into the complexity of the plant microbiome. The nature of plant-endophyte interactions ranges from mutualism to pathogenicity. This depends on a set of abiotic and biotic factors, including the genotypes of plants and microbes, environmental conditions, and the dynamic network of interactions within the plant biome. In this review, we address the concept of endophytism, considering the latest insights into evolution, plant ecosystem functioning, and multipartite interactions. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Alkaloids May Not be Responsible for Endophyte Associated Reductions in Tall Fescue Decomposition Rates

    1. Fungal endophyte - grass symbioses can have dramatic ecological effects, altering individual plant physiology, plant and animal community structure and function, and ecosystem processes such as litter decomposition and nutrient cycling. 2. Within the tall fescue (Schedonorus arundinaceus) - funga...

  1. Diversity of endophytic bacterial populations and their interaction with Xylella fastidiosa in citrus plants

    Araujo, W.L.; Marcon, J.; jr. Maccheroni, W.; Elsas, van J.D.; Vuurde, van J.W.L.; Azevedo, de J.L.

    2002-01-01

    Citrus variegated chlorosis (CVC) is caused by Xylella fastidiosa, a phytopathogenic bacterium that can infect all Citrus sinensis cultivars. The endophytic bacterial communities of healthy, resistant, and CVC-affected citrus plants were studied by using cultivation as well as

  2. Molecular detection of TasA gene in endophytic Bacillus species ...

    Molecular detection of TasA gene in endophytic Bacillus species and characterization of the gene in Bacillus amyloliquefaciens. ... African Journal of Biotechnology ... in Bacillus amyloliquefaciens PEBA20 and 7 strains of Bacillus subtilis, ...

  3. Managing the tall fescue-fungal endophyte symbiosis for optimum forage-animal production

    Alkaloids produced by the fungal endophyte (Neotyphodium coenophialum) that infects tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] are a paradox to cattle production. While certain alkaloids impart tall fescue with tolerances to environmental stresses, such as moisture, heat, and herbivory, e...

  4. Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination

    Lumactud, Rhea; Shen, Shu Yi; Lau, Mimas; Fulthorpe, Roberta

    2016-01-01

    The bacterial endophytic communities of four plants growing abundantly in soils highly contaminated by hydrocarbons were analyzed through culturable and and culture-independent means. Given their tolerance to the high levels of petroleum contamination at our study site, we sought evidence that Achillea millefolium, Solidago canadensis, Trifolium aureum and Dactylis glomerata support high levels of hydrocarbon degrading endophytes. A total of 190 isolates were isolated from four plant species....

  5. Munumbicins, wide-spectrum antibiotics produced by Streptomyces NRRL 30562, endophytic on Kennedia nigriscans

    Castillo, UF; Strobel, GA; Ford, EJ; Hess, WM; Porter, H; Jensen, JB; Albert, H; Robison, R; Condron, MAM; Teplow, DB; Stevens, D; Yaver, D

    2002-01-01

    Munumbicins A, B, C and D are newly described antibiotics with a wide spectrum of activity against many human as well as plant pathogenic fungi and bacteria, and a Plasmodium sp. These compounds were obtained from Streptomyces NRRL 3052, which is endophytic in the medicinal plant snakevine (Kennedia nigriscans), native to the Northern Territory of Australia. This endophyte was cultured, the broth was extracted with an organic solvent and the contents of the residue were purified by bioassay-g...

  6. Conservation and diversity of seed associated endophytes in Zea across boundaries of evolution, ethnography and ecology.

    Johnston-Monje, David; Raizada, Manish N

    2011-01-01

    Endophytes are non-pathogenic microbes living inside plants. We asked whether endophytic species were conserved in the agriculturally important plant genus Zea as it became domesticated from its wild ancestors (teosinte) to modern maize (corn) and moved from Mexico to Canada. Kernels from populations of four different teosintes and 10 different maize varieties were screened for endophytic bacteria by culturing, cloning and DNA fingerprinting using terminal restriction fragment length polymorphism (TRFLP) of 16S rDNA. Principle component analysis of TRFLP data showed that seed endophyte community composition varied in relation to plant host phylogeny. However, there was a core microbiota of endophytes that was conserved in Zea seeds across boundaries of evolution, ethnography and ecology. The majority of seed endophytes in the wild ancestor persist today in domesticated maize, though ancient selection against the hard fruitcase surrounding seeds may have altered the abundance of endophytes. Four TRFLP signals including two predicted to represent Clostridium and Paenibacillus species were conserved across all Zea genotypes, while culturing showed that Enterobacter, Methylobacteria, Pantoea and Pseudomonas species were widespread, with γ-proteobacteria being the prevalent class. Twenty-six different genera were cultured, and these were evaluated for their ability to stimulate plant growth, grow on nitrogen-free media, solubilize phosphate, sequester iron, secrete RNAse, antagonize pathogens, catabolize the precursor of ethylene, produce auxin and acetoin/butanediol. Of these traits, phosphate solubilization and production of acetoin/butanediol were the most commonly observed. An isolate from the giant Mexican landrace Mixteco, with 100% identity to Burkholderia phytofirmans, significantly promoted shoot potato biomass. GFP tagging and maize stem injection confirmed that several seed endophytes could spread systemically through the plant. One seed isolate

  7. Seed Endophyte Microbiome of Crotalaria pumila Unpeeled: Identification of Plant-Beneficial Methylobacteria

    Sánchez-López, Ariadna S.; Pintelon, Isabel; Stevens, Vincent; Imperato, Valeria; Timmermans, Jean-Pierre; González-Chávez, Carmen; Carrillo-González, Rogelio; Van Hamme, Jonathan; Vangronsveld, Jaco; Thijs, Sofie

    2018-01-01

    Metal contaminated soils are increasing worldwide. Metal-tolerant plants growing on metalliferous soils are fascinating genetic and microbial resources. Seeds can vertically transmit endophytic microorganisms that can assist next generations to cope with environmental stresses, through yet poorly understood mechanisms. The aims of this study were to identify the core seed endophyte microbiome of the pioneer metallophyte Crotalaria pumila throughout three generations, and to bet...

  8. Culturable bacterial endophytes isolated from Mangrove tree (Rhizophora apiculata Blume) enhance seedling growth in Rice.

    Deivanai, Subramanian; Bindusara, Amitraghata Santhanam; Prabhakaran, Guruswamy; Bhore, Subhash Janardhan

    2014-07-01

    Endophytic bacteria do have several potential applications in medicine and in other various sectors of biotechnology including agriculture. Bacterial endophytes need to be explored for their potential applications in agricultural biotechnology. One of the potential applications of bacterial endophytes in agricultural is to enhance the growth of the agricultural crops. Hence, this study was undertaken to explore the plant growth promoting potential application of bacterial endophytes. The objective of this study was to examine the effect of endophytic bacteria from mangrove tree (Rhizophora apiculata Blume) for their efficacy in promoting seedling growth in rice. Eight endophytic bacterial isolates (EBIs) isolated from twig and petiole tissues of the mangrove were identified based on their 16S ribosomal ribonucleic acid (rRNA) gene sequence homology. Separately, surface sterilized paddy seeds were treated with cell-free broth and cell suspension of the EBIs. Rice seedlings were analyzed by various bioassays and data was recorded. The gene sequences of the isolates were closely related to two genera namely, Bacillus and Pantoea. Inoculation of EBIs from R. apiculata with rice seeds resulted in accelerated root and shoot growth with significant increase in chlorophyll content. Among the isolates, Pantoea ananatis (1MSE1) and Bacillus amyloliquefaciens (3MPE1) had shown predominance of activity. Endophytic invasion was recognized by the non-host by rapid accumulation of reactive oxygen species (ROS) and was counteracted by the production of hydrogen peroxide (H2O2) and lipid peroxide. The results demonstrated that EBIs from mangrove tree can increase the fitness of the rice seedlings under controlled conditions. These research findings could be useful to enhance the seedling growth and could serve as foundation in further research on enhancing the growth of the rice crop using endophytic bacteria.

  9. Potency of Six Isolates of Biocontrol Agents Endophytic Trichoderma Against Fusarium Wilt on Banana

    Taribuka, J; Wibowo, A; Widyastuti, S M; Sumardiyono, C

    2017-01-01

    Fusarium wilt caused by F. oxysporum f.sp. cubense is one of very damaging banana plant diseases which can cause plant death. Disease control using intensive chemical fungicides will have negative impacts on the environment and humans. Endophytic Trichoderma is one of the biological control agents which can reduce the amount of inoculum of pathogens, so it can reduce disease intensity. The objectives of this study was to assess the ability of endophytic Trichoderma in inducing plant resistanc...

  10. Conservation and Diversity of Seed Associated Endophytes in Zea across Boundaries of Evolution, Ethnography and Ecology

    Johnston-Monje, David; Raizada, Manish N.

    2011-01-01

    Endophytes are non-pathogenic microbes living inside plants. We asked whether endophytic species were conserved in the agriculturally important plant genus Zea as it became domesticated from its wild ancestors (teosinte) to modern maize (corn) and moved from Mexico to Canada. Kernels from populations of four different teosintes and 10 different maize varieties were screened for endophytic bacteria by culturing, cloning and DNA fingerprinting using terminal restriction fragment length polymorphism (TRFLP) of 16S rDNA. Principle component analysis of TRFLP data showed that seed endophyte community composition varied in relation to plant host phylogeny. However, there was a core microbiota of endophytes that was conserved in Zea seeds across boundaries of evolution, ethnography and ecology. The majority of seed endophytes in the wild ancestor persist today in domesticated maize, though ancient selection against the hard fruitcase surrounding seeds may have altered the abundance of endophytes. Four TRFLP signals including two predicted to represent Clostridium and Paenibacillus species were conserved across all Zea genotypes, while culturing showed that Enterobacter, Methylobacteria, Pantoea and Pseudomonas species were widespread, with γ-proteobacteria being the prevalent class. Twenty-six different genera were cultured, and these were evaluated for their ability to stimulate plant growth, grow on nitrogen-free media, solubilize phosphate, sequester iron, secrete RNAse, antagonize pathogens, catabolize the precursor of ethylene, produce auxin and acetoin/butanediol. Of these traits, phosphate solubilization and production of acetoin/butanediol were the most commonly observed. An isolate from the giant Mexican landrace Mixteco, with 100% identity to Burkholderia phytofirmans, significantly promoted shoot potato biomass. GFP tagging and maize stem injection confirmed that several seed endophytes could spread systemically through the plant. One seed isolate

  11. Spatial and temporal variation in fungal endophyte communities isolated from cultivated cotton (Gossypium hirsutum.

    María J Ek-Ramos

    Full Text Available Studies of fungi in upland cotton (Gossypium hirsutum cultivated in the United States have largely focused on monitoring and controlling plant pathogens. Given increasing interest in asymptomatic fungal endophytes as potential biological control agents, surveys are needed to better characterize their diversity, distribution patterns and possible applications in integrated pest management. We sampled multiple varieties of cotton in Texas, USA and tested for temporal and spatial variation in fungal endophyte diversity and community composition, as well as for differences associated with organic and conventional farming practices. Fungal isolates were identified by morphological and DNA identification methods. We found members of the genera Alternaria, Colletotrichum and Phomopsis, previously isolated as endophytes from other plant species. Other recovered species such as Drechslerella dactyloides (formerly Arthrobotrys dactyloides and Exserohilum rostratum have not, to our knowledge, been previously reported as endophytes in cotton. We also isolated many latent pathogens, but some species such as Alternaria tennuissima, Epicoccum nigrum, Acremonium alternatum, Cladosporium cladosporioides, Chaetomium globosum and Paecilomyces sp., are known to be antagonists against plant pathogens, insects and nematode pests. We found no differences in endophyte species richness or diversity among different cotton varieties, but did detect differences over time and in different plant tissues. No consistent patterns of community similarity associated with variety, region, farming practice, time of the season or tissue type were observed regardless of the ecological community similarity measurements used. Results indicated that local fungal endophyte communities may be affected by both time of the year and plant tissue, but the specific community composition varies across sites. In addition to providing insights into fungal endophyte community structure, our survey

  12. Potential Use of Endophytic Bacteria to Control Pratylenchus Brachyurus on Patchouli

    Harni, Rita; Supramana, Supramana; Supriadi, Supriadi

    2012-01-01

    Pratylenchus brachyurus is an important parasitic nematode which significantly decreases quality and quantity of patchouli oil. One potential measure for controlling the nematode is by using endophytic bacteria. These bacteria also induce plant growth. This study aimed to evaluate the potential of endo-phytic bacteria to control P. brachyurus. The experiments were carried out in the Bacteriological Laboratory of the Plant Protection Department, Bogor Agricultural University, and the Laborator...

  13. Epicoccum nigrum P16, a Sugarcane Endophyte, Produces Antifungal Compounds and Induces Root Growth

    Fávaro, Léia Cecilia de Lima; Sebastianes, Fernanda Luiza de Souza; Araújo, Welington Luiz

    2012-01-01

    Background Sugarcane is one of the most important crops in Brazil, mainly because of its use in biofuel production. Recent studies have sought to determine the role of sugarcane endophytic microbial diversity in microorganism-plant interactions, and their biotechnological potential. Epicoccum nigrum is an important sugarcane endophytic fungus that has been associated with the biological control of phytopathogens, and the production of secondary metabolites. In spite of several studies carried out to define the better conditions to use E. nigrum in different crops, little is known about the establishment of an endophytic interaction, and its potential effects on plant physiology. Methodology/Principal Findings We report an approach based on inoculation followed by re-isolation, molecular monitoring, microscopic analysis, plant growth responses to fungal colonization, and antimicrobial activity tests to study the basic aspects of the E. nigrum endophytic interaction with sugarcane, and the effects of colonization on plant physiology. The results indicate that E. nigrum was capable of increasing the root system biomass and producing compounds that inhibit the in vitro growth of sugarcane pathogens Fusarium verticillioides, Colletotrichum falcatum, Ceratocystis paradoxa, and Xanthomomas albilineans. In addition, E. nigrum preferentially colonizes the sugarcane surface and, occasionally, the endophytic environment. Conclusions/Significance Our work demonstrates that E. nigrum has great potential for sugarcane crop application because it is capable of increasing the root system biomass and controlling pathogens. The study of the basic aspects of the interaction of E. nigrum with sugarcane demonstrated the facultative endophytism of E. nigrum and its preference for the phylloplane environment, which should be considered in future studies of biocontrol using this species. In addition, this work contributes to the knowledge of the interaction of this ubiquitous endophyte

  14. Natural Medium for Growing of Endophytic Bacteria from Solanaceae in Malang-Indonesia

    Purnawati Arika; Harijani Wiwik Sri; Windriyanti Wiwin

    2016-01-01

    Endophytic bacteria are important microorganisms having potential as biocontrol agents for many pathogens. Until now, the growth of it always uses semi-synthetic or synthetic medium so it was difficult to be used by farmers in the field and it was expensive to have its propagation as biocontrol agents. Based on the problem, this research will study the natural medium as propagation medium of Endophytic bacteria. It had natural ingredients such as soybean, chicken broth, egg, worms, snail, sor...

  15. Conservation and diversity of seed associated endophytes in Zea across boundaries of evolution, ethnography and ecology.

    David Johnston-Monje

    Full Text Available Endophytes are non-pathogenic microbes living inside plants. We asked whether endophytic species were conserved in the agriculturally important plant genus Zea as it became domesticated from its wild ancestors (teosinte to modern maize (corn and moved from Mexico to Canada. Kernels from populations of four different teosintes and 10 different maize varieties were screened for endophytic bacteria by culturing, cloning and DNA fingerprinting using terminal restriction fragment length polymorphism (TRFLP of 16S rDNA. Principle component analysis of TRFLP data showed that seed endophyte community composition varied in relation to plant host phylogeny. However, there was a core microbiota of endophytes that was conserved in Zea seeds across boundaries of evolution, ethnography and ecology. The majority of seed endophytes in the wild ancestor persist today in domesticated maize, though ancient selection against the hard fruitcase surrounding seeds may have altered the abundance of endophytes. Four TRFLP signals including two predicted to represent Clostridium and Paenibacillus species were conserved across all Zea genotypes, while culturing showed that Enterobacter, Methylobacteria, Pantoea and Pseudomonas species were widespread, with γ-proteobacteria being the prevalent class. Twenty-six different genera were cultured, and these were evaluated for their ability to stimulate plant growth, grow on nitrogen-free media, solubilize phosphate, sequester iron, secrete RNAse, antagonize pathogens, catabolize the precursor of ethylene, produce auxin and acetoin/butanediol. Of these traits, phosphate solubilization and production of acetoin/butanediol were the most commonly observed. An isolate from the giant Mexican landrace Mixteco, with 100% identity to Burkholderia phytofirmans, significantly promoted shoot potato biomass. GFP tagging and maize stem injection confirmed that several seed endophytes could spread systemically through the plant. One seed

  16. Endophytes from medicinal plants and their potential for producing indole acetic acid, improving seed germination and mitigating oxidative stress* #

    Khan, Abdul Latif; Gilani, Syed Abdullah; Waqas, Muhammad; Al-Hosni, Khadija; Al-Khiziri, Salima; Kim, Yoon-Ha; Ali, Liaqat; Kang, Sang-Mo; Asaf, Sajjad; Shahzad, Raheem; Hussain, Javid; Lee, In-Jung; Al-Harrasi, Ahmed

    2017-01-01

    Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31±0.21) µmol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses. PMID:28124841

  17. Endophytes from medicinal plants and their potential for producing indole acetic acid, improving seed germination and mitigating oxidative stress.

    Khan, Abdul Latif; Gilani, Syed Abdullah; Waqas, Muhammad; Al-Hosni, Khadija; Al-Khiziri, Salima; Kim, Yoon-Ha; Ali, Liaqat; Kang, Sang-Mo; Asaf, Sajjad; Shahzad, Raheem; Hussain, Javid; Lee, In-Jung; Al-Harrasi, Ahmed

    Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31±0.21) µmol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses.

  18. Endophytic bacterial effects on seed germination and mobilization of reserves in ammodendron biofolium

    Zhu, Y.; She, X.P.

    2017-01-01

    The main aim of this study was to analyze the mobilization of storage reserves during seed germination of Ammodendron bifolium by host plant-endophytic bacteria interaction and to determine the contribution of endophytic bacteria in plant establishment. The seeds were inoculated with three different endophytic bacteria from A. bifolium, Staphylococcus sp. AY3, Kocuria sp. AY9 and Bacillus sp. AG18, and they were germinated in the dark. Fresh weight changes and early seedling growth were assessed, and the content of storage compounds was quantified using biochemical assays in all germinated and non-germinated seeds. To understand the mechanism promoting seed germination, the activities of extracellular enzymes of bacterial isolates were also analyzed by the plate assay method. The results showed that treatment with endophytic bacteria accelerated seed germination; promoted further water absorption and radicle growth; and also promoted degradation of sucrose, protein and lipids during the germination process. At the same time, our results also showed that strain AG18 was able to produce protease and amylase, strain AY9 had only amylase activity, and strain AY3 had no extracellular enzyme activity. In summary, our current study showed that (i) endophytic bacteria improved seed germination and post-germination seedling growth of A. bifolium; (ii) inoculation with endophytic bacteria could promote storage reserve mobilization during or following germination; (iii) the degradation of protein, lipids and sucrose could provide essential energy for post-germination growth; and (iv) three bacterial isolates might have different action mechanisms on seed germination. (author)

  19. Endophytic Fungi Associated With Turmeric (Curcuma longa L. Can Inhibit Histamine-Forming Bacteria in Fish

    Eris Septiana

    2017-01-01

    Full Text Available Turmeric (Curcuma longa L. is a medicinal plant that is commonly used as spice and preservative. Many types of endophytic fungi have been reported as being associated with medicinal plants and able to synthesize secondary metabolites. In this study, endophytic fungi were isolated from all plant parts of turmeric plants. Identification of the endophytic fungi was done using morphological characteristics and sequencing of the internal transcribed spacer (ITS region of ribosomal DNA. The dual culture method was used for screening antibacterial activity of the endophytic fungi against Morganella morganii, a common histamine-producing bacteria. The disc diffusion method was used to test the ability of water fractions of selected endophytic fungi to inhibit M. morganii growth. Two-dimensional thin layer chromatography was used to determine the fungal extract inhibition activity on histamine formation. In total, 11 endophytic fungi were successfully isolated and identified as Arthrobotrys foliicola, Cochliobolus kusanoi, Daldinia eschscholzii, Fusarium oxysporum, Fusarium proliferatum, Fusarium solani, Fusarium verticillioides, Phanerochaete chrysosporium, and Phaeosphaeria ammophilae. Five isolates showed inhibition activity against M. morganii in the dual culture tests. Based on the disc diffusion assay, A. foliicola and F. verticillioides inhibited the growth of M. morganii as a histamine-producing bacteria, and inhibiting histamine formation in fish. The best effects in inhibiting growth of the histamine-producing bacteria and histamine formation inhibition in fish were produced with F. verticillioides water fraction at 0°C incubation.

  20. Distribution and antimicrobial potential of endophytic fungi associated with ethnomedicinal plant Melastoma malabathricum L.

    Mishra, Vineet Kumar; Singh, Garima; Passari, Ajit Kumar; Yadav, Mukesh Kumar; Gupta, Vijai Kumar; Singh, Bhim Pratap

    2016-03-01

    Distributions of endophytic fungi associated with ethnomedicinal plant Melastoma malabathricum L. was studied and 91 isolates belonging to 18 genera were recovered. The isolates were distributed to sordariomycetes (62.63%), dothideomycetes (19.78%), eurotiomycetes (7.69%), zygomycetes (4.19%), agaricomycetes (1.09%), and mycelia sterilia (4.39%). Based on colony morphology and examination of spores, the isolates were classified into 18 taxa, of which Colletotrichum, Phomopsis and Phoma were dominant, their relative frequencies were 23.07%, 17.58% and 12.08% respectively. The colonization rate of endophytic fungi was determined and found to be significantly higher in leaf segments (50.76%), followed by root (41.53%) and stem tissues (27.69%). All the isolates were screened for antimicrobial activity and revealed that 26.37% endophytic fungi were active against one or more pathogens. Twenty four isolates showing significant antimicrobial activity were identified by sequencing the ITS1-5.8S-ITS2 region of rRNA gene. Results indicated that endophytic fungi associated with leaf were functionally versatile as they showed antimicrobial activity against most of the tested pathogens. The endophytic fungi Diaporthe phaseolorum var. meridionalis (KF193982) inhibited all the tested bacterial pathogens, whereas, Penicillium chermesinum (KM405640) displayed most significant antifungal activity. This seems to be the first hand report to understand the distribution and antimicrobial ability of endophytic fungi from ethno-medicinal plant M. malabathricum.

  1. Glucanase and Chitinase from Some Isolates of Endophytic Fungus Trichoderma spp.

    Prasetyawan, Sasangka; Sulistyowati, Lilik; Aulanni'am

    2018-01-01

    Endophytic fungi are those fungi that are able to grow in plant tissue without causing symptoms of disease. It is thought that these fungi may confer on the host plants degree of resistance to parasitic invasion. Endophytic fungi have been isolated from stem tissue and these fungi are known to be antagonistic to pathogenic fungi. These endophytes produce chitinase and β-1,3-glucanase enzymes. Based on the fact that chitin and β-1,3-glucan are the main skeletal polysaccharides of the cell walls of fungal patogen. The aim of this research is to do potential test on some of isolates of Trichoderma’s endophytic (L-1, L-2, Is-1, Is-2 and Is-7) in the chitinase and β-1,3-glucanase activity in effort to determine endophytic which be chossen to be gene resource for the next research. The gene will be transformed to citrus plant japanese citroen in effort to make citrus plant transgenic resistance to phytopatogenic invasion. The result of this research is endofit namely L-1 is the most potential endophytic fungi with chitinase activities is 4,8 10-2 Unit and glucanase 24,2. 1012 Unit. The addition of chitin and cell wall of phytophtora causes chitinase activity significantly increase, and also addition of laminarin and cell wall of phytophtora makes glucanase activity increase.

  2. The Role of Endophytic Fungi in the Anticancer Activity of Morinda citrifolia Linn. (Noni

    Yougen Wu

    2015-01-01

    Full Text Available We hypothesize that the fungal endophytes of noni may possibly play a role in its overall pharmacological repertoire, especially since the perceived efficacy of the fruit in ethnomedicinal use is associated with the fermented juice. The foremost goal of this study is to explore the role of endophyte-derived secondary metabolites in the purported anticancer properties of noni. To that end, culturable endophytic fungi resident within the healthy leaves and fruit of the plant were isolated and identified by molecular sequence analysis of the 5.8S gene and internal transcribed spacers (ITS. Purified organisms were subjected to in vitro fermentation in malt extract broth for 8 weeks under anaerobic conditions at room temperature (25°C, in order to simulate the conditions under which traditional fermented noni juice is prepared. The cytotoxic potential of organic extracts derived from the fermented broths of individual endophytes was then tested against three major cancers that afflict humans. Twelve distinct endophytic fungal species were obtained from the leaves and 3 from the fruit. Three of the leaf endophytes inhibited the growth of human carcinoma cell lines LU-1 (lung, PC-3 (prostate, and MCF-7 (breast with IC50 values of ≤10 μg/mL.

  3. Azoarcus sp. CIB, an anaerobic biodegrader of aromatic compounds shows an endophytic lifestyle.

    Helga Fernández

    Full Text Available BACKGROUND: Endophytic bacteria that have plant growth promoting traits are of great interest in green biotechnology. The previous thought that the Azoarcus genus comprises bacteria that fit into one of two major eco-physiological groups, either free-living anaerobic biodegraders of aromatic compounds or obligate endophytes unable to degrade aromatics under anaerobic conditions, is revisited here. METHODOLOGY/PRINCIPAL FINDINGS: Light, confocal and electron microscopy reveal that Azoarcus sp. CIB, a facultative anaerobe β-proteobacterium able to degrade aromatic hydrocarbons under anoxic conditions, is also able to colonize the intercellular spaces of the rice roots. In addition, the strain CIB displays plant growth promoting traits such nitrogen fixation, uptake of insoluble phosphorus and production of indoleacetic acid. Therefore, this work demonstrates by the first time that a free-living bacterium able to degrade aromatic compounds under aerobic and anoxic conditions can share also an endophytic lifestyle. The phylogenetic analyses based on the 16S rDNA and nifH genes confirmed that obligate endophytes of the Azoarcus genus and facultative endophytes, such as Azoarcus sp. CIB, locate into different evolutionary branches. CONCLUSIONS/SIGNIFICANCE: This is the first report of a bacterium, Azoarcus sp. CIB, able to degrade anaerobically a significant number of aromatic compounds, some of them of great environmental concern, and to colonize the rice as a facultative endophyte. Thus, Azoarcus sp. CIB becomes a suitable candidate for a more sustainable agricultural practice and phytoremediation technology.

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

    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.

  5. Aboveground endophyte affects root volatile emission and host plant selection of a belowground insect.

    Rostás, Michael; Cripps, Michael G; Silcock, Patrick

    2015-02-01

    Plants emit specific blends of volatile organic compounds (VOCs) that serve as multitrophic, multifunctional signals. Fungi colonizing aboveground (AG) or belowground (BG) plant structures can modify VOC patterns, thereby altering the information content for AG insects. Whether AG microbes affect the emission of root volatiles and thus influence soil insect behaviour is unknown. The endophytic fungus Neotyphodium uncinatum colonizes the aerial parts of the grass hybrid Festuca pratensis × Lolium perenne and is responsible for the presence of insect-toxic loline alkaloids in shoots and roots. We investigated whether endophyte symbiosis had an effect on the volatile emission of grass roots and if the root herbivore Costelytra zealandica was able to recognize endophyte-infected plants by olfaction. In BG olfactometer assays, larvae of C. zealandica were more strongly attracted to roots of uninfected than endophyte-harbouring grasses. Combined gas chromatography-mass spectrometry and proton transfer reaction-mass spectrometry revealed that endophyte-infected roots emitted less VOCs and more CO2. Our results demonstrate that symbiotic fungi in plants may influence soil insect distribution by changing their behaviour towards root volatiles. The well-known defensive mutualism between grasses and Neotyphodium endophytes could thus go beyond bioactive alkaloids and also confer protection by being chemically less apparent for soil herbivores.

  6. From Concept to Commerce: Developing a Successful Fungal Endophyte Inoculant for Agricultural Crops.

    Murphy, Brian R; Doohan, Fiona M; Hodkinson, Trevor R

    2018-02-11

    The development of endophyte inoculants for agricultural crops has been bedevilled by the twin problems of a lack of reliability and consistency, with a consequent lack of belief among end users in the efficacy of such treatments. We have developed a successful research pipeline for the production of a reliable, consistent and environmentally targeted fungal endophyte seed-delivered inoculant for barley cultivars. Our approach was developed de novo from an initial concept to source candidate endophyte inoculants from a wild relative of barley, Hordeum murinum (wall barley). A careful screening and selection procedure and extensive controlled environment testing of fungal endophyte strains, followed by multi-year field trials has resulted in the validation of an endophyte consortium suitable for barley crops grown on relatively dry sites. Our approach can be adapted for any crop or environment, provided that the set of first principles we have developed is followed. Here, we report how we developed the successful pipeline for the production of an economically viable fungal endophyte inoculant for barley cultivars.

  7. Host and geographic structure of endophytic and endolichenic fungi at a continental scale.

    U'Ren, Jana M; Lutzoni, François; Miadlikowska, Jolanta; Laetsch, Alexander D; Arnold, A Elizabeth

    2012-05-01

    Endophytic and endolichenic fungi occur in healthy tissues of plants and lichens, respectively, playing potentially important roles in the ecology and evolution of their hosts. However, previous sampling has not comprehensively evaluated the biotic, biogeographic, and abiotic factors that structure their communities. Using molecular data we examined the diversity, composition, and distributions of 4154 endophytic and endolichenic Ascomycota cultured from replicate surveys of ca. 20 plant and lichen species in each of five North American sites (Madrean coniferous forest, Arizona; montane semideciduous forest, North Carolina; scrub forest, Florida; Beringian tundra and forest, western Alaska; subalpine tundra, eastern central Alaska). Endolichenic fungi were more abundant and diverse per host species than endophytes, but communities of endophytes were more diverse overall, reflecting high diversity in mosses and lycophytes. Endophytes of vascular plants were largely distinct from fungal communities that inhabit mosses and lichens. Fungi from closely related hosts from different regions were similar in higher taxonomy, but differed at shallow taxonomic levels. These differences reflected climate factors more strongly than geographic distance alone. Our study provides a first evaluation of endophytic and endolichenic fungal associations with their hosts at a continental scale. Both plants and lichens harbor abundant and diverse fungal communities whose incidence, diversity, and composition reflect the interplay of climatic patterns, geographic separation, host type, and host lineage. Although culture-free methods will inform future work, our study sets the stage for empirical assessments of ecological specificity, metabolic capability, and comparative genomics.

  8. Diversity and communities of foliar endophytic fungi from different agroecosystems of Coffea arabica L. in two regions of Veracruz, Mexico.

    Saucedo-García, Aurora; Anaya, Ana Luisa; Espinosa-García, Francisco J; González, María C

    2014-01-01

    Over the past 20 years, the biodiversity associated with shaded coffee plantations and the role of diverse agroforestry types in biodiversity conservation and environmental services have been topics of debate. Endophytic fungi, which are microorganisms that inhabit plant tissues in an asymptomatic manner, form a part of the biodiversity associated with coffee plants. Studies on the endophytic fungi communities of cultivable host plants have shown variability among farming regions; however, the variability in fungal endophytic communities of coffee plants among different coffee agroforestry systems is still poorly understood. As such, we analyzed the diversity and communities of foliar endophytic fungi inhabiting Coffea arabica plants growing in the rustic plantations and simple polycultures of two regions in the center of Veracruz, Mexico. The endophytic fungi isolates were identified by their morphological traits, and the majority of identified species correspond to species of fungi previously reported as endophytes of coffee leaves. We analyzed and compared the colonization rates, diversity, and communities of endophytes found in the different agroforestry systems and in the different regions. Although the endophytic diversity was not fully recovered, we found differences in the abundance and diversity of endophytes among the coffee regions and differences in richness between the two different agroforestry systems of each region. No consistent pattern of community similarity was found between the coffee agroforestry systems, but we found that rustic plantations shared the highest number of morphospecies. The results suggest that endophyte abundance, richness, diversity, and communities may be influenced predominantly by coffee region, and to a lesser extent, by the agroforestry system. Our results contribute to the knowledge of the relationships between agroforestry systems and biodiversity conservation and provide information regarding some endophytic fungi and

  9. Formulation of the endophytic fungus Cladosporium oxysporum Berk.

    Bensaci Oussama Ali

    2015-01-01

    Full Text Available Two formulations containing culture filtrates and conidial suspensions of the endophytic fungus Cladosporium oxysporum Berk. & M.A. Curtis, isolated previously from stems of Euphorbia bupleuroides subsp. luteola (Kralik Maire, were experimentally tested for their aphicid activity against the black bean aphid Aphis fabae Scop. found in Algeria. It was shown that invert emulsions are more effective against aphids, than using aqueous suspensions. This was especially true for formulations containing culture filtrates. The relatively insignificant mortalities obtained by formulations containing conidial suspensions indicated a low infectious potential towards the aphids. The proteolytic activity seemed to be more important than the chitinolytic activity of the fungus against the black bean aphid A. fabae

  10. Antimicrobial metabolites from the plant endophytic fungus Penicillium sp.

    Yang, Ming-Hua; Li, Tian-Xiao; Wang, Ying; Liu, Rui-Huan; Luo, Jun; Kong, Ling-Yi

    2017-01-01

    Five rare dichloro aromatic polyketides (1-5) were obtained from an endophytic fungus Penicillium sp., along with five known metabolites (6-10). Their structures were elucidated by extensive spectroscopic analysis, Mosher methods, as well as [Rh 2 (OCOCF 3 ) 4 ]-induced electronic circular dichroism (ECD) experiments. Compounds 2-4 and 6 structurally involved acyclic 1.3-diols, the uneasy configuration determinations of which were well carried out by double-derivation NMR methods. Compounds 1-10 were evaluated for their antibacterial and antifungal activities against five strains of human pathogenic microorganisms. Helvolic acid (7) showed potent inhibitory effects against Staphylococcus aureus and Pseudomonas aeruginosa with MIC (minimum inhibitory concentration) values of 5.8 and 4.6μg/mL, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Current perspectives on the volatile-producing fungal endophytes.

    Zhi-Lin, Yuan; Yi-Cun, Chen; Bai-Ge, Xu; Chu-Long, Zhang

    2012-12-01

    Microbial-derived volatiles are ubiquitous in the environment and actively engaged in bio-communication with other organisms. Recently, some volatile-producing endophytes (VPEs), cryptic fungal symbionts persisting in healthy plant tissues, have attracted great attention due to their strong antibiotic activity or production of carbon chains that are identical to many of those found in petroleum, while other fragrant volatiles can be used in the flavoring industries. From an application-oriented and biotechnological point of view, these findings show significant promise for sustainable development of agriculture, forestry, and industry, especially in the control of fruit postharvest diseases, soil-borne pathogen management, and bio-fuel production. In comparison, the ecological importance of VPEs has only rarely been addressed and warrants further exploration. In this review, we summarize the current knowledge and future directions in this fascinating research field, and also highlight the constraints and progresses towards commercialization of VPEs products.

  12. Laparoscopic partial nephrectomy for endophytic hilar tumors: feasibility and outcomes.

    Di Pierro, G B; Tartaglia, N; Aresu, L; Polara, A; Cielo, A; Cristini, C; Grande, P; Gentile, V; Grosso, G

    2014-06-01

    To analyze feasibility and outcomes of laparoscopic partial nephrectomy (LPN) for endophytic hilar tumors in low-intermediate (ASA I-II) risk patients. This is a single centre retrospective study. From May 2009 to September 2011, 208 LPNs were performed at our institution. Overall 11 (5.2%) elective LPNs were for hilar tumors not visible on kidney surface. Hilar tumor was defined as a mass located in the renal hilum and in contact with a major renal vessel on preoperative imaging. Procedures were carried out by a single experienced surgeon (G.G.) via retroperitoneal approach by clamping the only main renal artery. Mean (range) age of patients was 45.3 years (38.2-64.1), tumor size 1.6 cm (1.2-2.0), warm ischemia time 24 min (19-32), operative time 140 min (110-200) and estimated blood loss 270 ml (100-750). Two collecting system injuries were observed and repaired intraoperatively. No conversion to open surgery was required. Final pathological examination revealed 10 renal cell carcinomas and 1 oncocytoma. A negative surgical margin was obtained in 10/11 (91%) patients. Renal function and serum hemoglobin were nearly unaltered pre and post-surgery. No tumor recurrence was observed at mean (range) follow-up of 34 months (15-43). In experienced hands, LPN represents a feasible, safe and effective treatment for selected patients diagnosed with endophytic hilar masses. A larger number of patients and longer follow-up are required to draw definitive conclusions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Species diversity of culturable endophytic fungi from Brazilian mangrove forests.

    de Souza Sebastianes, Fernanda Luiza; Romão-Dumaresq, Aline Silva; Lacava, Paulo Teixeira; Harakava, Ricardo; Azevedo, João Lúcio; de Melo, Itamar Soares; Pizzirani-Kleiner, Aline Aparecida

    2013-08-01

    This study aimed to perform a comparative analysis of the diversity of endophytic fungal communities isolated from the leaves and branches of Rhizophora mangle, Avicennia schaueriana and Laguncularia racemosa trees inhabiting two mangroves in the state of São Paulo, Brazil [Cananeia and Bertioga (oil spill-affected and unaffected)] in the summer and winter. Three hundred and forty-three fungi were identified by sequencing the ITS1-5.8S-ITS2 region of rDNA. Differences were observed in the frequencies of fungi isolated from the leaves and branches of these three different plant species sampled from the Bertioga oil spill-affected and the oil-unaffected mangrove sites in the summer and winter; these differences indicate a potential impact on fungal diversity in the study area due to the oil spill. The molecular identification of the fungi showed that the fungal community associated with these mangroves is composed of at least 34 different genera, the most frequent of which were Diaporthe, Colletotrichum, Fusarium, Trichoderma and Xylaria. The Shannon and the Chao1 indices [H'(95 %) = 4.00, H'(97 %) = 4.22, Chao1(95 %) = 204 and Chao1(97 %) = 603] indicated that the mangrove fungal community possesses a vast diversity and richness of endophytic fungi. The data generated in this study revealed a large reservoir of fungal genetic diversity inhabiting these Brazilian mangrove forests and highlighted substantial differences between the fungal communities associated with distinct plant tissues, plant species, impacted sites and sampling seasons.

  14. Bioreactor with Ipomoea hederifolia adventitious roots and its endophyte Cladosporium cladosporioides for textile dye degradation

    Patil, Swapnil M.; Chandanshive, Vishal V.; Rane, Niraj R.; Khandare, Rahul V.; Watharkar, Anuprita D.; Govindwar, Sanjay P.

    2016-01-01

    In vitro grown untransformed adventitious roots (AR) culture of Ipomoea hederifolia and its endophytic fungus (EF) Cladosporium cladosporioides decolorized Navy Blue HE2R (NB-HE2R) at a concentration of 20 ppm up to 83.3 and 65%, respectively within 96 h. Whereas the AR-EF consortium decolorized the dye more efficiently and gave 97% removal within 36 h. Significant inductions in the enzyme activities of lignin peroxidase, tyrosinase and laccase were observed in roots, while enzymes like tyrosinase, laccase and riboflavin reductase activities were induced in EF. Metabolites of dye were analyzed using UV—vis spectroscopy, FTIR and gas chromatography-mass spectrometry. Possible metabolic pathways of NB-HE2R were proposed with AR, EF and AR-EF systems independently. Looking at the superior efficacy of AR-EF system, a rhizoreactor was developed for the treatment of NB-HE2R at a concentration of 1000 ppm. Control reactor systems with independently grown AR and EF gave 94 and 85% NB-HE2R removal, respectively within 36 h. The AR-EF rhizoreactor, however, gave 97% decolorization. The endophyte colonization additionally increased root and shoot lengths of candidate plants through mutualism. Combined bioreactor strategies can be effectively used for future eco-friendly remediation purposes. - Highlights: • Endophytic fungus on Ipomoea hederifolia promotes root growth and shoot development • Endophytic Cladosporium cladosporioides synergistically degrade Navy Blue-HE2R dye • Endophyte colonized I. hederifolia roots proved superior in dye decolorization • Dye stress and toxicity was efficiently dealt by root-endophyte consortium • Root-endophyte consortium can be used as a sustainable remediation strategy

  15. Bioreactor with Ipomoea hederifolia adventitious roots and its endophyte Cladosporium cladosporioides for textile dye degradation

    Patil, Swapnil M. [Department of Biotechnology, Shivaji University, Kolhapur-416004 (India); Chandanshive, Vishal V. [Department of Biochemistry, Shivaji University, Kolhapur-416004 (India); Rane, Niraj R.; Khandare, Rahul V. [Department of Biotechnology, Shivaji University, Kolhapur-416004 (India); Watharkar, Anuprita D. [Department of Biochemistry, Shivaji University, Kolhapur-416004 (India); Govindwar, Sanjay P., E-mail: spg_biochem@unishivaji.ac.in [Department of Biochemistry, Shivaji University, Kolhapur-416004 (India)

    2016-04-15

    In vitro grown untransformed adventitious roots (AR) culture of Ipomoea hederifolia and its endophytic fungus (EF) Cladosporium cladosporioides decolorized Navy Blue HE2R (NB-HE2R) at a concentration of 20 ppm up to 83.3 and 65%, respectively within 96 h. Whereas the AR-EF consortium decolorized the dye more efficiently and gave 97% removal within 36 h. Significant inductions in the enzyme activities of lignin peroxidase, tyrosinase and laccase were observed in roots, while enzymes like tyrosinase, laccase and riboflavin reductase activities were induced in EF. Metabolites of dye were analyzed using UV—vis spectroscopy, FTIR and gas chromatography-mass spectrometry. Possible metabolic pathways of NB-HE2R were proposed with AR, EF and AR-EF systems independently. Looking at the superior efficacy of AR-EF system, a rhizoreactor was developed for the treatment of NB-HE2R at a concentration of 1000 ppm. Control reactor systems with independently grown AR and EF gave 94 and 85% NB-HE2R removal, respectively within 36 h. The AR-EF rhizoreactor, however, gave 97% decolorization. The endophyte colonization additionally increased root and shoot lengths of candidate plants through mutualism. Combined bioreactor strategies can be effectively used for future eco-friendly remediation purposes. - Highlights: • Endophytic fungus on Ipomoea hederifolia promotes root growth and shoot development • Endophytic Cladosporium cladosporioides synergistically degrade Navy Blue-HE2R dye • Endophyte colonized I. hederifolia roots proved superior in dye decolorization • Dye stress and toxicity was efficiently dealt by root-endophyte consortium • Root-endophyte consortium can be used as a sustainable remediation strategy.

  16. Endophytic fungi associated with roots of date palm (Phoenix dactylifera) in coastal dunes.

    Mohamed Mahmoud, Fadila; Krimi, Zoulikha; Maciá-Vicente, Jose G; Brahim Errahmani, Mohamed; Lopez-Llorca, Luis V

    Symbiotic interactions with fungal endophytes are argued to be responsible for the tolerance of plants to some stresses and for their adaptation to natural conditions. In this study we aimed to examine the endophytic fungal diversity associated with roots of date palms growing in coastal dune systems, and to screen this collection of endophytes for potential use as biocontrol agents, for antagonistic activity and mycoparasitism, and as producers of antifungal compounds with potential efficacy against root diseases of date palm. Roots of nine individual date palms growing in three coastal locations in the South-East of Spain (Guardamar, El Carabassí, and San Juan) were selected to isolate endophytic fungi. Isolates were identified on the basis of morphological and/or molecular characters. Five hundred and fifty two endophytic fungi were isolated and assigned to thirty morphological taxa or molecular operational taxonomic units. Most isolates belonged to Ascomycota, and the dominant order was Hypocreales. Fusarium and Clonostachys were the most frequently isolated genera and were present at all sampling sites. Comparisons of the endophytic diversity with previous studies, and their importance in the management of the date palm crops are discussed. This is the first study on the diversity of endophytic fungi associated with roots of date palm. The isolates obtained might constitute a source of biological control agents and biofertilizers for use in crops of this plant. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. Bioactive Constituents from an Endophytic Fungus, Penicillium polonicum NFW9, Associated with Taxus fauna.

    Fatima, Nighat; Sripisut, Tawanun; Youn, Ui J; Ahmed, Safia; Ul-Haq, Ihsan; Munoz-Acuna, Ulyana; Simmons, Charles J; Qazi, Muneer A; Jadoon, Muniba; Tan, Ghee T; de Blanco, Esperanza J C; Chang, Leng C

    2017-01-01

    Endophytic fungi are being recognized as vital and untapped sources of a variety of structurally novel and unique bioactive secondary metabolites in the field of natural products drug discovery. Herein, this study reports the isolation and characterization of secondary metabolites from an endophytic fungus Penicillium polonicum (NFW9) associated with Taxus fuana. The extracts of the endophytic fungus cultured on potato dextrose agar were purified using several chromatographic techniques. Biological evaluation was performed based on their abilities to inhibit tumor necrosis factor-alpha (TNF-α)-induced nuclear factor-kappa B (NF-κB) and cytotoxicity assays. Bioactivity-directed fractionation of the ethyl acetate extract of a fermentation culture of an endophytic fungus, Penicillium polonicum led to the isolation of a dimeric anthraquinone, (R)- 1,1',3,3',5,5'-hexahydroxy-7,7'-dimethyl[2,2'-bianthracene]-9,9',10,10'-tetraone (1), a steroidal furanoid (-)-wortmannolone (2), along with three other compounds (3-4). Moreover, this is the first report on the isolation of compound 1 from an endophytic fungus. All purified metabolites were characterized by NMR and MS data analyses. The stereo structure of compound 1 was determined by the measurement of specific optical rotation and CD spectrum. The relative stereochemistry of 2 was confirmed by single-crystal X-ray diffraction. Compounds 2-3 showed inhibitory activities in the TNF-α-induced NF-κB assay with IC50 values in the range of 0.47-2.11 µM. Compounds 1, 4 and 5 showed moderate inhibition against NF-κB and cancer cell lines. The endophytic fungus Penicillium polonicum of Taxus fuana is capable of producing biologically active natural compounds. Our results provide a scientific rationale for further chemical investigations into endophyte-producing natural products, drug discovery and development. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  18. Colonization of Onions by Endophytic Fungi and Their Impacts on the Biology of Thrips tabaci

    Muvea, Alexander M.; Meyhöfer, Rainer; Subramanian, Sevgan; Poehling, Hans-Michael; Ekesi, Sunday; Maniania, Nguya K.

    2014-01-01

    Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant–herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci. PMID:25254657

  19. Endophytic fungi associated with Ziziphus species from mountainous area of Oman and new records

    SAIFELDIN A.F. EL-NAGERABI

    2013-04-01

    Full Text Available El-Nagerabi SAF, Elshafie AE, AlKhanjari SS. 2013. Endophytic fungi associated with Ziziphus species from mountainous area of Oman and new records. Biodiversitas 14: 10-16. Ziziphus species of the family Rhamnaceae grow extensively in arid and semi-arid regions. It is possible that the endophytic fungi associated with this plant might enhance the host resistance to the environmental impacts. The endophytic fungal population inhabiting the healthy leaves of Z. spina-christi and Z. hajanensis plants were determined from April 2008 to October 2011. The endophytic fungal communities varied between the two species, and 45 fungal species, 18 sterile mycelia and 12 yeasts were isolated from Z. spina-christi, whereas 35 fungi, 11 sterile mycelia and 5 yeasts were recovered from Z. hajanensis indicating tissue and species-specificity and without any seasonal variation among the endophytes. These endophytes are new to Ziziphus plants and 45 species are new to the mycoflora of Oman, whereas 27 species are new to Arabian Peninsula. The genus Alternaria was the most prevalent (19-81% followed by Aspergillus (19-78%, Rhizopus stolonifer (78%, Mycelia sterilia (69%, yeasts (47%, Cladosporium (11-56%, Drechslera (14-53%, Curvularia (8-50%, Fusarium (6-33%, Ulocladium (41-31%, Penicillium (3-22%, Alysidium resine (11%, Trichocladium (6-11%, Anguillospora longissima, Bactrodesmium rahmii, Catenularia (8%, Helminthosporium sorghi (7%, Dendryphiella infuscans (6%, Hansfordia biophila (3-6%, Arthrinium, Dissophora, and Phoma sorghina (3%. The recovery of many fungal isolates, morphologically various sterile mycelia and yeasts suggests the high biodiversity of the endophytes invading these plants with strong evidence for future isolation of numerous fungal species through adopting more advanced molecular and DNA identification methods.

  20. Plant genotype-specific archaeal and bacterial endophytes but similar Bacillus antagonists colonize Mediterranean olive trees

    Henry eMueller

    2015-03-01

    Full Text Available Endophytes have an intimate and often symbiotic interaction with their hosts. Less is known about the composition and function of endophytes in trees. In order to evaluate our hypothesis that plant genotype and origin have a strong impact on both, endophytes of leaves from 10 Olea europaea L. cultivars from the Mediterranean basin growing at a single agricultural site in Spain and from nine wild olive trees located in natural habitats in Greece, Cyprus and on Madeira Island were studied. The composition of the bacterial endophytic communities as revealed by 16S rRNA gene amplicon sequencing and the subsequent PCoA analysis showed a strong correlation to the plant genotypes. The bacterial distribution patterns were congruent with the plant origins in Eastern and Western areas of the Mediterranean basin. Subsequently, the endophytic microbiome of wild olives was shown to be closely related to those of cultivated olives of the corresponding geographic origins. The olive leaf endosphere harbored mostly Proteobacteria, followed by Firmicutes, Actinobacteria and Bacteroidetes. The detection of a high portion of archaeal taxa belonging to the phyla Thaumarchaeota, Crenarchaeota and Euryarchaeota in the amplicon libraries was an unexpected discovery, which was confirmed by quantitative real-time PCR revealing an archaeal portion of up to 35.8%. Although the function of these Archaea for their host plant remains speculative, this finding suggests a significant relevance of archaeal endophytes for plant-microbe interactions. In addition, the antagonistic potential of culturable endophytes was determined; all isolates with antagonistic activity against the olive-pathogenic fungus Verticillium dahliae Kleb. belong to Bacillus amyloliquefaciens. In contrast to the specific global structural diversity, BOX-fingerprints of the antagonistic Bacillus isolates were highly similar and independent of the olive genotype from which they were isolated.

  1. Arsenic transformation and plant growth promotion characteristics of As-resistant endophytic bacteria from As-hyperaccumulator Pteris vittata.

    Xu, Jia-Yi; Han, Yong-He; Chen, Yanshan; Zhu, Ling-Jia; Ma, Lena Q

    2016-02-01

    The ability of As-resistant endophytic bacteria in As transformation and plant growth promotion was determined. The endophytes were isolated from As-hyperaccumulator Pteris vittata (PV) after growing for 60 d in a soil containing 200 mg kg(-1) arsenate (AsV). They were isolated in presence of 10 mM AsV from PV roots, stems, and leaflets, representing 4 phyla and 17 genera. All endophytes showed at least one plant growth promoting characteristics including IAA synthesis, siderophore production and P solubilization. The root endophytes had higher P solubilization ability than the leaflet (60.0 vs. 18.3 mg L(-1)). In presence of 10 mM AsV, 6 endophytes had greater growth than the control, suggesting As-stimulated growth. Furthermore, root endophytes were more resistant to AsV while the leaflet endophytes were more tolerant to arsenite (AsIII), which corresponded to the dominant As species in PV tissues. Bacterial As resistance was positively correlated to their ability in AsV reduction but not AsIII oxidation. The roles of those endophytes in promoting plant growth and As resistance in P. vittata warrant further investigation. Published by Elsevier Ltd.

  2. Functional and molecular characterization of genes involved in antagonisms between two maize endophytes, Fusarium verticillioides and Sarocladium zeae

    Fusarium verticillioides (Fv) is a prevalent seed-borne maize endophyte capable of causing severe kernel rot and fumonisin mycotoxin contamination. Within maize kernels, Fv is primarily confined to the pedicel, while another seed-borne fungal endophyte, Sarocladium zeae (Sz), is observed in embryos....

  3. A Battle in a Kernel: Molecular Exploration of Antagonisms between Two Maize Endophytes, Fusarium verticillioides and Acremonium zeae

    Fusarium verticillioides (Fv) is a prevalent seed-borne maize endophyte capable of causing severe kernel rot and fumonisin mycotoxin contamination. Within maize kernels, Fv is primarily confined to the pedicel, while another seed-borne fungal endophyte, Acremonium zeae (Az), is observed in embryos. ...

  4. Quorum signaling mycotoxins: A new risk strategy for bacterial biocontrol of Fusarium verticillioides and other endophytic fungal species?

    Bacterial endophytes are used as biocontrol organisms for plant pathogens such as the maize endophyte Fusarium verticillioides and its production of fumonisin mycotoxins. However, such applications are not always predictable and efficient. All bacteria communicate via cell-dependent signals, which...

  5. Restructuring of Endophytic Bacterial Communities in Grapevine Yellows-Diseased and Recovered Vitis vinifera L. Plants ▿

    Bulgari, Daniela; Casati, Paola; Crepaldi, Paola; Daffonchio, Daniele; Quaglino, Fabio; Brusetti, Lorenzo; Bianco, Piero Attilio

    2011-01-01

    Length heterogeneity-PCR assays, combined with statistical analyses, highlighted that the endophytic bacterial community associated with healthy grapevines was characterized by a greater diversity than that present in diseased and recovered plants. The findings suggest that phytoplasmas can restructure the bacterial community by selecting endophytic strains that could elicit a plant defense response. PMID:21622794

  6. Molecular phylogeny, diversity and bioprospecting of endophytic fungi associated with wild ethnomedicinal North American plant Echinacea purpurea (Asteraceae)

    The endophytic fungal community associated with the wild ethnomedicinal North American plant Echinacea purpurea was investigated as well as its potential for providing antifungal compounds against plant pathogenic fungi. A total of 233 endophytic fungal isolates were obtained and classified into 42 ...

  7. Endophytic fungal communities associated with field-grown soybean roots and seeds in the Huang-Huai region of China

    Hongjun Yang

    2018-04-01

    Full Text Available Plants depend on beneficial interactions between roots and fungal endophytes for growth, disease suppression, and stress tolerance. In this study, we characterized the endophytic fungal communities associated with the roots and corresponding seeds of soybeans grown in the Huang-Huai region of China. For the roots, we identified 105 and 50 genera by culture-independent and culture-dependent (CD methods, respectively, and isolated 136 fungal strains (20 genera from the CD samples. Compared with the 52 soybean endophytic fungal genera reported in other countries, 28 of the genera we found were reported, and 90 were newly discovered. Even though Fusarium was the most abundant genus of fungal endophyte in every sample, soybean root samples from three cities exhibited diverse endophytic fungal communities, and the results between samples of roots and seeds were also significantly different. Together, we identified the major endophytic fungal genera in soybean roots and seeds, and revealed that the diversity of soybean endophytic fungal communities was influenced by geographical effects and tissues. The results will facilitate a better understanding of soybean–endophytic fungi interaction systems and will assist in the screening and utilization of beneficial microorganisms to promote healthy of plants such as soybean.

  8. Alterations in serotonin receptor-induced contractility of bovine lateral saphenous vein in cattle grazing endophyte-infected tall fescue

    As part of a large 2-year study documenting the physiologic impact of grazing endophyte-infected tall fescue on growing cattle, 2 experiments were conducted to characterize and evaluate the effects of grazing 2 levels of toxic endophyte-infected tall fescue pastures on vascular contractility and ser...

  9. Aspergillus oryzae NRRL 35191 from coffee, a non-toxigenic endophyte with the ability to synthesize kojic acid

    Aspergillus oryzae was isolated as an endophyte from coffee leaves and found to produce kojic acid in culture. When inoculated in cacao seedlings (Theobroma cacao L.), A. oryzae grew endophytically and synthesize kojic acid in planta. Cacao seedlings inoculated with A. oryzae produced higher levels...

  10. Inter- and intracellular colonization of Arabidopsis roots by endophytic actinobacteria and the impact of plant hormones on their antimicrobial activity

    Meij, van der Anne; Willemse, Joost; Schneijderberg, Martinus A.; Geurts, René; Raaijmakers, Jos M.; Wezel, van Gilles P.

    2018-01-01

    Many actinobacteria live in close association with eukaryotes such as fungi, insects, animals and plants. Plant-associated actinobacteria display (endo)symbiotic, saprophytic or pathogenic life styles, and can make up a substantial part of the endophytic community. Here, we characterised endophytic

  11. Inter- and intracellular colonization of Arabidopsis roots by endophytic actinobacteria and the impact of plant hormones on their antimicrobial activity

    Van der Meij, Anne; Willemse, Joost; Schneijderberg, Martinus A.; Geurts, Rene; Raaijmakers, Jos; van Wezel, Gilles

    2018-01-01

    Many actinobacteria live in close association with eukaryotes like fungi, insects, animals and plants. Plant-associated actinobacteria display (endo)symbiotic, saprophytic or pathogenic life styles, and can make up a substantial part of the endophytic community. Here, we characterised endophytic

  12. Antagonism of lateral saphenous vein serotonin receptors from steers grazing endophyte-free, wild-type, or novel endophyte-infected tall fescue

    Pharmacologic profiling of 5-hydroxytryptamine (5HT) receptors of bovine lateral saphenous vein has shown that cattle grazing endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum) have altered responses to ergovaline (ERV), 5HT, 5HT2A and 5HT7 agonists. To determine if 5HT...

  13. Contributions of North American endophytes to the phylogeny, ecology, and taxonomy of Xylariaceae (Sordariomycetes, Ascomycota).

    U'Ren, Jana M; Miadlikowska, Jolanta; Zimmerman, Naupaka B; Lutzoni, François; Stajich, Jason E; Arnold, A Elizabeth

    2016-05-01

    The Xylariaceae (Sordariomycetes) comprise one of the largest and most diverse families of Ascomycota, with at least 85 accepted genera and ca. 1343 accepted species. In addition to their frequent occurrence as saprotrophs, members of the family often are found as endophytes in living tissues of phylogenetically diverse plants and lichens. Many of these endophytes remain sterile in culture, precluding identification based on morphological characters. Previous studies indicate that endophytes are highly diverse and represent many xylariaceous genera; however, phylogenetic analyses at the family level generally have not included endophytes, such that their contributions to understanding phylogenetic relationships of Xylariaceae are not well known. Here we use a multi-locus, cumulative supermatrix approach to integrate 92 putative species of fungi isolated from plants and lichens into a phylogenetic framework for Xylariaceae. Our collection spans 1933 isolates from living and senescent tissues in five biomes across the continental United States, and here is analyzed in the context of previously published sequence data from described species and additional taxon sampling of type specimens from culture collections. We found that the majority of strains obtained in our surveys can be classified in the hypoxyloid and xylaroid subfamilies, although many also were found outside of these lineages (as currently circumscribed). Many endophytes were placed in lineages previously not known for endophytism. Most endophytes appear to represent novel species, but inferences are limited by potential gaps in public databases. By linking our data, publicly available sequence data, and records of ascomata, we identify many geographically widespread, host-generalist clades capable of symbiotic associations with diverse photosynthetic partners. Concomitant with such cosmopolitan host use and distributions, many xylariaceous endophytes appear to inhabit both living and non-living plant

  14. Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination.

    Lumactud, Rhea; Shen, Shu Yi; Lau, Mimas; Fulthorpe, Roberta

    2016-01-01

    The bacterial endophytic communities of four plants growing abundantly in soils highly contaminated by hydrocarbons were analyzed through culturable and culture-independent means. Given their tolerance to the high levels of petroleum contamination at our study site, we sought evidence that Achillea millefolium, Solidago canadensis, Trifolium aureum, and Dactylis glomerata support high levels of hydrocarbon degrading endophytes. A total of 190 isolates were isolated from four plant species. The isolates were identified by partial 16S rDNA sequence analysis, with class Actinobacteria as the dominant group in all species except S. canadensis, which was dominated by Gammaproteobacteria. Microbacterium foliorum and Plantibacter flavus were present in all the plants, with M. foliorum showing predominance in D. glomerata and both endophytic bacterial species dominated T. aureum. More than 50% of the isolates demonstrated degradative capabilities for octanol, toluene, naphthalene, kerosene, or motor oil based on sole carbon source growth screens involving the reduction of tetrazolium dye. P. flavus isolates from all the sampled plants showed growth on all the petroleum hydrocarbons (PHCs) substrates tested. Mineralization of toluene and naphthalene was confirmed using gas-chromatography. 16S based terminal restriction fragment length polymorphism analysis revealed significant differences between the endophytic bacterial communities showing them to be plant host specific at this site. To our knowledge, this is the first account of the degradation potential of bacterial endophytes in these commonly occurring pioneer plants that were not previously known as phytoremediating plants.

  15. A fungal endophyte helps plants to tolerate root herbivory through changes in gibberellin and jasmonate signaling.

    Cosme, Marco; Lu, Jing; Erb, Matthias; Stout, Michael Joseph; Franken, Philipp; Wurst, Susanne

    2016-08-01

    Plant-microbe mutualisms can improve plant defense, but the impact of root endophytes on below-ground herbivore interactions remains unknown. We investigated the effects of the root endophyte Piriformospora indica on interactions between rice (Oryza sativa) plants and its root herbivore rice water weevil (RWW; Lissorhoptrus oryzophilus), and how plant jasmonic acid (JA) and GA regulate this tripartite interaction. Glasshouse experiments with wild-type rice and coi1-18 and Eui1-OX mutants combined with nutrient, jasmonate and gene expression analyses were used to test: whether RWW adult herbivory above ground influences subsequent damage caused by larval herbivory below ground; whether P. indica protects plants against RWW; and whether GA and JA signaling mediate these interactions. The endophyte induced plant tolerance to root herbivory. RWW adults and larvae acted synergistically via JA signaling to reduce root growth, while endophyte-elicited GA biosynthesis suppressed the herbivore-induced JA in roots and recovered plant growth. Our study shows for the first time the impact of a root endophyte on plant defense against below-ground herbivores, adds to growing evidence that induced tolerance may be an important root defense, and implicates GA as a signal component of inducible plant tolerance against biotic stress. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  16. Diversity of endophytic fungi of Myricaria laxiflora grown under pre- and post-flooding conditions.

    Tian, W; Bi, Y H; Zeng, W; Jiang, W; Xue, Y H; Wang, G X; Liu, S P

    2015-09-09

    Myricaria laxiflora is distributed along the riverbanks of the Yangtze River valley. The Three Gorges Dam has dramatically changed the habitat of M. laxiflora, which has evolved to develop increased resistance to flooding stress. In order to elucidate the relationship between plant endophytic fungi and flooding stress, we isolated and taxonomically characterized the endophytic fungi of M. laxiflora. One hundred and sixty-three fungi were isolated from healthy stems, leaves and roots of M. laxiflora grown under pre- and post-flooding conditions. Culture and isolation were carried out under aerobic and anaerobic conditions. Based on internal transcribed spacer sequence analysis and morphological characteristics, the isolates exhibited abundant biodiversity; they were classified into 5 subphyla, 7 classes, 12 orders, 17 families, and 26 genera. Dominant endophytes varied between pre- and post-flooding plants, among different plant tissues, and between aerobic and anaerobic culture conditions. Aspergillus and Alternaria accounted for more than 55% of all isolates. Although the number of isolates from post-flooding plants was greater, endophytes from pre-flooding plants were more diverse and abundant. Endophytes were distributed preferentially in particular tissues; this affinity was constrained by both the host habitat and the oxygen availability of the host.

  17. Seaweed temporal distribution in southeast coast of Peninsular Malaysia and isolation of endophytic fungi

    Zainee, Nur Farah Ain; Ismail, Ahmad; Ibrahim, Nazlina; Ismail, Asmida

    2018-04-01

    Temporal study of seaweeds was carried out between on February 2015 and November 2015 at Kampung Jawa Darat and Kampung Sungai Buntu at Pengerang, Johor, Malaysia. The research objectives were to study the diversity of seaweed and to determine the presence of fungal endophyte in the seaweed. The diversity of seaweed in the sampling site was calculated by using quadrat with 25 meter line transect by 3 replication for each site. The specimen were identified and processed in laboratory and kept for reference in the Algae Herbarium, Universiti Kebangsaan Malaysia. The specimen for fungal endophyte isolation was collected randomly by choosing the complete thallus, transferred into sterile zip-lock plastic bag and kept in freezer until used. From this study, a total of 29 species have been successfully identified including 12 species of Chlorophyta, 2 species of Phaeophyta and 14 species of Rhodophyta. From February to November 2015, the number of species highly varied and a significant change in community structure was noted. Kampung Sungai Buntu shows the highest diversity throughout the study compared to Kampung Jawa Darat. Eighteen seaweed species were screened for the presence of fungal endophyte, Sargassum polycystum shows the highest number of fungal endophyte. This study documented the seaweed diversity in two sites at Pengerang, Johor that accommodates fungal endophytes.

  18. Isolation and enzyme bioprospection of endophytic bacteria associated with plants of Brazilian mangrove ecosystem.

    Castro, Renata A; Quecine, Maria Carolina; Lacava, Paulo T; Batista, Bruna D; Luvizotto, Danice M; Marcon, Joelma; Ferreira, Anderson; Melo, Itamar S; Azevedo, João L

    2014-01-01

    The mangrove ecosystem is a coastal tropical biome located in the transition zone between land and sea that is characterized by periodic flooding, which confers unique and specific environmental conditions on this biome. In these ecosystems, the vegetation is dominated by a particular group of plant species that provide a unique environment harboring diverse groups of microorganisms, including the endophytic microorganisms that are the focus of this study. Because of their intimate association with plants, endophytic microorganisms could be explored for biotechnologically significant products, such as enzymes, proteins, antibiotics and others. Here, we isolated endophytic microorganisms from two mangrove species, Rhizophora mangle and Avicennia nitida, that are found in streams in two mangrove systems in Bertioga and Cananéia, Brazil. Bacillus was the most frequently isolated genus, comprising 42% of the species isolated from Cananéia and 28% of the species from Bertioga. However, other common endophytic genera such as Pantoea, Curtobacterium and Enterobacter were also found. After identifying the isolates, the bacterial communities were evaluated for enzyme production. Protease activity was observed in 75% of the isolates, while endoglucanase activity occurred in 62% of the isolates. Bacillus showed the highest activity rates for amylase and esterase and endoglucanase. To our knowledge, this is the first reported diversity analysis performed on endophytic bacteria obtained from the branches of mangrove trees and the first overview of the specific enzymes produced by different bacterial genera. This work contributes to our knowledge of the microorganisms and enzymes present in mangrove ecosystems.

  19. Antiangiogenic, wound healing and antioxidant activity of Cladosporium cladosporioides (Endophytic Fungus isolated from seaweed (Sargassum wightii

    Manjunath M. Hulikere

    2016-10-01

    Full Text Available Endophytic fungi from marine seaweeds are the less studied group of organisms with vast medical applications. The aim of the present study was to evaluate antioxidant, antiangiogenic as well as wound healing potential of the endophytic fungus isolated from the seaweed Sargassum wightii. The morphological characters and the rDNA internal transcribed spacer sequence analysis (BLAST search in Gen Bank database was used for the identification of endophytic fungus. The antioxidant potential of the ethyl acetate extract of endophytic fungus was assessed by, 1,1-diphenyl-2-picryl-hydrazyl radical scavenging method. The fungal extract was also analysed for reducing power, total phenolic and flavonoid content. Antiangiogenic activity of the fungal extract was studied in vitro by inhibition of wound healing scratch assay and in vivo by Chick chorioallantoic membrane assay. The endophytic fungus was identified as Cladosporium cladosporioides (Gen Bank ID – KT384175. The ethyl acetate extract of C. cladosporioides showed a significant antioxidant and angiosuppressive activity. The ESI-LC-MS analysis of the extract revealed the presence of wide range of secondary metabolites. Results suggest that C. cladosporioides extract could be exploited as a potential source for angiogenic modulators.

  20. Anticestodal Activity of Endophytic Pestalotiopsis sp. on Protoscoleces of Hydatid Cyst Echinococcus granulosus

    Vijay C. Verma

    2013-01-01

    Full Text Available Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9, Colletotrichum (469 ± 25.8, Fusarium (355 ± 14.5, and Chaetomium (332 ± 28.3 showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0, which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity.

  1. Occurrence of Bacillus amyloliquefaciens as a systemic endophyte of vanilla orchids.

    White, James F; Torres, Mónica S; Sullivan, Raymond F; Jabbour, Rabih E; Chen, Qiang; Tadych, Mariusz; Irizarry, Ivelisse; Bergen, Marshall S; Havkin-Frenkel, Daphna; Belanger, Faith C

    2014-11-01

    We report the occurrence of Bacillus amyloliquefaciens in vanilla orchids (Vanilla phaeantha) and cultivated hybrid vanilla (V. planifolia × V. pompona) as a systemic bacterial endophyte. We determined with light microscopy and isolations that tissues of V. phaeantha and the cultivated hybrid were infected by a bacterial endophyte and that shoot meristems and stomatal areas of stems and leaves were densely colonized. We identified the endophyte as B. amyloliquefaciens using DNA sequence data. Since additional endophyte-free plants and seed of this orchid were not available, additional studies were performed on surrogate hosts Amaranthus caudatus, Ipomoea tricolor, and I. purpurea. Plants of A. caudatus inoculated with B. amyloliquefaciens demonstrated intracellular colonization of guard cells and other epidermal cells, confirming the pattern observed in the orchids. Isolations and histological studies suggest that the bacterium may penetrate deeply into developing plant tissues in shoot meristems, forming endospores in maturing tissues. B. amyloliquefaciens produced fungal inhibitors in culture. In controlled experiments using morning glory seedlings we showed that the bacterium promoted seedling growth and reduced seedling necrosis due to pathogens. We detected the gene for phosphopantetheinyl transferase (sfp), an enzyme in the pathway for production of antifungal lipopeptides, and purified the lipopeptide "surfactin" from cultures of the bacterium. We hypothesize that B. amyloliquefaciens is a robust endophyte and defensive mutualist of vanilla orchids. Whether the symbiosis between this bacterium and its hosts can be managed to protect vanilla crops from diseases is a question that should be evaluated in future research. © 2014 Wiley Periodicals, Inc.

  2. Endophytic bacterial diversity in the phyllosphere of Amazon Paullinia cupana associated with asymptomatic and symptomatic anthracnose.

    Bogas, Andréa Cristina; Ferreira, Almir José; Araújo, Welington Luiz; Astolfi-Filho, Spartaco; Kitajima, Elliot Watanabe; Lacava, Paulo Teixeira; Azevedo, João Lúcio

    2015-01-01

    Endophytes colonize an ecological niche similar to that of phytopathogens, which make them candidate for disease suppression. Anthracnose is a disease caused by Colletotrichum spp., a phytopathogen that can infect guarana (Paullinia cupana), an important commercial crop in the Brazilian Amazon. We investigated the diversity of endophytic bacteria inhabiting the phyllosphere of asymptomatic and symptomatic anthracnose guarana plants. The PCR-denaturation gradient gel electrophoresis (PCR-DGGE) fingerprints revealed differences in the structure of the evaluated communities. Detailed analysis of endophytic bacteria composition using culture-dependent and 16S rRNA clone libraries revealed the presence of Firmicutes, Proteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria phyla. Firmicutes comprised the majority of isolates in asymptomatic plants (2.40E(-4)). However, cloning and sequencing of 16S rRNA revealed differences at the genus level for Neisseria (1.4E(-4)), Haemophilus (2.1E(-3)) and Arsenophonus (3.6E(-5)) in asymptomatic plants, Aquicella (3.5E(-3)) in symptomatic anthracnose plants, and Pseudomonas (1.1E(-3)), which was mainly identified in asymptomatic plants. In cross-comparisons of the endophytic bacterial communities as a whole, symptomatic anthracnose plants contained higher diversity, as reflected in the Shannon-Weaver and Simpson indices estimation (P anthracnose can restructure endophytic bacterial communities by selecting certain strains in the phyllosphere of P. cupana. The understanding of these interactions is important for the development of strategies of biocontrol for Colletotrichum.

  3. Phytoremediation of Alberta oil sand tailings using native plants and fungal endophytes

    Repas, T.; Germida, J.; Kaminskyj, S.

    2012-04-01

    Fungal endophytes colonize host plants without causing disease. Some endophytes confer plant tolerance to harsh environments. One such endophyte, Trichoderma harzianum strain TSTh20-1, was isolated from a plant growing on Athabasca oil sand tailings. Tailing sands are a high volume waste product from oil sand extraction that the industry is required to remediate. Tailing sands are low in organic carbon and mineral nutrients, and are hydrophobic due to residual polyaromatic hydrocarbons. Typically, tailing sands are remediated by planting young trees in large quantities of mulch plus mineral fertilizer, which is costly and labour intensive. In greenhouse trials, TSTh20-1 supports growth of tomato seedlings on tailing sands without fertilizer. The potential use of TSTh20-1 in combination with native grasses and forbs to remediate under field conditions is being assessed. Twenty-three commercially available plant species are being screened for seed germination and growth on tailing sands in the presence of TSTh20-1. The best candidates from this group will be used in greenhouse and small scale field trials. Potential mechanisms that contribute to endophyte-induced plant growth promotion, such as plant hormone production, stress tolerance, mineral solubilization, and uptake are also being assessed. As well, TSTh20-1 appears to be remarkably frugal in its nutrient requirements and the possibility that this attribute is characteristic of other plant-fungal endophytes from harsh environments is under study.

  4. Fungal endophytes from Acer ginnala Maxim: isolation, identification and their yield of gallic acid.

    Qi, F-H; Jing, T-Z; Wang, Z-X; Zhan, Y-G

    2009-07-01

    The aim of the study was to isolate the endophytic fungi from Acer ginnala and screen isolates rich in gallic acid. After epiphytic sterilization, 145 fungal endophytes were isolated from the stem, annual twig and seed of Acer ginnala. The endophytes were grouped into ten different taxa, Phomopsis sp., Neurospora sp., Phoma sp., Epicoccum sp., Penicillium sp., Alternaria sp., Fusarium sp., Trichoderma sp., Cladosporium sp. and a species of Pleosporales Incertae Sedis, by their morphological traits and ITS-rDNA sequence analysis. The content and yield of gallic acid of 141 isolates were determined by HPLC. On average, the species of Pleosporales Incertae Sedis had the highest content and yield of gallic acid (13.28 mg g(-1) DW; 119.62 mg l(-1)), while Alternaria sp. had the lowest. Of 141 fungal endophytes from A. ginnala, Phomopsis sp. isolate SX10 showed both the highest content and the highest yield of gallic acid (29.25 mg g(-1) DW; 200.47 mg l(-1)). Endophytic fungi isolated from A. ginnala may be used as potential producers of gallic acid and other compounds with biological activities, or functioned as elicitors to produce natural compounds.

  5. Bacterial endophytes isolated from plants in natural oil seep soils with chronic hydrocarbon contamination

    Rhea eLumactud

    2016-05-01

    Full Text Available The bacterial endophytic communities of four plants growing abundantly in soils highly contaminated by hydrocarbons were analyzed through culturable and and culture-independent means. Given their tolerance to the high levels of petroleum contamination at our study site, we sought evidence that Achillea millefolium, Solidago canadensis, Trifolium aureum and Dactylis glomerata support high levels of hydrocarbon degrading endophytes. A total of 190 isolates were isolated from four plant species. The isolates were identified by partial 16S rDNA sequence analysis, with class Actinobacteria as the dominant group in all species except Solidago canadensis, which was dominated by Gammaproteobacteria. Microbacterium foliorum and Plantibacter flavus were present in all the plants, with M. foliorum showing predominance in D. glomerata and both endophytic bacterial species dominated T. aureum. More than 50% of the isolates demonstrated degradative capabilities for octanol, toluene, naphthalene, kerosene or motor oil based on sole carbon source growth screens involving the reduction of tetrazolium dye. P. flavus isolates from all the sampled plants showed growth on all the petroleum hydrocarbons substrates tested. Mineralization of toluene and naphthalene was confirmed using gas-chromatography. 16S based terminal restriction fragment length polymorphism analysis revealed significant differences between the endophytic bacterial communities showing them to be plant host specific at this site. To our knowledge, this is the first account of the degradation potential of bacterial endophytes in these commonly occurring pioneer plants that were not previously known as phytoremediating plants.

  6. Stimulate The Growth of Rice Using Endophytic Bacteria from Lowland Rice Plant Tissue

    Nuni Gofar

    2015-07-01

    Full Text Available Exploration and selection of endophytic bacteria from healthy food crops grown in lowland ecosystem is important to be conducted in order to get growth-stimulating endophytic bacteria at soil with low fertility level so that capable to optimize initial growth of food crops and subsequently can increase productivity level of lowland soil.The research objective was to isolate and to test the IAA-producing endophytic bacteria isolate in stimulating the rice crop growth at lowland area. Endophytic bacteria are isolated from tissues of rice, corn and peanut crops which grown at shallow swamp land in Ogan Ilir and Ogan Komering Ilir Districts, South Sumatra, Indonesia. There was nine isolates of nitrogen-fixer endophytic bacteria that capable to contribute IAA phytohormone into their growth media. The P31 isolate from rice crop tisssue of 2 months old produce the best rice sprouts than other isolates. This isolate can contribute of about 10 mg kg-1 IAA to its growth medium and increase the crowns dry weight and the roots dry weight respectively with magnitudes of 133% and 225% compared to control treatment. Concentration and absorbtion of N for rice crops innoculated with P31 isolates had increased by 169% and 400%, recpectively. The P31 isolates had been identified as Burkholderia pseudomallei (also known as Pseudomonas pseudomallei.

  7. Some fungal endophytes from vegetable crops and their anti-oomycete activities against tomato late blight.

    Kim, H-Y; Choi, G J; Lee, H B; Lee, S-W; Lim, H K; Jang, K S; Son, S W; Lee, S O; Cho, K Y; Sung, N D; Kim, J-C

    2007-03-01

    To isolate endophytic fungi from vegetable plants and examine their in vivo anti-oomycete activity against Phytophthora infestans in tomato plants. Endophytic fungi were isolated from surface-sterilized plant tissues and anti-oomycete activity was measured by in vivo assay using tomato seedlings. Endophytic fungi showing potent anti-oomycete activity were identified by morphological characteristics and nuclear ribosomal ITS1-5.8S-ITS2 sequence analysis. A total of 152 isolates were obtained from 66 healthy tissue samples of cucumber, red pepper, tomato, pumpkin and Chinese cabbage and the fermentation broths of 23 isolates showed potent in vivo anti-oomycete activity against tomato late blight with control values over 90%. The Fusarium oxysporum strain EF119, which was isolated from roots of red pepper, showed the most potent disease control efficacy against tomato late blight. In dual-culture tests, it inhibited the growth of Pythium ultimum, P. infestans and Phytophthora capsici. Among endophytic fungi isolated from healthy tissues of vegetable plants, F. oxysporum EF119 showed the most potent in vivo anti-oomycete activity against tomato late blight and in vitro anti-oomycete activity against several oomycete pathogens. Endophytic fungi showing anti-oomycete activity in vitro and in vivo may be used as biocontrol agents particularly of tomato late blight.

  8. Natural Medium for Growing of Endophytic Bacteria from Solanaceae in Malang-Indonesia

    Purnawati Arika

    2016-01-01

    Full Text Available Endophytic bacteria are important microorganisms having potential as biocontrol agents for many pathogens. Until now, the growth of it always uses semi-synthetic or synthetic medium so it was difficult to be used by farmers in the field and it was expensive to have its propagation as biocontrol agents. Based on the problem, this research will study the natural medium as propagation medium of Endophytic bacteria. It had natural ingredients such as soybean, chicken broth, egg, worms, snail, sorghum and they were easy to get by farmers. This study used endophytic bacteria from Solanaceae in Malang- Indonesia. Four isolates of endophytic bacteria were grown in agar and liquid medium with ingredients of corn flour, soybean flour, sorghum flour, snail flour, and worm flour. There is no difference in the incubation period, color, shape, and surface colony. The population in medium with snail flour ingredients at a concentration of 107 cfu/ml is the highest and snail flour is the best medium for growing endophytic bacteria.

  9. [Isolation, identification and characterization of ACC deaminase-containing endophytic bacteria from halophyte Suaeda salsa].

    Teng, Songshan; Liu, Yanping; Zhao, Lei

    2010-11-01

    We Isolated and characterized 1-aminocyclopropane-1-carboxylate (ACC) deaminase-containing endophytic bacteria from halophyte Suaeda salsa to understand the interactions between endophytes and halophyte. ACC deaminase-containing endophytic bacteria were isolated from root, stalk and leaf of Suaeda salsa and were identified based on morphological, physiological-biochemical properties, API and 16S rRNA sequence analysis. Isolates were evaluated for their ACC deaminase, antifungal, protease activity, siderophores and phytohormones, such as indole-3-acetic acid, gibberellic acid and abscisic acid production, as well as atmospheric nitrogen fixation and phosphate solubilization. Four ACC deaminase-containing endophytic bacteria strains named as LP11, SS12, TW1 and TW2 were isolated and identified as Pseudomonas oryzihabitans, Pseudomonas sp., Pantoea agglomerans and Pseudomonas putida respectively. All the strains possessed the phosphate-solubilizing ability and could produce siderophores and phytohormones more or less. None of them could fix atmospheric nitrogen or produce protease. Only strain SS12 showed antagonism against two phytopathogenic fungi viz Fusarium oxysporum f. sp. conglutinans and F. oxysporum f. sp. cucumerinum. ACC deaminase-containing endophytic bacteria of Pseudomonas sp. and Pantoea sp. isolated from halophyte Suaeda salsa have abundant biological characteristics related to plant growth promotion, stress homeostasis regulation and biocontrol activity.

  10. Isolation and characterization of endophytic huperzine A-producing fungi from Huperzia serrata.

    Wang, Ya; Zeng, Qing Gui; Zhang, Zhi Bin; Yan, Ri Ming; Wang, Ling Yun; Zhu, Du

    2011-09-01

    Huperzia serrata is a producer of huperzine A (HupA), a cholinesterase inhibitor (ChEI). Over 120 endophytic fungi were recovered from this plant and screened for Hup-A and nine were found. These nine represented seven different fungal genera with the most significant producer being Shiraia sp. A total of 127 endophytic fungi isolates obtained from the root, stem, and leaf segments of H. serrata were grouped into 19 genera based on their morphological traits and sequence analysis of the internal transcribed spacers (ITS1-5.8S-ITS2), indicating endophytic fungi in H. serrata are diverse and abundant. Aspergillus, Podospora, Penicillium, Colletotrichum, and Acremonium were the frequent genera, whereas the remaining genera were infrequent groups. Overall, 39 endophytic fungi isolates showed acetylcholinesterase (AChE) inhibition in vitro. Nine endophytic fungi isolates from seven distinct genera were capable of producing HupA verified by thin-layer chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC). Among the HupA-producing fungi, the yield of HupA produced by the Shiraia sp. Slf14 was 327.8 μg/l in potato dextrose broth, and the fungal HupA was further validated by mass spectrometry (ESI-MS). The present study demonstrated that H. serrata was a fascinating fungal reservoir for producing HupA and other ChEIs.

  11. Metabolic response induced by endophytic fungi and bacteria in H. marrubioides Epling in vitro micro plants

    Vitorino, Luciana Cristina; Silva, Fabiano Guimaraes, E-mail: fabianocefetrv@yahoo.com.br [Instituto Federal de Educacao, Ciencia e Tecnologia Goiano, Rio Verde, GO (Brazil); Lima, William Cardoso; Soares, Marcos Antonio [Universidade Federal de Mato Grosso (UFMT), Cuiaba, MT (Brazil). Dept. de Botanica e Ecologia; Pedroso, Rita Cassia Nascimento; Silva, Maroli Rodrigues; Dias, Herbert Junior; Crotti, Antonio Eduardo Miller; Silva, Marcio Luis Andrade e; Cunha, Wilson Roberto; Pauletti, Patricia Mendonca; Januario, Ana Helena [Universidade de Franca, SP (Brazil). Nucleo de Pesquisa em Ciencias Exatas e Tecnologicas

    2013-10-01

    Hyptis marrubioides Epling is a native plant from Brazilian Cerrado. In this paper, the response of in vitro micro plants of this species to inoculation with bacterial and fungal endophytic isolates is evaluated. HPLC-DAD analysis showed the presence of 3,4-O-(Z)-dicaffeoylquinic acid and quercetin-7-O-glucoside as the main components. GC/MS analysis demonstrated that the sesquiterpenes Greek-Small-Letter-Tau -cadinol and caryophyllene oxide were only produced in micro plants inoculated with endophytic bacteria, while methyl hexadecanoate, methyl heptadecanoate and methyl (Z,Z,Z) 9,12,15-octadecatrienoate and the triterpene methyl 3{beta}-hydroxy-urs-12-en-28-oate were over expressed only when the micro plant was treated with endophytic fungi. (author)

  12. Antibiotic Properties of the endophytic Streptomyces Spp. Isolated from the Leaves of Myanmar Medicinal Plants

    Aye Pe; Mar Mar Nyein; Win Maung

    2002-02-01

    Three medicinal plants of Myanmar are selected in the study of endophytic microorganisms and are taxonomically classified and identified to be Sa-ba-lin (Cymbopogon citratus Stapf.), Shazaungtinga- neah (Euphorbia splendens Bojer. ex Hooker) and Ma-shaw (Sauropus grandifolius Pax. and Hoffm.). The screening of endophytic microorganisms is performed according to the ISP method (International Streptomyces Projects 1993). The morphological and physicochemical properties of isolated strains are studied and identified to be the Genus Streptomyces. The test of apparent antimicrobial activity of isolated Streptomyces is done on 18 strains of pathogenic bacteria. It is found that the isolated endophytic Sireptomyces showed the significant antibacterial activity on most of the test organisms. (author)

  13. Metabolic response induced by endophytic fungi and bacteria in H. marrubioides Epling in vitro micro plants

    Vitorino, Luciana Cristina; Silva, Fabiano Guimaraes; Lima, William Cardoso; Soares, Marcos Antonio; Pedroso, Rita Cassia Nascimento; Silva, Maroli Rodrigues; Dias, Herbert Junior; Crotti, Antonio Eduardo Miller; Silva, Marcio Luis Andrade e; Cunha, Wilson Roberto; Pauletti, Patricia Mendonca; Januario, Ana Helena

    2013-01-01

    Hyptis marrubioides Epling is a native plant from Brazilian Cerrado. In this paper, the response of in vitro micro plants of this species to inoculation with bacterial and fungal endophytic isolates is evaluated. HPLC-DAD analysis showed the presence of 3,4-O-(Z)-dicaffeoylquinic acid and quercetin-7-O-glucoside as the main components. GC/MS analysis demonstrated that the sesquiterpenes τ-cadinol and caryophyllene oxide were only produced in micro plants inoculated with endophytic bacteria, while methyl hexadecanoate, methyl heptadecanoate and methyl (Z,Z,Z) 9,12,15-octadecatrienoate and the triterpene methyl 3β-hydroxy-urs-12-en-28-oate were over expressed only when the micro plant was treated with endophytic fungi. (author)

  14. EFFICACY OF ENDOPHYTIC BACTERIA IN REDUCING PLANT PARASITIC NEMATODE Pratylenchus brachyurus

    Rita Harni

    2014-04-01

    Full Text Available Pratylenchus brachyurus is a major parasitic nematode on patchouli that reduces plant production up to 85%. The use of endophytic bacteria is promising for controlling nematode and promoting plant growth through production of phytohormones and enhancing the availability of soil nutrients. The objective of the study was to evaluate the efficacy of endophytic bacteria to control P. brachyurus on patchouli plant and its influence on plant productions (plant fresh weight and patchouli oil. The study was conducted at Cimanggu Experimental Garden and Laboratory of the Indonesian Spice and Medicinal Crops Research Institute (ISMECRI, Bogor, West Java. The experi-ment was designed in a randomized block with seven treatments and eight replications; each replication consisted of 10 plants. The treatments evaluated were five isolates of endophytic bacteria (Achromobacter xylosoxidans TT2, Alcaligenes faecalis NJ16, Pseudomonas putida EH11, Bacillus cereus MSK and Bacillus subtilis NJ57, synthetic nematicide as a reference, and non-treated plant as a control.  Four-week old patchouli plants of cv. Sidikalang were treated by soaking the roots in suspension of endophytic bacteria (109 cfu  ml-1 for one hour before trans-planting to the field. At one month after planting, the plants were drenched with the bacterial suspension as much as 100 ml per plant. The results showed that applications of the endophytic bacteria could suppress the nematode populations (52.8-80% and increased plant weight (23.62-57.48% compared to the control. The isolate of endophytic bacterium Achromobacter xylosoxidans TT2 was the best and comparable with carbofuran.

  15. Endophytic bacterial community of grapevine leaves influenced by sampling date and phytoplasma infection process.

    Bulgari, Daniela; Casati, Paola; Quaglino, Fabio; Bianco, Piero A

    2014-07-21

    Endophytic bacteria benefit host plant directly or indirectly, e.g. by biocontrol of the pathogens. Up to now, their interactions with the host and with other microorganisms are poorly understood. Consequently, a crucial step for improving the knowledge of those relationships is to determine if pathogens or plant growing season influence endophytic bacterial diversity and dynamic. Four healthy, four phytoplasma diseased and four recovered (symptomatic plants that spontaneously regain a healthy condition) grapevine plants were sampled monthly from June to October 2010 in a vineyard in north-western Italy. Metagenomic DNA was extracted from sterilized leaves and the endophytic bacterial community dynamic and diversity were analyzed by taxon specific real-time PCR, Length-Heterogeneity PCR and genus-specific PCR. These analyses revealed that both sampling date and phytoplasma infection influenced the endophytic bacterial composition. Interestingly, in June, when the plants are symptomless and the pathogen is undetectable (i) the endophytic bacterial community associated with diseased grapevines was different from those in the other sampling dates, when the phytoplasmas are detectable inside samples; (ii) the microbial community associated with recovered plants differs from that living inside healthy and diseased plants. Interestingly, LH-PCR database identified bacteria previously reported as biocontrol agents in the examined grapevines. Of these, Burkholderia, Methylobacterium and Pantoea dynamic was influenced by the phytoplasma infection process and seasonality. Results indicated that endophytic bacterial community composition in grapevine is correlated to both phytoplasma infection and sampling date. For the first time, data underlined that, in diseased plants, the pathogen infection process can decrease the impact of seasonality on community dynamic. Moreover, based on experimental evidences, it was reasonable to hypothesize that after recovery the restructured

  16. Endophytic fungi associated with Monarda citriodora, an aromatic and medicinal plant and their biocontrol potential.

    Katoch, Meenu; Pull, Shipra

    2017-12-01

    The Food and Agriculture Organization has estimated that every year considerable losses of the food crops occur due to plant diseases. Although fungicides are extensively used for management of plant diseases, they are expensive and hazardous to the environment and human health. Alternatively, biological control is the safe way to overcome the effects of plant diseases and to sustain agriculture. Since Monarda citriodora Cerv. ex Lag. (Lamiaceae/Labiatae) is known for its antifungal properties, it was chosen for the study. The isolation of endophytic fungi from M. citriodora and assessing their biocontrol potential. The isolated endophytes were characterized using ITS-5.8 S rDNA sequencing. Their biocontrol potential was assessed using different antagonistic assays against major plant pathogens. Twenty-eight endophytes representing 11 genera were isolated, of which, around 82% endophytes showed biocontrol potential against plant pathogens. MC-2 L (Fusarium oxysporum), MC-14 F (F. oxysporum), MC-22 F (F. oxysporum) and MC-25 F (F. redolens) displayed significant antagonistic activity against all the tested pathogens. Interestingly, MC-10 L (Muscodor yucatanensis) completely inhibited the growth of Sclerotinia sp., Colletotrichum capsici, Aspergillus flavus and A. fumigatus in dual culture assay, whereas MC-8 L (A. oryzae) and MC-9 L (Penicillium commune) completely inhibited the growth of the Sclerotinia sp. in fumigation assay. Endophytes MC-2 L, MC-14 F, MC-22 F and MC-25 F could effectively be used to control broad range of phytopathogens, while MC-10 L, MC-8 L and MC-9 L could be used to control specific pathogens. Secondly, endophytes showing varying degrees of antagonism in different assays represented the chemo-diversity not only as promising biocontrol agents but also as a resource of defensive and bioactive metabolites.

  17. Existence of entomopathogen fungi, Beauveria bassiana as an endophyte in cacao seedlings

    Endang Sulistyowati

    2015-12-01

    Full Text Available Beauveria bassiana is one of the entomopathogen fungi which is known as biological control agent of cocoa pod borer and cocoa mirids (Helopeltis spp.. Because of its effectiveness in the fields is still not consistent, so we conduct a research with the objective to know the possibility of Beauveria bassiana to be established as a endophyte. Various fungal entomopathogens have already been reported as endophytes and the various methods used to inoculate the plants with B. bassiana were partially effective. The research has been conducted in laboratory of Plant Protection, Indonesian Coffee and Cocoa Research Institute by inoculating of cocoa seeds and cocoa nursery with B. bassiana suspension.  The trial was arranged  by randomized complete block design with a factorial arrangement. The factor were spore concentration of B. bassiana (0; 2; and 4 g/ 10 l and cocoa varieties (family of ICS 60, TSH858, and hybrid. The trial were use  four replications. The results showed that the fungal entomopathogen B. bassiana was established as an endophyte in cocoa seedling, both from cocoa seeds and nursery application. Percentage of existence of B. bassiana colonies as endophytes one month after seeds application were ICS 60 amounted to 93.3 % both on concentration treatments, while the families of TSH 858 by 80 % and 86.67 % respectively in 2 g and 4 g per 10 l of B. bassiana spores concentration treament.. The lowest percentage was in hybrids, which amounted to 66.67% and 50%. B. bassiana colonies was exixtence as an endophyte in culture from root, stem and leaves of cocoa seedling up to 5 months post inoculation. While the application on nursery by soil drenshing, leaf spraying, and stem injection , it was known that B. bassiana colonies were found in the tissues of leaves, stems, and roots until two months after application. Colonies of B. bassiana as endophytes still exsist until six weeks after nursery was planted in the field. 

  18. A novel endophytic Huperzine A-producing fungus, Shiraia sp. Slf14, isolated from Huperzia serrata.

    Zhu, D; Wang, J; Zeng, Q; Zhang, Z; Yan, R

    2010-10-01

    To characterize and identify a novel Huperzine A (HupA)-producing fungal strain Slf14 isolated from Huperzia serrata (Thunb. ex Murray) Trev. in China. The isolation, identification and characterization of a novel endophytic fungus producing HupA specifically and consistently from the leaves of H. serrata were investigated. The fungus was identified as Shiraia sp. Slf14 by molecular and morphological methods. The HupA produced by this endophytic fungus was shown to be identical to authentic HupA analysed by thin layer chromatographic, High-performance liquid chromatography (HPLC), LC-MS, (1) H NMR and acetylcholinesterase (AChE) inhibition activity in vitro. The amount of HupA produced by Shiraia sp. Slf14 was quantified to be 327.8 μg l(-1) by HPLC, which was far higher than that of the reported endophytic fungi, Acremonium sp., Blastomyces sp. and Botrytis sp. The production of HupA by endophyte Shiraia sp. Slf14 is an enigmatic observation. It would be interesting to further study the HupA production and regulation by the cultured endophyte in H. serrata and in axenic cultures. Although the current accumulation of HupA by the endophyte is not very high, it could provide a promising alterative approach for large-scale production of HupA. However, further strain improvement and the fermentation process optimization are required to result in the consistent and dependable production. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

  19. High-yielding Wheat Varieties Harbour Superior Plant Growth Promoting-Bacterial Endophytes

    Mehwish Yousaf

    2017-06-01

    Full Text Available Background and Objective: The purpose of this study was to compare the endophytic microbial flora of different wheat varieties to check whether a better yielding variety also harbours superior plant growth promoting bacteria. Such bacteria are helpful in food biotechnology as their application can enhance the yield of the crop.Material and Methods: Three wheat varieties (Seher, Faisalabad and Lasani were selected, Seher being the most superior variety. endophytic bacteria were isolated from the histosphere of the leaves and roots at different growth phases of the plants. The isolates were analyzed for plant growth promoting activities. Isolates giving best results were identified through 16S rRNA gene sequencing. Statistical analysis was done using Microsoft Excel 2013. All the experiments were conducted in triplicates.Results and Conclusion: The endophytes of Seher variety showed maximum plant growth promoting abilities. Among the shoot endophytes, the highest auxin production was shown by Seher isolate SHHP1-3 up to 51.9μg ml-1, whereas in the case of root endophytes, the highest auxin was produced by SHHR1-5 up to 36 μg ml-1. The bacteria showing significant plant growth promoting abilities were identified by 16S rRNA sequencing. Bacillus, Proteobacteria and Actinobacteria species were the dominant bacteria showing all the traits of plant growth promotion. It can be concluded that Seher variety harbours superior plant growth promoting endophytes that must be one of the reasons for its better growth and yield as compared to the other two varieties. The investigated results support possible utilization of the selected isolates in wheat growth promotion with respect to increase in agro-productivity. The application of such bacteria could be useful to enhance wheat yield and can help in food biotechnology.Conflict of interest: The authors declare no conflict of interest.

  20. Role of X-ray examination methods in the diagnosis of endophytic stomach carcinomas

    Gorshkov, A.N.; Akberov, R.F.

    1998-01-01

    Results of studying potentialities of radiographic methods in the diagnosis of stomach endophytic neoplasms (130 cases) are presented. All of patients were exposed to complex radiographic-endoscopic studies of stomach. X-ray computerized tomography is used as an additional method. It is shown that the complex approach to the diagnosis of endophytic neoplasms of stomach is necessary. Radiographic method is proposed to be used as an initial examination method. Endoscopic method with multiple biopsy is also used. X-ray computerized tomography is used for certain anatomic stomach section at the final stage [ru

  1. Identification of the fungal endophyte of Ammophila breviligulata (American beachgrass as Epichloë amarillans

    Ian Drake

    2018-01-01

    Full Text Available The grass Ammophila breviligulata (American beachgrass is known to host an endophyte of the genus Epichloë. Based on morphological characteristics it was originally identified as Acremonium typhinum var. ammophilae and is currently designated as Epichloë typhina var. ammophilae. However, the Epichloë species has not previously been identified based on DNA sequence data. Based on phylogenetic placement of beta-tubulin and translation elongation factor 1-alpha DNA sequences the endophyte is identified as a member of E. amarillans rather than E. typhina.

  2. The pathogenicity of Beauveria bassiana: what happens after an endophytic phase in plants?

    Akello, J; Dubois, T; Coyne, D; Kyamanywa, S

    2010-01-01

    The banana weevil Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae) is a serious constraint to banana (Musa spp.) production throughout the world. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) offers a potential weevil management option, but conventional delivery mechanisms have limited its success. As an endophyte, however, B. bassiana can be efficiently delivered to banana planting materials for the potential management of C. sordidus. However, entomopathogens can change morphology and efficacy against their target host when successively sub-cultured on artificial media or when exposed to certain physical and chemical environmental conditions. Whether such changes occur in B. bassiana after an endophytic phase inside a banana plant remains unknown. The primary aim of our study was to evaluate the viability, growth, sporulation and pathogenicity of endophytic B. bassiana. To attain this, two sets of experiments, namely morphological characterization and larval bioassays, were conducted under laboratory conditions. In these experiments, growth and pathogenicity of the wild-type B. bassiana strain G41, obtained originally from banana farms, was compared with the endophytic B. bassiana strain G41, re-isolated from the rhizome of B. bassiana-inoculated banana plants at one month post-inoculation. Morphological characterization, conidial germination, colony growth and sporulation rate was assessed on SDAY media while pathogenicity was determined 15 days after immersing the larvae of C. sordidus in different conidial doses. No differences were observed in colony appearance and growth rate between the endophytic and wild-type strain. Percentage conidial germination for the endophytic strain (91.4-94.0%) was higher than for the wild-type (86.6-89.7%). LD50 equated 1.76 x 10(5) and 0.71 x 10(5) conidia/ml for the wild-type and endophytic B. bassiana strains, respectively, but did not differ between strains. Our study

  3. Isolation and Molecular Identification of Endophytic Bacteria From Rambutan Fruits (Nephelium lappaceum L. Cultivar Binjai

    Sony Suhandono

    2016-01-01

    Full Text Available Interactions between plants and endophytic bacteria are mutualistic. Plant provides nutrient for bacteria, and bacteria will protect the plant from pathogen, help the phytohormone synthesis and nitrogen fixation, and also increase absorption of minerals. These bacteria called plant growth-promoting bacteria. The aim for this study is to identify endophytic bacteria on rambutan (Nephelium lappaceum L. cultivar Binjai with 16S rRNA. Sequencing results showed that the bacteria is derived from genus Corynebacterium, Bacillus, Chryseobacterium, Staphylococcus and Curtobacterium, which suspected play a role as plant growth-promoting bacteria.

  4. Diversity and seasonal variation of endophytic fungi isolated from three conifers in mt. Taehwa, Korea.

    Kim, Chang-Kyun; Eo, Ju-Kyeong; Eom, Ahn-Heum

    2013-06-01

    The needled leaves of three conifer species were collected in Mt. Taehwa during different seasons of the year. Total 59 isolates and 19 species of endophytic fungi were isolated from the leaves and identified using morphological and molecular characteristics. As a result, Shannon index was different in its host plant; Larix kaempferi had a highest value of species diversity. According to the sampling season, 9 species of 19 species were isolated during fall season. The results suggest that the existing of host plant and sampling season are major factors of distribution of endophytic fungi.

  5. Biocontrol for Rhizoctonia Stem Rot Disease by Using Combination of Specific Endophyte in Paddy Tidal Swamp

    Budi, Ismed Setya; Mariana, Mariana

    2013-01-01

    The use of combination of specific endophytic in tidal swamps to control stem root disease as biological control agents has not been done. It is expected that this combination is able to continuously protect plants from pathogen interference. The research was carried out in type C tidal swamp in Banjar regency of South Kalimantan, from March to November 2011, temperature 29-32oC, and pH 4.0-5.5. The method used was Split Plot design. Biocontrol preparation for both types of endophytic was ap...

  6. Antifungal Monoterpene Derivatives from the Plant Endophytic Fungus Pestalotiopsis foedan.

    Xu, Dan; Zhang, Bing-Yang; Yang, Xiao-Long

    2016-10-01

    A new monoterpene lactone, (1R,4R,5R,8S)-8-hydroxy-4,8-dimethyl-2-oxabicyclo[3.3.1]nonan-3-one (1), along with one related known compound, (2R)-2-[(1R)-4-methylcyclohex-3-en-1-yl]propanoic acid (2), were isolated from the liquid culture of the plant endophytic fungus Pestalotiopsis foedan obtained from the branch of Bruguiera sexangula. The structure and absolute configuration of 1 were determined on the basis of extensive analysis of NMR spectra combined with computational methods via calculation of the optical rotation (OR) and 13 C-NMR. Both compounds exhibited strong antifungal activities against Botrytis cinerea and Phytophthora nicotianae with MIC values of 3.1 and 6.3 μg/ml, respectively, which are comparable to those of the known antifungal drug ketoconazole. Compound 2 also showed modest antifungal activity against Candida albicans with a MIC value of 50 μg/ml. © 2016 Wiley-VHCA AG, Zürich.

  7. Volatile antimicrobials from Muscodor crispans, a novel endophytic fungus.

    Mitchell, Angela M; Strobel, Gary A; Moore, Emily; Robison, Richard; Sears, Joe

    2010-01-01

    Muscodor crispans is a recently described novel endophytic fungus of Ananas ananassoides (wild pineapple) growing in the Bolivian Amazon Basin. The fungus produces a mixture of volatile organic compounds (VOCs); some of the major components of this mixture, as determined by GC/MS, are propanoic acid, 2-methyl-, methyl ester; propanoic acid, 2-methyl-; 1-butanol, 3-methyl-;1-butanol, 3-methyl-, acetate; propanoic acid, 2-methyl-, 2-methylbutyl ester; and ethanol. The fungus does not, however, produce naphthalene or azulene derivatives as has been observed with many other members of the genus Muscodor. The mixture of VOCs produced by M. crispans cultures possesses antibiotic properties, as does an artificial mixture of a majority of the components. The VOCs of the fungus are effective against a wide range of plant pathogens, including the fungi Pythium ultimum, Phytophthora cinnamomi, Sclerotinia sclerotiorum and Mycosphaerella fijiensis (the black sigatoka pathogen of bananas), and the serious bacterial pathogen of citrus, Xanthomonas axonopodis pv. citri. In addition, the VOCs of M. crispans killed several human pathogens, including Yersinia pestis, Mycobacterium tuberculosis and Staphylococcus aureus. Artificial mixtures of the fungal VOCs were both inhibitory and lethal to a number of human and plant pathogens, including three drug-resistant strains of Mycobacterium tuberculosis. The gaseous products of Muscodor crispans potentially could prove to be beneficial in the fields of medicine, agriculture, and industry.

  8. Antagonistic Activity Of Endophytic Bacteria Isolated From Mentha Rotundifolia L.

    Elhartiti Abla

    2015-08-01

    Full Text Available Abstract This study is implemented for the isolation purification and identification of endophytic bacteria which produces antifungal substances from the roots of Mentha rotundifolia L. The 59 obtained bacterial isolates were tested for their antagonistic activity by the dual confrontation against the phytopathogenic fungi Fusarium oxysporum Aspergillus Niger and Botrytis cinerea. Eight bacterial strains were selected for their strong antifungal activity. These are strains M21 M23 M3a M4 M14d and M3c which belong to the family Bacillaceae M12 and M3b which belongs to the family of Pseudomonadaceae. Among these three bacterial strains namely M21 M23 and M12 induce 70 of inhibition of mycelial growth of phytopathogenic fungi Fusarium oxysporum and Aspergillus Niger while the five bacterial strains M3a M3c M3b M4 and M14d have proved to be effective in inhibiting more than 60 of mycelial growth of Botrytis cinerea.

  9. Endophytic Herbaspirillum seropedicae expresses nif genes in gramineous plants.

    Roncato-Maccari, Lauren D B; Ramos, Humberto J O; Pedrosa, Fabio O; Alquini, Yedo; Chubatsu, Leda S; Yates, Marshall G; Rigo, Liu U; Steffens, Maria Berenice R; Souza, Emanuel M

    2003-07-01

    Abstract The interactions between maize, sorghum, wheat and rice plants and Herbaspirillum seropedicae were examined microscopically following inoculation with the H. seropedicae LR15 strain, a Nif(+) (Pnif::gusA) mutant obtained by the insertion of a gusA-kanamycin cassette into the nifH gene of the H. seropedicae wild-type strain. The expression of the Pnif::gusA fusion was followed during the association of the diazotroph with the gramineous species. Histochemical analysis of seedlings of maize, sorghum, wheat and rice grown in vermiculite showed that strain LR15 colonized root surfaces and inner tissues. In early steps of the endophytic association, H. seropedicae colonized root exudation sites, such as axils of secondary roots and intercellular spaces of the root cortex; it then occupied the vascular tissue and there expressed nif genes. The expression of nif genes occurred in roots, stems and leaves as detected by the GUS reporter system. The expression of nif genes was also observed in bacterial colonies located in the external mucilaginous root material, 8 days after inoculation. Moreover, the colonization of plant tissue by H. seropedicae did not depend on the nitrogen-fixing ability, since similar numbers of cells were isolated from roots or shoots of the plants inoculated with Nif(+) or Nif(-) strains.

  10. [Chemical constituents from endophyte Chaetomium globosum in Imperata cylindrical].

    Shen, Li; Zhu, Li; Wei, Zhong-qi; Li, Xiao-wen; Li, Ming; Song, Yong-chun

    2015-12-01

    Isolation and purification of chemical constituents from solid culture of endophyte Chaetomium globosum in Imperata cylindrical was performed through silica gel column chromatography, gel filtration over Sephadex LH-20 and preparative HPLC. Nine compounds were obtained and their structures were determined as chaetoglobosin F(1), chaetoglobosin Fex(2), chaetoglobosin E(3) cytoglobosin A(4), penochalasin C(S), isochaetoglobosin D (6), N-benzoylphenylalaninyl-N-benzoyphenylalaninate(7), uracil(8) and 5-methyluracil(9), respectively, based on HR-MS and NMR data and comparison with literatures. Compound 7 was isolated from Chaeeomium sp. for the first time. In vitro cytotoxicity of compounds was evaluated using MTT mothed and 1,3,4 and 5 showed inhibition activity to the human cervical carcinoma cell HeLa with IC50 values of 99.43, 23.77, 97.92, 86.25 micromol x L(-1), while positive cotolocisnin Ad apno1ch alse IC50 24.33 micromol x L(-1).

  11. The endophyte Verticillium Vt305 protects cauliflower against Verticillium wilt.

    Tyvaert, L; França, S C; Debode, J; Höfte, M

    2014-06-01

    To investigate the interaction between cauliflower and the isolate VerticilliumVt305, obtained from a field suppressive to Verticillium wilt of cauliflower, and to evaluate the ability of VerticilliumVt305 to control Verticillium wilt of cauliflower caused by V. longisporum. Single and combined inoculations of VerticilliumVt305 and V. longisporum were performed on cauliflower seedlings. Symptom development was evaluated, and fungal colonization was measured in the roots, hypocotyl and stem with real-time PCR. No symptoms were observed after single inoculation of VerticilliumVt305, although it colonized the plant tissues. Pre-inoculation of VerticilliumVt305 reduced symptom development and colonization of plant tissues by V. longisporum. VerticilliumVt305 is an endophyte on cauliflower plants and showed effective biological control of V. longisporum in controlled conditions. This work can contribute to the development of a sustainable control measure of V. longisporum in Brassicaceae hosts, which is currently not available. Additionally, this study provides evidence for the different roles of Verticillium species present in the agro-ecosystem. © 2014 The Society for Applied Microbiology.

  12. Endophytic fungi and soil microbial community characteristics over different years of phytoremediation in a copper tailings dam of Shanxi, China.

    Tong, Jia; Miaowen, Cao; Juhui, Jing; Jinxian, Liu; Baofeng, Chai

    2017-01-01

    We conducted a survey of native grass species infected by endophytic fungi in a copper tailings dam over progressive years of phytoremediation. We investigated how endophytic fungi, soil microbial community structure and soil physiochemical properties and enzymatic activity varied in responses to heavy metal pollution over different stages of phytoremediation. endophyte infection frequency increased with years of phytoremediation. Rates of endophyte infection varied among different natural grass species in each sub-dam. Soil carbon content and soil enzymatic activity gradually increased through the years of phytoremediation. endophyte infection rates of Bothriochloa ischaemum and Festuca rubra were positively related to levels of cadmium (Cd) pollution levels, and fungal endophytes associated with Imperata cylindrical and Elymus dahuricus developed tolerance to lead (Pb). The structure and relative abundance of bacterial communities varied little over years of phytoremediation, but there was a pronounced variation in soil fungi types. Leotiomycetes were the dominant class of resident fungi during the initial phytoremediation period, but Pezizomycetes gradually became dominant as the phytoremediation period progressed. Fungal endophytes in native grasses as well as soil fungi and soil bacteria play different ecological roles during phytoremediation processes. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Genotypic variation in the response of chickpea to arbuscular mycorrhizal fungi and non-mycorrhizal fungal endophytes.

    Bazghaleh, Navid; Hamel, Chantal; Gan, Yantai; Tar'an, Bunyamin; Knight, Joan Diane

    2018-04-01

    Plant roots host symbiotic arbuscular mycorrhizal (AM) fungi and other fungal endophytes that can impact plant growth and health. The impact of microbial interactions in roots may depend on the genetic properties of the host plant and its interactions with root-associated fungi. We conducted a controlled condition experiment to investigate the effect of several chickpea (Cicer arietinum L.) genotypes on the efficiency of the symbiosis with AM fungi and non-AM fungal endophytes. Whereas the AM symbiosis increased the biomass of most of the chickpea cultivars, inoculation with non-AM fungal endophytes had a neutral effect. The chickpea cultivars responded differently to co-inoculation with AM fungi and non-AM fungal endophytes. Co-inoculation had additive effects on the biomass of some cultivars (CDC Corrine, CDC Anna, and CDC Cory), but non-AM fungal endophytes reduced the positive effect of AM fungi on Amit and CDC Vanguard. This study demonstrated that the response of plant genotypes to an AM symbiosis can be modified by the simultaneous colonization of the roots by non-AM fungal endophytes. Intraspecific variations in the response of chickpea to AM fungi and non-AM fungal endophytes indicate that the selection of suitable genotypes may improve the ability of crop plants to take advantage of soil ecosystem services.

  14. Fungal endophytes from seeds of invasive, non-native Phragmites australis and their potential role in germination and seedling growth

    Shearin, Zackery R. C.; Filipek, Matthew; Desai, Rushvi; Bickford, Wesley A.; Kowalski, Kurt P.; Clay, Keith

    2018-01-01

    Background and aimsWe characterized fungal endophytes of seeds of invasive, non-native Phragmites from three sites in the Great Lakes region to determine if fungal symbiosis could contribute to invasiveness through their effects on seed germination and seedling growth.MethodsField-collected seeds were surface sterilized and plated on agar to culture endophytes for ITS sequencing. Prevalence of specific endophytes from germinated and non-germinated seeds, and from seedlings, was compared.ResultsOne-third of 740 seeds yielded endophyte isolates. Fifteen taxa were identified with Alternaria sp. representing 54% of all isolates followed by Phoma sp. (21%) and Penicillium corylophilum (12%). Overall germination of seeds producing an isolate (36%) was significantly higher than seeds not producing an isolate (20%). Penicillium in particular was strongly associated with increased germination of seeds from one site. Sixty-three isolates and 11 taxa were also obtained from 30 seedlings where Phoma, Penicillium and Alternaria respectively were most prevalent. There was a significant effect of isolating an endophyte from the seed on seedling growth.ConclusionsThese results suggest that many endophyte taxa are transmitted in seeds and can increase seed germination and seedling growth of invasive Phragmites. The role of fungal endophytes in host establishment, growth and invasiveness in nature requires further research.

  15. Isolation and identification of local bacteria endophyte and screening of its antimicrobial property against pathogenic bacteria and fungi

    Fikri, Ahmad Syairazie Ibrahim; Rahman, Irman Abdul; Nor, Norefrina Shafinaz Md; Hamzah, Ainon

    2018-04-01

    Endophytes are organisms, often fungi and bacteria that live in living plant cells. These organisms reside in the living tissues of the host plant in a variety of relationships, ranging from symbiotic to slightly pathogenic. The endophytes may produce a plethora of substances that have potential to be used in modern medicine, agriculture and industry. The aims of this study are to isolate, identify and screening antimicrobial activity of bacterial endophytes. The endophytes were isolated using nutrient agar, incubated at 37°C for 48 hours. Identification of the isolates were done based on morphological characteristics, biochemical tests and 16S rDNA molecular analysis. Disk diffusion method was used to screen for antimicrobial activity of metabolites from endophytes against pathogenic bacteria. Screening for antifungal activity of selected endophytes was done using dual culture method againts pathogenic fungi followed by Kirby-Bauer method. Results showed endophytes designated as B2c and B7b have positive antimicrobial activity. The metabolites from isolate B2c showed antimicrobial activity against pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus and Staphylococcus epidermis, while isolate B7b have positive activities againts MRSA, S. aureus and Pseudomonas aeruginosa. Isolates B2c displayed antifungal activity against Fusarium oxysporum, Fusarium solani, Phytophthora palmivora and Colletotrichum gloeosporioides. Identification using biochemical tests and 16S rDNA sequences identified isolate B2c as Pseudomonas resinovorans with 97% homology and isolate B7b as Bacillus subtilis with 98% homology.

  16. Fungal endophyte (Epichloë festucae alters the nutrient content of Festuca rubra regardless of water availability.

    Beatriz R Vázquez-de-Aldana

    Full Text Available Festuca rubra plants maintain associations with the vertically transmitted fungal endophyte Epichloë festucae. A high prevalence of infected host plants in semiarid grasslands suggests that this association could be mutualistic. We investigated if the Epichloë-endophyte affects the growth and nutrient content of F. rubra plants subjected to drought. Endophyte-infected (E+ and non-infected (E- plants of two half-sib lines (PEN and RAB were subjected to three water availability treatments. Shoot and root biomass, nutrient content, proline, phenolic compounds and fungal alkaloids were measured after the treatments. The effect of the endophyte on shoot and root biomass and dead leaves depended on the plant line. In the PEN line, E+ plants had a greater S:R ratio than E-, but the opposite occurred in RAB. In both plant lines and all water treatments, endophyte-infected plants had greater concentrations of N, P and Zn in shoots and Ca, Mg and Zn in roots than E- plants. On average, E+ plants contained in their shoots more P (62%, Zn (58% and N (19% than E- plants. While the proline in shoots increased in response to water stress, the endophyte did not affect this response. A multivariate analysis showed that endophyte status and plant line impose stronger differences in the performance of the plants than the water stress treatments. Furthermore, differences between PEN and RAB lines seemed to be greater in E- than in E+ plants, suggesting that E+ plants of both lines are more similar than those of their non-infected version. This is probably due to the endophyte producing a similar effect in both plant lines, such as the increase in N, P and Zn in shoots. The remarkable effect of the endophyte in the nutrient balance of the plants could help to explain the high prevalence of infected plants in natural grasslands.

  17. Fungal endophytes of turmeric (Curcuma longa L.) and their biocontrol potential against pathogens Pythium aphanidermatum and Rhizoctonia solani.

    Vinayarani, G; Prakash, H S

    2018-03-14

    Endophytic fungi have been isolated from the healthy turmeric (Curcuma longa L.) rhizomes from South India. Thirty-one endophytes were identified based on morphological and ITS-rDNA sequence analysis. The isolated endophytes were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric respectively. Results revealed that only six endophytes showed > 70% suppression of test pathogens in antagonistic dual culture assays. The endophyte T. harzianum TharDOB-31 showed significant in vitro mycelial growth inhibition of P. aphanidermatum (76.0%) and R. solani (76.9%) when tested by dual culture method. The SEM studies of interaction zone showed morphological abnormalities like parasitism, shriveling, breakage and lysis of hyphae of the pathogens by endophyte TharDOB-31. Selected endophytic isolates recorded multiple plant growth promoting traits in in vitro studies. The rhizome bacterization followed by soil application of endophyte TharDOB-31 showed lowest Percent Disease Incidence of rhizome rot and leaf blight, 13.8 and 11.6% respectively. The treatment of TharDOB-31 exhibited significant increase in plant height (85 cm) and fresh rhizome yield/plant (425 g) in comparison with untreated control under greenhouse condition. The confocal microscopy validates the colonization of the TharDOB-31 in turmeric rhizomes. The secondary metabolites in ethyl acetate extract of TharDOB-31 were found to contain higher number of antifungal compounds by high resolution liquid chromatograph mass spectrometer analysis. Thereby, endophyte T. harzianum isolate can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

  18. Antibacterial, antifungal and cytotoxic activities exhibited by endophytic fungi from the Brazilian marine red alga Bostrychia tenella (Ceramiales

    Rafael de Felício

    Full Text Available Abstract Marine environment is one of the most important sources regarding natural products research. Besides, marine microorganisms have been denominated as a talented natural source for discovery of new leads. Although the association of macroalgae and fungi has been described regarding ecological issues, there is a lack of studies about marine seaweed endophytic fungi. In this context, the goal of this study was to evaluate cytotoxic, antifungal and antibacterial activities of endophytic fungi isolated from the Brazilian marine seaweed Bostrychia tenella (J.V. Lamouroux J. Agardh (Ceramiales, Rhodophyta. Forty-five endophytic microorganism strains were isolated from B. tenella. Crude extracts and organic fractions of ten selected strains were obtained after growth in rice medium. Samples were evaluated for cytotoxicity, antifungal and antibacterial assays. Penicillium strains showed positive results in a diversity of assays, and other five strains were active in at least one test. In addition, cytochalasin D was isolated from Xylaria sp. This alga is composed of a microbiological potential, since its endophytic strains exhibited remarkable biological properties. Moreover, cytochalasin D isolation has confirmed chemical potential of marine endophytic strains. This is the first study in which cultured fungi isolates from the Brazilian macroalga B. tenella were evaluated concerning biological properties. Results corroborated that this species could be a pharmaceutical source from marine environment. Furthermore, Acremonium implicatum is being firstly described as marine endophyte and Xylaria sp., Trichoderma atroviride and Nigrospora oryzae as marine seaweed endophytes. Thus, this work reports the first study relating detailed isolation, cultivation and biological evaluation (cytotoxic, antifungal and antibacterial of endophytes Penicillium decaturense and P. waksmanii from the Brazilian marine red alga B. tenella. We are also reporting the

  19. Identification of a New Uncompetitive Inhibitor of Adenosine Deaminase from Endophyte Aspergillus niger sp.

    Zhang, Xin-Guo; Liu, Jin-Wen; Tang, Peng; Liu, Zi-Yu; Guo, Guang-Jun; Sun, Qiao-Yun; Yin, Jian-Jun

    2018-05-01

    Adenosine deaminase (ADA) is an enzyme widely distributed from bacteria to humans. ADA is known as a potential therapeutic target for the treatment of lymphoproliferative disorders and cancer. Endophytes are endosymbionts, often bacteria or fungi, which live within plant tissues and internal organs or intercellular space. Endophytes have a broad variety of bioactive metabolites that are used for the identification of novel natural compounds. Here, 54 morphologically distinct endophyte strains were isolated from six plants such as Peganum harmala Linn., Rheum officinale Baill., Gentiana macrophylla Pall., Radix stephaniae tetrandrae, Myrrha, and Equisetum hyemale Linn. The isolated strains were used for the search of ADA inhibitors that resulted in the identification of the strain with the highest inhibition activity, Aspergillus niger sp. Four compounds were isolated from this strain using three-step chromatography procedure, and compound 2 was determined as the compound with the highest inhibition activity of ADA. Based on the results of 1 H and 13 C NMR spectroscopies, compound 2 was identified as 3-(4-nitrophenyl)-5-phenyl isoxazole. We showed that compound 2 was a new uncompetitive inhibitor of ADA with high cytotoxic effect on HepG2 and SMCC-7721 cells (the IC 50 values were 0.347 and 0.380 mM, respectively). These results suggest that endophyte strains serve as promising sources for the identification of ADA inhibitors, and compound 2 could be an effective drug in the cancer treatment.

  20. Chemical communication between the endophytic fungus Paraconiothyrium variabile and the phytopathogen Fusarium oxysporum.

    Audrey Combès

    Full Text Available Paraconiothyrium variabile, one of the specific endophytic fungi isolated from the host plant Cephalotaxus harringtonia, possesses the faculty to inhibit the growth of common phytopathogens, thus suggesting a role in its host protection. A strong antagonism between the endophyte P. variabile and Fusarium oxysporum was observed and studied using optic and electronic microscopies. A disorganization of the mycelium of F. oxysporum was thus noticed. Interestingly, the biological effect of the main secondary metabolites isolated from P. variabile against F. oxysporum did not account for this strong antagonism. However, a metabolomic approach of pure fungal strains and confrontation zones using the data analysis tool XCMS were analyzed and pointed out a competition-induced metabolite production by the endophyte in the presence of the phytopathogen. Subsequent MS/MS fragmentations permitted to identify one of the induced metabolites as 13-oxo-9,11-octadecadienoic acid and highlighted a negative modulation of the biosynthesis of beauvericin, one of the most potent mycotoxin of F. oxysporum, during the competition with the endophyte.

  1. Endophytic fungi from medicinal plant Bauhinia forficata: Diversity and biotechnological potential.

    Bezerra, Jadson D P; Nascimento, Carlos C F; Barbosa, Renan do N; da Silva, Dianny C V; Svedese, Virgínia M; Silva-Nogueira, Eliane B; Gomes, Bruno S; Paiva, Laura M; Souza-Motta, Cristina M

    2015-03-01

    Bauhinia forficata is native to South America and used with relative success in the folk medicine in Brazil. The diversity, antibacterial activity, and extracellular hydrolytic enzymes of endophytic fungi associated with this plant were studied. Plant samples, which included leaves, sepals, stems, and seeds, were used. Ninety-five endophytic fungal were isolated (18 from leaves, 22 from sepals, 46 from stems, and nine from seeds), comprising 28 species. The most frequently isolated species were Acremonium curvulum (9.5%), Aspergillus ochraceus (7.37%), Gibberella fujikuroi (10.53%), Myrothecium verrucaria (10.53%) and Trichoderma piluliferum (7.37%). Diversity and species richness were higher in stem tissues, and Sorensen's index of similarity between the tissues was low. Eleven fungi showed antibacterial activity. Aspergillus ochraceus , Gibberella baccata , Penicillium commune , and P. glabrum were those with the greatest antibacterial activity against Staphylococcus aureus and/or Streptococcus pyogenes . Thirteen species showed proteolytic activity, particularly Phoma putaminum . Fourteen species were cellulase positive, particularly the Penicillium species and Myrmecridium schulzeri . All isolates tested were xylanase positive and 10 showed lipolytic activity, especially Penicillium glabrum . It is clear that the endophytic fungi from B. forficata have potential for the production of bioactive compounds and may be a source of new therapeutic agents for the effective treatment of diseases in humans, other animals, and plants. To our knowledge, this is the first study of endophytic fungi from different tissues of B. forficata and their biotechnological potential.

  2. Fungal endophytes which invade insect galls: insect pathogens, benign saprophytes, or fungal inquilines?

    Wilson, Dennis

    1995-08-01

    Fungi are frequently found within insect galls. However, the origin of these fungi, whether they are acting as pathogens, saprophytes invading already dead galls, or fungal inquilines which invade the gall but kill the gall maker by indirect means, is rarely investigated. A pathogenic role for these fungi is usually inferred but never tested. I chose the following leaf-galling-insect/host-plant pairs (1) a cynipid which forms two-chambered galls on the veins of Oregon white oak, (2) a cynipid which forms single-chambered galls on California coast live oak, and (3) an aphid which forms galls on narrowleaf cottonwood leaves. All pairs were reported to have fungi associated with dead insects inside the gall. These fungi were cultured and identified. For the two cynipids, all fungi found inside the galls were also present in the leaves as fungal endophytes. The cottonwood leaves examined did not harbor fungal endophytes. For the cynipid on Oregon white oak, the fungal endophyte grows from the leaf into the gall and infects all gall tissue but does not directly kill the gall maker. The insect dies as a result of the gall tissue dying from fungal infection. Therefore, the fungus acts as an inquiline. Approximately 12.5% of these galls die as a result of invasion by the fungal endophyte.

  3. Characterization of Antifungal Natural Products Isolated from Endophytic Fungi of Finger Millet (Eleusine coracana).

    Mousa, Walaa Kamel; Schwan, Adrian L; Raizada, Manish N

    2016-09-03

    Finger millet is an ancient African-Indian crop that is resistant to many pathogens including the fungus, Fusarium graminearum. We previously reported the first isolation of putative fungal endophytes from finger millet and showed that the crude extracts of four strains had anti-Fusarium activity. However, active compounds were isolated from only one strain. The objectives of this study were to confirm the endophytic lifestyle of the three remaining anti-Fusarium isolates, to identify the major underlying antifungal compounds, and to initially characterize the mode(s) of action of each compound. Results of confocal microscopy and a plant disease assay were consistent with the three fungal strains behaving as endophytes. Using bio-assay guided fractionation and spectroscopic structural elucidation, three anti-Fusarium secondary metabolites were purified and characterized. These molecules were not previously reported to derive from fungi nor have antifungal activity. The purified antifungal compounds were: 5-hydroxy 2(3H)-benzofuranone, dehydrocostus lactone (guaianolide sesquiterpene lactone), and harpagoside (an iridoide glycoside). Light microscopy and vitality staining were used to visualize the in vitro interactions between each compound and Fusarium; the results suggested a mixed fungicidal/fungistatic mode of action. We conclude that finger millet possesses fungal endophytes that can synthesize anti-fungal compounds not previously reported as bio-fungicides against F. graminearum.

  4. Characterization of Antifungal Natural Products Isolated from Endophytic Fungi of Finger Millet (Eleusine coracana

    Walaa Kamel Mousa

    2016-09-01

    Full Text Available Finger millet is an ancient African-Indian crop that is resistant to many pathogens including the fungus, Fusarium graminearum. We previously reported the first isolation of putative fungal endophytes from finger millet and showed that the crude extracts of four strains had anti-Fusarium activity. However, active compounds were isolated from only one strain. The objectives of this study were to confirm the endophytic lifestyle of the three remaining anti-Fusarium isolates, to identify the major underlying antifungal compounds, and to initially characterize the mode(s of action of each compound. Results of confocal microscopy and a plant disease assay were consistent with the three fungal strains behaving as endophytes. Using bio-assay guided fractionation and spectroscopic structural elucidation, three anti-Fusarium secondary metabolites were purified and characterized. These molecules were not previously reported to derive from fungi nor have antifungal activity. The purified antifungal compounds were: 5-hydroxy 2(3H-benzofuranone, dehydrocostus lactone (guaianolide sesquiterpene lactone, and harpagoside (an iridoide glycoside. Light microscopy and vitality staining were used to visualize the in vitro interactions between each compound and Fusarium; the results suggested a mixed fungicidal/fungistatic mode of action. We conclude that finger millet possesses fungal endophytes that can synthesize anti-fungal compounds not previously reported as bio-fungicides against F. graminearum.

  5. [Isolation of endophytic fungi from medicinal plant Brucea javanica and their microbial inhibition activity].

    Liang, Zi-Ning; Zhu, Hua; Lai, Kai-Ping; Chen, Long

    2014-04-01

    To isolate and identify endophytic fungi from Brucea javanica, and to detect the antimicrobial activity of these strains. Endophytic fungi were isolated by tissue inoculation culture and identified by conventional morphological characteristic method. Seven kinds of pathogenic fungi and three kinds of bacteria were used as targeting microbes to test microbial inhibition activities by agar plate antagonistic action and modified agar gel diffusion methods, respectively. A total of 83 endophytic fungi strains were isolated from the root, stem, leaf and fruit of Brucea javanica. 34 strains were obtained from the stem, 32 strains were obtained from the leaf, 15 strains were isolated from the root and 2 strains came from the fruit. These 73 strains which had been identified attribute to 5 orders, 6 families and 12 genera. For the isolated strains, 14 strains had antifungal activities against at least one pathogenic fungi, 9 strains showed antibacterial activities against one or more bacteria. Especially, the strain YJ-17 which belonged to Phomopsis genus showed the best inhibitory effect on the targeting microbes. The endophytic fungi from Brucea javanica show diversity and microbial inhibition activity, and are worthy for further study on plant disease controlling.

  6. [Screening and identification of endophytic fungi with growth promoting effect on Dendrobium officinale].

    Hou, Xiao-qiang; Guo, Shun-xing

    2014-09-01

    The endophytic fungi with plant growth promoting effects were screened by co-culture of each endophytic fungus and seedlings of Dendrobium officinale. Anatomical features of the inoculated roots were studied by paraffin sectioning. Morphological characteristics and rDNA ITS1-5. 8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. The results showed that 8 strains inoculated to D. officinale seedlings greatly enhanced plant height, stem diameter, new roots number and biomass. According to the anatomical features of the inoculated roots, each fungus could infect the velamina of seedlings. The hyphae or pelotons were existed in the exodermis passage cells and cortex cells. The effective fungi could not infect the endodermis and vascular bundle sheath, but which was exception for other fungi with harmful to seedlings. Combined with classic morphologic classification, 2 effective strains were identified which were subjected to Pestalotiopsis and Eurotium. Six species of fungi without conidiophore belonged to Pyrenochaeta, Coprinellus, Pholiota, Alternaria, Helotiales, which were identified by sequencing the PCR-amplified rDNA ITS1-5. 8S-ITS2 regions. The co-culture technology of effective endophytic fungi and plant can apply to cultivate the seedlings of D. officinale. It is feasible to shorten growth cycle of D. officinale and increase the resource of Chinese herbs.

  7. Seed Endophyte Microbiome of Crotalaria pumila Unpeeled: Identification of Plant-Beneficial Methylobacteria.

    Sánchez-López, Ariadna S; Pintelon, Isabel; Stevens, Vincent; Imperato, Valeria; Timmermans, Jean-Pierre; González-Chávez, Carmen; Carrillo-González, Rogelio; Van Hamme, Jonathan; Vangronsveld, Jaco; Thijs, Sofie

    2018-01-19

    Metal contaminated soils are increasing worldwide. Metal-tolerant plants growing on metalliferous soils are fascinating genetic and microbial resources. Seeds can vertically transmit endophytic microorganisms that can assist next generations to cope with environmental stresses, through yet poorly understood mechanisms. The aims of this study were to identify the core seed endophyte microbiome of the pioneer metallophyte Crotalaria pumila throughout three generations, and to better understand the plant colonisation of the seed endophyte Methylobacterium sp. Cp3. Strain Cp3 was detected in C. pumila seeds across three successive generations and showed the most dominant community member. When inoculated in the soil at the time of flowering, strain Cp3 migrated from soil to seeds. Using confocal microscopy, Cp3-mCherry was demonstrated to colonise the root cortex cells and xylem vessels of the stem under metal stress. Moreover, strain Cp3 showed genetic and in planta potential to promote seed germination and seedling development. We revealed, for the first time, that the seed microbiome of a pioneer plant growing in its natural environment, and the colonisation behaviour of an important plant growth promoting systemic seed endophyte. Future characterization of seed microbiota will lead to a better understanding of their functional contribution and the potential use for seed-fortification applications.

  8. Evaluating a novel endophytic grass for suppressing invertebrates that contribute to bird strike risk at airports

    BACKGROUND: Tall fescue containing a selected strain of the fungal endophyte Epichlöe coenophiala purported to express high levels of bioactive alkaloids (Avanex®) was recently commercialized for reducing airport bird strike hazard. We compared bioactivity of Avanex and KY 31, a ubiquitous cultivar...

  9. Antimicrobial activity of saponins produced by two novel endophytic fungi from Panax notoginseng.

    Jin, Zhaoxia; Gao, Lin; Zhang, Lin; Liu, Tianyi; Yu, Fang; Zhang, Zongshen; Guo, Qiong; Wang, Biying

    2017-11-01

    Endophytes in plants may be co-producer of the bioactive compounds of their hosts. We conducted a study to bioprospect for saponin-producing endophytic fungi from Panax notoginseng and evaluate the antimicrobial activity of saponins. Two novel fungal endophytes, Fusarium sp. PN8 and Aspergillus sp. PN17, were isolated from traditional Chinese medicinal herb P. notoginseng. After eight days of fermentation, the total saponins produced in the culture broth of PN8 and PN17 were 1.061 and 0.583 mg mL -1 , respectively. The saponin extracts exhibited moderate to high (inhibition zone diameter 15.7-28.4 mm, MIC 1.6-12.5 mg mL -1 ) antimicrobial activity against pathogens tested. Further analysis showed that triterpenoid saponins produced by Fusarium PN8 were Rb 1 , Rd and 20(S)-Rg 3 , while Aspergillus PN17 had the ability to synthesise ginsenoside Re, Rd and 20(S)-Rg 3 . The isolated endophytes may be used as potential sources for microbial production of plant secondary metabolites and for antimicrobial agents.

  10. Metabolism of carbamazepine in plant roots and endophytic rhizobacteria isolated from Phragmites australis.

    Sauvêtre, Andrés; May, Robert; Harpaintner, Rudolf; Poschenrieder, Charlotte; Schröder, Peter

    2018-01-15

    Carbamazepine (CBZ) is a pharmaceutical frequently categorized as a recalcitrant pollutant in the aquatic environment. Endophytic bacteria previously isolated from reed plants have shown the ability to promote growth of their host and to contribute to CBZ metabolism. In this work, a horseradish (Armoracia rusticana) hairy root (HR) culture has been used as a plant model to study the interactions between roots and endophytic bacteria in response to CBZ exposure. HRs could remove up to 5% of the initial CBZ concentration when they were grown in spiked Murashige and Skoog (MS) medium. Higher removal rates were observed when HRs were inoculated with the endophytic bacteria Rhizobium radiobacter (21%) and Diaphorobacter nitroreducens (10%). Transformation products resulting from CBZ degradation were identified using liquid chromatography-ultra high-resolution quadrupole time of flight mass spectrometry (LC-UHR-QTOF-MS). CBZ metabolism could be divided in four pathways. Metabolites involving GSH conjugation and 2,3-dihydroxylation, as well as acridine related compounds are described in plants for the first time. This study presents strong evidence that xenobiotic metabolism and degradation pathways in plants can be modulated by the interaction with their endophytic community. Hence it points to plausible applications for the elimination of recalcitrant compounds such as CBZ from wastewater in CWs. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Activity of Scottish plant, lichen and fungal endophyte extracts against Mycobacterium aurum and Mycobacterium tuberculosis.

    Gordien, Andréa Y; Gray, Alexander I; Ingleby, Kevin; Franzblau, Scott G; Seidel, Véronique

    2010-05-01

    With tuberculosis the leading bacterial killer worldwide and other mycobacterial diseases on the increase, the search for new antimycobacterial agents is timely. In this study, extracts from plants, lichens and fungal endophytes of Scottish provenance were screened for activity against Mycobacterium aurum and M. tuberculosis H(37)Rv. The best activity against M. aurum was observed for extracts of Juniperus communis roots and Cladonia arbuscula (MIC = 4 microg/mL), and a fungal endophyte isolated from Vaccinium myrtillus (MIC = 8 microg/mL). The best activity against M. tuberculosis was observed for extracts of C. arbuscula, Empetrum nigrum, J. communis roots, Calluna vulgaris aerial parts, Myrica gale roots and stems (93 to 99% inhibition at 100 microg/mL). Potent antitubercular activity (90 to 96% inhibition at 100 microg/mL) was also observed for the ethanol extracts of Xerocomus badius, Chalciporus piperatus, Suillus luteus and of endophytes isolated from C. vulgaris, E. nigrum, Vaccinium vitis-idaea and V. myrtillus. The results obtained this study provide, in part, some scientific basis for the traditional use of some of the selected plants in the treatment of tuberculosis. They also indicate that fungal endophytes recovered from Scottish plants are a source of antimycobacterial agents worthy of further investigation. Copyright (c) 2009 John Wiley & Sons, Ltd.

  12. The place of roentgen computesized tomography in the diagnosis of endophytic carcinoma of the stomach

    Gorshkov, A.N.; Akberov, R.F.

    1996-01-01

    The possibilities of roentgen computer tomography in the diagnosis of infiltrative carcinoma of the stomach, especially in the determination of stomach wall enlargement, process occurrence, are estimated positively using the investigations of 45 patients with endophytic carcinoma of the stomach. It should be used after usual roentgen and endoscopic methods. Refs. 5, figs. 5

  13. Monitoring endophyte populations in pine plantations and native oak forests in Northern Spain

    Martinez-Alvarez, P.; Martin-Garcia, J.; Rodriguez-Ceinos, S.; Diez, J. J.

    2012-07-01

    The replacement of native forest with plantations of other species may have important impacts on ecosystems. Some of these impacts have been widely studied, but very little is known about the effects on fungal communities and specifically endo phytic fungi. In this study, endophyte assemblages in pine plantations (Pinus sylvestris, P. nigra and P. pinaster) and native oak forests (Quercus pyrenaica) in the north of the province of Palencia (Spain) were analyzed. For this purpose, samples of needles/leaves and twigs were collected from three trees in each of three plots sampled per host species. The samples were later processed in the laboratory to identify all of the endo phytic species present. In addition, an exhaustive survey was carried out of the twelve sites to collect data on the environmental, crown condition, dendrometric and soil variables that may affect the distribution of the fungi. The endophyte assemblages isolated from P. sylvestris and P. nigra were closely related to each other, but were different from those isolated from P. pinaster. The endophytes isolated from Q. pyrenaica were less closely related to those from the other hosts, and therefore preservation of oak stands is important to prevent the loss of fungal diversity. Finally, the distribution of the endophyte communities was related to some of the environmental variables considered. (Author) 42 refs.

  14. Adenosine deaminase production by an endophytic bacterium (Lysinibacillus sp.) from Avicennia marina.

    Kathiresan, Kandasamy; Saravanakumar, Kandasamy; Sahu, Sunil Kumar; Sivasankaran, Muthu

    2014-06-01

    The present study was carried out with the following objectives: (1) to isolate the endophytic bacilli strains from the leaves of mangrove plant Avicennia marina, (2) to screen the potential strains for the production of adenosine deaminase, (3) to statistically optimize the factors that influence the enzyme activity in the potent strain, and (4) to identify the potent strain using 16S rRNA sequence and construct its phylogenetic tree. The bacterial strains isolated from the fresh leaves of a mangrove A. marina were assessed for adenosine deaminase activity by plating method. Optimization of reaction process was carried out using response surface methodology of central composite design. The potent strain was identified based on 16S rRNA sequencing and phylogeny. Of five endophytic strains, EMLK1 showed a significant deaminase activity over other four strains. The conditions for maximum activity of the isolated adenosine deaminase are described. The potent strain EMLK1 was identified as Lysinibacillus sp. (JQ710723) being the first report as a mangrove endophyte. Mangrove-derived endophytic bacillus strain Lysinibacillus sp. EMLK1 is proved to be a promising source for the production of adenosine deaminase and this enzyme deserves further studies for purification and its application in disease diagnosis.

  15. The relationship between an endangered North American tree and an endophytic fungus.

    Lee, J C; Yang, X; Schwartz, M; Strobel, G; Clardy, J

    1995-11-01

    The Florida torreya (Torreya taxifolia) began a catastrophic decline in the late 1950s and is now the rarest tree in North America for which a full species designation has been established. The trees have common plant disease symptoms, but the reason for the decline has never been identified. T. taxifolia's imminent extinction gains special poignancy through its close relationship to the Pacific yew (Taxus brevifolia), which produces the potent anticancer agent, taxol. An examination of the endophytic fungal communities of wild torreyas consistently found a filamentous fungus, Pestalotiopsis microspora, associated with diseased trees and also with most symptomless trees. P. microspora can be cultured in the laboratory, and when it is introduced into greenhouse-grown torreyas, it causes disease symptoms similar to those seen in the field. The fungus can then be reisolated from these deliberately infected trees. The phytotoxins pestalopyrone, hydroxypestalopyrone and pestaloside have been isolated and characterized from axenic fungal cultures, and both pestalopyrone and hydroxypestalopyrone can be isolated from artificially infected torreyas. In addition, pestaloside has antifungal activity against other fungal endophytes of T. taxifolia. The filamentous fungus, P. microspora, has an endophytic-pathologic relationship with T. taxifolia. The fungus resides in the inner bark of symptomless trees, and physiological or environmental factors could trigger its pathological activity. P. microspora produces the phytotoxins pestalopyrone, hydroxypestalopyrone, and pestaloside which give rise to the disease. Pestaloside, which also has antifungal activity, could reduce competition from other fungal endophytes within the host.

  16. Fluorescence in situ hybridization for phytoplasma and endophytic bacteria localization in plant tissues.

    Bulgari, Daniela; Casati, Paola; Faoro, Franco

    2011-11-01

    In the present study, we developed a rapid and efficient fluorescence in situ hybridization assay (FISH) in non-embedded tissues of the model plant Catharanthus roseus for co-localizing phytoplasmas and endophytic bacteria, opening new perspectives for studying the interaction between these microorganisms. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Bacteria in oral secretions of an endophytic insect inhibit antagonistic fungi

    Yasmin J. Cardoza; Kier D. Klepzig; Kenneth F. Raffa

    2006-01-01

    1. Colonisation of host trees by an endophytic herbivore, the spruce beetle, Dendroctonus rufipennis , is accompanied by invasion of its galleries by a number of fungal species. Four of these associated species were identified as Leptographium abietinum , Aspergillus fumigatus , Aspergillus nomius , and ...

  18. Highly Diverse Endophytic and Soil Fusarium oxysporum Populations Associated with Field-Grown Tomato Plants

    Demers, Jill E.; Gugino, Beth K.

    2014-01-01

    The diversity and genetic differentiation of populations of Fusarium oxysporum associated with tomato fields, both endophytes obtained from tomato plants and isolates obtained from soil surrounding the sampled plants, were investigated. A total of 609 isolates of F. oxysporum were obtained, 295 isolates from a total of 32 asymptomatic tomato plants in two fields and 314 isolates from eight soil cores sampled from the area surrounding the plants. Included in this total were 112 isolates from the stems of all 32 plants, a niche that has not been previously included in F. oxysporum population genetics studies. Isolates were characterized using the DNA sequence of the translation elongation factor 1α gene. A diverse population of 26 sequence types was found, although two sequence types represented nearly two-thirds of the isolates studied. The sequence types were placed in different phylogenetic clades within F. oxysporum, and endophytic isolates were not monophyletic. Multiple sequence types were found in all plants, with an average of 4.2 per plant. The population compositions differed between the two fields but not between soil samples within each field. A certain degree of differentiation was observed between populations associated with different tomato cultivars, suggesting that the host genotype may affect the composition of plant-associated F. oxysporum populations. No clear patterns of genetic differentiation were observed between endophyte populations and soil populations, suggesting a lack of specialization of endophytic isolates. PMID:25304514

  19. Screenhouse and field persistence of nonpathogenic endophytic Fusarium oxysporum in Musa tissue culture plants.

    Paparu, Pamela; Dubois, Thomas; Gold, Clifford S; Niere, Björn; Adipala, Ekwamu; Coyne, Daniel

    2008-04-01

    Two major biotic constraints to highland cooking banana (Musa spp., genome group AAA-EA) production in Uganda are the banana weevil Cosmopolites sordidus and the burrowing nematode Radopholus similis. Endophytic Fusarium oxysporum strains inoculated into tissue culture banana plantlets have shown control of the banana weevil and the nematode. We conducted screenhouse and field experiments to investigate persistence in the roots and rhizome of two endophytic Fusarium oxysporum strains, V2w2 and III4w1, inoculated into tissue-culture banana plantlets of highland cooking banana cultivars Kibuzi and Nabusa. Re-isolation of F. oxysporum showed that endophyte colonization decreased faster from the rhizomes than from the roots of inoculated plants, both in the screenhouse and in the field. Whereas rhizome colonization by F. oxysporum decreased in the screenhouse (4-16 weeks after inoculation), root colonization did not. However, in the field (17-33 weeks after inoculation), a decrease was observed in both rhizome and root colonization. The results show a better persistence in the roots than rhizomes of endophytic F. oxysporum strains V2w2 and III4w1.

  20. Endophytic l-asparaginase-producing fungi from plants associated with anticancer properties

    YiingYng Chow

    2015-11-01

    Full Text Available Endophytes are novel sources of natural bioactive compounds. This study seeks endophytes that produce the anticancer enzyme l-asparaginase, to harness their potential for mass production. Four plants with anticancer properties; Cymbopogon citratus, Murraya koenigii, Oldenlandia diffusa and Pereskia bleo, were selected as host plants. l-Asparaginase-producing endophytes were detected by the formation of pink zones on agar, a result of hydrolyzes of asparagine into aspartic acid and ammonia that converts the phenol red dye indicator from yellow (acidic condition to pink (alkaline condition. The anticancer enzyme asparaginase was further quantified via Nesslerization. Results revealed that a total of 89 morphotypes were isolated; mostly from P. bleo (40, followed by O. diffusa (25, C. citratus (14 and M. koenigii (10. Only 25 of these morphotypes produced l-asparaginase, mostly from P. bleo and their asparaginase activities were between 0.0069 and 0.025 μM mL−1 min−1. l-Asparaginase producing isolates were identified as probable species of the genus Colletotrichum, Fusarium, Phoma and Penicillium. Studies here revealed that endophytes are good alternative sources for l-asparaginase production and they can be sourced from anticancer plants, particularly P. bleo.

  1. Identification of New Lactone Derivatives Isolated from Trichoderma sp., An Endophytic Fungus of Brotowali (Tinaspora crispa

    ELFITA

    2014-03-01

    Full Text Available Endophytic fungi is a rich source of novel organic compounds with interesting biological activities and a high level of structural diversity. As a part of our systematic search for new bioactive lead structures and specific profiles from endophytic fungi, an endophytic fungus was isolated from roots of brotowali (Tinaspora crispa, an important medicinal plant. Colonial morphological trait and microscopic observation revealed that the endophytic fungus was Trichoderma sp. The pure fungal strain was cultivated on 7 L Potatos Dextose Broth (PDB medium under room temperature (no shaking for 8 weeks. The ethyl acetate were added to cultur medium and left overnight to stop cell growth. The culture filtrates were collected and extracted with EtOAc and then taken to evaporation. Two new lactone derivatives, 5-hydroxy-4-hydroxymethyl-2H-pyran-2-one (1 and (5-hydroxy-2-oxo-2H pyran-4-yl methyl acetate (2 were obtained from the EtOAc extracts of Trichoderma sp. Their structures were determined on the basic of spectroscopic methods including UV, IR, 1H-NMR, 13C-NMR, HMQC, and HMBC.

  2. New endophytic Toxicocladosporium species from cacti in Brazil, and description of Neocladosporium gen. nov.

    Bezerra, Jadson D.P.; Sandoval-Denis, Marcelo; Paiva, Laura M.; Silva, Gladstone A.; Groenewald, Johannes Z.; Souza-Motta, Cristina M.; Crous, Pedro W.

    2017-01-01

    Brazil harbours a unique ecosystem, the Caatinga, which belongs to the tropical dry forest biome. This region has an important diversity of organisms, and recently several new fungal species have been described from different hosts and substrates within it. During a survey of fungal endophyte

  3. New Eudesmane-Type Sesquiterpenoids from the Mangrove-Derived Endophytic Fungus Penicillium sp. J-54

    Liuming Qiu; Pei Wang; Ge Liao; Yanbo Zeng; Caihong Cai; Fandong Kong; Zhikai Guo; Peter Proksch; Haofu Dai; Wenli Mei

    2018-01-01

    Four new eudesmane-type sesquiterpenoids, penicieudesmol A–D (1–4), were isolated from the fermentation broth of the mangrove-derived endophytic fungus Penicillium sp. J-54. Their structures were determined by spectroscopic methods, the in situ dimolybdenum CD method, and modified Mosher’s method. The bioassays results showed that 2 exhibited weak cytotoxicity against K-562 cells.

  4. New Eudesmane-Type Sesquiterpenoids from the Mangrove-Derived Endophytic Fungus Penicillium sp. J-54.

    Qiu, Liuming; Wang, Pei; Liao, Ge; Zeng, Yanbo; Cai, Caihong; Kong, Fandong; Guo, Zhikai; Proksch, Peter; Dai, Haofu; Mei, Wenli

    2018-03-28

    Four new eudesmane-type sesquiterpenoids, penicieudesmol A-D ( 1 - 4 ), were isolated from the fermentation broth of the mangrove-derived endophytic fungus Penicillium sp. J-54. Their structures were determined by spectroscopic methods, the in situ dimolybdenum CD method, and modified Mosher's method. The bioassays results showed that 2 exhibited weak cytotoxicity against K-562 cells.

  5. Whole-Genome Sequence and Classification of 11 Endophytic Bacteria from Poison Ivy (Toxicodendron radicans).

    Tran, Phuong N; Tan, Nicholas E H; Lee, Yin Peng; Gan, Han Ming; Polter, Steven J; Dailey, Lucas K; Hudson, André O; Savka, Michael A

    2015-11-19

    Here, we report the whole-genome sequences and annotation of 11 endophytic bacteria from poison ivy (Toxicodendron radicans) vine tissue. Five bacteria belong to the genus Pseudomonas, and six single members from other genera were found present in interior vine tissue of poison ivy. Copyright © 2015 Tran et al.

  6. Whole-Genome Sequence and Classification of 11 Endophytic Bacteria from Poison Ivy (Toxicodendron radicans)

    Tran, Phuong N.; Tan, Nicholas E. H.; Lee, Yin Peng; Gan, Han Ming; Polter, Steven J.; Dailey, Lucas K.; Hudson, Andr? O.; Savka, Michael A.

    2015-01-01

    Here, we report the whole-genome sequences and annotation of 11 endophytic bacteria from poison ivy (Toxicodendron radicans) vine tissue. Five bacteria belong to the genus Pseudomonas, and six single members from other genera were found present in interior vine tissue of poison ivy.

  7. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell (Scrophulariaceae).

    Katoch, Meenu; Singh, Gurpreet; Sharma, Sadhna; Gupta, Nidhi; Sangwan, Payare Lal; Saxena, Ajit Kumar

    2014-02-11

    Endophytes, which reside in plant tissues, have the potential to produce novel metabolites with immense benefits for health industry. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell were investigated. Endophytic fungi were isolated from the Bacopa monnieri. Extracts from liquid cultures were tested for cytotoxicity against a number of cancer cell lines using the MTT assay. Antimicrobial activity was determined using the micro dilution method. 22% of the examined extracts showed potent (IC50 of <20 μg/ml) cytotoxic activity against HCT-116 cell line. 5.5%, 11%, 11% of the extracts were found to be cytotoxic for MCF-7, PC-3, and A-549 cell lines respectively. 33% extracts displayed antimicrobial activity against at least one test organism with MIC value 10-100 μg/ml. The isolate B9_Pink showed the most potent cytotoxic activity for all the cell lines examined and maximum antimicrobial activity against the four pathogens examined which was followed by B19. Results indicated the potential for production of bioactive agents from endophytes of Bacopa monnieri.

  8. The contribution of endophytic bacteria to Albizia lebbeck-mediated phytoremediation of tannery effluent contaminated soil.

    Manikandan, Muthu; Kannan, Vijayaraghavan; Mendoza, Ordetta Hannah; Kanimozhi, Mahalingam; Chun, Sechul; Pašić, Lejla

    2016-01-01

    Toxicity of chromium often impairs the remediation capacity of plants used in phytoremediation of polluted soils. In this study, we have identified Albizia lebbeck as a prospective chromium hyperaccumulator and examined cultivable diversity of endophytes present in chromium-treated and control saplings. High numbers (22-100%) of endophytic bacteria, isolated from root, stem, and leaf tissues, could tolerate elevated (1-3 mM) concentrations of K2CrO7. 16S rRNA gene sequence-based phylogenetic analysis showed that the 118 isolates obtained comprised of 17 operational taxonomic units affiliated with the proteobacterial genera Rhizobium (18%), Marinomonas (1%), Pseudomonas (16%), and Xanthomonas (7%) but also with members of Firmicutes genera, such as Bacillus (35%) and Salinococcus (3%). The novel isolates belonging to Salinococcus and Bacillus could tolerate high K2CrO7 concentrations (3 mM) and also showed elevated activity of chromate reductase. In addition, majority (%) of the endophytic isolates also showed production of indole-3-acetic acid. Taken together, our results indicate that the innate endophytic bacterial community assists plants in reducing heavy metal toxicity.

  9. The Genetic Diversity of Endophytic and Phyllosphere Bacteria from Several Indonesian Herbal Plants

    Devi Rachelia

    2012-04-01

    Full Text Available Herbal plants have been believed by Indonesians to be an alternative medicine to treat illnesses. The bioactivecompounds in the plant can be derived from secondary metabolites or from endophytic and phyllosphere bacteria whichcoexist within medicinal plants. A total of 18 endophytic bacteria and 32 phyllosphere bacteria were isolated from theherbal plants of Citrus sp., Pluchea indica, Curcuma longa, Nothopanax scuttelarium, Piper crocatum, andAndrographis paniculata. About 72% of endophytic bacteria isolates have proteolytic activity and about 11% havelipolytic activity. On the other hand, about 59% of phyllosphere bacteria isolates have proteolytic activity and about19% have lipolytic activity. Phylogenetic diversity analysis was conducted by using the amplified ribosomal DNArestriction analysis (ARDRA method and the sequence of 16S rDNA was digested with endonuclease restrictionenzymes: MspI, RsaI, and Sau961. The diversity of endophytic and phyllosphere bacterium from the samples of herbalplants was high. Bacteria isolated from the same herbal plant does not always have a close genetic relationship exceptfor the bacteria isolated from the P. indica plant which showed a close genetic relationship with each other.

  10. Life without a cell membrane: Challenging the specificity of bacterial endophytes within Bryopsis (Bryopsidales, Chlorophyta

    Hollants Joke

    2011-11-01

    Full Text Available Abstract Background The siphonous green macroalga Bryopsis has some remarkable characteristics. Besides hosting a rich endophytic bacterial flora, Bryopsis also displays extraordinary wound repair and propagation mechanisms. This latter feature includes the formation of protoplasts which can survive in the absence of a cell membrane for several minutes before regenerating into new individuals. This transient 'life without a membrane' state, however, challenges the specificity of the endophytic bacterial communities present and raises the question whether these bacteria are generalists, which are repeatedly acquired from the environment, or if there is some specificity towards the Bryopsis host. Results To answer this question, we examined the temporal stability and the uniqueness of endobiotic bacterial communities within Bryopsis samples from the Mexican west coast after prolonged cultivation. DGGE analysis revealed that Bryopsis endophytic bacterial communities are rather stable and clearly distinct from the epiphytic and surrounding cultivation water bacterial communities. Although these endogenous communities consist of both facultative and obligate bacteria, results suggest that Bryopsis owns some intrinsic mechanisms to selectively maintain and/or attract specific bacteria after repeated wounding events in culture. Conclusions This suggests that Bryopsis algae seem to master transient stages of life without a cell membrane well as they harbor specific - and possibly ecological significant - endophytic bacteria.

  11. Crosstalk between endophytes and a plant host within information-processing networks

    Kozyrovska N. O.

    2013-05-01

    Full Text Available Plants are heavily populated by pro- and eukaryotic microorganisms and represent therefore the tremendous complexity as a biological system. This system exists as an information-processing entity with rather complex processes of communication, occurring throughout the individual plant. The plant cellular information-proces- sing network constitutes the foundation for processes like growth, defense, and adaptation to the environment. Up to date, the molecular mechanisms, underlying perception, transfer, analysis, and storage of the endogenous and environmental information within the plant, remain to be fully understood. The associated microorganisms and their investment in the information conditioning are often ignored. Endophytes as plant partners are indispen- sable integrative part of the plant system. Diverse endophytic microorganisms comprise «normal» microbiota that plays a role in plant immunity and helps the plant system to survive in the environment (providing assistance in defense, nutrition, detoxification etc.. The role of endophytic microbiota in the processing of information may be presumed, taking into account a plant-microbial co-evolution and empirical data. Since the literature are be- ginning to emerge on this topic, in this article, I review key works in the field of plant-endophytes interactions in the context of information processing and represent the opinion on their putative role in plant information web under defense and the adaptation to changed conditions.

  12. A polyphasic approach for the characterization of endophytic Alternaria strains isolated from grapevines

    Polizzotto, Rachele; Andersen, Birgitte; Martini, Marta

    2012-01-01

    A polyphasic approach was set up and applied to characterize 20 fungal endophytes belonging to the genus Alternaria, recovered from grapevine in different Italian regions.Morphological, microscopical, molecular and chemical investigations were performed and the obtained results were combined in a...

  13. Fungal Endophytes of Wild Barley and their Effects on Diuraphis noxia Population Development

    S.L. Clement; A. Dan Wilson; D.G. Lester; C.M. Davitt

    1997-01-01

    Laboratory experiments were conducted to compare the expression of Diuraphis noxia (Mordvilko) (Homoptera: Aphididae) resistance in four plant introduction (PI) lines of wild barley (Hordeum) infected with different species or strains of endophytic fungi (tribe Balansieae, family Clavicipitaceae, Neotyphodium gen. nov. [formerly...

  14. Comparative genomics provides insights into the lifestyle and reveals functional heterogeneity of dark septate endophytic fungi.

    Knapp, Dániel G; Németh, Julianna B; Barry, Kerrie; Hainaut, Matthieu; Henrissat, Bernard; Johnson, Jenifer; Kuo, Alan; Lim, Joanne Hui Ping; Lipzen, Anna; Nolan, Matt; Ohm, Robin A; Tamás, László; Grigoriev, Igor V; Spatafora, Joseph W; Nagy, László G; Kovács, Gábor M

    2018-04-20

    Dark septate endophytes (DSE) are a form-group of root endophytic fungi with elusive functions. Here, the genomes of two common DSE of semiarid areas, Cadophora sp. and Periconia macrospinosa were sequenced and analyzed with another 32 ascomycetes of different lifestyles. Cadophora sp. (Helotiales) and P. macrospinosa (Pleosporales) have genomes of 70.46 Mb and 54.99 Mb with 22,766 and 18,750 gene models, respectively. The majority of DSE-specific protein clusters lack functional annotation with no similarity to characterized proteins, implying that they have evolved unique genetic innovations. Both DSE possess an expanded number of carbohydrate active enzymes (CAZymes), including plant cell wall degrading enzymes (PCWDEs). Those were similar in three other DSE, and contributed a signal for the separation of root endophytes in principal component analyses of CAZymes, indicating shared genomic traits of DSE fungi. Number of secreted proteases and lipases, aquaporins, and genes linked to melanin synthesis were also relatively high in our fungi. In spite of certain similarities between our two DSE, we observed low levels of convergence in their gene family evolution. This suggests that, despite originating from the same habitat, these two fungi evolved along different evolutionary trajectories and display considerable functional differences within the endophytic lifestyle.

  15. Trichoderma amazonicum, a new endophytic species on Hevea brasiliensis and H. guianensis from the Amazon basin

    A new species of Trichoderma (teleomorph Hypocrea, Ascomycota, Sordariomycetes, Hypocreales, Hypocreaceae), T. amazonicum, endophytic on the living sapwood and leaves of Hevea spp. trees is described. Trichoderma amazonicum is distinguished from closely related species in the Harzianum clade (e.g. ...

  16. Diversity and taxonomy of endophytic xylariaceous fungi from medicinal plants of Dendrobium (Orchidaceae).

    Chen, Juan; Zhang, Li-Chun; Xing, Yong-Mei; Wang, Yun-Qiang; Xing, Xiao-Ke; Zhang, Da-Wei; Liang, Han-Qiao; Guo, Shun-Xing

    2013-01-01

    Dendrobium spp. are traditional Chinese medicinal plants, and the main effective ingredients (polysaccharides and alkaloids) have pharmacologic effects on gastritis infection, cancer, and anti-aging. Previously, we confirmed endophytic xylariaceous fungi as the dominant fungi in several Dendrobium species of tropical regions from China. In the present study, the diversity, taxonomy, and distribution of culturable endophytic xylariaceous fungi associated with seven medicinal species of Dendrobium (Orchidaceae) were investigated. Among the 961 endophytes newly isolated, 217 xylariaceous fungi (morphotaxa) were identified using morphological and molecular methods. The phylogenetic tree constructed using nuclear ribosomal internal transcribed spacer (ITS), large subunit of ribosomal DNA (LSU), and beta-tubulin sequences divided these anamorphic xylariaceous isolates into at least 18 operational taxonomic units (OTUs). The diversity of the endophytic xylariaceous fungi in these seven Dendrobium species was estimated using Shannon and evenness indices, with the results indicating that the dominant Xylariaceae taxa in each Dendrobium species were greatly different, though common xylariaceous fungi were found in several Dendrobium species. These findings implied that different host plants in the same habitats exhibit a preference and selectivity for their fungal partners. Using culture-dependent approaches, these xylariaceous isolates may be important sources for the future screening of new natural products and drug discovery.

  17. Interaction between a fungal endophyte and root herbivores of Ammophila arenaria

    Hol, W.H.G.; de la Peña, E.; Moens, M.; Cook, R.

    2007-01-01

    The effect of an endophytic fungus (Acremonium strictum) on plant-growth related parameters of marram grass (Ammophila arenaria), and its potential as a protective agent against root herbivores (Pratylenchus dunensis and Pratylenchus penetrans, root-lesion nematodes) was investigated in two

  18. Seed Endophyte Microbiome of Crotalaria pumila Unpeeled: Identification of Plant-Beneficial Methylobacteria

    Ariadna S. Sánchez-López

    2018-01-01

    Full Text Available Metal contaminated soils are increasing worldwide. Metal-tolerant plants growing on metalliferous soils are fascinating genetic and microbial resources. Seeds can vertically transmit endophytic microorganisms that can assist next generations to cope with environmental stresses, through yet poorly understood mechanisms. The aims of this study were to identify the core seed endophyte microbiome of the pioneer metallophyte Crotalaria pumila throughout three generations, and to better understand the plant colonisation of the seed endophyte Methylobacterium sp. Cp3. Strain Cp3 was detected in C. pumila seeds across three successive generations and showed the most dominant community member. When inoculated in the soil at the time of flowering, strain Cp3 migrated from soil to seeds. Using confocal microscopy, Cp3-mCherry was demonstrated to colonise the root cortex cells and xylem vessels of the stem under metal stress. Moreover, strain Cp3 showed genetic and in planta potential to promote seed germination and seedling development. We revealed, for the first time, that the seed microbiome of a pioneer plant growing in its natural environment, and the colonisation behaviour of an important plant growth promoting systemic seed endophyte. Future characterization of seed microbiota will lead to a better understanding of their functional contribution and the potential use for seed-fortification applications.

  19. Fungal endophytes isolated from Protium heptaphyllum and Trattinnickia rhoifolia as antagonists of Fusarium oxysporum.

    Fierro-Cruz, Juan E; Jiménez, Pedro; Coy-Barrera, Ericsson

    Control of fungal pathogens is mainly addressed by the use of chemically synthesized fungicides which result in environmental pollution, developing resistance after prolonged use. In this context, endophytes have been recognized as potential biocontrollers, and also as a promising source of antifungal metabolites. Therefore, as part of our research on phytopathogen controllers, 355 fungal endophytes were isolated from Protium heptaphyllum and Trattinnickia rhoifolia (Burseraceae), both ethnobotanically important tree species that produce secondary metabolites of agronomic and industrial interest. Endophytes were tested by in vitro dual culture against Fusarium oxysporum, a phytopathogen of agronomic importance. Five endophytes exerted at least 40% inhibition on F. oxysporum growth. Ethyl acetate (EtOAc) extracts were obtained from the most active antagonistic fungi, after growing them in three different liquid media. The extracts were tested against a conidial suspension of F. oxysporum by direct bioautography. Two extracts derived from fungi identified as Chaetomium globosum, F211_UMNG and Meyerozima sp. F281_UMNG showed inhibition of pathogen growth. Isolate C. globosum, F211_UMNG was selected for a chemical analysis by RP-HPLC-DAD-ESI-MS and antifungal molecules such as cladosporin, chaetoatrosin A and chaetoviridin A were annotated and identified based on their MS data. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  20. Antimicrobial biosynthetic potential and genetic diversity of endophytic actinomycetes associated with medicinal plants.

    Gohain, Anwesha; Gogoi, Animesh; Debnath, Rajal; Yadav, Archana; Singh, Bhim P; Gupta, Vijai K; Sharma, Rajeev; Saikia, Ratul

    2015-10-01

    Endophytic actinomycetes are one of the primary groups that share symbiotic relationships with medicinal plants and are key reservoir of biologically active compounds. In this study, six selective medicinal plants were targeted for the first time for endophytic actinomycetes isolation from Gibbon Wild Life Sanctuary, Assam, India, during winter and summer and 76 isolates were obtained. The isolates were found to be prevalent in roots followed by stem and leaves. 16S rRNA gene sequence analysis revealed 16 genera, including rare genera, Verrucosispora, Isoptericola and Kytococcus, which have never been previously reported as endophytic. The genus Streptomyces (66%) was dominant in both seasons. Shannon's diversity index showed that Azadirachta indica (1.49), Rauwolfia serpentina (1.43) and Emblica officinalis (1.24) were relatively good habitat for endophytic actinomycetes. Antimicrobial strains showed prevalence of polyketide synthase (PKS) type-II (85%) followed by PKS type-I (14%) encoded in the genomes. Expression studies showed 12-fold upregulation of PKSII gene in seventh day of incubation for Streptomyces antibioticus (EAAG90). Our results emphasize that the actinomycetes assemblages within plant tissue exhibited biosynthetic systems encoding for important biologically active compounds. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Priming effects of the endophytic fungus Phomopsis liquidambari on soil mineral N transformations.

    Chen, Yan; Ren, Cheng-Gang; Yang, Bo; Peng, Yao; Dai, Chuan-Chao

    2013-01-01

    Nitrogen (N) is a crucial nutrient for soil biota, and its cycling is determined by the organic carbon decomposing process. Some endophytic fungi are latent saprotrophs that trigger their saprotrophic metabolism to promote litter organic matter cycling as soon as the host tissue senesces or dies. However, the effects of endophytic fungi on litter and soil N dynamics in vitro have rarely been investigated. In this study, we investigated N dynamics (total and mineral N) in both litter and soil in incubations of a pure culture of an endophytic fungus Phomopsis liquidambari with litter and following soil burial of the litter. Soil enzymes and microbial communities participating in the N transformations were also investigated. A pure culture of P. liquidambari released litter NH (4) (+) -N in the initial stages (10 days) of the incubation. However, following soil burial, the presence of both P. liquidambari and soil ammonia-oxidizing bacteria (AOB) resulted in an increase in soil NO (3) (-) -N. These results indicate that the endophytic fungus P. liquidambari in vitro stimulates organic mineralization and promote NH (4) (+) -N release. Such effects triggered soil AOB-driven nitrification process.

  2. Endophytic actinobacteria: Diversity, secondary metabolism and mechanisms to unsilence biosynthetic gene clusters.

    Dinesh, Raghavan; Srinivasan, Veeraraghavan; T E, Sheeja; Anandaraj, Muthuswamy; Srambikkal, Hamza

    2017-09-01

    Endophytic actinobacteria, which reside in the inner tissues of host plants, are gaining serious attention due to their capacity to produce a plethora of secondary metabolites (e.g. antibiotics) possessing a wide variety of biological activity with diverse functions. This review encompasses the recent reports on endophytic actinobacterial species diversity, in planta habitats and mechanisms underlying their mode of entry into plants. Besides, their metabolic potential, novel bioactive compounds they produce and mechanisms to unravel their hidden metabolic repertoire by activation of cryptic or silent biosynthetic gene clusters (BGCs) for eliciting novel secondary metabolite production are discussed. The study also reviews the classical conservative techniques (chemical/biological/physical elicitation, co-culturing) as well as modern microbiology tools (e.g. next generation sequencing) that are being gainfully employed to uncover the vast hidden scaffolds for novel secondary metabolites produced by these endophytes, which would subsequently herald a revolution in drug engineering. The potential role of these endophytes in the agro-environment as promising biological candidates for inhibition of phytopathogens and the way forward to thoroughly exploit this unique microbial community by inducing expression of cryptic BGCs for encoding unseen products with novel therapeutic properties are also discussed.

  3. Metabolomic Tools to Assess the Chemistry and Bioactivity of Endophytic Aspergillus Strain.

    Tawfike, Ahmed F; Tate, Rothwelle; Abbott, Gráinne; Young, Louise; Viegelmann, Christina; Schumacher, Marc; Diederich, Marc; Edrada-Ebel, RuAngelie

    2017-10-01

    Endophytic fungi associated with medicinal plants are a potential source of novel chemistry and biology that may find applications as pharmaceutical and agrochemical drugs. In this study, a combination of metabolomics and bioactivity-guided approaches were employed to isolate secondary metabolites with cytotoxicity against cancer cells from an endophytic Aspergillus aculeatus. The endophyte was isolated from the Egyptian medicinal plant Terminalia laxiflora and identified using molecular biological methods. Metabolomics and dereplication studies were accomplished by utilizing the MZmine software coupled with the universal Dictionary of Natural Products database. Metabolic profiling, with aid of multivariate data analysis, was performed at different stages of the growth curve to choose the optimized method suitable for up-scaling. The optimized culture method yielded a crude extract abundant with biologically-active secondary metabolites. Crude extracts were fractionated using different high-throughput chromatographic techniques. Purified compounds were identified by HR-ESI-MS, 1D- and 2D-NMR. This study introduced a new method of dereplication utilizing both high-resolution mass spectrometry and NMR spectroscopy. The metabolites were putatively identified by applying a chemotaxonomic filter. We also present a short review on the diverse chemistry of terrestrial endophytic strains of Aspergillus, which has become a part of our dereplication work and this will be of wide interest to those working in this field. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  4. Antifungal and antibacterial activity of endophytic penicillium species isolated from salvadora species

    Korejo, F.; Shafique, H.A.; Haque, S.E.; Ali, S.A.

    2014-01-01

    Salvadora persica and S. S.oleoides are facultative holophytic plants, well known as miswak, are traditionally used to ensure oral hygiene among Muslim people in Asian and African counties. Species of Salvadora have a number of proven pharmacological importance. Besides, terrestrial fungi endophytic fungi are also gaining importance for the isolation of bioactive compounds. In this study 74 samples (root, shoot and leaves) from S. persica and S. oleoides were examined for endophytic fungi, 22 samples showed presence of Penicillium spp., 48 were found positive for aspergilli, whereas 10 samples showed infection of Fusarium solani, 4 were found infected with Macrophomina phaseolina and one with Rhizoctonia solani. Most of the Penicillium isolated were identified as P. restrictum, P. citrinum and P. canescens. In dual culture plate assay out of four Penicillium isolates tested, P. citrinum and one isolate of P. restrictum caused growth inhibition of all four test root rotting fungi, Fusarium solani, F. oxysporum, Macrophomina phaseolina and Rhizoctonia solani. Culture filtrates of Penicillium spp., were also evaluated against four common laboratory bacteria namely Bacillus subtilis, Staphylococcus aureus, Salmonella typhimurium and Escherichia coli and above mentioned root rotting fungi. Culture filtrates of endophytic Penicillium spp., also showed significant antibacterial and antifungal activity. Secondary metabolites of endophytic Penicillium spp., offer an exciting area of research for the discovery of novel antimicrobial compounds. (author)

  5. Concentration of petroleum-hydrocarbon contamination shapes fungal endophytic community structure in plant roots

    Guillaume eBourdel

    2016-05-01

    Full Text Available Plant-root inhabiting fungi are a universal phenomenon found in all ecosystems where plants are able to grow, even in harsh environments. Interactions between fungi and plant roots can vary widely from mutualism to parasitism depending on many parameters. The role of fungal endophytes in phytoremediation of polluted sites, and characterization of the endophytic diversity and community assemblages in contaminated areas remain largely unexplored. In this study, we investigated the composition of endophytic fungal communities in the roots of two plant species growing spontaneously in petroleum-contaminated sedimentation basins of a former petro-chemical plant. The three adjacent basins showed a highly heterogeneous patterns of pollutant concentrations. We combined a culture-based isolation approach with the pyrosequencing of fungal ITS ribosomal DNA. We selected two species, Eleocharis erythropoda Steud. and Populus balsamifera L., and sampled three individuals of each species from each of three adjacent basins, each with a different concentration of petroleum hydrocarbons. We found that contamination level significantly shaped endophytic fungal diversity and community composition in E. erythropoda, with only 9.9% of these fungal Operational Taxonomic Units (OTUs retrieved in all three basins. However, fungal community structure associated with P. balsamifera remained unaffected by the contamination level with 28.2% of fungal OTUs shared among all three basins. This could be explained by the smaller differences of pollutant concentrations in the soil around our set of P. balsamifera sampless compared to that around our set of E. erythropoda samples. Our culture-based approach allowed isolation of 11 and 30 fungal endophytic species from surface-sterilized roots of E. erythropoda and P. balsamifera, respectively. These isolates were ribotyped using ITS, and all were found in pyrosequensing datasets. Our results demonstrate that extreme levels of

  6. Use of rhizobacteria and endophytes for biological control of weeds

    Trognitz, Friederike

    2014-02-01

    Full Text Available Weeds cause severe yield losses in agriculture, with a maximum estimate of 34% of yield loss worldwide due to competition between the crops and the weeds for nutrition, light and humidity (OERKE, 2006. Invasive plants contribute partially to other problems. The pollen of common ragweed, Ambrosia artemisiifolia L., for example, is five times more allergenic than grass pollen; already ten pollen grains per m3 air can trigger allergy in sensitized patients, including rhinitis, conjunctivitis and asthma. This neophyte from America has extended the season of allergy in European patients to October. Common ragweed is currently most frequent in Hungary, France and Italy. In Austria, ragweed populations along roads have increased dramatically since 2000. The effective means to control this weed of the Asteraceae family are limited; a single plant can produce up to 6000 seeds which stay in the soil for 40 years. Control using selective herbicides is not possible within stands of the Asteraceae member sunflower. Efforts to use herbivore insects as biological control agents also failed due to the unavailability of insects specializing on this ragweed. The use of plant-associated rhizobacteria and endophytes as bio-herbicides offers a novel alternative to conventional methods. By analogy to experiences from other plant-microbe systems, the chances to find microbes of the desired characteristics are highest when isolating and testing specimens directly from ragweed plants. These organisms often have an extremely narrow host range that permits their use for the control of among several even closely related plant species growing together in a field.

  7. Hypoxylon pulicicidum sp. nov. (Ascomycota, Xylariales, a pantropical insecticide-producing endophyte.

    Gerald F Bills

    Full Text Available BACKGROUND: Nodulisporic acids (NAs are indole diterpene fungal metabolites exhibiting potent systemic efficacy against blood-feeding arthropods, e.g., bedbugs, fleas and ticks, via binding to arthropod specific glutamate-gated chloride channels. Intensive medicinal chemistry efforts employing a nodulisporic acid A template have led to the development of N-tert-butyl nodulisporamide as a product candidate for a once monthly treatment of fleas and ticks on companion animals. The source of the NAs is a monophyletic lineage of asexual endophytic fungal strains that is widely distributed in the tropics, tentatively identified as a Nodulisporium species and hypothesized to be the asexual state of a Hypoxylon species. METHODS AND RESULTS: Inferences from GenBank sequences indicated that multiple researchers have encountered similar Nodulisporium endophytes in tropical plants and in air samples. Ascomata-derived cultures from a wood-inhabiting fungus, from Martinique and closely resembling Hypoxylon investiens, belonged to the same monophyletic clade as the NAs-producing endophytes. The hypothesis that the Martinique Hypoxylon collections were the sexual state of the NAs-producing endophytes was tested by mass spectrometric analysis of NAs, multi-gene phylogenetic analysis, and phenotypic comparisons of the conidial states. We established that the Martinique Hypoxylon strains produced an ample spectrum of NAs and were conspecific with the pantropical Nodulisporium endophytes, yet were distinct from H. investiens. A new species, H. pulicicidum, is proposed to accommodate this widespread organism. CONCLUSIONS AND SIGNIFICANCE: Knowledge of the life cycle of H. pulicicidum will facilitate an understanding of the role of insecticidal compounds produced by the fungus, the significance of its infections in living plants and how it colonizes dead wood. The case of H. pulicicidum exemplifies how life cycle studies can consolidate disparate observations of a

  8. Elimination and molecular identification of endophytic bacterial contaminants during in vitro propagation of Bambusa balcooa.

    Ray, Syandan Sinha; Ali, Md Nasim; Mukherjee, Shibasis; Chatterjee, Gautam; Banerjee, Maitreyi

    2017-02-01

    Bambusa balcooa is an economically important, multipurpose bamboo species, decidedly used in construction industry. Availability of natural bamboo is depleting very rapidly due to accelerated deforestation and its unrestrained use. The large number and timely supply of saplings are the need of the hour for the restoration of bamboo stands. Micropropagation, being the potent alternative for season independent rapid regeneration, is restricted in bamboo because of endophytic contamination. An in vitro attempt has been taken to overcome the endophytic contamination by using broad spectrum antibiotics as surface sterilant as well as a media component. Ampicillin sodium salt (5 mg/ml for 30 min) as a surface sterilant was found as the best treatment for high bud breaking (80%) coupled with high branching and low contamination (20%) but it was found ineffective to control the contamination during multiplication stage. Then, two endophytes were isolated and minimum inhibitory concentration was determined through antibiotic susceptibility test for successful eradication at multiplication stage. Finally, contamination free cultures were obtained when streptocycline (100 μg/ml) and gentamicin sulphate (75 μg/ml) were added into the medium. The two isolated endophytes, BB1 and BB2, were identified through 16S rDNA techniques and NCBI-BLAST algorithm with 99% sequence similarity with those of Janibacter sp. (KX423734) and Serratia marcescens strain (KX423735). To our knowledge, this is the first report for B. balcooa where antibiotics were used as surface sterilant as well as medium component, to control endophytic bacterial contaminants, followed by their identification.

  9. Melanised endophytic fungi may increase stores of organic carbon in soil

    McGee, Peter; Mukasa Mugerwa, Tendo

    2013-04-01

    The processes underlying the carbon cycle in soil, especially sequestration of organic carbon (OC), are poorly understood. Hydrolysis and oxidation reduce organic matter. Hydrolysis degrades linear organic molecules in aerobic and anaerobic conditions, though it is slower in anaerobic conditions. Aromatic compounds are only degraded by oxidation. Oxygen is by far the most common electron acceptor in soil. Anaerobic conditions preclude oxidation in soil and will result in the preservation of aromatic compounds so long as the conditions remain anaerobic. We experimentally tested this model using melanised endophytic fungi. Melanin is a polyaromatic compound that can be readily visualised, though is difficult to quantify. An endophytic association provides the fungus with an ongoing source of energy. Fungal hyphae elongate considerable distances in soil where they may colonise aggregates, the core of which may be anaerobic. The hypothesis we tested is that melanised endophytic fungi increase OC in soil. Seedlings of subterranean clover inoculated with single isolates were grown in split pots where the impact of the fungus could be quantified in the hyphal chamber, separated from the roots by a steel mesh. We found that melanised endophytic fungi significantly increased OC and aromatic carbon in a well-aggregated carbon-rich soil. OC increased by up to 17% within 14 weeks. Twenty out of 24 isolates statistically significantly increased and none decreased OC. Increases differed between fungal isolates. Increases in the hyphal chamber were independent of any change in OC associated with the roots of the host plant. The storage of OC in field soils is being explored. Inoculation of plant roots with melanised endophytic fungi offers one means whereby OC may be increased in field soils.

  10. Enhanced degradation activity by endophytic bacteria of plants growing in hydrocarbon contaminated soil

    Phillips, L.; Germida, J.J. [Saskatchewan Univ., Saskatoon, SK (Canada); Greer, C.W. [National Research Council of Canada, Montreal, PQ (Canada). Biotechnology Research Inst.

    2006-07-01

    The feasibility of using phytoremediation for cleaning soils contaminated with petroleum hydrocarbons was discussed. Petroleum hydrocarbons are problematic because of their toxicity, mobility and persistence in the environment. Appropriate clean-up methods are needed, given that 60 per cent of Canada's contaminated sites contain these compounds. Phytoremediation is an in situ biotechnology in which plants are used to facilitate contaminant removal. The approach relies on a synergistic relationship between plants and their root-associated microbial communities. Previous studies on phytoremediation have focussed on rhizosphere communities. However, it is believed that endophytic microbes may also play a vital role in organic contaminant degradation. This study investigated the structural and functional dynamics of both rhizosphere and endophytic microbial communities of plants from a phytoremediation field site in south-eastern Saskatchewan. The former flare pit contains up to 10,000 ppm of F3 to F4 hydrocarbon fractions. Root samples were collected from tall wheatgrass, wild rye, saltmeadow grass, perennial ryegrass, and alfalfa. Culture-based and culture-independent methods were used to evaluate the microbial communities associated with these roots. Most probable number assays showed that the rhizosphere communities contained more n-hexadecane, diesel fuel, and PAH degraders. However, mineralization assays with 14C labelled n-hexadecane, naphthalene, and phenanthrene showed that endophytic communities had more degradation activities per standardized initial degrader populations. Total community DNA samples taken from bulk, rhizosphere, and endophytic samples, were analyzed by denaturing gradient gel electrophoresis. It was shown that specific bacteria increased in endophytic communities compared to rhizosphere communities. It was suggested plants may possibly recruit specific bacteria in response to hydrocarbon contamination, thereby increasing degradation

  11. Ethanol and methanol can improve huperzine A production from endophytic Colletotrichum gloeosporioides ES026.

    Zhao, Xin-Mei; Wang, Zhang-Qian; Shu, Shao-Hua; Wang, Wen-Juan; Xu, Hai-Jie; Ahn, Young-Joon; Wang, Mo; Hu, Xuebo

    2013-01-01

    Huperzine A (HupA) is a plant alkaloid that is of great interest as a therapeutic candidate for the treatment of Alzheimer's disease. However, the current production of HupA from plants in large quantity is unsustainable because the plant resource is scarce and the content of HupA in plants is extremely low. Surprisingly, this compound was recently found to be produced by various endophytic fungi, which are much more controllable than the plants due to simpler genetics and ease of manipulation. However, it might be due to the innate properties of endophytic symbiosis, that production of this chemical in large quantity from endophytes has not yet been put into practice. Endophytic Colletotrichum gloeosporioides ES026 was previously isolated from a HupA producing plant and the fungi also proved to produce HupA. In this study, various fermentation conditions were tried to optimize the production of HupA from C. gloeosporioides ES026. Optimization of these parameters resulted in a 25.58% increase in HupA yield. Potato extracts supplemented with glucose or sucrose but not maltose facilitated HupA producing from the fungi. A final concentration of 0.5-2% ethanol stimulated the growth of fungi while methanol with the same treatment slightly inhibited the growth. However, both methanol and ethanol greatly increased the HupA production with the highest yield of HupA (51.89% increment) coming from ethanol treatment. Further analysis showed that both ethanol and methanol were strong inducers of HupA production, while ethanol was partially used as a carbon source during fermentation. It was noticed that the color of that ethanol treated mycelia gradually became dark while methanol treated ones stayed grey during fermentation. The present study sheds light on the importance of optimizing the fermentation process, which, combined with effective inducers, maximizes production of chemicals of important economic interest from endophytic fungi.

  12. Ethanol and methanol can improve huperzine A production from endophytic Colletotrichum gloeosporioides ES026.

    Xin-Mei Zhao

    Full Text Available Huperzine A (HupA is a plant alkaloid that is of great interest as a therapeutic candidate for the treatment of Alzheimer's disease. However, the current production of HupA from plants in large quantity is unsustainable because the plant resource is scarce and the content of HupA in plants is extremely low. Surprisingly, this compound was recently found to be produced by various endophytic fungi, which are much more controllable than the plants due to simpler genetics and ease of manipulation. However, it might be due to the innate properties of endophytic symbiosis, that production of this chemical in large quantity from endophytes has not yet been put into practice. Endophytic Colletotrichum gloeosporioides ES026 was previously isolated from a HupA producing plant and the fungi also proved to produce HupA. In this study, various fermentation conditions were tried to optimize the production of HupA from C. gloeosporioides ES026. Optimization of these parameters resulted in a 25.58% increase in HupA yield. Potato extracts supplemented with glucose or sucrose but not maltose facilitated HupA producing from the fungi. A final concentration of 0.5-2% ethanol stimulated the growth of fungi while methanol with the same treatment slightly inhibited the growth. However, both methanol and ethanol greatly increased the HupA production with the highest yield of HupA (51.89% increment coming from ethanol treatment. Further analysis showed that both ethanol and methanol were strong inducers of HupA production, while ethanol was partially used as a carbon source during fermentation. It was noticed that the color of that ethanol treated mycelia gradually became dark while methanol treated ones stayed grey during fermentation. The present study sheds light on the importance of optimizing the fermentation process, which, combined with effective inducers, maximizes production of chemicals of important economic interest from endophytic fungi.

  13. In Vitro Morphogenesis of Arabidopsis to Search for Novel Endophytic Fungi Modulating Plant Growth.

    Francesco Dovana

    Full Text Available Fungal endophytes have shown to affect plant growth and to confer stress tolerance to the host; however, effects of endophytes isolated from water plants have been poorly investigated. In this study, fungi isolated from stems (stem-E and roots (root-E of Mentha aquatica L. (water mint were identified, and their morphogenetic properties analysed on in vitro cultured Arabidopsis (L. Heynh., 14 and 21 days after inoculation (DAI. Nineteen fungi were analysed and, based on ITS analysis, 17 isolates showed to be genetically distinct. The overall effect of water mint endophytes on Arabidopsis fresh (FW and dry weight (DW was neutral and positive, respectively, and the increased DW, mainly occurring 14 DAI, was possibly related to plant defence mechanism. Only three fungi increased both FW and DW of Arabidopsis at 14 and 21 DAI, thus behaving as plant growth promoting (PGP fungi. E-treatment caused a reduction of root depth and primary root length in most cases and inhibition-to-promotion of root area and lateral root length, from 14 DAI. Only Phoma macrostoma, among the water mint PGP fungi, increased both root area and depth, 21 DAI. Root depth and area 14 DAI were shown to influence DWs, indicating that the extension of the root system, and thus nutrient uptake, was an important determinant of plant dry biomass. Reduction of Arabidopsis root depth occurred to a great extent when plants where treated with stem-E while root area decreased or increased under the effects of stem-E and root-E, respectively, pointing to an influence of the endophyte origin on root extension. M. aquatica and many other perennial hydrophytes have growing worldwide application in water pollution remediation. The present study provided a model for directed screening of endophytes able to modulate plant growth in the perspective of future field applications of these fungi.

  14. In Vitro Morphogenesis of Arabidopsis to Search for Novel Endophytic Fungi Modulating Plant Growth.

    Dovana, Francesco; Mucciarelli, Marco; Mascarello, Maurizio; Fusconi, Anna

    2015-01-01

    Fungal endophytes have shown to affect plant growth and to confer stress tolerance to the host; however, effects of endophytes isolated from water plants have been poorly investigated. In this study, fungi isolated from stems (stem-E) and roots (root-E) of Mentha aquatica L. (water mint) were identified, and their morphogenetic properties analysed on in vitro cultured Arabidopsis (L.) Heynh., 14 and 21 days after inoculation (DAI). Nineteen fungi were analysed and, based on ITS analysis, 17 isolates showed to be genetically distinct. The overall effect of water mint endophytes on Arabidopsis fresh (FW) and dry weight (DW) was neutral and positive, respectively, and the increased DW, mainly occurring 14 DAI, was possibly related to plant defence mechanism. Only three fungi increased both FW and DW of Arabidopsis at 14 and 21 DAI, thus behaving as plant growth promoting (PGP) fungi. E-treatment caused a reduction of root depth and primary root length in most cases and inhibition-to-promotion of root area and lateral root length, from 14 DAI. Only Phoma macrostoma, among the water mint PGP fungi, increased both root area and depth, 21 DAI. Root depth and area 14 DAI were shown to influence DWs, indicating that the extension of the root system, and thus nutrient uptake, was an important determinant of plant dry biomass. Reduction of Arabidopsis root depth occurred to a great extent when plants where treated with stem-E while root area decreased or increased under the effects of stem-E and root-E, respectively, pointing to an influence of the endophyte origin on root extension. M. aquatica and many other perennial hydrophytes have growing worldwide application in water pollution remediation. The present study provided a model for directed screening of endophytes able to modulate plant growth in the perspective of future field applications of these fungi.

  15. Selenium hyperaccumulators harbor a diverse endophytic bacterial community characterized by high selenium resistance and plant growth promoting properties

    Sura-de Jong, M.; Reynolds, R.B.J.; Richterová, K.; Musilová, L.; Staicu, L. C.; Chocholatá, I.; Cappa, J. J.; Taghavi, S.; van der Lelie, D.; Frantík, Tomáš; Dolinová, I.; Strejček, M.; Cochran, A. T.; Lovecká, P.; Pilon-Smits, E. A. H.

    2015-01-01

    Roč. 6, č. 113 (2015), s. 1-17 ISSN 1664-462X Institutional support: RVO:67985939 Keywords : endophyte * bacteria * phytoremediation Subject RIV: EE - Microbiology, Virology Impact factor: 4.495, year: 2015

  16. Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae).

    Feitosa, André de O; Dias, Amanda Cristina S; Ramos, Gisele da C; Bitencourt, Heriberto R; Siqueira, José Edson S; Marinho, Patrícia Santana B; Barison, Andersson; Ocampos, Fernanda M M; Marinho, Andrey Moacir do R

    Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. A new endophytic insect-associated Daldinia species, recognised from a comparison of secondary metabolite profiles and molecular phylogeny

    Pažoutová, Sylvie; Follert, S.; Bitzer, J.; Keck, M.; Surup, F.; Šrůtka, P.; Holuša, J.; Stadler, M.

    2013-01-01

    Roč. 60, č. 1 (2013), s. 107-123 ISSN 1560-2745 Institutional support: RVO:61388971 Keywords : Fungi * Endophytes * Evolution Subject RIV: EE - Microbiology, Virology Impact factor: 6.938, year: 2013

  18. The endophytic fungus Piriformospora indica enhances Arabidopsis thaliana growth and modulates Na + /K + homeostasis under salt stress conditions

    Abdelaziz, Mohamed Ewis; Kim, Dongjin; Ali, Shawkat; Fedoroff, Nina V.; Al-Babili, Salim

    2017-01-01

    The mutualistic, endophytic fungus Piriformospora indica has been shown to confer biotic and abiotic stress tolerance to host plants. In this study, we investigated the impact of P. indica on the growth of Arabidopsis plants under normal and salt

  19. Microbial conversion of curcumin into colorless hydroderivatives by the endophytic fungus Diaporthe sp. associated with Curcuma longa.

    Maehara, Shoji; Ikeda, Michiteru; Haraguchi, Hiroyuki; Kitamura, Chinami; Nagoe, Tetsuro; Ohashi, Kazuyoshi; Shibuya, Hirotaka

    2011-01-01

    We investigated the microbial conversion of curcumin (1) using endophytic fungi associated with the rhizome of Curcuma longa (Zingiberaceae). We found that Diaporthe sp., an endophytic filamentous fungus, converts curcumin (1) into four colorless derivatives, namely (3R,5R)-tetrahydrocurcumin (2), a novel (3R,5S)-hexahydrocurcumin (3) named neohexahydrocurcumin, (3S,5S)-octahydrocurcumin (4) and meso-octahydrocurcumin (5).

  20. Indigenous endophytic seed bacteria promote seedling development and defend against fungal disease in browntop millet (Urochloa ramosa L.).

    Verma, S K; White, J F

    2018-03-01

    This study was conducted to investigate indigenous seed endophyte effects on browntop millet seedling development. We report that seed-inhabiting bacterial endophytes are responsible for promoting seedling development, including stimulation of root hair formation, increasing root and shoot length growth and increasing photosynthetic pigment content of seedlings. Bacterial endophytes also improved resistance of seedlings to disease. A total of four endophytic bacteria were isolated from surface-sterilized seeds and identified by 16S rDNA sequencing as Curtobacterium sp. (M1), Microbacterium sp. (M2), Methylobacterium sp. (M3) and Bacillus amyloliquefaciens (M4). Removal of bacteria with streptomycin treatment from the seeds compromised seedling growth and development. When endophytes were reinoculated onto seeds, seedlings recovered normal development. Strains M3 and M4 were found to be most potent in promoting growth of seedlings. Bacteria were found to produce auxin, solubilize phosphate and inhibit fungal pathogens. Significant protection of seedlings from Fusarium infection was found using strain M4 in microcosm assays. The antifungal lipopeptide genes for surfactin and iturin were detected in M4; culture extracts of M4 showed a positive drop collapse result for surfactins. This study demonstrates that browntop millet seeds vector indigenous endophytes that are responsible for modulation of seedling development and protection of seedlings from fungal disease. This study is significant and original in that it is the first report of seed-inhabiting endophytes of browntop millet that influence seedling development and function in defence against soilborne pathogens. This study suggests that conservation and management of seed-vectored endophytes may be important in development of more sustainable agricultural practices. © 2017 The Society for Applied Microbiology.

  1. The Biological Diversity and Production of Volatile Organic Compounds by Stem-Inhabiting Endophytic Fungi of Ecuador

    Rundell, Susan; Spakowicz, Daniel; Narváez-Trujillo, Alexandra; Strobel, Scott

    2015-01-01

    Fungal endophytes colonize every major lineage of land plants without causing apparent harm to their hosts. Despite their production of interesting and potentially novel compounds, endophytes—particularly those inhabiting stem tissues—are still a vastly underexplored component of microbial diversity. In this study, we explored the diversity of over 1500 fungal endophyte isolates collected from three Ecuadorian ecosystems: lowland tropical forest, cloud forest, and coastal dry forest. We sough...

  2. Influences of Plant Species, Season and Location on Leaf Endophytic Bacterial Communities of Non-Cultivated Plants.

    Ding, Tao; Melcher, Ulrich

    2016-01-01

    Bacteria are known to be associated endophytically with plants. Research on endophytic bacteria has identified their importance in food safety, agricultural production and phytoremediation. However, the diversity of endophytic bacterial communities and the forces that shape their compositions in non-cultivated plants are largely uncharacterized. In this study, we explored the diversity, community structure, and dynamics of endophytic bacteria in different plant species in the Tallgrass Prairie Preserve of northern Oklahoma, USA. High throughput sequencing of amplified segments of bacterial rDNA from 81 samples collected at four sampling times from five plant species at four locations identified 335 distinct OTUs at 97% sequence similarity, representing 16 phyla. Proteobacteria was the dominant phylum in the communities, followed by the phyla Bacteriodetes and Actinobacteria. Bacteria from four classes of Proteobacteria were detected with Alphaproteobacteria as the dominant class. Analysis of molecular variance revealed that host plant species and collecting date had significant influences on the compositions of the leaf endophytic bacterial communities. The proportion of Alphaproteobacteria was much higher in the communities from Asclepias viridis than from other plant species and differed from month to month. The most dominant bacterial groups identified in LDA Effect Size analysis showed host-specific patterns, indicating mutual selection between host plants and endophytic bacteria and that leaf endophytic bacterial compositions were dynamic, varying with the host plant's growing season in three distinct patterns. In summary, next generation sequencing has revealed variations in the taxonomic compositions of leaf endophytic bacterial communities dependent primarily on the nature of the plant host species.

  3. Influences of Plant Species, Season and Location on Leaf Endophytic Bacterial Communities of Non-Cultivated Plants.

    Tao Ding

    Full Text Available Bacteria are known to be associated endophytically with plants. Research on endophytic bacteria has identified their importance in food safety, agricultural production and phytoremediation. However, the diversity of endophytic bacterial communities and the forces that shape their compositions in non-cultivated plants are largely uncharacterized. In this study, we explored the diversity, community structure, and dynamics of endophytic bacteria in different plant species in the Tallgrass Prairie Preserve of northern Oklahoma, USA. High throughput sequencing of amplified segments of bacterial rDNA from 81 samples collected at four sampling times from five plant species at four locations identified 335 distinct OTUs at 97% sequence similarity, representing 16 phyla. Proteobacteria was the dominant phylum in the communities, followed by the phyla Bacteriodetes and Actinobacteria. Bacteria from four classes of Proteobacteria were detected with Alphaproteobacteria as the dominant class. Analysis of molecular variance revealed that host plant species and collecting date had significant influences on the compositions of the leaf endophytic bacterial communities. The proportion of Alphaproteobacteria was much higher in the communities from Asclepias viridis than from other plant species and differed from month to month. The most dominant bacterial groups identified in LDA Effect Size analysis showed host-specific patterns, indicating mutual selection between host plants and endophytic bacteria and that leaf endophytic bacterial compositions were dynamic, varying with the host plant's growing season in three distinct patterns. In summary, next generation sequencing has revealed variations in the taxonomic compositions of leaf endophytic bacterial communities dependent primarily on the nature of the plant host species.

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

    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...

  5. Distribution of endophytic bacteria in Alopecurus aequalis Sobol and Oxalis corniculata L. from soils contaminated by polycyclic aromatic hydrocarbons.

    Anping Peng

    Full Text Available The distributions of endophytic bacteria in Alopecurus aequalis Sobol and Oxalis corniculata L. grown in soils contaminated with different levels of polycyclic aromatic hydrocarbons (PAHs were investigated with polymerase chain reaction followed by denaturing gradient gel electrophoresis technology (PCR-DGGE and cultivation methods. Twelve types of PAHs, at concentrations varying from 0.16 to 180 mg·kg(-1, were observed in the roots and shoots of the two plants. The total PAH concentrations in Alopecurus aequalis Sobol obtained from three different PAH-contaminated stations were 184, 197, and 304 mg·kg(-1, and the total PAH concentrations in Oxalis corniculata L. were 251, 346, and 600 mg·kg(-1, respectively. The PCR-DGGE results showed that the endophytic bacterial communities in the roots and shoots of the two plants were quite different, although most bacteria belonged to Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. A total of 68 endophytic bacterial strains were isolated from different tissues of the two plants and classified into three phyla: Firmicutes, Proteobacteria and Bacteroidetes. In both plants, Bacillus spp. and Pseudomonas spp. were the dominant cultivable populations. With an increase in the PAH pollution level, the diversity and distribution of endophytic bacteria in the two plants changed correspondingly, and the number of cultivable endophytic bacterial strains decreased rapidly. Testing of the isolated endophytic bacteria for tolerance to each type of PAH showed that most isolates could grow well on Luria-Bertani media in the presence of different PAHs, and some isolates were able to grow rapidly on a mineral salt medium with a single PAH as the sole carbon and energy source, indicating that these strains may have the potential to degrade PAHs in plants. This research provides the first insight into the characteristics of endophytic bacterial populations under different PAH pollution levels and provides a

  6. Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds.

    Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

    2016-01-01

    Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents.

  7. Understanding colonization and proliferation potential of endophytes and pathogen in planta via plating, polymerase chain reaction and ergosterol assay.

    Chow, Yiing Yng; Rahman, Sadequr; Ting, Adeline Su Yien

    2017-01-01

    This study aimed to establish the colonization behavior and proliferation potential of three endophytes and one pathogen Ganoderma boninense (Gb) introduced into oil palm ramets (host model). The endophytes selected were Diaporthe phaseolorum (WAA02), Trichoderma asperellum (T2), and Penicillium citrinum (BTF08). Ramets were first inoculated with 100 mL of fungal cells (10 6  cfu mL - 1 ) via soil drenching. For the next 7 days, ramets were sampled and subjected to three different assays to detect and identify fungal colonization, and establish their proliferation potential in planta . Plate assay revealed the presence of endophytes in root, stem and leaf tissues within 7 days after inoculation. Polymerase Chain Reaction (PCR) detected and identified the isolates from the plant tissues. The ergosterol assay (via high performance liquid chromatography, HPLC) confirmed the presence of endophytes and Gb in planta . The increase in ergosterol levels throughout 49 days was however insignificant, suggesting that proliferation may be absent or may occur very slowly in planta . This study strongly suggests that the selected endophytes could colonize the host upon inoculation, but proliferation occurs at a slower rate, which may subsequently influence the biocontrol expression of endophytes against the pathogen.

  8. Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure

    Hao Zhang

    2017-11-01

    Full Text Available Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi (Brassica chinensis L. to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg−1 were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tetX, blaCTX-M, and sul1 and sul2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.

  9. Understanding colonization and proliferation potential of endophytes and pathogen in planta via plating, polymerase chain reaction, and ergosterol assay

    Yiing Yng Chow

    2017-01-01

    Full Text Available This study aimed to establish the colonization behavior and proliferation potential of three endophytes and one pathogen Ganoderma boninense (Gb introduced into oil palm ramets (host model. The endophytes selected were Diaporthe phaseolorum (WAA02, Trichoderma asperellum (T2, and Penicillium citrinum (BTF08. Ramets were first inoculated with 100 mL of fungal cells (106 cfu mL−1 via soil drenching. For the next 7 days, ramets were sampled and subjected to three different assays to detect and identify fungal colonization, and establish their proliferation potential in planta. Plate assay revealed the presence of endophytes in root, stem and leaf tissues within 7 days after inoculation. Polymerase Chain Reaction (PCR detected and identified the isolates from the plant tissues. The ergosterol assay (via high-performance liquid chromatography, HPLC confirmed the presence of endophytes and Gb in planta. The increase in ergosterol levels throughout 49 days was however insignificant, suggesting that proliferation may be absent or may occur very slowly in planta. This study strongly suggests that the selected endophytes could colonize the host upon inoculation, but proliferation occurs at a slower rate, which may subsequently influence the biocontrol expression of endophytes against the pathogen.

  10. Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure.

    Zhang, Hao; Li, Xunan; Yang, Qingxiang; Sun, Linlin; Yang, Xinxin; Zhou, Mingming; Deng, Rongzhen; Bi, Linqian

    2017-11-03

    Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs) in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi ( Brassica chinensis L.) to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC) levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg -1 were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tet X, bla CTX-M , and sul 1 and sul 2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.

  11. Growth, photosynthesis and antioxidant responses of endophyte infected and non-infected rice under lead stress conditions.

    Li, Xuemei; Bu, Ning; Li, Yueying; Ma, Lianju; Xin, Shigang; Zhang, Lihong

    2012-04-30

    An endophytic fungus was tested in rice (Oryza sativa L.) exposed to four levels of lead (Pb) stress (0, 50, 100 and 200 μM) to assess effects on plant growth, photosynthesis and antioxidant enzyme activity. Under Pb stress conditions, endophyte-infected seedlings had greater shoot length but lower root length compared to non-infected controls, and endophyte-infected seedlings had greater dry weight in the 50 and 100 μM Pb treatments. Under Pb stress conditions, chlorophyll and carotenoid levels were significantly higher in the endophyte-infected seedlings. Net photosynthetic rate, transpiration rate and water use efficiency were significantly higher in endophyte-infected seedlings in the 50 and 100 μM Pb treatments. In addition, chlorophyll fluorescence parameters Fv/Fm and Fv/Fo were higher in the infected seedlings compared to the non-infected seedlings under Pb stress. Malondialdehyde accumulation was induced by Pb stress, and it was present in higher concentration in non-infected seedlings under higher concentrations of Pb (100 and 200 μM). Antioxidant activity was either higher or unchanged in the infected seedlings due to responses to the different Pb concentrations. These results suggest that the endophytic fungus improved rice growth under moderate Pb levels by enhancing photosynthesis and antioxidant activity relative to non-infected rice. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Endophytic Fungi from Frankincense Tree Improves Host Growth and Produces Extracellular Enzymes and Indole Acetic Acid.

    Khan, Abdul Latif; Al-Harrasi, Ahmed; Al-Rawahi, Ahmed; Al-Farsi, Zainab; Al-Mamari, Aza; Waqas, Muhammad; Asaf, Sajjad; Elyassi, Ali; Mabood, Fazal; Shin, Jae-Ho; Lee, In-Jung

    2016-01-01

    Boswellia sacra, an economically important frankincense-producing tree found in the desert woodlands of Oman, is least known for its endophytic fungal diversity and the potential of these fungi to produce extracellular enzymes and auxins. We isolated various fungal endophytes belonging to Eurotiales (11.8%), Chaetomiaceae (17.6%), Incertae sadis (29.5%), Aureobasidiaceae (17.6%), Nectriaceae (5.9%) and Sporomiaceae (17.6%) from the phylloplane (leaf) and caulosphere (stem) of the tree. Endophytes were identified using genomic DNA extraction, PCR amplification and sequencing the internal transcribed spacer regions, whereas a detailed phylogenetic analysis of the same gene fragment was made with homologous sequences. The endophytic colonization rate was significantly higher in the leaf (5.33%) than the stem (0.262%). The Shannon-Weiner diversity index was H' 0.8729, while Simpson index was higher in the leaf (0.583) than in the stem (0.416). Regarding the endophytic fungi's potential for extracellular enzyme production, fluorogenic 4-methylumbelliferone standards and substrates were used to determine the presence of cellulases, phosphatases and glucosidases in the pure culture. Among fungal strains, Penicillum citrinum BSL17 showed significantly higher amounts of glucosidases (62.15±1.8 μM-1min-1mL) and cellulases (62.11±1.6 μM-1min-1mL), whereas Preussia sp. BSL10 showed significantly higher secretion of glucosidases (69.4±0.79 μM-1min-1mL) and phosphatases (3.46±0.31μM-1min-1mL) compared to other strains. Aureobasidium sp. BSS6 and Preussia sp. BSL10 showed significantly higher potential for indole acetic acid production (tryptophan-dependent and independent pathways). Preussia sp. BSL10 was applied to the host B. sacra tree saplings, which exhibited significant improvements in plant growth parameters and accumulation of photosynthetic pigments. The current study concluded that endophytic microbial resources producing extracellular enzymes and auxin could

  13. Endophytic Fungi from Frankincense Tree Improves Host Growth and Produces Extracellular Enzymes and Indole Acetic Acid.

    Abdul Latif Khan

    Full Text Available Boswellia sacra, an economically important frankincense-producing tree found in the desert woodlands of Oman, is least known for its endophytic fungal diversity and the potential of these fungi to produce extracellular enzymes and auxins. We isolated various fungal endophytes belonging to Eurotiales (11.8%, Chaetomiaceae (17.6%, Incertae sadis (29.5%, Aureobasidiaceae (17.6%, Nectriaceae (5.9% and Sporomiaceae (17.6% from the phylloplane (leaf and caulosphere (stem of the tree. Endophytes were identified using genomic DNA extraction, PCR amplification and sequencing the internal transcribed spacer regions, whereas a detailed phylogenetic analysis of the same gene fragment was made with homologous sequences. The endophytic colonization rate was significantly higher in the leaf (5.33% than the stem (0.262%. The Shannon-Weiner diversity index was H' 0.8729, while Simpson index was higher in the leaf (0.583 than in the stem (0.416. Regarding the endophytic fungi's potential for extracellular enzyme production, fluorogenic 4-methylumbelliferone standards and substrates were used to determine the presence of cellulases, phosphatases and glucosidases in the pure culture. Among fungal strains, Penicillum citrinum BSL17 showed significantly higher amounts of glucosidases (62.15±1.8 μM-1min-1mL and cellulases (62.11±1.6 μM-1min-1mL, whereas Preussia sp. BSL10 showed significantly higher secretion of glucosidases (69.4±0.79 μM-1min-1mL and phosphatases (3.46±0.31μM-1min-1mL compared to other strains. Aureobasidium sp. BSS6 and Preussia sp. BSL10 showed significantly higher potential for indole acetic acid production (tryptophan-dependent and independent pathways. Preussia sp. BSL10 was applied to the host B. sacra tree saplings, which exhibited significant improvements in plant growth parameters and accumulation of photosynthetic pigments. The current study concluded that endophytic microbial resources producing extracellular enzymes and auxin

  14. Plant Bioactive Metabolites and Drugs Produced by Endophytic Fungi of Spermatophyta

    Rosario Nicoletti

    2015-09-01

    Full Text Available It is known that plant-based ethnomedicine represented the foundation of modern pharmacology and that many pharmaceuticals are derived from compounds occurring in plant extracts. This track still stimulates a worldwide investigational activity aimed at identifying novel bioactive products of plant origin. However, the discovery that endophytic fungi are able to produce many plant-derived drugs has disclosed new horizons for their availability and production on a large scale by the pharmaceutical industry. In fact, following the path traced by the blockbuster drug taxol, an increasing number of valuable compounds originally characterized as secondary metabolites of plant species belonging to the Spermatophyta have been reported as fermentation products of endophytic fungal strains. Aspects concerning sources and bioactive properties of these compounds are reviewed in this paper.

  15. Endophytic bacteria improve phytoremediation of Ni and TCE co-contamination.

    Weyens, Nele; Croes, Sarah; Dupae, Joke; Newman, Lee; van der Lelie, Daniel; Carleer, Robert; Vangronsveld, Jaco

    2010-07-01

    The aim of this work was to investigate if engineered endophytes can improve phytoremediation of co-contaminations by organic pollutants and toxic metals. As a model system, yellow lupine was inoculated with the endophyte Burkholderia cepacia VM1468 possessing (a) the pTOM-Bu61 plasmid, coding for constitutive trichloroethylene (TCE) degradation, and (b) the ncc-nre Ni resistance/sequestration system. Plants were exposed to Ni and TCE and (a) Ni and TCE phytotoxicity, (b) TCE degradation and evapotranspiration, and (c) Ni concentrations in the roots and shoots were determined. Inoculation with B. cepacia VM1468 resulted in decreased Ni and TCE phytotoxicity, as measured by 30% increased root biomass and up to 50% decreased activities of enzymes involved in anti-oxidative defence in the roots. In addition, TCE evapotranspiration showed a decreasing trend and a 5 times higher Ni uptake was observed after inoculation. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  16. Molecular characterisation of fungal endophytic morphospecies associated with the indigenous forest tree, Theobroma gileri, in Ecuador.

    Thomas, Sarah E; Crozier, Jayne; Catherine Aime, M; Evans, Harry C; Holmes, Keith A

    2008-07-01

    Fungal endophytes were isolated from healthy stems and pods of Theobroma gileri, an alternative host of the frosty pod rot pathogen of cacao. Non-sporulating isolates were grouped into 46 different morphological species according to their colony morphology. Many of these morphospecies were assumed to be basidiomycetes and, therefore, were of particular interest. Basidiomycetous endophytes have received far less attention than ascomycetes and also have potential as biological control agents of the basidiomycetous pathogens of T. cacao: Moniliophthora roreri (frosty pod rot pathogen) and M. perniciosa (witches' broom disease). The morphospecies were further characterised by molecular analyses. Amplification of the nuLSU was undertaken for phylogenetic placement of these non-sporulating cultures and revealed a total of 31 different taxa of which 15 were basidiomycetes belonging to the class Agaricomycetes, and 16 ascomycetes primarily belonging to the Sordariomycetes.

  17. Amylase production by endophytic fungi Cylindrocephalum sp. isolated from medicinal plant Alpinia calcarata (Haw. Roscoe

    V. H. Sunitha.

    2012-09-01

    Full Text Available Amylases are among the most important enzymes used in modern biotechnology particularly in the process involving starch hydrolysis. Fungal amylase has large applications in food and pharmaceutical industries. Considering these facts, endophytic fungi isolated from the plant Alpinia calcarata (Haw. Roscoe were screened for amylolytic activity on glucose yeast extract peptone agar (GYP medium. Among thirty isolates of endophytic fungi, isolate number seven identified as Cylindrocephalum sp. (Ac-7 showed highest amylolytic activity and was taken for further study. Influence of various physical and chemical factors such as pH, temperature, carbon and nitrogen sources on amylase production in liquid media were studied. The maximal amylase production was found to be at 30ºC and at pH 7.0 of the growth medium. Among the various carbon and nitrogen sources tested, maltose at 1.5% and Sodium nitrate at 0.3% respectively gave optimum amylase production.

  18. Stelliosphaerols A and B, Sesquiterpene-Polyol Conjugates from an Ecuadorian Fungal Endophyte.

    Forcina, Giovanni C; Castro, Amaya; Bokesch, Heidi R; Spakowicz, Daniel J; Legaspi, Michelle E; Kucera, Kaury; Villota, Stephany; Narváez-Trujillo, Alexandra; McMahon, James B; Gustafson, Kirk R; Strobel, Scott A

    2015-12-24

    Endophytic fungi are plant tissue-associated fungi that represent a rich resource of unexplored biological and chemical diversity. As part of an ongoing effort to characterize Amazon rainforest-derived endophytes, numerous fungi were isolated and cultured from plants collected in the Yasuní National Park in Ecuador. Of these samples, phylogenetic and morphological data revealed a previously undescribed fungus in the order Pleosporales that was cultured from the tropical tree Duroia hirsuta. Extracts from this fungal isolate displayed activity against Staphylococcus aureus and were thus subjected to detailed chemical studies. Two compounds with modest antibacterial activity were isolated, and their structures were elucidated using a combination of NMR spectroscopic analysis, LC-MS studies, and chemical degradation. These efforts led to the identification of stelliosphaerols A (1) and B (2), new sesquiterpene-polyol conjugates that are responsible, at least in part, for the S. aureus inhibitory activity of the fungal extract.

  19. Metagenome analysis of the root endophytic microbial community of Indian rice (O. sativa L.

    Subhadipa Sengupta

    2017-06-01

    Full Text Available This study reports the root endophytic microbial community profile in rice (Oryza sativa L., the largest food crop of Asia, using 16S rRNA gene amplicon sequencing. Metagenome of OS01 and OS04 consisted of 11,17,900 sequences with 300 Mbp size and average 55.6% G + C content. Data of this study are available at NCBI Bioproject (PRJNA360379. The taxonomic analysis of 843 OTU's showed that the sequences belonged to four major phyla revealing dominance of Proteobacteria, Firmicutes, Cyanobacteria and Actinobacteria. Results reveal the dominance of Bacillus as major endophytic genera in rice roots, probably playing a key role in Nitrogen fixation.

  20. Larvicidal potential of Asteraceae family endophytic actinomycetes against Culex quinquefasciatus mosquito larvae.

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2014-01-01

    Pakistan is blessed with plants of Asteraceae family with known medicinal background used for centuries by Hakims (traditional physicians). Keeping in mind the background of their anti-larval potential, a total of 21 endophytic actinomycetes were isolated from four Asteraceae plants and screened against the first and fourth instar stages of Culex quinquefasciatus Say mosquito larvae. Of the 21 isolates, 6 of them gave strong larvicidal activity (80-100% mortality) in the screening results and 4 isolates gave a potent larvicidal activity (100% mortality) at the fourth instar stage. These isolates belonged to different species within the actinomycetes group, namely Streptomyces albovinaceus and Streptomyces badius. This communication reports the larvicidal potential of endophytic actinomycetes residing within the native Asteraceae plants in Pakistan. The study suggests further exploration through large-scale productions leading to the identification of the larvicidal compounds.

  1. Bioactive secondary metabolites from the endophytic fungus Chaetomium sp. isolated from Salvia officinalis growing in Morocco

    Ebel R.

    2009-01-01

    Full Text Available This study reports the chemical investigation and cytotoxic activity of the secondary metabolites produced by the endophytic fungus Chaetomium sp. isolated from Salvia officinalis growing in Morocco. This plant was collected from the Beni-Mellal Mountain in Morocco and belongs to the Lamiaceae family and is named in Morocco “Salmia”. The endophytic fungus Chaetomium sp. was isolated from the tissues of the stem of this plant. The fungal strain was identified by PCR. The crude organic extract of the fungal strain was proven to be active when tested for cytotoxicity against L5178Y mouse lymphoma cells. Chemical investigation of the secondary metabolites showed that cochliodinol is the main component beside isocochliodinol. The structures of the isolated compounds were determined on the basis of NMR analysis (1H, 13C, COSY and HMBC as well as by mass spectrometry using ESI (Electron Spray Ionisation as source.

  2. Laccase production by Monotospora sp., an endophytic fungus in Cynodon dactylon.

    Wang, J W; Wu, J H; Huang, W Y; Tan, R X

    2006-03-01

    The effects of the carbon and nitrogen sources, initial pH and incubation temperature on laccase production by the endophytic fungus Monotospora sp. were evaluated. The optimal temperature and initial pH for laccase production by Monotospora sp. in submerged culture were found to be 30 degrees C and 8.5, respectively. Maltose (2 g l(-1)) and ammonium tartrate (10 g l(-1)) were the most suitable carbon and nitrogen source for laccase production. Under optimal culture medium, the maximum laccase activity was determined to be 13.55 U ml(-1), which was approximately four times higher than that in basal medium. This is the first report on laccase production by an endophytic fungus.

  3. Huperzine A production by Paecilomyces tenuis YS-13, an endophytic fungus isolated from Huperzia serrata.

    Su, Jingqian; Yang, Minhe

    2015-01-01

    Huperzine A (HupA), a naturally occurring alkaloid in the plant family Huperziaceae, has drawn great interest for its potential application in Alzheimer disease therapy. Our primary objective was to identify alkaloid- and HupA-producing fungi from the Chinese folk herb, Huperzia serrata. We established a rapid and efficient model for screening HupA-producing endophytic fungal strains. The presence of HupA in Paecilomyces tenuis YS-13 was analysed by thin-layer chromatography, high-performance liquid chromatography and mass spectrometry. The fermentation yield of HupA was 21.0 μg/L, and the IC50 of the crude extract of YS-13 fermentation broth was 1.27 ± 0.04 mg/mL. This is the first report of P. tenuis as a HupA-producing endophyte isolated from Huperziaceae.

  4. Bacillus subtilis and Enterobacter cloacae endophytes from healthy Theobroma cacao L. trees can systemically colonize seedlings and promote growth.

    Leite, Hianna Almeida Câmara; Silva, Anderson Barbosa; Gomes, Fábio Pinto; Gramacho, Karina Peres; Faria, José Cláudio; de Souza, Jorge Teodoro; Loguercio, Leandro Lopes

    2013-03-01

    Clonal genotypes resistant to fungal diseases are an important component of the cocoa production system in southeastern Bahia state (Brazil), so that technologies for faster production of stronger and healthier plantlets are highly desirable. In this study, the effects of inoculated bacterial endophytes isolated from healthy adult cacao plants on seedlings, and aspects related to inoculation methods, colonization patterns, and photosynthesis were investigated. Sequencing of 16S rRNA, hsp-60, and rpo-B genes placed the wild-type isolates within the species Enterobacter cloacae (isolates 341 and 344) and Bacillus subtilis (isolate 629). Spontaneous rifampicin-resistant (rif(R)) variants for 344 were also produced and tested. Endophytic application was either by immersion of surface sterilized seeds in bacterial suspensions or direct inoculation into soil, 20 days after planting non-inoculated seeds into pots. Results from in vitro recovery of inoculated isolates showed that the wild-type endophytes and rif(R) variants systemically colonized the entire cacao seedlings in 15-20 days, regardless of the inoculation method. Some endophytic treatments showed significant increases in seedlings' height, number of leaves, and dry matter. Inoculation methods affected the combined application of endophytes, which maintained the growth-promotion effects, but not in the same manner as in single applications. Interestingly, the 344-3.2 rif(R) variant showed improved performance in relation to both the wild type and another related variant. Photosynthetic rates and stomatal conductance increased significantly for some endophytic treatments, being partially associated with effects on growth and affected by the inoculation method. The results suggest that E. cloacae and B. subtilis endophytes from healthy adult plants (not transmitted by seeds) were able to promote vegetative growth on cacao seedlings. The development of products for large-scale use in seedlings

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

    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

  6. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09

    Cai, Xunchao; Kang, Xingxing; Xi, Huan; Liu, Changhong; Xue, Yarong

    2016-01-01

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B.?velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and vo...

  7. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09.

    Cai, Xunchao; Kang, Xingxing; Xi, Huan; Liu, Changhong; Xue, Yarong

    2016-09-29

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome. Copyright © 2016 Cai et al.

  8. Genome Sequence of the Plant Growth Promoting Endophytic Bacterium Enterobacter sp. 638

    Taghavi, Safiyh; van der Lelie, Daniel; Hoffman, Adam; Zhang, Yian-Biao; Walla, Michael D.; Vangronsveld, Jaco; Newman, Lee; Monchy, Sébastien

    2010-01-01

    Enterobacter sp. 638 is an endophytic plant growth promoting gamma-proteobacterium that was isolated from the stem of poplar (Populus trichocarpa×deltoides cv. H11-11), a potentially important biofuel feed stock plant. The Enterobacter sp. 638 genome sequence reveals the presence of a 4,518,712 bp chromosome and a 157,749 bp plasmid (pENT638-1). Genome annotation and comparative genomics allowed the identification of an extended set of genes specific to the plant niche adaptation of this bacterium. This includes genes that code for putative proteins involved in survival in the rhizosphere (to cope with oxidative stress or uptake of nutrients released by plant roots), root adhesion (pili, adhesion, hemagglutinin, cellulose biosynthesis), colonization/establishment inside the plant (chemiotaxis, flagella, cellobiose phosphorylase), plant protection against fungal and bacterial infections (siderophore production and synthesis of the antimicrobial compounds 4-hydroxybenzoate and 2-phenylethanol), and improved poplar growth and development through the production of the phytohormones indole acetic acid, acetoin, and 2,3-butanediol. Metabolite analysis confirmed by quantitative RT–PCR showed that, the production of acetoin and 2,3-butanediol is induced by the presence of sucrose in the growth medium. Interestingly, both the genetic determinants required for sucrose metabolism and the synthesis of acetoin and 2,3-butanediol are clustered on a genomic island. These findings point to a close interaction between Enterobacter sp. 638 and its poplar host, where the availability of sucrose, a major plant sugar, affects the synthesis of plant growth promoting phytohormones by the endophytic bacterium. The availability of the genome sequence, combined with metabolome and transcriptome analysis, will provide a better understanding of the synergistic interactions between poplar and its growth promoting endophyte Enterobacter sp. 638. This information can be further exploited to

  9. Impacts of endophyte infection of ryegrass on rhizosphere metabolome and microbial community

    Wakelin, S.; Harrison, Scott James; Mander, C.

    2015-01-01

    37, within a genetically uniform breeding line of perennial ryegrass (Lolium perenne cv. Samson 11104) on the rhizosphere metabolome and the composition of the fungal, bacterial, and Pseudomonas communities. There were strong differences in the rhizosphere metabolomes between infested and non......-infested ryegrass strains (P=0.06). These were attributed to shifts in various n-alkane hydrocarbon compounds. The endophyte-associated alteration in rhizosphere metabolome was linked to changes in the total bacterial (P

  10. Genome Sequence of an Endophytic Fungus, Fusarium solani JS-169, Which Has Antifungal Activity.

    Kim, Jung A; Jeon, Jongbum; Park, Sook-Young; Kim, Ki-Tae; Choi, Gobong; Lee, Hyun-Jung; Kim, Yangsun; Yang, Hee-Sun; Yeo, Joo-Hong; Lee, Yong-Hwan; Kim, Soonok

    2017-10-19

    An endophytic fungus, Fusarium solani strain JS-169, isolated from a mulberry twig, showed considerable antifungal activity. Here, we report the draft genome sequence of this strain. The assembly comprises 17 scaffolds, with an N 50 value of 4.93 Mb. The assembled genome was 45,813,297 bp in length, with a G+C content of 49.91%. Copyright © 2017 Kim et al.

  11. Anti-mycobacterial activity of polyketides from Penicillium sp. endophyte isolated from Garcinia nobilis against Mycobacteriumsmegmatis.

    Jouda, Jean Bosco; Mawabo, Isabelle Kamga; Notedji, Augustin; Mbazoa, Céline Djama; Nkenfou, Jean; Wandji, Jean; Nkenfou, Céline Nguefeu

    2016-06-01

    According to estimates by the World Health Organization, there were 9.6 million new tuberculosis (TB) cases in 2014: 5.4 million among men, 3.2 million among women, and 1.0 million among children. There were also 1.5 million TB deaths. Although there are potent anti-TB molecules, the misuse of these drugs in addition to inconsistent or partial treatment have led to the development of multidrug-resistant TB and extensively drug-resistant TB. It is established that plants harbor microorganisms, collectively known as endophytes, which also produce metabolites. Exploring the as-yet untapped natural products from the endophytes increases the chances of finding novel and active compounds. The present study was aimed to investigate the antimycobacterial activity of the crude extract and compounds isolated from Penicillium sp. endophyte associated with Garcinia nobilis against Mycobacterium smegmatis. Liquid culture obtained from the fermentation of Penicillium sp. was extracted using ethylacetate and the liquid chromatography-mass spectrometry monitored fractionation of crude extracts yielded six compounds. Their structures were elucidated with spectroscopic analyses including two-dimensional nuclear magnetic resonance, high resolution mass spectrometry by dereplication using Antibase, and by comparison to literature data. All compounds and the crude extract from the liquid medium were evaluated for their antimycobacterial activity against M. smegmatis. In this study, the activity of penialidins A-C (1-3), citromycetin (4), p-hydroxy phenyl glyoxalaldoxime (5), and Brefeldin A (6) were tested against nonpathogenic M. smegmatis. Penialidin C was the most active compound with a minimum inhibitory concentration of 15.6μg/mL. Isolated compounds from Penicillium sp. harbored in G. nobilis exhibited promising antimycobacterial activity against M. smegmatis thus supporting the immensity of the potential of antimycobacterial drug discovery from endophytes from medicinal plants

  12. Ocurrence and diversity of endophytic colonization of taraxacum officinale by botrytis species: a preliminary study

    Yánez Navarrete, Zoila Elizabeth

    2014-01-01

    The genus Botrytis (sexual form Botryotinia Whetzel) contains renowned plant pathogenic species with a necrotrophic lifestyle in which kill and feed on its hosts. Recent reports of symptomless endophytic colonization by Botrytis cinerea in cultivated plants of lettuce (Lactuca sativa) and Botrytis spp. in the wild plants dandelion (Taraxacum agg) and spotted knapweed (Centaura stoebe) reveal a different “side” of Botrytis spp. These findings, expose additional strategies of Botrytis species t...

  13. Genetic Diversity of Nostoc Symbionts Endophytically Associated with Two Bryophyte Species

    Costa, José-Luis; Paulsrud, Per; Rikkinen, Jouko; Lindblad, Peter

    2001-01-01

    The diversity of the endophytic Nostoc symbionts of two thalloid bryophytes, the hornwort Anthoceros fusiformis and the liverwort Blasia pusilla, was examined using the tRNALeu (UAA) intron sequence as a marker. The results confirmed that many different Nostoc strains are involved in both associations under natural conditions in the field. The level of Nostoc diversity within individual bryophyte thalli varied, but single DNA fragments were consistently amplified from individual symbiotic col...

  14. The Endophytic Symbiont—Pseudomonas aeruginosa Stimulates the Antioxidant Activity and Growth of Achyranthes aspera L.

    Khaidem A. Devi

    2017-09-01

    Full Text Available A plant growth promoting bacterial endophyte designated as AL2-14B isolated from the leaves of Achyranthes aspera L. was identified as Pseudomonas aeruginosa based on its phenotypic and physiological features, and 16S rRNA gene sequence analysis. AL2-14B had plant growth stimulating attributes including siderophore and indole acetic acid release, inorganic phosphate solubilization, along with nitrogenase, ammonification, and protease activities. It also exhibited antifungal property against Rhizoctonia solani. The plantlets grown in germ-free condition were inoculated with AL2-14B and studied for the colonization of endophyte. Significant increase in population of AL2-14B between 3rd and 5th days after inoculation was recorded. The treatment of plants with endophytic P. aeruginosa AL2-14B increased nitrogen, phosphorus, potassium (NPK contents in plant by 3.8, 12.59, and 19.15%, respectively. Significant enhancement of shoot and root length, dry leaf, dry shoot and dry root weight, and leaf surface area as compared to control (P < 0.05 was recorded in AL2-14B inoculated plants. The antioxidant activities increased in plants grown in germ-free conditions and inoculated with AL2-14B. The present study emphasizes on the role of diazotrophic endophyte P. aeruginosa AL2-14B in stimulating growth of A. aspera L. and improvement of its medicinal properties. Significant increase in growth and antioxidant content of P. aeruginosa AL2-14B treated plants suggests the possibility of an economical and eco-friendly mean of achieving antioxidants rich, healthier A. aspera plants.

  15. Isolation and enzyme bioprospection of endophytic bacteria associated with plants of Brazilian mangrove ecosystem

    Castro, Renata A; Quecine, Maria Carolina; Lacava, Paulo T; Batista, Bruna D; Luvizotto, Danice M; Marcon, Joelma; Ferreira, Anderson; Melo, Itamar S; Azevedo, João L

    2014-01-01

    The mangrove ecosystem is a coastal tropical biome located in the transition zone between land and sea that is characterized by periodic flooding, which confers unique and specific environmental conditions on this biome. In these ecosystems, the vegetation is dominated by a particular group of plant species that provide a unique environment harboring diverse groups of microorganisms, including the endophytic microorganisms that are the focus of this study. Because of their intimate associatio...

  16. A new biphenyl derivative from the mangrove endophytic fungus Phomopsis longicolla HL-2232.

    Li, Xiao-Bao; Chen, Guang-Ying; Liu, Rui-Jie; Zheng, Cai-Juan; Song, Xin-Ming; Han, Chang-Ri

    2017-10-01

    A new biphenyl derivative 5,5'-dimethoxybiphenyl-2,2'-diol (1), together with five known compounds (2-5), was isolated from the mangrove endophytic fungus Phomopsis longicolla HL-2232. The structures of these compounds were elucidated using comprehensive spectroscopic methods. The absolute configuration of 4 was determined by single-crystal X-ray diffraction for the first time. The inhibitory activities of all compounds against two Vibrio bacteria were evaluated.

  17. Novel anthraquinone derivatives produced by Pestalotiopsis guepinii, an endophytic of the medicinal plant Virola michelii (Myristicaceae)

    Oliveira, Marilene N.; Santos, Lourivaldo S.; Guilhon, Giselle M.S.P; Santos, Alberdan S.; Ferreira, Isabel C.S.; Lopes-Junior, Manoel L.; Arruda, Mara Silvia P.; Marinho, Andrey M.R.; Silva, Milton N. da, E-mail: lss@ufpa.b [Universidade Federal do Para (UFPA), Belem, PA (Brazil). Inst. de Ciencias Exatas e Naturais. Programa de Pos-Graduacao em Quimica; Rodrigues-Filho, Edson [Universidade Federal de Sao Carlos (DQ/UFSCar), SP (Brazil). Dept. de Quimica; Oliveira, Maria C.F. [Universidade Federal do Ceara (UFC), Fortaleza (Brazil). Dept. de Quimica Organica e Inorganica

    2011-07-01

    A new anthraquinone derivative, named guepinone (1), along with the known substances isosulochrin (2) and chloroisosulochrin (3), were isolated from a rice culture of Pestalotiopsis guepinii, an endophytic fungus of Virola michelii. The compounds were identified by analysis of 1D and 2D NMR and MS spectral data. The antimicrobial activity of these compounds was evaluated and chloroisosulchrin (3) was the most active. (author)

  18. Diversity of endophytic fungal and bacterial communities in Ilex paraguariensis grown under field conditions.

    Pérez, María Laura; Collavino, Mónica Mariana; Sansberro, Pedro Alfonso; Mroginski, Luis Amado; Galdeano, Ernestina

    2016-04-01

    The composition and diversity of the endophytic community associated with yerba mate (Ilex paraguariensis) was investigated using culture-depending methods. Fungi were identified based on their micromorphological characteristics and internal transcribed spacer rDNA sequence analysis; for bacteria 16S rDNA sequence analysis was used. Fungal and bacterial diversity did not show significant differences between organ age. The highest fungal diversity was registered during fall season and the lowest in winter. Bacterial diversity was higher in stems and increased from summer to winter, in contrast with leaves, which decreased. The most frequently isolated fungus was Fusarium, followed by Colletotrichum; they were both present in all the sampling seasons and organ types assayed. Actinobacteria represented 57.5 % of all bacterial isolates. The most dominant bacterial taxa were Curtobacterium and Microbacterium. Other bacteria frequently found were Methylobacterium, Sphingomonas, Herbiconiux and Bacillus. Nitrogen fixation and phosphate solubilization activity, ACC deaminase production and antagonism against plant fungal pathogens were assayed in endophytic bacterial strains. In the case of fungi, strains of Trichoderma, Penicillium and Aspergillus were assayed for antagonism against pathogenic Fusarium sp. All microbial isolates assayed showed at least one growth promoting activity. Strains of Bacillus, Pantoea, Curtobacterium, Methylobacterium, Brevundimonas and Paenibacillus had at least two growth-promoting activities, and Bacillus, Paenibacillus and the three endophytic fungi showed high antagonistic activity against Fusarium sp. In this work we have made a wide study of the culturable endophytic community within yerba mate plants and found that several microbial isolates could be considered as potential inoculants useful for improving yerba mate production.

  19. Potency of six isolates of biocontrol agents endophytic Trichoderma against fusarium wilt on banana

    J Taribuka

    2017-01-01

    Full Text Available Fusarium wilt caused by F. oxysporum f.sp. cubense is one of very damaging banana plant diseases which can cause plant death. Disease control using intensive chemical fungicides will have negative impacts on the environment and humans. Endophytic Trichoderma is one of the biological control agents which can reduce the amount of inoculum of pathogens, so it can reduce disease intensity. The objectives of this study was to assess the ability of endophytic Trichoderma in inducing plant resistance against fusarium wilt. Endophytic Trichoderma was obtained from healthy roots of banana from three regencies in Yogyakarta, namely Trichoderma harzianum.swn-1, T. harzianum.swn-2, T. harzianum.psr-1, T. asperrellum, T. gamsii, and T. koningiopsis. Research on induced resintance was conducted in the greenhouse with polybag using Completely Randomized Design with 14 treatments and 3 replications. The results showed that the ability of Trichoderma gamsii antagonism against F. oxysporum f.sp. cubense was 60.61%. T. asperellum and T. harzianum.swn-2 could suppress this disease resulted in disease intensity of 8.33% which categorize as resistant. Trichoderma harzianum.psr-1 was significantly different in stimulating plant vegetative growth. Induced resistance by using endophytic Trichoderma spp. against  F. oxysporum f.sp. cubense showed increase in total phenolic compounds on the third and fourth weeks as well as peroxidase activity on the third, fourth and fifth weeks.  Observation of lignification on  the fifth week  showed that lignification occurred in root xylem

  20. [Diversity of Bacillus species inhabiting on the surface and endophyte of lichens collected from Wuyi Mountain].

    Ge, Cibin; Liu, Bo; Che, Jianmei; Chen, Meichun; Liu, Guohong; Wei, Jiangchun

    2015-05-04

    The present work reported the isolation, identification and diversity of Bacillus species colonizing on the surface and endophyte in lichens collected from Wuyi Mountain. Nine lichen samples of Evernia, Stereocaulon, Menegazzia and other 6 genera belonging to 7 families were collected from Wuyi mountain nature reserve. The bacillus-like species colonizing on the surface and endophyte in these lichens were isolated and identified by 16S rRNA gene sequence analysis. There was no bacillus-like species isolated from Evernia, Ramalina and Lecarona. A total of 34 bacillus-like bacteria were isolated from another 6 lichen samples. These bacteria were identified as 24 species and were classified into Bacillus, Paenibacillus, Brevibacillus, Lysinibacillus and Viridiibacillus. Paenibacillus and Bacillus are the dominant genera, and accounting for 41. 2% and 35. 3% of all isolated bacteria respectively. Brevibacillus, Lysinibacillus and Viridiibacillu were first reported being isolated from lichens. There were different species and quantity of bacillus colonizing on the surface and endophyte in different lichens. The quantity of bacillus colonizing on the surface of Physcia was more than 3.85 x 10(6) cfu/g and was the largest in the isolated bacteria, while the species of bacillus colonizing on the surface and endophyte in Stereocaulon was the most abundant. Most of the isolated bacteria were colonizing on (in) one lichen genera, but Paenibacillus taichungensis, Paenibacillus odorifer, Brevibacillus agri, Lysinibacillus xylanilyticus was respectively colonizing on (in) 2-3 lichen genera and Bacillus mycoides was colonizing on (in) Menegazzia, Cladonia Physcia, and Stereocaulon. There are species and quantity diversity of bacillus colonizing on (in) lichens.

  1. Identification of Endophytic Fungi of Medicinal Herbs of Lauraceae and Rutaceae with Antimicrobial Property

    Min-Yuan Ho

    2012-09-01

    Full Text Available This study was conducted to determine taxonomical features and antimicrobial activities of 156 isolates of endophytic fungi collected from twigs of medicinal plants of Lauraceae (67 isolates and Rutaceae (89 isolates in central and northern Taiwan. The 156 isolates of fungi were classified into 35 genera in 19 families based on morphological characteristics of mycelia and asexual/sexual spores, as well as molecular phylogenetic analysis of rDNA LSU D1/D2 and ITS regions. The most common endophytes were in the taxa of Colletotrichum, Guignardia, Hypoxylon, Nigrospora, Phomopsis and Xylaria, and the most common hosts were Citrus and Zanthoxylum of Rutaceae and Cinnamomum of Lauraceae. Molecular phylogenetic analysis showed that xylariaceous isolates could be separated into Xylaria and Hypoxylon groups based on rDNA of LSU D1/D2 and ITS regions. Four isolates of endophytic fungi including Lasmenia sp. isolate CB10, Ophioceras tenuisporum isolate CI02, Xylaria cubensis isolate LA04 and Cyanodermella sp. isolate TR09 were tested for antimicrobial activities using a dual culture method and Lasmenia sp. isolate CB10 and Cyanodermella sp. isolate TR09 showed better antimicrobial activity against 12 plant pathogens including 9 fungi and 3 bacteria. Spraying Chinese cabbage (Brassica rapa plants with culture filtrates of the endophytic fungus Lasmenia sp. isolate CB10 significantly reduced severity of anthracnose of Chinese cabbage caused by Colletotrichum higginsianum under greenhouse conditions. This study suggests that the Lasmenia sp. isolate CB10 may be of potential for management of anthracnose of Chinese cabbage.

  2. A New Terminal Cyano Group-containing Benzodiazepine Alkaloid from the Mangrove Endophytic Fungus Penicillium sp. .

    Li, Jing; Zhong, Yi-sheng; Yuan, Jie; Zhu, Xun; Lu, Yong-jun; Lin, Yong-cheng; Liu, Lan

    2015-09-01

    A new benzodiazepine alkaloid containing terminal cyano group has been isolated from a mangrove endophytic fungus, Penicillium 299#. Structure elucidation was determined by 1D and 2D NMR spectroscopy and the absolute configuration was determined by electronic circular dichroism (ECD). The new compound showed no cytotoxic activities in vitro against human cancer lines MDA-MB-435, HepG2, HCT-116, and Calu-3.

  3. Metabolites of the endophytic fungus Penicillium sp. FJ-1 of Acanthus ilicifolius.

    Liu, Jian-Fang; Chen, Wei-Jie; Xin, Ben-Ru; Lu, Jie

    2014-06-01

    Two new compounds, named as (2R,3S)-pinobanksin-3-cinnamate (1), and 15alpha-hydroxy-(22E,24R)-ergosta-3,5,8(14),22-tetraen-7-one (2), were isolated from the endophytic fungus Penicillium sp. FJ-1 of Acanthus ilicifolius Linn. Their structures were elucidated on the basis of spectroscopic analysis. Additionally, compound 1 exhibited potent neuroprotective effects on corticosterone-damaged PC12 cells, and compound 2 showed potent cytotoxicity on glioma cell lines.

  4. Polyketides with Immunosuppressive Activities from Mangrove Endophytic Fungus Penicillium sp. ZJ-SY₂.

    Liu, Hongju; Chen, Senhua; Liu, Weiyang; Liu, Yayue; Huang, Xishan; She, Zhigang

    2016-11-25

    Nine polyketides, including two new benzophenone derivatives, peniphenone ( 1 ) and methyl peniphenone ( 2 ), along with seven known xanthones ( 3 - 9 ) were obtained from mangrove endophytic fungus Penicillium sp. ZJ-SY₂ isolated from the leaves of Sonneratia apetala . Their structures were elucidated on the basis of MS, 1D, and 2D NMR data. Compounds 1 , 3 , 5 , and 7 showed potent immunosuppressive activity with IC 50 values ranging from 5.9 to 9.3 μg/mL.

  5. New Eudesmane-Type Sesquiterpenoids from the Mangrove-Derived Endophytic Fungus Penicillium sp. J-54

    Liuming Qiu

    2018-03-01

    Full Text Available Four new eudesmane-type sesquiterpenoids, penicieudesmol A–D (1–4, were isolated from the fermentation broth of the mangrove-derived endophytic fungus Penicillium sp. J-54. Their structures were determined by spectroscopic methods, the in situ dimolybdenum CD method, and modified Mosher’s method. The bioassays results showed that 2 exhibited weak cytotoxicity against K-562 cells.

  6. Endophytic Streptomyces spp. as Biocontrol Agents of Rice Bacterial Leaf Blight Pathogen (Xanthomonas oryzae pv. oryzae

    RATIH DEWI HASTUTI

    2012-12-01

    Full Text Available Xanthomonas oryzae pv. oryzae (Xoo, a causal agent of bacterial leaf blight (BLB, is one of the most important pathogens of rice. The effectiveness of ten Streptomyces spp. isolates in suppressing Xoo disease was assessed in planta and in vitro. In planta experiments were carried out in a greenhouse and arranged in a randomized completely block design (RCBD with three replications. Twenty treatments were tested which included plants inoculated with both Streptomyces spp. and Xoo, and plants inoculated with only Streptomyces spp. Plants inoculated with Xoo and sprayed with a chemical bactericide, and plants inoculated with only Xoo served as positive controls, whereas plants not inoculated with either Streptomyces spp. or Xoo were used as negative controls. The results showed that the effect of endophytic Streptomyces spp. on BLB disease expressed as area under disease progress curve (AUDPC was not significantly different to that on control plants (P > 0.05. However, plants inoculated with endophytic Streptomyces spp. were significantly taller and produced higher tiller number than control plants (P < 0.05. Streptomyces spp. isolate AB131-1 gave the highest plant height. In vitro studies on biocontrol mechanisms of selected Streptomyces spp. isolates showed that isolate LBR02 gave the highest inhibition activity on Xoo growth, followed by AB131-1 and AB131-2. Two isolates (AB131-1 and LBR02 were able to produce chitinase, phosphatase, and siderophore which included biocontrol characteristics. Morphological and colonization studies under SEM and light microscopy confirmed that the three isolates were endophytic Streptomyces spp. from different species. These studies found that the paddy plant which was inoculated with endophytic Streptomyces spp. AB131-1 and infected by Xoo could increase the height of plant and number of tillers.

  7. Levels of rhizome endophytic fungi fluctuate in Paris polyphylla var. yunnanensis as plants age

    Tao Liu

    2017-02-01

    Full Text Available Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plants age. We speculated that the concentrations of those components in the rhizomes are mediated by fungal endophytes. To test this hypothesis, we took both culture-dependent and -independent (metagenomics approaches to analyze the communities of endophytic fungi that inhabit those rhizomes in plants of different age classes (four, six, and eight years old. In all, 147 isolates representing 18 fungal taxa were obtained from 270 segments (90 per age class. Based on morphological and genetic characteristics, Fusarium oxysporum (46.55% frequency of occurrence was the predominant endophyte, followed by Leptodontidium sp. (8.66% and Trichoderma viride (6.81%. Colonization of endophytic fungi was maximized in the eight-year-old rhizomes (33.33% when compared with four-year-old (21.21% and six-year-old (15.15% rhizomes. Certain fungal species were present only at particular ages. For example, Alternaria sp., Cylindrocarpon sp., Chaetomium sp., Paraphaeosphaeria sporulosa, Pyrenochaeta sp., Penicillium swiecickii, T. viride, and Truncatella angustata were found only in the oldest plants. Analysis of (metagenomics community DNA extracted from different-aged samples revealed that, at the class level, the majority of fungi had the highest sequence similarity to members of Sordariomycetes, followed by Eurotiomycetes and Saccharomycetes. These results were mostly in accord with those we obtained using culture methods. Fungal diversity and richness also changed over time. Our investigation is the first to show that the diversity of fungi in rhizomes of P. polyphylla var. yunnanensis is altered as plants age, and our findings provide a foundation for future examinations of useful compounds.

  8. Biotechnological and Agronomic Potential of Endophytic Pink-Pigmented Methylotrophic Methylobacterium spp.

    Dourado, Manuella Nóbrega; Aparecida Camargo Neves, Aline; Santos, Daiene Souza; Araújo, Welington Luiz

    2015-01-01

    The genus Methylobacterium is composed of pink-pigmented facultative methylotrophic (PPFM) bacteria, which are able to synthesize carotenoids and grow on reduced organic compounds containing one carbon (C1), such as methanol and methylamine. Due to their high phenotypic plasticity, these bacteria are able to colonize different habitats, such as soil, water, and sediment, and different host plants as both endophytes and epiphytes. In plant colonization, the frequency and distribution may be in...

  9. Novel anthraquinone derivatives produced by Pestalotiopsis guepinii, an endophytic of the medicinal plant Virola michelii (Myristicaceae)

    Oliveira, Marilene N.; Santos, Lourivaldo S.; Guilhon, Giselle M.S.P; Santos, Alberdan S.; Ferreira, Isabel C.S.; Lopes-Junior, Manoel L.; Arruda, Mara Silvia P.; Marinho, Andrey M.R.; Silva, Milton N. da; Rodrigues-Filho, Edson; Oliveira, Maria C.F.

    2011-01-01

    A new anthraquinone derivative, named guepinone (1), along with the known substances isosulochrin (2) and chloroisosulochrin (3), were isolated from a rice culture of Pestalotiopsis guepinii, an endophytic fungus of Virola michelii. The compounds were identified by analysis of 1D and 2D NMR and MS spectral data. The antimicrobial activity of these compounds was evaluated and chloroisosulchrin (3) was the most active. (author)

  10. Diversity and Seasonal Variation of Endophytic Fungi Isolated from Three Conifers in Mt. Taehwa, Korea

    Kim, Chang-Kyun; Eo, Ju-Kyeong; Eom, Ahn-Heum

    2013-01-01

    The needled leaves of three conifer species were collected in Mt. Taehwa during different seasons of the year. Total 59 isolates and 19 species of endophytic fungi were isolated from the leaves and identified using morphological and molecular characteristics. As a result, Shannon index was different in its host plant; Larix kaempferi had a highest value of species diversity. According to the sampling season, 9 species of 19 species were isolated during fall season. The results suggest that th...

  11. Petroleum degradation by endophytic Streptomyces spp. isolated from plants grown in contaminated soil of southern Algeria.

    Baoune, Hafida; Ould El Hadj-Khelil, Aminata; Pucci, Graciela; Sineli, Pedro; Loucif, Lotfi; Polti, Marta Alejandra

    2018-01-01

    Petroleum hydrocarbons are well known by their high toxicity and recalcitrant properties. Their increasing utilization around worldwide led to environmental contamination. Phytoremediation using plant-associated microbe is an interesting approach for petroleum degradation and actinobacteria have a great potential for that. For this purpose, our study aimed to isolate, characterize, and assess the ability of endophytic actinobacteria to degrade crude petroleum, as well as to produce plant growth promoting traits. Seventeen endophytic actinobacteria were isolated from roots of plants grown naturally in sandy contaminated soil. Among them, six isolates were selected on the basis of their tolerance to petroleum on solid minimal medium and characterized by 16S rDNA gene sequencing. All petroleum-tolerant isolates belonged to the Streptomyces genus. Determination by crude oil degradation by gas chromatorgraph-flame ionization detector revealed that five strains could use petroleum as sole carbon and energy source and the petroleum removal achieved up to 98% after 7 days of incubation. These isolates displayed an important role in the degradation of the n-alkanes (C 6 -C 30 ), aromatic and polycyclic aromatic hydrocarbons. All strains showed a wide range of plant growth promoting features such as siderophores, phosphate solubilization, 1-aminocyclopropane-1-carboxylate deaminase, nitrogen fixation and indole-3-acetic acid production as well as biosurfactant production. This is the first study highlighting the petroleum degradation ability and plant growth promoting attributes of endophytic Streptomyces. The finding suggests that the endophytic actinobacteria isolated are promising candidates for improving phytoremediation efficiency of petroleum contaminated soil. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Rapid Discovery and Functional Characterization of Terpene Synthases from Four Endophytic Xylariaceae.

    Weihua Wu

    Full Text Available Endophytic fungi are ubiquitous plant endosymbionts that establish complex and poorly understood relationships with their host organisms. Many endophytic fungi are known to produce a wide spectrum of volatile organic compounds (VOCs with potential energy applications, which have been described as "mycodiesel". Many of these mycodiesel hydrocarbons are terpenes, a chemically diverse class of compounds produced by many plants, fungi, and bacteria. Due to their high energy densities, terpenes, such as pinene and bisabolene, are actively being investigated as potential "drop-in" biofuels for replacing diesel and aviation fuel. In this study, we rapidly discovered and characterized 26 terpene synthases (TPSs derived from four endophytic fungi known to produce mycodiesel hydrocarbons. The TPS genes were expressed in an E. coli strain harboring a heterologous mevalonate pathway designed to enhance terpene production, and their product profiles were determined using Solid Phase Micro-Extraction (SPME and GC-MS. Out of the 26 TPS's profiled, 12 TPS's were functional, with the majority of them exhibiting both monoterpene and sesquiterpene synthase activity.

  13. Biotransformation of limonene by an endophytic fungus using synthetic and orange residue-based media.

    Bier, Mário Cesar Jucoski; Medeiros, Adriane Bianchi Pedroni; Soccol, Carlos Ricardo

    2017-02-01

    Aroma and fragrances have high commercial value for use in food, cosmetics and perfumes. The biotransformation of terpenes by microorganisms represents an attractive alternative method for production of flavourings. Endophytic fungi offer a great potential for the production of several groups of compounds; however, few studies have evaluated the biotransformation of limonene. Following preliminary studies on the biotransformation of limonene, submerged fermentation was carried out using an endophytic fungus isolated from Pinus taeda and identified as Phomopsis sp. The presence of several biotransformation products was detected and identified by mass spectrometry (GC-MS). The studied strain showed a divergent metabolic behaviour, as compounds of interest such as α-terpineol, carvone, and limoneno-1,2-diol were produced under different conditions. In addition to the minor metabolites terpinen-4-ol, menthol and carveol, this strain also produced major metabolites, including 0.536 g L -1 carvone and 2.08 g L -1 limonene-1,2-diol in synthetic medium and 2.10 g L -1 limonene-1,2-diol in a natural orange extract medium with single fed-batch, while the cyclic fed-batch resulted in concentrations less than 1 g L -1 . Therefore, our study produced a wide variety of limonene derivatives at a high concentration using a natural medium and a newly isolated endophytic fungal strain. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  14. Potential of Endophytic Bacterial to Control Lesion Nematode (Pratylenchus brachyurus on Patchouli

    RITA HARNI

    2007-03-01

    Full Text Available Root lesion nematode (Pratylenchus brachyurus is one of the most important pathogens of patchouli that caused significant losses. Studies on the potential of endophytic bacterial to control P. brachyurus on patchouli had been conducted. To evaluate the effectiveness of endophytic bacterial against to P. brachyurus on patchouli, nine isolates of bacteria ( NJ2, NJ25, NJ41, NJ46, NJ57, NA22, ERB21, ES32, and E26 were applied by deeping root seedling into bacterial suspension. A study of the physiological characteristics of nine isolates was conducted by using specific medium. The results showed that endophytic bacterial was significantly reduced the population of P. brachyurus and all isolates bacterial promoted growth of patchouli (shoot weight, root weight, and root length. Four isolates, i.e. Bacillus NJ46, Bacillus Na22, Bacillus NJ2, and Bacillus NJ57 were among the potential control agents that reduced nematode populations as much as 68.1-73.9%. Almost all of the isolated bacteria from patchouli roots were able to solubilizing phosphate, while some of them had the ability to produce chitinase, cellulase, protease, HCN, and fluorescency.

  15. Endophytic fungi from Myrcia guianensis at the Brazilian Amazon: distribution and bioactivity.

    Dos Banhos, Elissandro Fonseca; de Souza, Antonia Queiroz Lima; de Andrade, Juliano Camurça; de Souza, Afonso Duarte Leão; Koolen, Hector Henrique Ferreira; Albuquerque, Patrícia Melchionna

    2014-01-01

    Beneficial interactions between plants and microorganisms have been investigated under different ecological, physiological, biochemical, and genetic aspects. However, the systematic exploration of biomolecules with potential for biotechnological products from this interaction still is relatively scarce. Therefore, this study aimed the evaluation of the diversity and antimicrobial activity of the endophytic fungi obtained from roots, stems and leafs of Myrcia guianensis (Myrtaceae) from the Brazilian Amazon. 156 endophytic fungi were isolated and above 80% were identified by morphological examination as belonging to the genera Pestalotiopsis, Phomopsis, Aspergillus, Xylaria, Nectria, Penicillium and Fusarium. Fermented broth of those fungi were assayed for antimicrobial activity and four inhibited the growth of Staphylococcus aureus, Enterococcus faecalis, Candida albicans and Penicillium avellaneum. As the strain named MgRe2.2.3B (Nectria haematococca) had shown the most promising results against those pathogenic strains, its fermented broth was fractioned and only its two low polar fractions demonstrated to be active. Both fractions exhibited a minimum bactericidal concentration of 50 μg.mL(-1) against S. aureus and a minimum fungicidal concentration of 100 μg.mL(-1) against P. avellaneum. These results demonstrate the diversity of fungal genera in M. guianensis and the potential of these endophytic fungi for the production of new antibiotics.

  16. Endophytic bacteria improve phytoremediation of Ni and TCE co-contamination

    Weyens, Nele; Croes, Sarah; Dupae, Joke; Newman, Lee; Lelie, Daniel van der; Carleer, Robert; Vangronsveld, Jaco

    2010-01-01

    The aim of this work was to investigate if engineered endophytes can improve phytoremediation of co-contaminations by organic pollutants and toxic metals. As a model system, yellow lupine was inoculated with the endophyte Burkholderia cepacia VM1468 possessing (a) the pTOM-Bu61 plasmid, coding for constitutive trichloroethylene (TCE) degradation, and (b) the ncc-nre Ni resistance/sequestration system. Plants were exposed to Ni and TCE and (a) Ni and TCE phytotoxicity, (b) TCE degradation and evapotranspiration, and (c) Ni concentrations in the roots and shoots were determined. Inoculation with B. cepacia VM1468 resulted in decreased Ni and TCE phytotoxicity, as measured by 30% increased root biomass and up to 50% decreased activities of enzymes involved in anti-oxidative defence in the roots. In addition, TCE evapotranspiration showed a decreasing trend and a 5 times higher Ni uptake was observed after inoculation. - Engineered endophytes can improve phytoremediation of mixed contaminations via enhanced degradation of organic contaminants and improved metal uptake and translocation.

  17. Diversity of Endophytic Actinomycetes from Wheat and its Potential as Plant Growth Promoting and Biocontrol Agents

    M. Gangwar

    2012-01-01

    Full Text Available A total of 35 endophytic actinomycetes strains was isolated from the roots, stems and leaves tissues of healthy wheat plants and identified as Streptomyces sp. (24, Actinopolyspora sp. (3, Nocardia sp. (4, Saccharopolyspora sp. (2 Pseudonocardia (1 and Micromonospora sp. (1. Seventeen endophytic actinomycetes isolate showed abilities to solubilize phosphate and produce IAA in the range of 5 to 42mg/100ml and 18-42µg/ml respectively. Nineteen isolates produced catechol-type of siderophore ranging between 1.3-20.32µg/ml. Also, hydroxamate-type siderophore produced by 9 isolates in the range of 13.33-50.66µg/ml. Maximum catechol-type of siderophore production was observed in Streptomyces roseosporus W9 (20.32µg/ml which was also displaying maximum antagonistic activity against ten different pathogenic fungi. The results indicated that internal tissues of healthy wheat plants exhibited endophytic actinomycetes diversity not only in terms of different types of isolates but also in terms of functional diversity.

  18. Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava roots following soil drench inoculation

    Greenfield, Melinda; Gómez-Jiménez, María I.; Ortiz, Viviana; Vega, Fernando E.; Kramer, Matthew; Parsa, Soroush

    2016-01-01

    We investigated the fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae to determine if endophytic colonization could be achieved in cassava. An inoculation method based on drenching the soil around cassava stem cuttings using conidial suspensions resulted in endophytic colonization of cassava roots by both entomopathogens, though neither was found in the leaves or stems of the treated cassava plants. Both fungal entomopathogens were detected more often in the proximal end of the root than in the distal end. Colonization levels of B. bassiana were higher when plants were sampled at 7–9 days post-inoculation (84%) compared to 47–49 days post-inoculation (40%). In contrast, the colonization levels of M. anisopliae remained constant from 7–9 days post-inoculation (80%) to 47–49 days post-inoculation (80%), which suggests M. anisopliae is better able to persist in the soil, or as an endophyte in cassava roots over time. Differences in colonization success and plant growth were found among the fungal entomopathogen treatments. PMID:27103778

  19. Enzyme Inhibitory Radicinol Derivative from Endophytic fungus Bipolaris sorokiniana LK12, Associated with Rhazya stricta

    Abdul Latif Khan

    2015-07-01

    Full Text Available Endophytes, living inside plant tissues, play an essential role in plant growth and development, whilst producing unique bioactive secondary metabolites. In the current study, the endophytic fungus Bipolaris sorokiniana LK12 was isolated from the leaves of ethno-medicinal and alkaloidal rich Rhazya stricta. The bulk amount of ethyl acetate extract of fungus was subjected to advance column chromatographic techniques, which resulted in the isolation of a new radicinol derivative, bipolarisenol (1. It was found to be a derivative of radicinol. The structure elucidation was carried out by the combined use of 1D and 2D nuclear magnetic resonance, infrared spectroscopy, mass, and UV spectrometric analyses. The bipolarisenol was assessed for its potential role in enzyme inhibition of urease and acetyl cholinesterase (AChE. Results showed that bipolarisenol significantly inhibited the AChE activity with low IC50 (67.23 ± 5.12 µg·mL−1. Bipolarisenol inhibited urease in a dose-dependent manner with high IC50 (81.62 ± 4.61 µg·mL−1. The new compound also showed a moderate anti-lipid peroxidation potential (IC50 = 168.91 ± 4.23 µg·mL−1. In conclusion, endophytes isolated from medicinal plants possess a unique potential to be considered for future drug discovery.

  20. Characterization of New Bioactive Enzyme Inhibitors from Endophytic Bacillus amyloliquefaciens RWL-1

    Raheem Shahzad

    2018-01-01

    Full Text Available Endophytic bacteria are known to produce a wide array of bioactive secondary metabolites with beneficial effects on human health. In the current study, a novel endophytic bacterial strain, Bacillus amyloliquefaciens RWL-1, was isolated from the seeds of Oryza sativa. Initially, the crude extract of RWL-1 was assessed for potential biological effects of enzyme inhibition and cytotoxicity and was found to exhibit a broad spectrum inhibition for α-glucosidase (37 ± 0.09% and urease (49.4 ± 0.53%. The screening results were followed by bioassay-guided isolation of secondary metabolite(s from RWL-1. Extensive chromatographic and spectrophotometry analyses revealed the presence of compound 1 (S-2-hydroxy-N-((S-1-((S-8-hydroxy-1-oxoisochroman-3-yl-3-methylbutyl-2-((S-5-oxo-2,5-dihydrofuran-2-ylacetamide. Further bioassays of compound 1 showed significant inhibition of α-glucosidase (52.98 ± 0.8% and urease (51.27 ± 1.0%, compared with positive control values of 79.14 ± 1.9% and 88.24 ± 2.2%, and negative controls (0.08 ± 0.1% and 0.05 ± 0.01%, respectively. The current study suggests that bacterial endophytes are a rich source of novel bioactive compounds with high therapeutic value.

  1. Use of the Endophytic Fungus Daldinia cf. concentrica and Its Volatiles as Bio-Control Agents.

    Orna Liarzi

    Full Text Available Endophytic fungi are organisms that spend most of their life cycle within plant tissues without causing any visible damage to the host plant. Many endophytes were found to secrete specialized metabolites and/or emit volatile organic compounds (VOCs, which may be biologically active and assist fungal survival inside the plant as well as benefit their hosts. We report on the isolation and characterization of a VOCs-emitting endophytic fungus, isolated from an olive tree (Olea europaea L. growing in Israel; the isolate was identified as Daldinia cf. concentrica. We found that the emitted VOCs were active against various fungi from diverse phyla. Results from postharvest experiments demonstrated that D. cf. concentrica prevented development of molds on organic dried fruits, and eliminated Aspergillus niger infection in peanuts. Gas chromatography-mass spectrometry analysis of the volatiles led to identification of 27 VOCs. On the basis of these VOCs we prepared two mixtures that displayed a broad spectrum of antifungal activity. In postharvest experiments these mixtures prevented development of molds on wheat grains, and fully eliminated A. niger infection in peanuts. In light of these findings, we suggest use of D. cf. concentrica and/or its volatiles as an alternative approach to controlling phytopathogenic fungi in the food industry and in agriculture.

  2. BIOCONTROL FOR RHIZOCTONIA STEM ROT DISEASE BY USING COMBINATION OF SPECIFIC ENDOPHYTE IN PADDY TIDAL SWAMP

    Ismed Setya Budi

    2013-10-01

    Full Text Available The use of combination of specific endophytic in tidal swamps to control stem root disease as biological control agents has not been done. It is expected that this combination is able to continuously protect plants from pathogen interference. The research was carried out in type C tidal swamp in Banjar regency of South Kalimantan, from March to November 2011, temperature 29-32oC, and pH 4.0-5.5. The method used was Split Plot design. Biocontrol preparation for both types of endophytic was applied in seeds in 7 days after planting (DAP. Observation on high intensity and plant diseases of planting stage on tidal swamps (taradak, ampak and lacak was conducted. The results showed that there was a reduction of disease ranging from 58.70 to 87.29%. The application of combination of two biocontrol agents (T. viride PS-2.1 + P. fluorescent PS-4.8, (Fusarium non-pathogenic PS-1.5 + P. fluorescent PS-4.8 and (T. viride PS-2.1+ FNP PS-1.5 isolate gave the best inhibition result, reduced disease intensity, and increased plant height. The result of soil analysis before and after application of endophytic showed that there was an increase in soil fertility with the element addition of N, P, K and pH.

  3. Cholinesterase inhibitor (Altenuene) from an endophytic fungus Alternaria alternata: optimization, purification and characterization.

    Bhagat, J; Kaur, A; Kaur, R; Yadav, A K; Sharma, V; Chadha, B S

    2016-10-01

    The aim of this study was to screen endophytic fungi isolated from Vinca rosea for their potential to produce acetylcholinesterase (AChE) inhibitors. Endophytic fungi isolated from V. rosea (Catharanthus roseus), were screened for AChE inhibitor production using Ellman's method. Maximum inhibition against AChE (78%) was observed in an isolate VS-10, identified to be Alternaria alternata on morphological and molecular basis. The isolate also inhibited butyrylcholinesterase (73%). Significant increase (1·3 fold) was achieved after optimization of process parameters using one variable at time approach. The inhibitor was purified using chromatographic techniques. The structure elucidation of the inhibitor was carried out using spectroscopic techniques and was identified to be 'altenuene'. The purified inhibitor possessed antioxidant potential as revealed by dot blot assay. The insecticidal potential of purified inhibitor was evaluated by feeding Spodoptora litura on diet amended with inhibitor. It evinced significant larval mortality. Endophytic A. alternata can serve as a source of dual cholinesterase inhibitor 'altenuene' with significant antioxidant and insecticidal activity. This is the first report on acetylcholinestearse inhibitory activity of altenuene. Alternaria alternata has the potential to produce a dual ChE inhibitor with antioxidant activity useful in the treatment of neurodegenerative disorders and in agriculture as biocontrol agent. © 2016 The Society for Applied Microbiology.

  4. Selection and Characterization of Endophytic Bacteria as Biocontrol Agents of Tomato Bacterial Wilt Disease

    ABDJAD ASIH NAWANGSIH

    2011-06-01

    Full Text Available Biological control of bacterial wilt pathogen (Ralstonia solanacearum of tomato using endophytic bacteria is one of the alternative control methods to support sustainable agriculture. This study was conducted to select and characterize endophytic bacteria isolated from healthy tomato stems and to test their ability to promote plant growth and suppress bacterial wilt disease. Among 49 isolates successfully isolated, 41 were non-plant pathogenic. Green house test on six selected isolates based on antagonistic effect on R. solanacearum or ability to suppress R. solanacearum population in dual culture assays obtained BC4 and BL10 isolates as promising biocontrol agents. At six weeks after transplanting, plants treated with BC4 isolate showed significantly lower disease incidence (33% than that of control (83%. Plants height was not significantly affected by endophytic bacterial treatments. Based on 16S rRNA sequence, BC4 isolate had 97% similarity with Staphylococcus epidermidis (accession number EU834240.1, while isolate BL10 had 98% similarity with Bacillus amyloliquefaciens strain JK-SD002 (accession number AB547229.1.

  5. Enhancement of Thiamine Biosynthesis in Oil Palm Seedlings by Colonization of Endophytic Fungus Hendersonia toruloidea

    Kamarudin, Amirah N.; Lai, Kok S.; Lamasudin, Dhilia U.; Idris, Abu S.; Balia Yusof, Zetty N.

    2017-01-01

    Thiamine, or vitamin B1 plays an indispensable role as a cofactor in crucial metabolic reactions including glycolysis, pentose phosphate pathway and the tricarboxylic acid cycle in all living organisms. Thiamine has been shown to play a role in plant adaptation toward biotic and abiotic stresses. The modulation of thiamine biosynthetic genes in oil palm seedlings was evaluated in response to root colonization by endophytic Hendersonia toruloidea. Seven-month-old oil palm seedlings were inoculated with H. toruloidea and microscopic analyses were performed to visualize the localization of endophytic H. toruloidea in oil palm roots. Transmission electron microscopy confirmed that H. toruloidea colonized cortical cells. The expression of thiamine biosynthetic genes and accumulation of total thiamine in oil palm seedlings were also evaluated. Quantitative real-time PCR was performed to measure transcript abundances of four key thiamine biosynthesis genes (THI4, THIC, TH1, and TPK) on days 1, 7, 15, and 30 in response to H. toruloidea colonization. The results showed an increase of up to 12-fold in the expression of all gene transcripts on day 1 post-inoculation. On days 7, 15, and 30 post-inoculation, the relative expression levels of these genes were shown to be downregulated. Thiamine accumulation was observed on day 7 post-colonization and subsequently decreased until day 30. This work provides the first evidence for the enhancement of thiamine biosynthesis by endophytic colonization in oil palm seedlings. PMID:29089959

  6. Combined endophytic inoculants enhance nickel phytoextraction from serpentine soil in the hyperaccumulator Noccaea caerulescens.

    Visioli, Giovanna; Vamerali, Teofilo; Mattarozzi, Monica; Dramis, Lucia; Sanangelantoni, Anna M

    2015-01-01

    This study assesses the effects of specific bacterial endophytes on the phytoextraction capacity of the Ni-hyperaccumulator Noccaea caerulescens, spontaneously growing in a serpentine soil environment. Five metal-tolerant endophytes had already been selected for their high Ni tolerance (6 mM) and plant growth promoting ability. Here we demonstrate that individual bacterial inoculation is ineffective in enhancing Ni translocation and growth of N. caerulescens in serpentine soil, except for specific strains Ncr-1 and Ncr-8, belonging to the Arthrobacter and Microbacterium genera, which showed the highest indole acetic acid production and 1-aminocyclopropane-1-carboxylic acid-deaminase activity. Ncr-1 and Ncr-8 co-inoculation was even more efficient in promoting plant growth, soil Ni removal, and translocation of Ni, together with that of Fe, Co, and Cu. Bacteria of both strains densely colonized the root surfaces and intercellular spaces of leaf epidermal tissue. These two bacterial strains also turned out to stimulate root length, shoot biomass, and Ni uptake in Arabidopsis thaliana grown in MS agar medium supplemented with Ni. It is concluded that adaptation of N. caerulescens in highly Ni-contaminated serpentine soil can be enhanced by an integrated community of bacterial endophytes rather than by single strains; of the former, Arthrobacter and Microbacterium may be useful candidates for future phytoremediation trials in multiple metal-contaminated sites, with possible extension to non-hyperaccumulator plants.

  7. Combined endophytic inoculants enhance nickel phytoextraction from serpentine soil in the hyperaccumulator Noccaea caerulescens

    Giovanna eVisioli

    2015-08-01

    Full Text Available This study assesses the effects of specific bacterial endophytes on the phytoextraction capacity of the Ni-hyperaccumulator Noccaea caerulescens, spontaneously growing in a serpentine soil environment. Five metal-tolerant endophytes had already been selected for their high Ni tolerance (6 mM and plant growth promoting ability. Here we demonstrate that individual bacterial inoculation is ineffective in enhancing Ni translocation and growth of N. caerulescens in serpentine soil, except for specific strains Ncr-1 and Ncr-8, belonging to the Arthrobacter and Microbacterium genera, which showed the highest IAA production and ACC-deaminase activity. Ncr-1 and Ncr-8 co-inoculation was even more efficient in promoting plant growth, soil Ni removal and translocation of Ni, together with that of Fe, Co and Cu. Bacteria of both strains densely colonised the root surfaces and intercellular spaces of leaf epidermal tissue. These two bacterial strains also turned out to stimulate root length, shoot biomass and Ni uptake in Arabidopsis thaliana grown in MS agar medium supplemented with Ni. It is concluded that adaptation of N. caerulescens in highly Ni-contaminated serpentine soil can be enhanced by an integrated community of bacterial endophytes rather than by single strains; of the former, Arthrobacter and Microbacterium may be useful candidates for future phytoremediation trials

  8. Endophytic fungi from leaves of Centella asiatica: occurrence and potential interactions within leaves.

    Rakotoniriana, E F; Munaut, F; Decock, C; Randriamampionona, D; Andriambololoniaina, M; Rakotomalala, T; Rakotonirina, E J; Rabemanantsoa, C; Cheuk, K; Ratsimamanga, S U; Mahillon, J; El-Jaziri, M; Quetin-Leclercq, J; Corbisier, A M

    2008-01-01

    Fungal endophytes were isolated from leaves of Centella asiatica (Apiaceae) collected at Mangoro (middle eastern region of Madagascar, 200 km from Antananarivo). Forty- five different taxa were recovered. The overall foliar colonization rate was 78%. The most common endophytes were the non-sporulating species 1 (isolation frequency IF 19.2%) followed by Colletotrichum sp.1 (IF 13.2%), Guignardia sp. (IF 8.5%), Glomerella sp. (IF 7.7%), an unidentified ascomycete (IF 7.2%), the non-sporulating species 2 (IF 3.7%) and Phialophora sp. (IF 3.5%). Using sequences of the ribosomal DNA internal transcribed spacer (ITS) regions, major endophytes (IF > 7%) were identified as xylariaceous taxa or as Colletotrichum higginsianum, Guignardia mangiferae and Glomerella cingulata. Results from in vitro fungal disk experiments showed a strong inhibitory activity of the xylariaceous non-sporulating species 1 against G. mangiferae and C. higginsianum and of C. higginsianum against G. mangiferae. This can be explained by antagonism between dominant taxa.

  9. Endophytic fungi occurring in fennel, lettuce, chicory, and celery--commercial crops in southern Italy.

    D'Amico, Margherita; Frisullo, Salvatore; Cirulli, Matteo

    2008-01-01

    The occurrence of endophytic fungi in fennel, lettuce, chicory, and celery crops was investigated in southern Italy. A total of 186 symptomless plants was randomly collected and sampled at the stage of commercial ripeness. Fungal species of Acremonium, Alternaria, Fusarium, and Plectosporium were detected in all four crops; Plectosporium tabacinum was the most common in all crop species and surveyed sites. The effect of eight endophytic isolates (five belonging to Plectosporium tabacinum and three to three species of Acremonium) inoculated on lettuce plants grown in gnotobiosis was assessed by recording plant height, root length and dry weight, collar diameter, root necrosis, and leaf yellowing. P. tabacinum and three species of Acremonium, inoculated on gnotobiotically grown lettuce plants, showed pathogenic activity that varied with the fungal isolate. Lettuce plants inoculated with the isolates Ak of Acremonium kiliense, Ac of Acremonium cucurbitacearum, and P35 of P. tabacinum showed an increased root growth, compared to the non-inoculated control. The high frequency of P. tabacinum isolation recorded in lettuce plants collected in Bari and Metaponto, and in fennel plants from Foggia agricultural districts, suggests a relationship not only between a crop species and P. tabacinum, but also between the occurrence of the endophyte and the crop rotation history of the soil.

  10. Alkaloid (Meleagrine and Chrysogine) from endophytic fungi (Penicillium sp.) of Annona squamosa L.

    Yunianto, Prasetyawan; Rusman, Yudi; Saepudin, Endang; Suwarso, Wahyudi Priyono; Sumaryono, Wahono

    2014-05-01

    Several endophytic fungal strains from Srikaya plants (Annona squamosa L.) have been isolated and one of them was identified as Penicillium sp. Penicillium has been proven as an established source for a wide array of unique bioactive secondary metabolites that exhibit a variety of biological activities. The aim of this study is isolation of secondary metabolite from Penicillium, an endophytic of A. squamosa L. Penicillium sp. from endophytic of A. squamosa L. was fermented in Wicherham media. The whole extract from both liquid media and mycelium was partitioned by ethyl acetate and evaporated to obtain crude ethyl acetate extract. The ethyl acetate extract was then brokedown using column chromatography with silica as stationary phase and mixture of ethyl acetate/methanol (98%:2%) as mobile phase and then was separated by sephadex column. Structure elucidation of isolated compounds were mainly done by analysis of one and two dimensional NMR (Nuclear Magnetic Resonance) data and supported by HPLC (High performance Liquid Chromatography) and MS-TOF (Mass Spectrometer-Time of Flight). Isolated secondary metabolites were tested using in vitro assays for anticancer and antimicrobial activity. For anticancer activity, the metabolites were tested against breast cancer cells (MCF-7) using MTT assay, while for antimicrobial activity was performed using disk diffusion assays. From these physical, chemical and spectral evidences that the secondary metabolites were confirmed as Chrysogine and Meleagrine. Chrysogine and Meleagrine have no activity as anticancer and antimicrobial.

  11. Isolation and evaluation of endophytic fungi with antimicrobial ability from Phyllostachys edulis

    Xiaoye Shen

    2012-12-01

    Full Text Available Endophytic fungi (30 isolates from bamboo branches were categorized into 12 genera, based on the blast analyses of ITS nrDNA sequence in GenBank and microscopic examination. The aim of this work was to investigate the antibacterial and antifungal activities of endophytic fungi. Inhibitory effects against clinical pathogens and phytopathogens have been screened for all the isolates preliminarily and strains tentatively identified as Cladosporium sphaerospermum (PE106, Simplicillium lanosoniveum (PE120, Curvularia sp. (PE127, Didymella sp. (PE128 and Penicillium cf. raistrickii (PE130 presented bioactivity against at least four tested pathogens using the agar diffusion method. Crude extracts of PE106, PE120, PE127 and PE130 displayed broad-spectrum activity against plant pathogenic fungi by mycelial radial growth test. All of the four isolates were found to have high bioactivity against the frequent plant pathogenic fungus Botryotinia fuckeliana, and two of the isolates (PE120 and PE130 also inhibited the growth of phytopathogen Thanatephorus cucumeris noteworthily. This study is the first report on the antimicrobial activity of endophytic fungi associated with branches of Ph. edulis.

  12. Influence of Culturing Conditions on Bioprospecting and the Antimicrobial Potential of Endophytic Fungi from Schinus terebinthifolius.

    Tonial, Fabiana; Maia, Beatriz H L N S; Gomes-Figueiredo, Josiane A; Sobottka, Andrea M; Bertol, Charise D; Nepel, Angelita; Savi, Daiani C; Vicente, Vânia A; Gomes, Renata R; Glienke, Chirlei

    2016-02-01

    In this study, we analyzed the antimicrobial activity of extracts harvested from 17 endophytic fungi isolated from the medicinal plant Schinus terebinthifolius. Morphological and molecular analyses indicated that these fungal species belonged to the genera Alternaria, Bjerkandera, Colletotrichum, Diaporthe, Penicillium, and Xylaria. Of the endophytes analyzed, 64.7 % produced antimicrobial compounds under at least one of the fermentation conditions tested. Nine isolates produced compounds that inhibited growth of Staphylococcus aureus, four produced compounds that inhibited Candida albicans, and two that inhibited Pseudomonas aeruginosa. The fermentation conditions of the following endophytes were optimized: Alternaria sp. Sect. Alternata-LGMF626, Xylaria sp.-LGMF673, and Bjerkandera sp.-LGMF713. Specifically, the carbon and nitrogen sources, initial pH, temperature, and length of incubation were varied. In general, production of antimicrobial compounds was greatest when galactose was used as a carbon source, and acidification of the growth medium enhanced the production of compounds that inhibited C. albicans. Upon large-scale fermentation, Alternaria sp. Sect. Alternata-LGMF626 produced an extract containing two fractions that were active against methicillin-resistant S. aureus. One of the extracts exhibited high activity (minimum inhibitory concentration of 18.52 µg/mL), and the other exhibited moderate activity (minimum inhibitory concentration of 55.55 µg/mL). The compounds E-2-hexyl-cinnamaldehyde and two compounds of the pyrrolopyrazine alkaloids class were identified in the active fractions by gas chromatography-mass spectrometry.

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

    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.

  14. Isolation, taxonomic analysis, and phenotypic characterization of bacterial endophytes present in alfalfa (Medicago sativa) seeds.

    López, José Luis; Alvarez, Florencia; Príncipe, Analía; Salas, María Eugenia; Lozano, Mauricio Javier; Draghi, Walter Omar; Jofré, Edgardo; Lagares, Antonio

    2018-02-10

    A growing body of evidence has reinforced the central role of microbiomes in the life of sound multicellular eukaryotes, thus more properly described as true holobionts. Though soil was considered a main source of plant microbiomes, seeds have been shown to be endophytically colonized by microorganisms thus representing natural carriers of a selected microbial inoculum to the young seedlings. In this work we have investigated the type of culturable endophytic bacteria that are carried within surface-sterilized alfalfa seeds. MALDI-TOF analysis revealed the presence of bacteria that belonged to 40 separate genera, distributed within four taxa (Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes). Nonsymbiotic members of the Rhizobiaceae family were also found. The evaluation of nine different in-vitro biochemical activities demonstrated isolates with complex combinations of traits that, upon a Principal-Component-Analysis, could be classified into four phenotypic groups. That isolates from nearly half of the genera identified had been able to colonize alfalfa plants grown under axenic conditions was remarkable. Further analyses should be addressed to investigating the colonization mechanisms of the alfalfa seeds, the evolutionary significance of the alfalfa-seed endophytes, and also how after germination the seed microbiome competes with spermospheric and rhizospheric soil bacteria to colonize newly emerging seedlings. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Endophytic Actinomycetes: A Novel Source of Potential Acyl Homoserine Lactone Degrading Enzymes

    Surang Chankhamhaengdecha

    2013-01-01

    Full Text Available Several Gram-negative pathogenic bacteria employ N-acyl-L-homoserine lactone (HSL quorum sensing (QS system to control their virulence traits. Degradation of acyl-HSL signal molecules by quorum quenching enzyme (QQE results in a loss of pathogenicity in QS-dependent organisms. The QQE activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high HSL-degrading activity. Among 344 rhizospheric and 132 endophytic isolates, 127 (36.9% and 68 (51.5% of them, respectively, possessed the QQE activity. The highest HSL-degrading activity was at 151.30±3.1 nmole/h/mL from an endophytic actinomycetes isolate, LPC029. The isolate was identified as Streptomyces based on 16S  rRNA gene sequence similarity. The QQE from LPC029 revealed HSL-acylase activity that was able to cleave an amide bond of acyl-side chain in HSL substrate as determined by HPLC. LPC029 HSL-acylase showed broad substrate specificity from C6- to C12-HSL in which C10HSL is the most favorable substrate for this enzyme. In an in vitro pathogenicity assay, the partially purified HSL-acylase efficiently suppressed soft rot of potato caused by Pectobacterium carotovorum ssp. carotovorum as demonstrated. To our knowledge, this is the first report of HSL-acylase activity derived from an endophytic Streptomyces.

  16. The endophytic bacterium Serratia sp. PW7 degrades pyrene in wheat.

    Zhu, Xuezhu; Wang, Wanqing; Crowley, David E; Sun, Kai; Hao, Shupeng; Waigi, Michael Gatheru; Gao, Yanzheng

    2017-03-01

    This research was conducted to isolate polycyclic aromatic hydrocarbon-degrading (PAH-degrading) endophytic bacteria and investigate their potential in protecting plants against PAH contamination. Pyrene-degrading endophytic bacteria were isolated from plants grown in PAH-contaminated soil. Among these endophytic bacteria, strain PW7 (Serratia sp.) isolated from Plantago asiatica was selected to investigate the suppression of pyrene accumulation in Triticum aestivum L. In the in vitro tests, strain PW7 degraded 51.2% of the pyrene in the media within 14 days. The optimal biodegradation conditions were pH 7.0, 30 °C, and MS medium supplemented with additional glucose, maltose, sucrose, and peptones. In the in vivo tests, strain PW7 successfully colonized the roots and shoots of inoculated (E + ) wheat plants, and its colonization decreased pyrene accumulation and pyrene transportation from roots to shoots. Remarkably, the concentration of pyrene in shoots decreased much more than that in roots, suggesting that strain PW7 has the potential for protecting wheat against pyrene contamination and mitigating the threat of pyrene to human health via food consumption.

  17. Initial growth of maize in response to application of rock phosphate, vermicompost and endophytic bacteria

    Lílian Estrela Borges Baldotto

    2012-04-01

    Full Text Available Due to the high energy requirement and demand for non-renewable resources for the production of chemical fertilizers, added also to the environmental impact caused by the use of such products, it is important to intensify research on bio-based agricultural inputs. The use of nitrogen-fixing endophytic and phosphate solubilizing bacteria can provide these nutrients to the plants from the air and poorly soluble phosphorus sources, such as phosphate rock. The objective of this study was to evaluate the nutrition and initial growth of maize (Zea mays L. in response to the inoculation of nitrogen-fixing and rock phosphate solubilizing endophytic bacteria, in single or mixed formulation, applied with vermicompost. The treatments containing bacteria, both diazotrophic and phosphate solubilizing, when compared to controls, showed higher levels of leaf nitrogen and phosphorus in maize, as well as higher growth characteristics. The application of vermicompost showed synergistic effect when combined with endophytic bacteria. Thus, the innovation of the combination of the studied factors may contribute to the early development of maize.

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

    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.

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

    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.

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

    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.

  1. Inhibition of Bacterial Quorum Sensing by Extracts from Aquatic Fungi: First Report from Marine Endophytes

    Alberto J. Martín-Rodríguez

    2014-11-01

    Full Text Available In our search for quorum-sensing (QS disrupting molecules, 75 fungal isolates were recovered from reef organisms (endophytes, saline lakes and mangrove rhizosphere. Their QS inhibitory activity was evaluated in Chromobacterium violaceum CVO26. Four strains of endophytic fungi stood out for their potent activity at concentrations from 500 to 50 μg mL−1. The molecular characterization, based on the internal transcribed spacer (ITS region sequences (ITS1, 5.8S and ITS2 between the rRNA of 18S and 28S, identified these strains as belonging to four genera: Sarocladium (LAEE06, Fusarium (LAEE13, Epicoccum (LAEE14, and Khuskia (LAEE21. Interestingly, three came from coral species and two of them came from the same organism, the coral Diploria strigosa. Metabolic profiles obtained by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS suggest that a combination of fungal secondary metabolites and fatty acids could be the responsible for the observed activities. The LC-HRMS analysis also revealed the presence of potentially new secondary metabolites. This is, to the best of our knowledge, the first report of QS inhibition by marine endophytic fungi.

  2. ITS2 sequence-structure phylogeny reveals diverse endophytic Pseudocercospora fungi on poplars.

    Yan, Dong-Hui; Gao, Qian; Sun, Xiaoming; Song, Xiaoyu; Li, Hongchang

    2018-04-01

    For matching the new fungal nomenclature to abolish pleomorphic names for a fungus, a genus Pseudocercospora s. str. was suggested to host holomorphic Pseudocercosproa fungi. But the Pseudocercosproa fungi need extra phylogenetic loci to clarify their taxonomy and diversity for their existing and coming species. Internal transcribed spacer 2 (ITS2) secondary structures have been promising in charactering species phylogeny in plants, animals and fungi. In present study, a conserved model of ITS2 secondary structures was confirmed on fungi in Pseudocercospora s. str. genus using RNAshape program. The model has a typical eukaryotic four-helix ITS2 secondary structure. But a single U base occurred in conserved motif of U-U mismatch in Helix 2, and a UG emerged in UGGU motif in Helix 3 to Pseudocercospora fungi. The phylogeny analyses based on the ITS2 sequence-secondary structures with compensatory base change characterizations are able to delimit more species for Pseudocercospora s. str. than phylogenic inferences of traditional multi-loci alignments do. The model was employed to explore the diversity of endophytic Pseudocercospora fungi in poplar trees. The analysis results also showed that endophytic Pseudocercospora fungi were diverse in species and evolved a specific lineage in poplar trees. This work suggested that ITS2 sequence-structures could become as additionally significant loci for species phylogenetic and taxonomic studies on Pseudocerospora fungi, and that Pseudocercospora endophytes could be important roles to Pseudocercospora fungi's evolution and function in ecology.

  3. Nodulation-dependent communities of culturable bacterial endophytes from stems of field-grown soybeans.

    Okubo, Takashi; Ikeda, Seishi; Kaneko, Takakazu; Eda, Shima; Mitsui, Hisayuki; Sato, Shusei; Tabata, Satoshi; Minamisawa, Kiwamu

    2009-01-01

    Endophytic bacteria (247 isolates) were randomly isolated from surface-sterilized stems of non-nodulated (Nod(-)), wild-type nodulated (Nod(+)), and hypernodulated (Nod(++)) soybeans (Glycine max [L.] Merr) on three agar media (R2A, nutrient agar, and potato dextrose agar). Their diversity was compared on the basis of 16S rRNA gene sequences. The phylogenetic composition depended on the soybean nodulation phenotype, although diversity indexes were not correlated with nodulation phenotype. The most abundant phylum throughout soybean lines tested was Proteobacteria (58-79%). Gammaproteobacteria was the dominant class (21-72%) with a group of Pseudomonas sp. significantly abundant in Nod(+) soybeans. A high abundance of Alphaproteobacteria was observed in Nod(-) soybeans, which was explained by the increase in bacterial isolates of the families Rhizobiaceae and Sphingomonadaceae. A far greater abundance of Firmicutes was observed in Nod(-) and Nod(++) mutant soybeans than in Nod(+) soybeans. An impact of culture media on the diversity of isolated endophytic bacteria was also observed: The highest diversity indexes were obtained on the R2A medium, which enabled us to access Alphaproteobacteria and other phyla more frequently. The above results indicated that the extent of nodulation changes the phylogenetic composition of culturable bacterial endophytes in soybean stems.

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

    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.

  5. A potent feed preservative candidate produced by Calcarisporium sp., an endophyte residing in stargrass (Cynodon dactylon).

    Ji, L L; Song, Y C; Tan, R X

    2004-01-01

    The cultures of an endophytic fungus Calcarisporium sp. were screened for inhibitors on the growth of feed-associated moulds and on the aflatoxin biosynthesis to find a safe and effective feed preservative. Eight test fungi were isolated from the spoiled poultry feed. The endophytic fungus Calcarisporium sp. was separated from the Chinese coastal grass Cynodon dactylon. The antifungal action concerning the endophytic culture extract (ECE) was performed with propionic acid (PPA) as the corresponding reference. The ECE had a similar antifungal efficacy to PPA in a concentration-dependent manner. The susceptibility order of the ECE to the test fungi was found to be Fusarium sp. > Aspergillus spp. > Penicillium spp. Furthermore, the application of the ECE in pelleted-layer duck feed as a preservative was carried out at a humidity of 10, 15 and 20%. It has been discerned that mould growth and aflatoxin biosynthesis could be co-inhibited almost completely by ECE at concentrations higher than 1.0% (w/w). The LD50 of the ECE on mice was shown to be higher than 28 g kg-1. The ECE can be selected as an inhibitor to preserve poultry feed on inhibiting the growth of mould and aflatoxin biosynthesis during feed storage. The ECE may be an effective and biosafe antifungal ingredient for poultry feed and holds a potential market prospect in feed industry.

  6. Indentification of huperzine A-producing endophytic fungi isolated from Huperzia serrata.

    Dong, Li-Hui; Fan, San-Wei; Ling, Qing-Zhi; Huang, Bei-Bei; Wei, Zhao-Jun

    2014-03-01

    This present study was designed to investigate the production of huperzine A (HupA), an acetylcholine inhibitor, which was produced by an endophytic fungi isolated from Huperzia serrata. Screening of 94 endophytic fungal isolates obtained from plant H. serrata was carried out for the production of HupA. Their morphological characteristics were studied and rDNA sequence analysis was carried out. The cultures were grown in liquid culture medium and the extracted metabolites were analyzed by thin layer chromatography and high performance liquid chromatograph for the presence of HupA. The DPPH scavenging ratio and inhibition ratio of acetylcholinesterase (AchE) of the same were determined. 3 out of 94 strains i.e. S29, L44 and S94 showed significant AchE-inhibitory activity and antioxidant activity. Strain L44 which exhibited maximum yield of HupA (37.63 μg/g on dry weight basis) was identified as Trichoderma species by ITS sequence analysis. In conclusion, endophytic fungi from H. serrata can be used as a new resource of HupA.

  7. Bacterial endophytes enhance phytostabilization in soils contaminated with uranium and lead.

    Ahsan, Muhammad Tayyab; Najam-Ul-Haq, Muhammad; Idrees, Muhammad; Ullah, Inayat; Afzal, Muhammad

    2017-10-03

    The combined use of plants and bacteria is a promising approach for the remediation of polluted soil. In the current study, the potential of bacterial endophytes in partnership with Leptochloa fusca (L.) Kunth was evaluated for the remediation of uranium (U)- and lead (Pb)-contaminated soil. L. fusca was vegetated in contaminated soil and inoculated with three different endophytic bacterial strains, Pantoea stewartii ASI11, Enterobacter sp. HU38, and Microbacterium arborescens HU33, individually as well as in combination. The results showed that the L. fusca can grow in the contaminated soil. Bacterial inoculation improved plant growth and phytoremediation capacity: this manifested in the form of a 22-51% increase in root length, 25-62% increase in shoot height, 10-21% increase in chlorophyll content, and 17-59% more plant biomass in U- and Pb-contaminated soils as compared to plants without bacterial inoculation. Although L. fusca plants showed potential to accumulate U and Pb in their root and shoot on their own, bacterial consortia further enhanced metal uptake capacity by 53-88% for U and 58-97% for Pb. Our results indicate that the combination of L. fusca and endophytic bacterial consortia can effectively be used for the phytostabilization of both U- and Pb-contaminated soils.

  8. Endophytic Actinobacteria Associated with Dracaena cochinchinensis Lour.: Isolation, Diversity, and Their Cytotoxic Activities.

    Salam, Nimaichand; Khieu, Thi-Nhan; Liu, Min-Jiao; Vu, Thu-Trang; Chu-Ky, Son; Quach, Ngoc-Tung; Phi, Quyet-Tien; Narsing Rao, Manik Prabhu; Fontana, Angélique; Sarter, Samira; Li, Wen-Jun

    2017-01-01

    Dracaena cochinchinensis Lour. is an ethnomedicinally important plant used in traditional Chinese medicine known as dragon's blood. Excessive utilization of the plant for extraction of dragon's blood had resulted in the destruction of the important niche. During a study to provide a sustainable way of utilizing the resources, the endophytic Actinobacteria associated with the plant were explored for potential utilization of their medicinal properties. Three hundred and four endophytic Actinobacteria belonging to the genera Streptomyces , Nocardiopsis , Brevibacterium , Microbacterium , Tsukamurella , Arthrobacter , Brachybacterium , Nocardia , Rhodococcus , Kocuria , Nocardioides , and Pseudonocardia were isolated from different tissues of D. cochinchinensis Lour. Of these, 17 strains having antimicrobial and anthracyclines-producing activities were further selected for screening of antifungal and cytotoxic activities against two human cancer cell lines, MCF-7 and Hep G2. Ten of these selected endophytic Actinobacteria showed antifungal activities against at least one of the fungal pathogens, of which three strains exhibited cytotoxic activities with IC 50 -values ranging between 3 and 33  μ g·mL -1 . Frequencies for the presence of biosynthetic genes, polyketide synthase- (PKS-) I, PKS-II, and nonribosomal peptide synthetase (NRPS) among these 17 selected bioactive Actinobacteria were 29.4%, 70.6%, and 23.5%, respectively. The results indicated that the medicinal plant D. cochinchinensis Lour. is a good niche of biologically important metabolites-producing Actinobacteria.

  9. Cellular composition and expression of potential stem cell markers in mammary tissue of cows consuming endophyte-infected fescue seed during the dry period and early lactation

    We evaluated the impact of consuming endophyte-infected fescue during late pregnancy on parameters of mammary development in Holstein cows. Cows (N = 16) were fed 10% of their ration as tall fescue seed that was free from (CON) or infected with endophyte (INF) from 90d before expected calving until ...

  10. Diversity and antifungal activity of the endophytic fungi associated with the native medicinal cactus Opuntia humifusa (Cactaceae) from the United States

    The endophytic fungal community associated with the native cactus Opuntia humifusa in the United States was investigated and its potential for providing antifungal compounds. A total of 108 endophytic fungal isolates were obtained and identified by molecular methods into 17 different taxa of the gen...

  11. Symbiotic lifestyle expression by fungal endophytes and the adaptation of plants to stress: unraveling the complexities of intimacy

    Redman, Regina S.; Henson, Joan M.; Rodriguez, Russell J.

    2005-01-01

    The fossil record indicates that fungal symbionts have been associated with plants since the Ordovician period (approximately 400 million years ago), when plants first became established on land (Pirozynski and Malloch, 1975; Redecker et al., 2000; Remy et al., 1994; Simon et al., 1993). Transitioning from aquatic to terrestrial habitats likely presented plants with new stresses, including periods of desiccation. Since symbiotic fungi are known to confer drought tolerance to plants (Bacon, 1993; Read and Camp, 1986), it has been suggested that fungal symbiosis was involved with or responsible for the establishment of land plants (Pirozynski and Malloch, 1975). Symbiosis was first defined by De Bary in 1879, and since that time, all plants in natural ecosystems have been found to be colonized with fungal and bacterial symbionts. It is clear that individual plants represent symbiotic communities with microorganisms associated in or on tissues below- and aboveground.There are two major classes of fungal symbionts associated with internal plant tissues: fungal endophytes that reside entirely within plants and may be associated with roots, stems leaves, or flowers; and mycorrhizal fungi that reside only in roots but extend out into the rhizosphere. In addition, fungal endophytes may be divided into two classes: (1) a relatively small number of fastidious species that are limited to a few monocot hosts (Clay and Schardl, 2002), and (2) a large number of tractable species with broad host ranges, including both monocots and eudicots (Stone et al., 2000). While significant resources and research have been invested in mycorrhizae and class 1 endophytes, comparatively little is known about class 2 endophytes, which may represent the largest group of fungal symbionts. This is partially because the symbiotic functionalities of class 2 endophytes have only recently been elucidated and shown to be responsible for the adaptation of some plants to high-stress environments (Redman

  12. Production of Gentisyl Alcohol from Phoma herbarum Endophytic in Curcuma longa L. and Its Antagonistic Activity Towards Leaf Spot Pathogen Colletotrichum gloeosporioides.

    Gupta, Suruchi; Kaul, Sanjana; Singh, Baljinder; Vishwakarma, Ram A; Dhar, Manoj K

    2016-11-01

    Endophytes from medicinal plants represent a potential source of bioactive compounds. During the present investigation, fungal endophytes were isolated from turmeric (Curcuma longa), an important medicinal plant. A total of 207 endophytic fungal isolates were obtained from the rhizome of C. longa L. They were grouped into seven genera based on morphological and molecular data. The fungal endophytes of C. longa were evaluated for antifungal activity against Colletotrichum gloeosporioides, the causal organism of leaf spot of turmeric. The disease is a major cause for economic loss in turmeric cultivation. Endophytic Phoma herbarum showed significant activity against C. gloeosporioides and was therefore selected for further studies. A compound gentisyl alcohol was isolated from P. herbarum which showed effective antagonism against C. gloeosporioides. The organism could therefore be used as a biocontrol agent against C. gloeosporioides.

  13. Study of endophytic Xylariaceae in Thailand: diversity and taxonomy inferred from rDNA sequence analyses with saprobes forming fruit bodies in the field

    Okane, Izumi; Srikitikulchai, Prasert; Toyama, Kyoko

    2008-01-01

    to reveal the diversity and taxonomy of endophytes and the relationships between those endophytes and saprobic Xylariaceae in Thailand that have been recorded according to fruit-body formation on decayed plant materials. Analysis of 28S rDNA D1/D2 sequences revealed 21 xylariaceous species inhabiting......A study of the diversity, taxonomy, and ecology of endophytic Xylariaceae (Ascomycota) was carried out. In this study, we obtained isolates of Xylariaceae from healthy, attached leaves and teleomorphic stromata on decayed plant materials in a permanent plot at Khao Yai National Park (Thailand......). In addition, strains deposited beforehand were selected in which both endophytic strains isolated from living plant tissues and saprobic strains from fruit bodies were included. Consequently, 405 strains of Xylariaceae (273 endophytic and 132 saprobic strains, including identified strains) were studied...

  14. Endophytic Paraconiothyrium sp. from Zingiber officinale Rosc. Displays Broad-Spectrum Antimicrobial Activity by Production of Danthron.

    Anisha, C; Sachidanandan, P; Radhakrishnan, E K

    2018-03-01

    The bioactivity spectrum of fungal endophytes isolated from Zingiber officinale was analyzed against clinical pathogens and against the phytopathogen Pythium myriotylum, which causes Pythium rot in ginger. One of the isolates GFM13 showed broad bioactivity against various pathogens tested including P. myriotylum. The spore suspension as well as the culture filtrate of the endophytic fungal isolate was found to effectively protect ginger rhizomes from Pythium rot. By molecular identification, the fungal endophyte was identified as Paraconiothyrium sp. The bioactive compound produced by the isolate was separated by bioactivity-guided fractionation and was identified by GC-MS as danthron, an anthraquinone derivative. PCR amplification showed the presence of non-reducing polyketide synthase gene (NR-PKS) in the endophyte GFM13, which is reported to be responsible for the synthesis of anthraquinones in fungi. This is the first report of danthron being produced as the biologically active component of Paraconiothyrium sp. Danthron is reported to have wide pharmaceutical and agronomic applications which include its use as a fungicide in agriculture. The broad-spectrum antimicrobial activity of danthron and the endophytic origin of Paraconiothyrium sp. offer immense applications of the study.

  15. Combined genetic and bioactivity-based prioritization leads to the isolation of an endophyte-derived antimycobacterial compound.

    Alvin, A; Kalaitzis, J A; Sasia, B; Neilan, B A

    2016-05-01

    To initiate a genetic and bioactivity-based screening programme of culturable endophytes to identify micro-organisms capable of producing bioactive polyketides and peptides. Fungal endophytes were isolated from flowers, leaves and roots of Rhoeo spathacea, revealing a community consisting of Colletotrichum sp., Fusarium sp., Guignardia sp., Phomopsis sp., Phoma sp. and Microdochium sp. Genetic screening showed that all isolates had polyketide synthase (PKS) genes and most had nonribosomal peptide synthetase (NRPS) genes. Ethyl acetate extracts of the fungal isolates exhibited antiproliferative activity against at least one of the seven bacterial and mycobacterial test strains. Nuclear Magnetic Resonance -guided fractionation of the crude extract from a Fusarium sp. strain which exhibited strong antiproliferative activity against Mycobacterium tuberculosis resulted in the isolation of the polyketide javanicin. This compound was active against Myco. tuberculosis (MIC = 25 μg ml(-1)) and Mycobacterium phlei (MIC = 50 μg ml(-1)). The medicinal plant R. spathacea hosts a variety of fungal endophytes capable of producing antibacterial and antimycobacterial compounds. There is a positive correlation between the presence of PKS and/or NRPS encoding genes in endophytes and the bioactivity of their respective organic extracts. This is the first report on the fungal endophytic diversity of R. spathacea, and the isolation of an antimycobacterial compound from the plant which has been traditionally used for the treatment of tuberculosis symptoms. © 2016 The Society for Applied Microbiology.

  16. Screening for Endophytic Fungi from Turmeric Plant (Curcuma longa L.) of Sukabumi and Cibinong with Potency as Antioxidant Compounds Producer.

    Bustanussalam; Rachman, Fauzy; Septiana, Eris; Lekatompessy, Sylvia J R; Widowati, Tiwit; Sukiman, Harmastini I; Simanjuntak, Partomuan

    2015-01-01

    Potency of medicinal plant is related to microorganisms lived in the plant tissue. Those microorganisms are known as endophytic microbes that live and form colonies in the plant tissue without harming its host. Each plant may contains several endophytic microbes that produce biological compounds or secondary metabolites due to co-evolution or genetic transfer from the host plant to endophytic microbes. Endophytic fungi research done for turmeric plant (Curcuma longa L.) gave 44 isolated fungi as results. Those 44 fungi isolated were fermented in Potato Dextrose Broth (PDB) media, filtered, extracted with ethylacetate and then were analyzed by Thin Layer Chromatography (TLC) method and tested for their antioxidant activity by radical scavenging method. The antioxidant activity of the ethylacetate filtrate extracts either from Sukabumi or Cibinong were higher than the biomass extracts. There were 6 fungi that showed antioxidant activities over 65%, i.e., with code name K.Cl.Sb.R9 (93.58%), K.Cl.Sb.A11 (81.49%), KCl.Sb.B1 (78.81%), KCl.Sb.R11 (71.67%) and K.Cl.Sb.A12 (67.76%) from Sukabumi and K.Cl.Cb.U1 (69.27%) from Cibinong. These results showed that bioproduction by endophytic microbes can gave potential antioxidant compounds.

  17. Oasis desert farming selects environment-specific date palm root endophytic communities and cultivable bacteria that promote resistance to drought

    Cherif, Hanene; Marasco, Ramona; Rolli, Eleonora; Ferjani, Raoudha; Fusi, Marco; Soussi, Asma; Mapelli, Francesca; Blilou, Ikram; Borin, Sara; Boudabous, Abdellatif; Cherif, Ameur; Daffonchio, Daniele; Ouzari, Hadda

    2015-01-01

    Oases are desert-farming agro-ecosystems, where date palm (Phoenix dactyliferaL.) plays a keystone role in offsetting the effects of drought and maintaining a suitable microclimate for agriculture. At present, abundance, diversity and plant growth promotion (PGP) of date palm root-associated bacteria remain unknown. Considering the environmental pressure determined by the water scarcity in the desert environments, we hypothesized that bacteria associated with date palm roots improve plant resistance to drought. Here, the ecology of date palm root endophytes from oases in the Tunisian Sahara was studied with emphasis on their capacity to promote growth under drought. Endophytic communities segregated along a north-south gradient in correlation with geo-climatic parameters. Screening of 120 endophytes indicated that date palm roots select for bacteria with multiple PGP traits. Bacteria rapidly cross-colonized the root tissues of different species of plants, including the original Tunisian date palm cultivar, Saudi Arabian cultivars and Arabidopsis. Selected endophytes significantly increased the biomass of date palms exposed to repeated drought stress periods during a 9-month greenhouse experiment. Overall, results indicate that date palm roots shape endophytic communities that are capable to promote plant growth under drought conditions, thereby contributing an essential ecological service to the entire oasis ecosystem. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. Richness of endophytic fungi isolated from Opuntia ficus-indica Mill. (Cactaceae) and preliminary screening for enzyme production.

    Bezerra, J D P; Santos, M G S; Svedese, V M; Lima, D M M; Fernandes, M J S; Paiva, L M; Souza-Motta, C M

    2012-05-01

    Opuntia ficus-indica Mill. (forage cactus) is farmed with relative success in the semi-arid region of the Brazilian northeast for commercial purposes, particularly as forage and food. Endophytic microorganisms are those that can be isolated inside plant tissues and can be a new source to production of enzymes with different potentialities. The objective of this study was to describe the richness of endophytic fungi from O. ficus-indica and to detect the capacity of these species to produce extracellular hydrolytic enzymes. Forty-four endophytic fungi species were isolated. Among them, the most commonly found were Cladosporium cladosporioides (20.43%) and C. sphaerospermum (15.99%). Acremonium terricola, Monodictys castaneae, Penicillium glandicola, Phoma tropica and Tetraploa aristata are being reported for the first time as endophytic fungi for Brazil. The majority of isolated fungi exhibited enzymatic potential. Aspergillus japonicus and P. glandicola presented pectinolytic activity. Xylaria sp. was the most important among the other 14 species with positive cellulase activity. All 24 isolates analysed were xylanase-positive. Protease was best produced by isolate PF103. The results indicate that there is a significant richness of endophytic fungi in O. ficus-indica, and that these isolates indicate promising potential for deployment in biotechnological processes involving production of pectinases, cellulases, xylanases and proteases.

  19. Allelochemical effects of volatile compounds and organic extracts from Muscodor yucatanensis, a tropical endophytic fungus from Bursera simaruba.

    Macías-Rubalcava, Martha L; Hernández-Bautista, Blanca E; Oropeza, Fabiola; Duarte, Georgina; González, María C; Glenn, Anthony E; Hanlin, Richard T; Anaya, Ana Luisa

    2010-10-01

    Muscodor yucatanensis, an endophytic fungus, was isolated from the leaves of Bursera simaruba (Burseraceae) in a dry, semideciduous tropical forest in the Ecological Reserve El Eden, Quintana Roo, Mexico. We tested the mixture of volatile organic compounds (VOCs) produced by M. yucatanensis for allelochemical effects against other endophytic fungi, phytopathogenic fungi and fungoids, and plants. VOCs were lethal to Guignardia mangifera, Colletotrichum sp., Phomopsis sp., Alternaria solani, Rhizoctonia sp., Phytophthora capsici, and P. parasitica, but had no effect on Fusarium oxysporum, Xylaria sp., the endophytic isolate 120, or M. yucatanensis. VOCs inhibited root elongation in amaranth, tomato, and barnyard grass, particularly those produced during the first 15 days of fungal growth. VOCs were identified by gas chromatography/mass spectrometry and included compounds not previously reported from other Muscodor species and the previously reported compounds octane, 2-methyl butyl acetate, 2-pentyl furan, caryophyllene, and aromadendrene. We also evaluated organic extracts from the culture medium and mycelium of M. yucatanensis on the same endophytes, phytopathogens, and plants. In general, extracts inhibited plants more than endophytic or phytopathogens fungi. G. mangifera was the only organism that was significantly stimulated by both extracts regardless of concentration. Compounds in both organic extracts were identified by gas chromatography/mass spectrometry. We discuss the possible allelopathic role that metabolites of M. yucatanensis play in its ecological interactions with its host plant and other organisms.

  20. Endophytic bacterial community living in roots of healthy and 'Candidatus Phytoplasma mali'-infected apple (Malus domestica, Borkh.) trees.

    Bulgari, Daniela; Bozkurt, Adem I; Casati, Paola; Cağlayan, Kadriye; Quaglino, Fabio; Bianco, Piero A

    2012-11-01

    'Candidatus Phytoplasma mali', the causal agent of apple proliferation (AP) disease, is a quarantine pathogen controlled by chemical treatments against insect vectors and eradication of diseased plants. In accordance with the European Community guidelines, novel strategies should be developed for sustainable management of plant diseases by using resistance inducers (e.g. endophytes). A basic point for the success of this approach is the study of endophytic bacteria associated with plants. In the present work, endophytic bacteria living in healthy and 'Ca. Phytoplasma mali'-infected apple trees were described by cultivation-dependent and independent methods. 16S rDNA sequence analysis showed the presence of the groups Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria, Chlamydiae, and Firmicutes. In detail, library analyses underscored 24 and 17 operational taxonomic units (OTUs) in healthy and infected roots, respectively, with a dominance of Betaproteobacteria. Moreover, differences in OTUs number and in CFU/g suggested that phytoplasmas could modify the composition of endophytic bacterial communities associated with infected plants. Intriguingly, the combination of culturing methods and cloning analysis allowed the identification of endophytic bacteria (e.g. Bacillus, Pseudomonas, and Burkholderia) that have been reported as biocontrol agents. Future research will investigate the capability of these bacteria to control 'Ca. Phytoplasma mali' in order to develop sustainable approaches for managing AP.