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Sample records for azospirillum brasilense tarrand

  1. Reclassification of Roseomonas fauriae Rihs et al. 1998 as a later heterotypic synonym of Azospirillum brasilense Tarrand et al. 1979.

    Science.gov (United States)

    Helsel, Leta O; Hollis, Dannie G; Steigerwalt, Arnold G; Levett, Paul N

    2006-12-01

    The relatedness of Roseomonas fauriae and Azospirillum brasilense was investigated using phenotypic methods and DNA-DNA hybridization. Conventional biochemical tests did not differentiate between the two taxa. DNA-DNA hybridization experiments revealed high values for relatedness between the type strains of these species and suggest that these two taxa constitute a single species. Strains previously identified as R. fauriae should be reclassified as A. brasilense, with the name Roseomonas fauriae as a later heterotypic synonym of Azospirillum brasilense.

  2. Hydrogen oxidation in Azospirillum brasilense

    Energy Technology Data Exchange (ETDEWEB)

    Tibelius, K.

    1984-01-01

    Hydrogen oxidation by Azospirillum brasilense Sp7 was studied in N/sub 2/-fixing and NH/sub 4//sup +/-grown batch cultures. The K/sub m/ for H/sub 2/ of O/sub 2/-dependent H/sup 3/H oxidation in whole cells was 9 uM. The rates of H/sup 3/H and H/sub 2/ oxidation were very similar, indicating that the initial H/sub 2/ activation step in the overall H/sub 2/ oxidation reaction was not rate-limiting and that H/sup 3/H oxidation was a valid measure of H/sub 2/-oxidation activity. Hydrogen-oxidation activity was inhibited irreversibly by air. In N-free cultures the O/sub 2/ optima for O/sub 2/-dependent H/sub 2/ oxidation, ranging from 0.5-1.25% O/sub 2/ depending on the phase of growth, were significantly higher than those of C/sub 2/H/sub 2/ reduction, 0.15-0.35%, suggesting that the H/sub 2/-oxidation system may have a limited ability to aid in the protection of nitrogenase against inactivation by O/sub 2/. Oxygen-dependent H/sub 2/ oxidation was inhibited by NO/sub 2//sup +/, NO, CO, and C/sub 2/H/sub 2/ with apparent K/sub 1/ values of 20, 0.4, 28, and 88 uM, respectively. Hydrogen-oxidation activity was 50 to 100 times higher in denitrifying cultures when the terminal electron acceptor for growth was N/sub 2/O rather than NO/sub 3//sup -/, possibly due to the irreversible inhibition of hydrogenase by NO/sub 2//sup -/ and NO in NO/sub 3//sup -/-grown cultures.

  3. [Biosynthesis of gold nanoparticles by Azospirillum brasilense].

    Science.gov (United States)

    Kupriashina, M A; Vetchinkina, E P; Burov, A M; Ponomareva, E G; Nikitina, V E

    2014-01-01

    Plant-associated nitrogen-fixing soil bacteria Azospirillum brasilense were shown to reduce the gold of chloroauric acid to elemental gold, resulting in formation of gold nanoparicles. Extracellular phenoloxidizing enzymes (laccases and Mn peroxidases) were shown to participate in reduction of Au+3 (HAuCl4) to Au(0). Transmission electron microscopy revealed accumulation of colloidal gold nanoparticles of diverse shape in the culture liquid of A. brasilense strains Sp245 and Sp7. The size of the electron-dense nanospheres was 5 to 50 nm, and the size of nanoprisms varied from 5 to 300 nm. The tentative mechanism responsible for formation of gold nanoparticles is discussed.

  4. Migration of Azospirillum brasilense Yu62 from Root to Stem and Leaves Inside Rice and Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    CHIFeng; SHENShi-Hua; CHENSan-Feng; JINGYu-Xiang

    2004-01-01

    Azospirillum brasilense Tarrand, Krieg et Doebereiner is one of the important plant growthpromotion endophytes. A. brasilense Yu62 tagged with gfp gene was inoculated into roots of rice and tobacco seedlings, which were then, cultured in gnotobiotic condition. At a certain days after inoculation the different portions of the seedling were observed under laser confocal microscope, resulting in that A.brasilense Yu62 bacteria were colonized in epidermal and cortical cells, intercellular spaces and vascular system of stem and leaf tissue interiors besides in roots. Higher populations of the bacteria isolated from roots, stems and leaves indicated that A. brasilense Yu62 bacteria could ascend themselves from roots to stems and leaves of rice and tobacco. This observation lays down the foundation for ecology and cell morphology of bacterial migration inside plants, interaction between A. brasilense Yu62 bacteria and host cells as well as the plant-growth promotion, provides scientific basis for further application, and is of importance in science and practice.

  5. Biosíntesis de ácido antranílico y ácido indolacético a partir de triptófano en una cepa de Azospirillum brasilense nativa de Tamaulipas, México

    OpenAIRE

    J. L. Hernández-Mendoza; J. D. Quiroz-Velásquez; V. R. Moreno-Medina; N Mayek-Pérez

    2008-01-01

    La bacteria Azospirillum brasilense [Tarrand et al., 1979] metaboliza el triptófano y produce diversos metabolitos; entre ellos, el ácido indolacético. En este trabajo se utilizó la cepa CBG 497 de A. brasilense aislada de maíz para identificar los metabolitos producidos a partir de triptófano en laboratorio. Entre los metabolitos identificados destacan los ácidos antranílico e indolacético. El ácido antranílico es un compuesto intermediario en la síntesis y degradación del triptófano y, junt...

  6. SEED INOCULATION WITH Azospirillum brasilense, ASSOCIATED WITH THE USE OF BIOREGULATORS IN MAIZE

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    ALESSANDRO DE LUCCA E BRACCINI

    2012-01-01

    Full Text Available The inoculation of seeds with the bacterium Azospirillum has been carried out in maize culture and other grasses. The application of growth bio-regulators is another technology whose results in maize culture have yet to become more widespread. Current study evaluates the agronomic effectiveness of seed inoculation with Azospirillum brasilense in maize, associated with the use of the growth regulator Stimulate ®. Triple hybrid maize CD 304 underwent the following treatments: 1 - control without nitrogen and without Azospirillum brasilense; 2 - Treatment without nitrogen but with Azospirillum brasilense; 3 - Treatment without nitrogen but with Azospirillum brasilense + Stimulate ®; 4 - Treatment with 50% of nitrogen dose recommended for maize culture; 5 - Treatment with 50% of nitrogen dose and inoculation with Azospirillum brasilense; 6 - Same as 5 but with Stimulate ®; 7 - Total N recommended; 8 - Total N recommended + Azospirillum brasilense ; 9 - Total N recommended + Azospirillum brasilense + Stimulate ®. The inoculation of maize seeds with Azospirillum brasilense increases plant height and grain yield when compared with rates in control. The use of 50% of N dose in sowing, associated with the inoculation of maize seeds with Azospirillum brasilense at 200 mL ha-1 (mixed to the seeds and associated with Stimulate ® (in foliar application, is viable.

  7. Inoculation of maize with Azospirillum brasilense in the seed furrow

    OpenAIRE

    Tâmara Prado de Morais; Césio Humberto de Brito; Afonso Maria Brandão; Wender Santos Rezende

    2016-01-01

    ABSTRACT Several studies addressing the inoculation of cereals with diazotrophic microorganisms can be found in the literature. However, in many experiments, investigators have overlooked the feasibility of applying these microorganisms to the furrow together with the seed, and the effect of bacterial concentration on phytostimulation. The aim of this work was to evaluate the effect of doses of an inoculant based on Azospirillum brasilense, applied to the seed furrow when planting maize, comb...

  8. Network Analysis of Plasmidomes: The Azospirillum brasilense Sp245 Case.

    Science.gov (United States)

    Orlandini, Valerio; Emiliani, Giovanni; Fondi, Marco; Maida, Isabel; Perrin, Elena; Fani, Renato

    2014-01-01

    Azospirillum brasilense is a nitrogen-fixing bacterium living in association with plant roots. The genome of the strain Sp245, isolated in Brazil from wheat roots, consists of one chromosome and six plasmids. In this work, the A. brasilense Sp245 plasmids were analyzed in order to shed some light on the evolutionary pathways they followed over time. To this purpose, a similarity network approach was applied in order to identify the evolutionary relationships among all the A. brasilense plasmids encoded proteins; in this context a computational pipeline specifically devoted to the analysis and the visualization of the network-like evolutionary relationships among different plasmids molecules was developed. This information was supplemented with a detailed (in silico) functional characterization of both the connected (i.e., sharing homology with other sequences in the dataset) and the unconnected (i.e., not sharing homology) components of the network. Furthermore, the most likely source organism for each of the genes encoded by A. brasilense plasmids was checked, allowing the identification of possible trends of gene loss/gain in this microorganism. Data obtained provided a detailed description of the evolutionary landscape of the plasmids of A. brasilense Sp245, suggesting some of the molecular mechanisms responsible for the present-day structure of these molecules.

  9. Network Analysis of Plasmidomes: The Azospirillum brasilense Sp245 Case

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    Valerio Orlandini

    2014-01-01

    Full Text Available Azospirillum brasilense is a nitrogen-fixing bacterium living in association with plant roots. The genome of the strain Sp245, isolated in Brazil from wheat roots, consists of one chromosome and six plasmids. In this work, the A. brasilense Sp245 plasmids were analyzed in order to shed some light on the evolutionary pathways they followed over time. To this purpose, a similarity network approach was applied in order to identify the evolutionary relationships among all the A. brasilense plasmids encoded proteins; in this context a computational pipeline specifically devoted to the analysis and the visualization of the network-like evolutionary relationships among different plasmids molecules was developed. This information was supplemented with a detailed (in silico functional characterization of both the connected (i.e., sharing homology with other sequences in the dataset and the unconnected (i.e., not sharing homology components of the network. Furthermore, the most likely source organism for each of the genes encoded by A. brasilense plasmids was checked, allowing the identification of possible trends of gene loss/gain in this microorganism. Data obtained provided a detailed description of the evolutionary landscape of the plasmids of A. brasilense Sp245, suggesting some of the molecular mechanisms responsible for the present-day structure of these molecules.

  10. Inoculation of maize with Azospirillum brasilense in the seed furrow

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    Tâmara Prado de Morais

    2016-06-01

    Full Text Available ABSTRACT Several studies addressing the inoculation of cereals with diazotrophic microorganisms can be found in the literature. However, in many experiments, investigators have overlooked the feasibility of applying these microorganisms to the furrow together with the seed, and the effect of bacterial concentration on phytostimulation. The aim of this work was to evaluate the effect of doses of an inoculant based on Azospirillum brasilense, applied to the seed furrow when planting maize, combined with different doses of nitrogen fertiliser. The experiment was carried out in the field, in soil of the cerrado region of Brazil. An experimental design of randomised blocks in bands was adopted, comprising nitrogen (40, 100, 200 and 300 kg ha-1 and doses of an A. brasilense-based liquid inoculant applied to the seed furrow (0, 100, 200, 300 and 400 mL ha-1. The dose of 200 mL ha-1Azospirillum was noteworthy for grain production. This is the first report of the effective application of Azospirillum in the seed furrow when planting maize in the cerrado region of Brazil.

  11. Phenotypic variation in Azospirillum brasilense exposed to starvation.

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    Lerner, Anat; Valverde, Angel; Castro-Sowinski, Susana; Lerner, Hadas; Okon, Yaacov; Burdman, Saul

    2010-08-01

    Bacteria have developed mechanisms that allow them maintaining cell viability during starvation and resuming growth when nutrients become available. Among these mechanisms are adaptive mutations and phase variation, which are often associated with DNA rearrangements. Azospirillum brasilense is a Gram-negative, nitrogen-fixing, plant growth-promoting rhizobacterium. Here we report phenotypic variants of A. brasilense that were collected after exposure to prolonged starvation or after re-isolation from maize roots. The variants differed in several features from the parental strains, including pigmentation, aggregation ability, EPS amount and composition and LPS structure. One of the phenotypic variants, overproducing EPS and showing an altered LPS structure, was further characterized and showed differential response to several stresses and antibiotics relative to its parental strain. Characterization of the variants by repetitive-PCR revealed that phenotypic variation was often associated with DNA rearrangements.

  12. Identification of the glutamine synthetase adenylyltransferase of Azospirillum brasilense.

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    Van Dommelen, Anne; Spaepen, Stijn; Vanderleyden, Jozef

    2009-04-01

    Glutamine synthetase, a key enzyme in nitrogen metabolism of both prokaryotes and eukaryotes, is strictly regulated. One means of regulation is the modulation of activity through adenylylation catalyzed by adenylyltransferases. Using PCR primers based on conserved sequences in glutamine synthetase adenylyltransferases, we amplified part of the glnE gene of Azospirillum brasilense Sp7. The complete glnE sequence of A. brasilense Sp245 was retrieved from the draft genome sequence of this organism (http://genomics.ornl.gov/research/azo/). Adenylyltransferase is a bifunctional enzyme consisting of an N-terminal domain responsible for deadenylylation activity and a C-terminal domain responsible for adenylylation activity. Both domains are partially homologous to each other. Residues important for catalytic activity were present in the deduced amino acid sequence of the A. brasilense Sp245 glnE sequence. A glnE mutant was constructed in A. brasilense Sp7 by inserting a kanamycin resistance cassette between the two active domains of the enzyme. The resulting mutant was unable to adenylylate the glutamine synthetase enzyme and was impaired in growth when shifted from nitrogen-poor to nitrogen-rich medium.

  13. Directed mutagenesis affects recombination in Azospirillum brasilense nif genes

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    C.P. Nunes

    2000-12-01

    Full Text Available In order to improve the gene transfer/mutagenesis system for Azospirillum brasilense, gene-cartridge mutagenesis was used to replace the nifD gene with the Tn5 kanamycin resistance gene. The construct was transferred to A. brasilense by electrotransformation. Of the 12 colonies isolated using the suicide plasmid pSUP202 as vector, only four did not show vector integration into the chromosome. Nevertheless, all 12 colonies were deficient in acetylene reduction, indicating an Nif- phenotype. Four Nif- mutants were analyzed by Southern blot, using six different probes spanning the nif and Km r genes and the plasmid vector. Apparently, several recombination events occurred in the mutant genomes, probably caused mainly by gene disruption owing to the mutagenesis technique used: resistance gene-cartridge mutagenesis combined with electrotransformation.Com o objetivo de melhorar os sistemas de transferência gênica e mutagênese para Azospirillum brasilense, a técnica de mutagênese através do uso de um gene marcador ("gene-cartridge mutagenesis" foi utilizada para substituir a região genômica de A. brasilense correspondente ao gene nifD por um segmento de DNA do transposon Tn5 contendo o gene que confere resistência ao antibiótico canamicina. A construção foi transferida para a linhagem de A. brasilense por eletrotransformação. Doze colônias transformantes foram isoladas com o plasmídeo suicida pSUP202 servindo como vetor. Dessas, somente quatro não possuíam o vetor integrado no cromossomo da bactéria. Independentemente da integração ou não do vetor, as 12 colônias foram deficientes na redução do gás acetileno, evidenciando o fenótipo Nif -. Quatro mutantes Nif - foram analisados através da técnica de Southern blot, utilizando-se seis diferentes fragmentos contendo genes nif, de resistência à canamicina e do vetor como sondas. Os resultados sugerem a ocorrência de eventos recombinacionais variados no genoma dos mutantes. A

  14. Azospirillum brasilense siderophores with antifungal activity against Colletotrichum acutatum.

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    Tortora, María L; Díaz-Ricci, Juan C; Pedraza, Raúl O

    2011-04-01

    Anthracnose, caused by the fungus Colletotrichum acutatum is one of the most important diseases in strawberry crop. Due to environmental pollution and resistance produced by chemical fungicides, nowadays biological control is considered a good alternative for crop protection. Among biocontrol agents, there are plant growth-promoting bacteria, such as members of the genus Azospirillum. In this work, we demonstrate that under iron limiting conditions different strains of A. brasilense produce siderophores, exhibiting different yields and rates of production according to their origin. Chemical assays revealed that strains REC2 and REC3 secrete catechol type siderophores, including salicylic acid, detected by thin layer chromatography coupled with fluorescence spectroscopy and gas chromatography-mass spectrometry analysis. Siderophores produced by them showed in vitro antifungal activity against C. acutatum M11. Furthermore, this latter coincided with results obtained from phytopathological tests performed in planta, where a reduction of anthracnose symptoms on strawberry plants previously inoculated with A. brasilense was observed. These outcomes suggest that some strains of A. brasilense could act as biocontrol agent preventing anthracnose disease in strawberry.

  15. Aggregation of the rhizospheric bacterium Azospirillum brasilense in response to oxygen

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    Abdoun, Hamid; McMillan, Mary; Pereg, Lily

    2016-04-01

    Azospirillum brasilense spp. have ecological, scientific and agricultural importance. As model plant growth promoting rhizobacteria they interact with a large variety of plants, including important food and cash crops. Azospirillum strains are known for their production of plant growth hormones that enhance root systems and for their ability to fix nitrogen. Azospirillum cells transform in response to environmental cues. The production of exopolysaccharides and cell aggregation during cellular transformation are important steps in the attachment of Azospirillum to roots. We investigate signals that induce cellular transformation and aggregation in the Azospirillum and report on the importance of oxygen to the process of aggregation in this rhizospheric bacterium.

  16. Accessing inoculation methods of maize and wheat with Azospirillum brasilense.

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    Fukami, Josiane; Nogueira, Marco Antonio; Araujo, Ricardo Silva; Hungria, Mariangela

    2016-03-01

    The utilization of inoculants containing Azospirillum is becoming more popular due to increasing reports of expressive gains in grain yields. However, incompatibility with pesticides used in seed treatments represents a main limitation for a successful inoculation. Therefore, in this study we searched for alternatives methods for seed inoculation of maize and wheat, aiming to avoid the direct contact of bacteria with pesticides. Different doses of inoculants containing Azospirillum brasilense were employed to perform inoculation in-furrow, via soil spray at sowing and via leaf spray after seedlings had emerged, in comparison to seed inoculation. Experiments were conducted first under greenhouse controlled conditions and then confirmed in the field at different locations in Brazil. In the greenhouse, most parameters measured responded positively to the largest inoculant dose used in foliar sprays, but benefits could also be observed from both in-furrow and soil spray inoculation. However, our results present evidence that field inoculation with plant-growth promoting bacteria must consider inoculant doses, and point to the need of fine adjustments to avoid crossing the threshold of growth stimulation and inhibition. All inoculation techniques increased the abundance of diazotrophic bacteria in plant tissues, and foliar spray improved colonization of leaves, while soil inoculations favored root and rhizosphere colonization. In field experiments, inoculation with A. brasilense allowed for a 25 % reduction in the need for N fertilizers. Our results have identified alternative methods of inoculation that were as effective as the standard seed inoculation that may represent an important strategy to avoid the incompatibility between inoculant bacteria and pesticides employed for seed treatment.

  17. Seed quality, chlorophyll content index and leaf nitrogen levels in maize inoculated with Azospirillum brasilense

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    Lívia de Matos Pereira; Elise de Matos Pereira; Lucas Tadeu Mazza Revolti; Sonia Marli Zingaretti; Gustavo Vitti Môro

    2015-01-01

    The aim of this study was to evaluate differences between maize genotypes in relation to the germination response of the seeds and the growth of seedlings inoculated with Azospirillum brasilense, as well as the effect of inoculation on nitrogen levels and the chlorophyll content index of the leaves. The physiological seeds quality from the single-cross hybrids AG7098 and 2B707, and from the experimental synthetic varieties V2 and V4, inoculated with A. brasilense, was tested for germination, ...

  18. [Chemical composition and immunochemical characteristics of the lipopolysaccharide of nitrogen-fixing rhizobacterium Azospirillum brasilense CD].

    Science.gov (United States)

    Konnova, O N; Burygin, G L; Fedonenko, Iu P; Matora, L Iu; Pankin, K E; Konnova, S A; Ignatov, V V

    2006-01-01

    The chemical composition of the lipopolysaccharide of the associative diazotrophic rhizobacterium Azospirillum brasilense Cd has been studied. Among the main components of the hydrophobic part of the lipopolysaccharide, we identified 3-hydroxytetradecanoic, hexadecenoic, 3-hydroxyhexadecanoic, hexadecanoic, octadecenoic, and nanodecanoic fatty acids; the carbohydrate part contained rhamnose, galactose, and mannose. Polyclonal antibodies against the preparation under study were raised in rabbits. Serological relations between A. brasilense Cd and other strains of Azospirillum spp. were studied using double radial immunodiffusion and enzyme-linked immunosorbent assay.

  19. SEED INOCULATION WITH Azospirillum brasilense, ASSOCIATED WITH THE USE OF BIOREGULATORS IN MAIZE

    OpenAIRE

    Alessandro de Lucca e Braccini; LILIAN GOMES DE MORAES DAN; GLEBERSON GUILLEN PICCININ; LEANDRO PAIOLA ALBRECHT; MAURO CEZAR BARBOSA; ALEX HENRIQUE TIENE ORTIZ

    2012-01-01

    The inoculation of seeds with the bacterium Azospirillum has been carried out in maize culture and other grasses. The application of growth bio-regulators is another technology whose results in maize culture have yet to become more widespread. Current study evaluates the agronomic effectiveness of seed inoculation with Azospirillum brasilense in maize, associated with the use of the growth regulator Stimulate ®. Triple hybrid maize CD 304 underwent the following treatments: 1 - control withou...

  20. Oxidative and antioxidative responses in the wheat-Azospirillum brasilense interaction.

    Science.gov (United States)

    Méndez-Gómez, Manuel; Castro-Mercado, Elda; Alexandre, Gladys; García-Pineda, Ernesto

    2016-03-01

    Azospirillum is a plant growth-promoting rhizobacteria (PGPR) able to enhance the growth of wheat. The aim of this study was to test the effect of Azospirillum brasilense cell wall components on superoxide (O2·(-)) production in wheat roots and the effect of oxidative stress on A. brasilense viability. We found that inoculation with A. brasilense reduced O2·(-) levels by approx. 30 % in wheat roots. Inoculation of wheat with papain-treated A. brasilense, a Cys protease, notably increased O2·(-) production in all root tissues, as was observed by the nitro blue tetrazolium (NBT) reduction. However, a 24-h treatment with rhizobacteria lipopolysaccharides (50 and 100 μg/mL) alone did not affect the pattern of O2·(-) production. Analysis of the effect of plant cell wall components on A. brasilense oxidative enzyme activity showed no changes in catalase activity but a decrease in superoxide dismutase activity in response to polygalacturonic acid treatment. Furthermore, A. brasilense growth was only affected by high concentrations of H2O2 or paraquat, but not by sodium nitroprusside. Our results suggest that rhizobacterial cell wall components play an important role in controlling plant cell responses and developing tolerance of A. brasilense to oxidative stress produced by the plant.

  1. Trehalose accumulation in Azospirillum brasilense improves drought tolerance and biomass in maize plants.

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    Rodríguez-Salazar, Julieta; Suárez, Ramón; Caballero-Mellado, Jesús; Iturriaga, Gabriel

    2009-07-01

    Bacteria of the genus Azospirillum increase the grain yield of several grass crops. In this work the effect of inoculating maize plants with genetically engineered Azospirillum brasilense for trehalose biosynthesis was determined. Transformed bacteria with a plasmid harboring a trehalose biosynthesis gene-fusion from Saccharomyces cerevisiae were able to grow up to 0.5 M NaCl and to accumulate trehalose, whereas wild-type A. brasilense did not tolerate osmotic stress or accumulate significant levels of the disaccharide. Moreover, 85% of maize plants inoculated with transformed A. brasilense survived drought stress, in contrast with only 55% of plants inoculated with the wild-type strain. A 73% increase in biomass of maize plants inoculated with transformed A. brasilense compared with inoculation with the wild-type strain was found. In addition, there was a significant increase of leaf and root length in maize plants inoculated with transformed A. brasilense. Therefore, inoculation of maize plants with A. brasilense containing higher levels of trehalose confers drought tolerance and a significant increase in leaf and root biomass. This work opens the possibility that A. brasilense modified with a chimeric trehalose biosynthetic gene from yeast could increase the biomass, grain yield and stress tolerance in other relevant crops.

  2. Electrophysical characteristics of Azospirillum brasilense Sp245 during interaction with antibodies to various cell surface epitopes.

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    Guliy, Olga I; Matora, Larisa Y; Burygin, Gennady L; Dykman, Lev A; Ostudin, Nikolai A; Bunin, Viktor D; Ignatov, Vladimir V; Ignatov, Oleg V

    2007-11-15

    This work was undertaken to examine the electrooptical characteristics of cells of Azospirillum brasilense Sp245 during their interaction with antibodies developed to various cell surface epitopes. We used the dependences of the cell suspension optical density changes induced by electroorientation on the orienting field frequency (740, 1000, 1450, 2000, and 2800kHz). Cell interactions with homologous strain-specific antibodies to the A. brasilense Sp245 O antigen and with homologous antibodies to whole bacterial cells brought about considerable changes in the electrooptical properties of the bacterial suspension. When genus-specific antibodies to the flagellin of the Azospirillum sheathed flagellum and antibodies to the serologically distinct O antigen of A. brasilense Sp7 were included in the A. brasilense Sp245 suspension, the changes caused in the electrooptical signal were slight and had values close to those for the above changes. These findings agree well with the immunochemical characteristics of the Azospirillum O antigens and with the data on the topographical distribution of the Azospirillum major cell surface antigens. The obtained results can serve as a basis for the development of a rapid test for the intraspecies detection of microorganisms.

  3. Repressor mutant forms of the Azospirillum brasilense NtrC protein.

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    Huergo, Luciano F; Assumpção, Marcelo C; Souza, Emanuel M; Steffens, M Berenice R; Yates, M Geoffrey; Chubatsu, Leda S; Pedrosa, Fábio O

    2004-10-01

    The Azospirillum brasilense mutant strains FP8 and FP9, after treatment with nitrosoguanidine, showed a null Nif phenotype and were unable to use nitrate as their sole nitrogen source. Sequencing of the ntrC genes revealed single nucleotide mutations in the NtrC nucleotide-binding site. The phenotypes of these strains are discussed in relation to their genotypes.

  4. Interaction of Azospirillum brasilense and Glomus intrarradix in Sugar Cane Roots.

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    Bellone, Carlos H; de Bellone Silvia, Carrizo

    2012-03-01

    Fifteen-day-old variety NA 56-79 sugar cane seedlings were inoculated with Azospirillum brasilense and Glomus intrarradix. This article aims at examining changes in sugar cane root seedlings inoculated with Glomus intrarradix and Azospirillum brasilense, the increase in microbial biomass and the acetylene reduction process as well. The internal root colonization was studied 20 days after inoculation using scanning and a transmission electron microscope. Both microorganisms entered the sugar cane root through the emergent lateral roots. The microorganisms were capable of coexisting both intra and intercellularly, producing changes in the cell wall, thus allowing colonization and interaction between the organisms. These changes increased the number of microorganisms inside the root as well as acetylene nitrogen reduction. Sugar cane plant biomass increased with joint-inoculation. The number of endophytic microorganisms and nitrogen fixing activity increased when they were colonized by Azospirillum and Glomus together.

  5. Azospirillum brasilense ameliorates the response of Arabidopsis thaliana to drought mainly via enhancement of ABA levels.

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    Cohen, Ana C; Bottini, Rubén; Pontin, Mariela; Berli, Federico J; Moreno, Daniela; Boccanlandro, Hernán; Travaglia, Claudia N; Piccoli, Patricia N

    2015-01-01

    Production of phytohormones is one of the main mechanisms to explain the beneficial effects of plant growth-promoting rhizobacteria (PGPR) such as Azospirillum sp. The PGPRs induce plant growth and development, and reduce stress susceptibility. However, little is known regarding the stress-related phytohormone abscisic acid (ABA) produced by bacteria. We investigated the effects of Azospirillum brasilense Sp 245 strain on Arabidopsis thaliana Col-0 and aba2-1 mutant plants, evaluating the morphophysiological and biochemical responses when watered and in drought. We used an in vitro-grown system to study changes in the root volume and architecture after inoculation with Azospirillum in Arabidopsis wild-type Col-0 and on the mutant aba2-1, during early growth. To examine Arabidopsis development and reproductive success as affected by the bacteria, ABA and drought, a pot experiment using Arabidopsis Col-0 plants was also carried out. Azospirillum brasilense augmented plant biomass, altered root architecture by increasing lateral roots number, stimulated photosynthetic and photoprotective pigments and retarded water loss in correlation with incremented ABA levels. As well, inoculation improved plants seed yield, plants survival, proline levels and relative leaf water content; it also decreased stomatal conductance, malondialdehyde and relative soil water content in plants submitted to drought. Arabidopsis inoculation with A. brasilense improved plants performance, especially in drought.

  6. Bacteriophage prevalence in the genus Azospirillum and analysis of the first genome sequence of an Azospirillum brasilense integrative phage.

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    Boyer, Mickaël; Haurat, Jacqueline; Samain, Sylvie; Segurens, Béatrice; Gavory, Frédérick; González, Víctor; Mavingui, Patrick; Rohr, René; Bally, René; Wisniewski-Dyé, Florence

    2008-02-01

    The prevalence of bacteriophages was investigated in 24 strains of four species of plant growth-promoting rhizobacteria belonging to the genus Azospirillum. Upon induction by mitomycin C, the release of phage particles was observed in 11 strains from three species. Transmission electron microscopy revealed two distinct sizes of particles, depending on the identity of the Azospirillum species, typical of the Siphoviridae family. Pulsed-field gel electrophoresis and hybridization experiments carried out on phage-encapsidated DNAs revealed that all phages isolated from A. lipoferum and A. doebereinerae strains had a size of about 10 kb whereas all phages isolated from A. brasilense strains displayed genome sizes ranging from 62 to 65 kb. Strong DNA hybridizing signals were shown for most phages hosted by the same species whereas no homology was found between phages harbored by different species. Moreover, the complete sequence of the A. brasilense Cd bacteriophage (phiAb-Cd) genome was determined as a double-stranded DNA circular molecule of 62,337 pb that encodes 95 predicted proteins. Only 14 of the predicted proteins could be assigned functions, some of which were involved in DNA processing, phage morphogenesis, and bacterial lysis. In addition, the phiAb-Cd complete genome was mapped as a prophage on a 570-kb replicon of strain A. brasilense Cd, and a region of 27.3 kb of phiAb-Cd was found to be duplicated on the 130-kb pRhico plasmid previously sequenced from A. brasilense Sp7, the parental strain of A. brasilense Cd.

  7. brasilense nativa de Tamaulipas, México

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    J. L. Hernández-Mendoza

    2008-01-01

    Full Text Available La bacteria Azospirillum brasilense [Tarrand et al., 1979] metaboliza el triptófano y produce diversos metabolitos; entre ellos, el ácido indolacético. En este trabajo se utilizó la cepa CBG 497 de A. brasilense aislada de maíz para identificar los metabolitos producidos a partir de triptófano en laboratorio. Entre los metabolitos identificados destacan los ácidos antranílico e indolacético. El ácido antranílico es un compuesto intermediario en la síntesis y degradación del triptófano y, junto con el ácido indolacético, estaría involucrado en la capacidad promotora del crecimiento vegetal de A. brasilense

  8. Characterization of the flagellar biosynthesis regulatory geneflbD in Azospirillum brasilense

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A flagellar gene cluster fragment includingflbD of Azospirillum brasilense was cloned and sequenced, The flbD mutant strain was found to be nonmotile-losing both polar and lateral flagella (Fla-Laf-), Motility and flagella were regained by complementation with plasmid-borne multicopy flbD, but altered with larger swarming circle and fewer lateral flagella on the semisolid plate, This result indicated that FIbD plays an important role in the regulation of both polar and lateral flagellar biosynthesis in A.brasilense.

  9. Structure of the polysaccharides from the lipopolysaccharide of Azospirillum brasilense Jm125A2.

    Science.gov (United States)

    Sigida, Elena N; Fedonenko, Yuliya P; Shashkov, Alexander S; Zdorovenko, Evelina L; Konnova, Svetlana A; Ignatov, Vladimir V; Knirel, Yuriy A

    2015-10-30

    Two polysaccharides were obtained by mild acid degradation of the lipopolysaccharide of associative nitrogen-fixing bacteria Azospirillum brasilense Jm125A2 isolated from the rhizosphere of a pearl millet. The following structures of the polysaccharides were established by sugar and methylation analyses, Smith degradation, and (1)H and (13)C NMR spectroscopy: [Formula: see text] Structure 1 has been reported earlier for a polysaccharide from A. brasilense S17 (Fedonenko YP, Konnova ON, Zdorovenko EL, Konnova SA, Zatonsky GV, Shaskov AS, Ignatov VV, Knirel YA. Carbohydr Res 2008;343:810-6), whereas to our knowledge structure 2 has not been hitherto found in bacterial polysaccharides.

  10. GlnB is specifically required for Azospirillum brasilense NifA activity in Escherichia coli.

    Science.gov (United States)

    Araújo, Luiza M; Monteiro, Rose A; Souza, Emanuel M; Steffens, M Berenice R; Rigo, Liu U; Pedrosa, Fábio O; Chubatsu, Leda S

    2004-01-01

    The Azospirillum brasilense transcription regulator NifA and the nitrogen-status signaling proteins GlnB, GlnZ and GlnK were expressed in Escherichia coli and analyzed for their ability to activate nif gene expression. When expressed separately, none of the proteins were able to activate nifH promoter expression in any tested conditions; in contrast, nifH expression was observed in cells grown in the absence of ammonium and oxygen and when expressing simultaneously NifA and GlnB proteins, but not when expressing NifA and GlnZ or GlnK. Our results show that the GlnB protein is required for transcription activation by Azospirillum brasilense NifA and it cannot be replaced by GlnZ or GlnK.

  11. [Effect of lectins from Azospirillum brasilense to peroxidase and oxalate oxidase activity regulation in wheat roots].

    Science.gov (United States)

    Alen'kina, S A; Nikitina, V E

    2010-01-01

    Lectins were extracted from the surface of nitrogen-fixing soil bacteria Azospirillum brasilense Sp7 and from its mutant A. brasilense Sp7.2.3 defective in lectin activity. The ability oflectins to stimulate the rapid formation of hydrogen peroxide related to increase of oxalate oxidase and peroxidase activity in the roots of wheat seedlings has been demonstrated. The most rapid induced pathway of hydrogen peroxide formation in the roots of wheat seedlings was the oxalic acid oxidation by oxalate oxidase which is the effect oflectin in under 10 min in a concentration of 10 microg/ml. The obtained results show that lectins from Azospirillum are capable of inducing the adaptation processes in the roots of wheat seedlings.

  12. Co-inoculation of Azospirillum brasilense and Herbaspirillum seropedicae in maize

    OpenAIRE

    Janaína Dartora; Deniele Marini; Edilaine. D. V. Gonçalves; Vandeir F. Guimarães

    2016-01-01

    ABSTRACT Bacteria from the genera Azospirillum and Herbaspirillum have been associated with increments in maize yield. The aim of this study was to evaluate the maize yield and nutritional content in response to inoculation with A. brasilense and H. seropediceae in association with nitrogen (N) fertilization. The experimental design was randomized blocks in a 4 x 5 factorial scheme, with four replicates. The treatments consisted of seed inoculation (control without N and inoculation, A. brasi...

  13. Fourier transform infrared spectroscopic study of intact cells of the nitrogen-fixing bacterium Azospirillum brasilense

    Science.gov (United States)

    Kamnev, A. A.; Ristić, M.; Antonyuk, L. P.; Chernyshev, A. V.; Ignatov, V. V.

    1997-06-01

    The data of Fourier transform infrared (FTIR) spectroscopic measurements performed on intact cells of the soil nitrogen-fixing bacterium Azospirillum brasilense grown in a standard medium and under the conditions of an increased metal uptake are compared and discussed. The structural FTIR information obtained is considered together with atomic absorption spectrometry (AAS) data on the content of metal cations in the bacterial cells. Some methodological aspects concerning preparation of bacterial cell samples for FTIR measurements are also discussed.

  14. Transcriptome analysis of the rhizosphere bacterium Azospirillum brasilense reveals an extensive auxin response.

    Science.gov (United States)

    Van Puyvelde, Sandra; Cloots, Lore; Engelen, Kristof; Das, Frederik; Marchal, Kathleen; Vanderleyden, Jos; Spaepen, Stijn

    2011-05-01

    The rhizosphere bacterium Azospirillum brasilense produces the auxin indole-3-acetic acid (IAA) through the indole-3-pyruvate pathway. As we previously demonstrated that transcription of the indole-3-pyruvate decarboxylase (ipdC) gene is positively regulated by IAA, produced by A. brasilense itself or added exogenously, we performed a microarray analysis to study the overall effects of IAA on the transcriptome of A. brasilense. The transcriptomes of A. brasilense wild-type and the ipdC knockout mutant, both cultured in the absence and presence of exogenously added IAA, were compared.Interfering with the IAA biosynthesis/homeostasis in A. brasilense through inactivation of the ipdC gene or IAA addition results in much broader transcriptional changes than anticipated. Based on the multitude of changes observed by comparing the different transcriptomes, we can conclude that IAA is a signaling molecule in A. brasilense. It appears that the bacterium, when exposed to IAA, adapts itself to the plant rhizosphere, by changing its arsenal of transport proteins and cell surface proteins. A striking example of adaptation to IAA exposure, as happens in the rhizosphere, is the upregulation of a type VI secretion system (T6SS) in the presence of IAA. The T6SS is described as specifically involved in bacterium-eukaryotic host interactions. Additionally, many transcription factors show an altered regulation as well, indicating that the regulatory machinery of the bacterium is changing.

  15. Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilense

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    L.M.P. Passaglia

    1998-11-01

    Full Text Available NifA protein activates transcription of nitrogen fixation operons by the alternative sigma54 holoenzyme form of RNA polymerase. This protein binds to a well-defined upstream activator sequence (UAS located at the -200/-100 position of nif promoters with the consensus motif TGT-N10-ACA. NifA of Azospirillum brasilense was purified in the form of a glutathione-S-transferase (GST-NifA fusion protein and proteolytic release of GST yielded inactive and partially soluble NifA. However, the purified NifA was able to induce the production of specific anti-A. brasilense NifA-antiserum that recognized NifA from A. brasilense but not from K. pneumoniae. Both GST-NifA and NifA expressed from the E. coli tac promoter are able to activate transcription from the nifHDK promoter but only in an A. brasilense background. In order to investigate the mechanism that regulates NifA binding capacity we have used E. coli total protein extracts expressing A. brasilense nifA in mobility shift assays. DNA fragments carrying the two overlapping, wild-type or mutated UAS motifs present in the nifH promoter region revealed a retarded band of related size. These data show that the binding activity present in the C-terminal domain of A. brasilense NifA protein is still functional even in the presence of oxygen.

  16. Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilense.

    Science.gov (United States)

    Passaglia, L M; Van Soom, C; Schrank, A; Schrank, I S

    1998-11-01

    NifA protein activates transcription of nitrogen fixation operons by the alternative sigma 54 holoenzyme form of RNA polymerase. This protein binds to a well-defined upstream activator sequence (UAS) located at the -200/-100 position of nif promoters with the consensus motif TGT-N10-ACA. NifA of Azospirillum brasilense was purified in the form of a glutathione-S-transferase (GST)-NifA fusion protein and proteolytic release of GST yielded inactive and partially soluble NifA. However, the purified NifA was able to induce the production of specific anti-A. brasilense NifA-antiserum that recognized NifA from A. brasilense but not from K. pneumoniae. Both GST-NifA and NifA expressed from the E. coli tac promoter are able to activate transcription from the nifHDK promoter but only in an A. brasilense background. In order to investigate the mechanism that regulates NifA binding capacity we have used E. coli total protein extracts expressing A. brasilense nifA in mobility shift assays. DNA fragments carrying the two overlapping, wild-type or mutated UAS motifs present in the nifH promoter region revealed a retarded band of related size. These data show that the binding activity present in the C-terminal domain of A. brasilense NifA protein is still functional even in the presence of oxygen.

  17. [Determination of Azospirillum Brasilense Cells With Bacteriophages via Electrooptical Analysis of Microbial Suspensions].

    Science.gov (United States)

    Gulii, O I; Karavayeva, O A; Pavlii, S A; Sokolov, O I; Bunin, V D; Ignatov, O V

    2015-01-01

    The dependence-of changes in the electrooptical properties of Azospirillum brasilense cell suspension Sp7 during interaction with bacteriophage ΦAb-Sp7 on the number and time of interactions was studied. Incubation of cells with bacteriophage significantly changed the electrooptical signal within one minute. The selective effect of bacteriophage ΦAb on 18 strains of bacteria of the genus Azospirillum was studied: A. amazonense Ami4, A. brasilense Sp7, Cd, Sp107, Sp245, Jm6B2, Brl4, KR77, S17, S27, SR55, SR75, A. halopraeferans Au4, A. irakense KBC1, K A3, A. lipoferum Sp59b, SR65 and RG20a. We determined the limit of reliable determination of microbial cells infected with bacteriophage: - 10(4) cells/mL. The presence of foreign cell cultures of E. coli B-878 and E. coli XL-1 did not complicate the detection of A brasilense Sp7 cells with the use of bacteriophage ΦAb-Sp7. The results demonstrated that bacteriophage (ΦAb-Sp7 can be used for the detection of Azospirillum microbial cells via t electrooptical analysis of cell suspensions.

  18. [Biofilm Formation by the Nonflagellated flhB1 Mutant of Azospirillum brasilense Sp245].

    Science.gov (United States)

    Shelud'ko, A V; Filip'echeva, Yu A; Shumiliva, E M; Khlebtsov, B N; Burov, A M; Petrova, L P; Katsy, E I

    2015-01-01

    Azospirillum brasilense Sp245 with mixed flagellation are able to form biofilms on various surfaces. A nonflagellated mutant of this strain with inactivated chromosomal copy of the flhB gene (flhB1) was shown to exhibit specific traits at the later stages of biofilm formation on a hydrophilic (glass) surface. Mature biofilms of the flhB1::Omegon-Km mutant Sp245.1063 were considerably thinner than those of the parent strain Sp245. The biofilms of the mutant were more susceptible to the forces of hydrodynamic shear. A. brasilense Sp245 cells in biofilms were not found to possess lateral flagella. Cells with polar flagella were, however, revealed by atomic force microscopy of mature native biofilms of strain Sp245. Preservation of a polar flagellum (probably nonmotile) on the cells of A. brasilense Sp245 may enhance the biofilm stability.

  19. [Factors inducing transition from growth to dormancy in rhizobacteria Azospirillum brasilense].

    Science.gov (United States)

    Kushneruk, M A; Tugarova, A V; Il'chukova, A V; Slavkina, E A; Starichkova, N I; Bogatyrev, V A; Antoniuk, L P

    2013-01-01

    The factors suppressing division of the cells of the rhizobacterium Azospirillum brasilense and inducing their transition to a dormant state were analyzed. These included the presence of hexylresorcinol or heavy metals (Cu and Co) in the medium, oxygen stress, and transfer of the cells into the physiological saline or phosphate buffer solution. The results were used to develop a protocol for obtaining of uncultured cells of A. brasilense Sp245, a natural symbiont of wheat. The cells lost their ability to grow on synthetic agar medium, but could revert to growth when incubated in freshly prepared liquid medium. Needle-shaped crystals differing from struvite, which has been previously reported for this strain, were found in the dormant culture of A. brasilense Sp245.

  20. Denitrification-derived nitric oxide modulates biofilm formation in Azospirillum brasilense.

    Science.gov (United States)

    Arruebarrena Di Palma, Andrés; Pereyra, Cintia M; Moreno Ramirez, Lizbeth; Xiqui Vázquez, María L; Baca, Beatriz E; Pereyra, María A; Lamattina, Lorenzo; Creus, Cecilia M

    2013-01-01

    Azospirillum brasilense is a rhizobacterium that provides beneficial effects on plants when they colonize roots. The formation of complex bacterial communities known as biofilms begins with the interaction of planktonic cells with surfaces in response to appropriate signals. Nitric oxide (NO) is a signaling molecule implicated in numerous processes in bacteria, including biofilm formation or dispersion, depending on genera and lifestyle. Azospirillum brasilense Sp245 produces NO by denitrification having a role in root growth promotion. We analyzed the role of endogenously produced NO on biofilm formation in A. brasilense Sp245 and in a periplasmic nitrate reductase mutant (napA::Tn5; Faj164) affected in NO production. Cells were statically grown in media with nitrate or ammonium as nitrogen sources and examined for biofilm formation using crystal violet and by confocal laser microscopy. Both strains formed biofilms, but the mutant produced less than half compared with the wild type in nitrate medium showing impaired nitrite production in this condition. NO measurements in biofilm confirmed lower values in the mutant strain. The addition of a NO donor showed that NO influences biofilm formation in a dose-dependent manner and reverses the mutant phenotype, indicating that Nap positively regulates the formation of biofilm in A. brasilense Sp245.

  1. Key physiological properties contributing to rhizosphere adaptation and plant growth promotion abilities of Azospirillum brasilense.

    Science.gov (United States)

    Fibach-Paldi, Sharon; Burdman, Saul; Okon, Yaacov

    2012-01-01

    Azospirillum brasilense is a plant growth promoting rhizobacterium (PGPR) that is being increasingly used in agriculture in a commercial scale. Recent research has elucidated key properties of A. brasilense that contribute to its ability to adapt to the rhizosphere habitat and to promote plant growth. They include synthesis of the auxin indole-3-acetic acid, nitric oxide, carotenoids, and a range of cell surface components as well as the ability to undergo phenotypic variation. Storage and utilization of polybetahydroxyalkanoate polymers are important for the shelf life of the bacteria in production of inoculants, products containing bacterial cells in a suitable carrier for agricultural use. Azospirillum brasilense is able to fix nitrogen, but despite some controversy, as judging from most systems evaluated so far, contribution of fixed nitrogen by this bacterium does not seem to play a major role in plant growth promotion. In this review, we focus on recent advances in the understanding of physiological properties of A. brasilense that are important for rhizosphere performance and successful interactions with plant roots.

  2. Quantification of Azospirillum brasilense FP2 Bacteria in Wheat Roots by Strain-Specific Quantitative PCR.

    Science.gov (United States)

    Stets, Maria Isabel; Alqueres, Sylvia Maria Campbell; Souza, Emanuel Maltempi; Pedrosa, Fábio de Oliveira; Schmid, Michael; Hartmann, Anton; Cruz, Leonardo Magalhães

    2015-10-01

    Azospirillum is a rhizobacterial genus containing plant growth-promoting species associated with different crops worldwide. Azospirillum brasilense strains exhibit a growth-promoting effect by means of phytohormone production and possibly by N2 fixation. However, one of the most important factors for achieving an increase in crop yield by plant growth-promoting rhizobacteria is the survival of the inoculant in the rhizosphere, which is not always achieved. The objective of this study was to develop quantitative PCR protocols for the strain-specific quantification of A. brasilense FP2. A novel approach was applied to identify strain-specific DNA sequences based on a comparison of the genomic sequences within the same species. The draft genome sequences of A. brasilense FP2 and Sp245 were aligned, and FP2-specific regions were filtered and checked for other possible matches in public databases. Strain-specific regions were then selected to design and evaluate strain-specific primer pairs. The primer pairs AzoR2.1, AzoR2.2, AzoR5.1, AzoR5.2, and AzoR5.3 were specific for the A. brasilense FP2 strain. These primer pairs were used to monitor quantitatively the population of A. brasilense in wheat roots under sterile and nonsterile growth conditions. In addition, coinoculations with other plant growth-promoting bacteria in wheat were performed under nonsterile conditions. The results showed that A. brasilense FP2 inoculated into wheat roots is highly competitive and achieves high cell numbers (∼10(7) CFU/g [fresh weight] of root) in the rhizosphere even under nonsterile conditions and when coinoculated with other rhizobacteria, maintaining the population at rather stable levels for at least up to 13 days after inoculation. The strategy used here can be applied to other organisms whose genome sequences are available.

  3. A cytochrome cbb3 (cytochrome c) terminal oxidase in Azospirillum brasilense Sp7 supports microaerobic growth.

    Science.gov (United States)

    Marchal, K; Sun, J; Keijers, V; Haaker, H; Vanderleyden, J

    1998-11-01

    Spectral analysis indicated the presence of a cytochrome cbb3 oxidase under microaerobic conditions in Azospirillum brasilense Sp7 cells. The corresponding genes (cytNOQP) were isolated by using PCR. These genes are organized in an operon, preceded by a putative anaerobox. The phenotype of an A. brasilense cytN mutant was analyzed. Under aerobic conditions, the specific growth rate during exponential phase (mu(e)) of the A. brasilense cytN mutant was comparable to the wild-type specific growth rate (m(e) of approximately 0.2 h-1). In microaerobic NH4+-supplemented conditions, the low respiration of the A. brasilense cytN mutant affected its specific growth rate (mu(e) of approximately 0.02 h-1) compared to the wild-type specific growth rate (mu(e) of approximately 0.2 h-1). Under nitrogen-fixing conditions, both the growth rates and respiration of the wild type were significantly diminished in comparison to those under NH4+-supplemented conditions. Differences in growth rates and respiration between the wild type and the A. brasilense cytN mutant were less pronounced under these nitrogen-fixing conditions (mu(e) of approximately 0.03 h-1 for the wild type and 0.02 h-1 for the A. brasilense cytN mutant). The nitrogen-fixing capacity of the A. brasilense cytN mutant was still approximately 80% of that determined for the wild-type strain. This leads to the conclusion that the A. brasilense cytochrome cbb3 oxidase is required under microaerobic conditions, when a high respiration rate is needed, but that under nitrogen-fixing conditions the respiration rate does not seem to be a growth-limiting factor.

  4. The Azospirillum brasilense rpoN gene is involved in nitrogen fixation, nitrate assimilation, ammonium uptake, and flagellar biosynthesis.

    Science.gov (United States)

    Milcamps, A; Van Dommelen, A; Stigter, J; Vanderleyden, J; de Bruijn, F J

    1996-05-01

    The rpoN (ntrA) gene (encoding sigma 54) of Azospirillum brasilense Sp7 was isolated by using conserved rpoN primers and the polymerase chain reaction, and its nucleotide sequence was determined. The deduced amino acid sequence of the RpoN protein was found to share a high degree of homology with other members of the sigma 54 family. Two additional open reading frames were found in the Azospirillum brasilense rpoN region, with significant similarity to equivalent regions surrounding the rpoN locus in other bacteria. An rpoN mutant of Azospirillum brasilense Sp7 was constructed by gene replacement and found to be defective in nitrogen fixation, nitrate assimilation, and ammonium uptake. Lack of ammonium uptake was also found in previously isolated Azospirillum brasilense ntrB and ntrC mutants, further supporting the role of the ntr system in this process. In addition, the rpoN mutant was found to be nonmotile, suggesting a role of RpoN in Azospirillum brasilense flagellar biosynthesis.

  5. Seed quality, chlorophyll content index and leaf nitrogen levels in maize inoculated with Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    Lívia de Matos Pereira

    Full Text Available ABSTRACTThe aim of this study was to evaluate differences between maize genotypes in relation to the germination response of the seeds and the growth of seedlings inoculated with Azospirillum brasilense, as well as the effect of inoculation on nitrogen levels and the chlorophyll content index of the leaves. The physiological seeds quality from the single-cross hybrids AG7098 and 2B707, and from the experimental synthetic varieties V2 and V4, inoculated with A. brasilense, was tested for germination, percentage and rate of emergence, and dry matter weight of the shoots and roots. Nitrogen levels and chlorophyll content index were evaluated in leaves of the same four genotypes grown in a greenhouse under different nitrogen supply systems and methods of inoculation with A. brasilense. The genotypes differ with regard to inoculation with A. brasilense. The hybrids were responsive to inoculation with A. brasilense for root dry matter weight. The V2 variety had a lower performance as regards root dry matter weight, with the opposite being seen for the dry matter weight of the shoots. V4 displayed no significant differences when inoculated. The results of the chlorophyll content index were not significant. Each genotype under evaluation displayed a different response for leaf nitrogen levels. It is possible to infer that the hybrids responded better to inoculation with the bacteria, with the greater root development leading to a better utilisation of water and nutrients.

  6. Strain-specific salt tolerance and osmoregulatory mechanisms in Azospirillum brasilense.

    Science.gov (United States)

    Chowdhury, Soumitra Paul; Nagarajan, Thirunavukkarasu; Tripathi, Rachna; Mishra, Mukti Nath; Le Rudulier, Daniel; Tripathi, Anil Kumar

    2007-02-01

    Salinity stress inhibits the growth and nitrogen fixation ability of the plant growth-promoting rhizobacterium Azospirillum brasilense. Five strains of A. brasilense were isolated from the rhizosphere of Indian cereals and grasses and identified on the basis of their phenotypic features and 16S rRNA gene sequence. The five Indian isolates and two standard strains of A. brasilense, Sp7 and Cd, showed notable differences in growth, acetylene-reducing activity under salt stress, and ability to take up and use glycine betaine for the restoration of growth and acetylene-reducing activity under salt stress. Salt stress also enhanced the production of exopolysaccharides and cell aggregates, the extent of which varied in different strains of A. brasilense at different carbon to nitrogen ratios in the culture medium. It can be concluded that the production of exopolysaccharides and cell aggregates is a more consistent physiological response of A. brasilense to salt stress than is the uptake and osmoprotection by glycine betaine.

  7. Evaluation of reference genes for gene expression analysis using quantitative RT-PCR in Azospirillum brasilense.

    Directory of Open Access Journals (Sweden)

    Mary McMillan

    Full Text Available Azospirillum brasilense is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions A. brasilense exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for A. brasilense. Here we evaluated the expression of ten candidate reference genes (16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA in wild-type and mutant A. brasilense strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that gyrA, glyA and recA are the most stably expressed reference genes in A. brasilense. The results also suggested that the use of two reference genes (gyrA and glyA is sufficient for effective normalisation of qRT-PCR data.

  8. Evaluation of reference genes for gene expression analysis using quantitative RT-PCR in Azospirillum brasilense.

    Science.gov (United States)

    McMillan, Mary; Pereg, Lily

    2014-01-01

    Azospirillum brasilense is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions A. brasilense exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for A. brasilense. Here we evaluated the expression of ten candidate reference genes (16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA) in wild-type and mutant A. brasilense strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that gyrA, glyA and recA are the most stably expressed reference genes in A. brasilense. The results also suggested that the use of two reference genes (gyrA and glyA) is sufficient for effective normalisation of qRT-PCR data.

  9. (Methyl)ammonium transport in the nitrogen-fixing bacterium Azospirillum brasilense.

    Science.gov (United States)

    Van Dommelen, A; Keijers, V; Vanderleyden, J; de Zamaroczy, M

    1998-05-01

    An ammonium transporter of Azospirillum brasilense was characterized. In contrast to most previously reported putative prokaryotic NH4+ transporter genes, A. brasilense amtB is not part of an operon with glnB or glnZ which, in A. brasilense, encode nitrogen regulatory proteins PII and PZ, respectively. Sequence analysis predicts the presence of 12 transmembrane domains in the deduced AmtB protein and classifies AmtB as an integral membrane protein. Nitrogen regulates the transcription of the amtB gene in A. brasilense by the Ntr system. amtB is the first gene identified in A. brasilense whose expression is regulated by NtrC. The observation that ammonium uptake is still possible in mutants lacking the AmtB protein suggests the presence of a second NH4+ transport mechanism. Growth of amtB mutants at low ammonium concentrations is reduced compared to that of the wild type. This suggests that AmtB has a role in scavenging ammonium at low concentrations.

  10. Residual effect of nitrogen fertilization and Azospirillum brasilense inoculation in the maize culture
    Efeito residual da adubação nitrogenada e inoculação de Azospirillum brasilense na cultura do milho

    OpenAIRE

    Jackson Huzar Novakowiski; Aníbal de Moraes; Margarete Kimie Falbo; Itacir Eloi Sandini; Jaqueline Huzar Novakowiski; Nicole Colombari Cheng

    2011-01-01

    The diazotrophic bacteria Azospirillum brasilense is an organism able to fix nitrogen of atmosphere and produce plant hormones. Nevertheless, there is lack of information with regard to use in field conditions, especially in production systems that have presence of animals in a determined year period. The objective of paper was to evaluate the association of the nitrogen residual effect of fertilization in pasture winter and the inoculation with A. brasilense in the maize culture. Were carrie...

  11. Reduction of selenite by Azospirillum brasilense with the formation of selenium nanoparticles.

    Science.gov (United States)

    Tugarova, Anna V; Vetchinkina, Elena P; Loshchinina, Ekaterina A; Burov, Andrei M; Nikitina, Valentina E; Kamnev, Alexander A

    2014-10-01

    The ability to reduce selenite (SeO(3)(2-)) ions with the formation of selenium nanoparticles was demonstrated in Azospirillum brasilense for the first time. The influence of selenite ions on the growth of A. brasilense Sp7 and Sp245, two widely studied wild-type strains, was investigated. Growth of cultures on both liquid and solid (2 % agar) media in the presence of SeO(3)(2-) was found to be accompanied by the appearance of the typical red colouration. By means of transmission electron microscopy (TEM), electron energy loss spectroscopy (EELS) and X-ray fluorescence analysis (XFA), intracellular accumulation of elementary selenium in the form of nanoparticles (50 to 400 nm in diameter) was demonstrated for both strains. The proposed mechanism of selenite-to-selenium (0) reduction could involve SeO(3)(2-) in the denitrification process, which has been well studied in azospirilla, rather than a selenite detoxification strategy. The results obtained point to the possibility of using Azospirillum strains as endophytic or rhizospheric bacteria to assist phytoremediation of, and cereal cultivation on, selenium-contaminated soils. The ability of A. brasilense to synthesise selenium nanoparticles may be of interest to nanobiotechnology for "green synthesis" of bioavailable amorphous red selenium nanostructures.

  12. Nitrogenase switch-off by ammonium ions in Azospirillum brasilense requires the GlnB nitrogen signal-transducing protein.

    Science.gov (United States)

    Klassen, Giseli; Souza, Emanuel M; Yates, M Geoffrey; Rigo, Liu Un; Costa, Roberta M; Inaba, Juliana; Pedrosa, Fábio O

    2005-09-01

    Nitrogenase activity in several diazotrophs is switched off by ammonium and reactivated after consumption. The signaling pathway to this system in Azospirillum brasilense is not understood. We show that ammonium-dependent switch-off through ADP-ribosylation of Fe protein was partial in a glnB mutant of A. brasilense but absent in a glnB glnZ double mutant. Triggering of inactivation by anaerobic conditions was not affected in either mutant. The results suggest that glnB is necessary for full ammonium-dependent nitrogenase switch-off in A. brasilense.

  13. Preparation of miniantibodies to Azospirillum brasilense Sp245 surface antigens and their use for bacterial detection.

    Science.gov (United States)

    Dykman, Lev A; Staroverov, Sergei A; Guliy, Olga I; Ignatov, Oleg V; Fomin, Alexander S; Vidyasheva, Irina V; Karavaeva, Olga A; Bunin, Viktor D; Burygin, Gennady L

    2012-01-01

    This article reports the first preparation of miniantibodies to Azospirillum brasilense Sp245 surface antigens by using a combinatorial phage library of sheep antibodies. The prepared phage antibodies were used for the first time for lipopolysaccharide and flagellin detection by dot assay, electro-optical analysis of cell suspensions, and transmission electron microscopy. Interaction of A. brasilense Sp245 with antilipopolysaccharide and antiflagellin phage-displayed miniantibodies caused the magnitude of the electro-optical signal to change considerably. The electro-optical results were in good agreement with the electron microscopic data. This is the first reported possibility of employing phage-displayed miniantibodies in bacterial detection aided by electro-optical analysis of cell suspensions.

  14. [Electrooptical properties of soil nitrogen-fixing bacterium Azospirillum brasilense: effect of copper ions].

    Science.gov (United States)

    Ignatov, O V; Kamnev, A A; Markina, L N; Antoniuk, L P; Kolina, M; Ignatov, V V

    2001-01-01

    The effects of copper ions on the uptake of some essential metals in the biomass and the electrooptical properties of cell suspensions of the nitrogen-fixing soil bacterium Azospirillum brasilense sp. 245 were studied. Copper cations were shown to be effectively taken up by the cell biomass from the culture medium. The addition of copper ions increased the rate of uptake of some other metals present in the culture medium. This was accompanied by changes in the electrooptical characteristics of cell suspension as measured within the orienting electric field frequency range of 10 to 10,000 kHz. The effects observed during short-term incubation of A. brasilense in the presence of copper cations were less significant than during long-term incubation. These results can be used for rapid screening of microbial cultures for enhanced efficiency of sorption and uptake of metals.

  15. Rhizobial Inoculation, Alone or Coinoculated with Azospirillum brasilense, Promotes Growth of Wetland Rice

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    Leandro Hahn

    Full Text Available ABSTRACT Rhizobia and associative bacteria promote growth in rice plants (Oryza sativa L. through a series of mechanisms, but most studies on inoculation have been performed based on inoculation with these bacteria in a separate or singular manner. The objective of this study was to assess the efficiency of single/isolated inoculation and inoculation combined with symbiotic rhizobia from forage legume and with Azospirillum brasilense on promoting growth and the root colonization process in wetland rice. Two rhizobia among four isolates from a greenhouse and a laboratory experiment were selected that efficiently promoted seed germination and rice plant growth in a sterilized substrate and in soil. The two most efficient isolates (UFRGS Vp16 and UFRGS Lc348 were inoculated alone or in combination with a commercial product containing A. brasilense in two field experiments using two wetland rice cultivars over two growing seasons. In the field experiments, these isolates coinoculated with A. brasilense promoted larger increases in the agronomic variables of wetland rice compared to the control without inoculation. Confocal laser microscopy confirmed the presence of inoculated bacteria tagged with gfp (UFRGS Vp16, UFRGS Lc348, and A. brasilense colonizing the root surface of the rice seedlings, mainly in the root hairs and lateral roots.

  16. Potential roles for the glnB and ntrYX genes in Azospirillum brasilense.

    Science.gov (United States)

    Vitorino, J C; Steffens, M B; Machado, H B; Yates, M G; Souza, E M; Pedrosa, F O

    2001-07-24

    Three Azospirillum brasilense mutants constitutive for nitrogen fixation (Nif(C)) in the presence of NH4(+) and deficient in nitrate-dependent growth were used as tools to define the roles of the glnB and ntrYX genes in this organism. Mutant HM14 was complemented for nitrate-dependent growth and NH4(+) regulation of nitrogenase by plasmid pL46 which contains the ntrYX genes of A. brasilense. Mutant HM26 was restored for NH4(+) regulation and nitrate-dependent growth by plasmid pJC1, carrying the A. brasilense glnB gene expressed from a constitutive promoter. Mutant HM053, on the other hand, was not complemented for NH4(+) regulation of nitrogenase and nitrate-dependent growth by both plasmids pJCI and pL46. The levels and control of glutamine synthetase activity of all mutants were not affected by both plasmids pL46 (ntrYX) and pJC1 (glnB). These results support the characterization of strains HM14 as an ntrYX mutant and strain HM26 as a glnB mutant and the involvement of ntrYX and glnB in the regulation of the general nitrogen metabolism in A. brasilense.

  17. Corn Yield and Foliar Diagnosis Affected by Nitrogen Fertilization and Inoculation with Azospirillum brasilense

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    Fernando Shintate Galindo

    2016-01-01

    Full Text Available ABSTRACT The biological nitrogen fixation (BNF process in grasses is caused by diazotrophic bacteria, particularly Azospirillum brasilense. However, studies are lacking on BNF efficiency to define how much mineral nitrogen (N can be applied to achieve more sustainable high yields. Furthermore, there should be an analysis of whether urea with the urease enzyme inhibitor NBPT is less harmful, benefiting BNF in grasses. The objective of this study was to evaluate the effect of N sources and N rates associated with inoculation with Azospirillum brasilense regarding foliar diagnosis and leaf chlorophyll index (LCI, agronomic efficiency (AE, and corn grain yield in the Cerrado (Brazilian tropical savanna region. The experiment was conducted in a no-tillage system in a Latossolo Vermelho Distroférrico (Oxisol. A randomized block experimental design was used with four replications in a 2 × 5 × 2 factorial arrangement as follows: two N sources - urea and Super N, urea with urease enzyme inhibitor NBPT [N - (n-butyl thiophosphoric triamide]; five N rates (0, 50, 100, 150, and 200 kg ha-1 applied in topdressing; and two seed inoculation treatments, one with and one without A. brasilense. N rate positively influenced the LCI and concentrations of N, S, and Mn in leaves, and may increase the concentrations of P, Cu, and Fe; however, higher N rates can reduce AE. The N sources had similar effects, and therefore urea is recommended for N fertilization. Inoculation with A. brasilense decreased leaf concentration of Fe and increased LCI, leaf concentration of P, AE, and corn grain yield; the use of this diazotrophic bacterium is therefore viable even when high rates of N are applied.

  18. Increased acidification in the rhizosphere of cactus seedlings induced by Azospirillum brasilense

    Science.gov (United States)

    Carrillo, Angel; Li, Ching; Bashan, Yoav

    2002-08-01

    Acidification of the rhizosphere of cactus seedlings (giant cardon, Pachycereus pringlei) after inoculation with the plant growth-promoting bacterium Azospirillum brasilense Cd, in the presence or absence of ammonium and nitrate, was studied to understand how to increase growth of cardon seedlings in poor desert soils. While ammonium enhanced rhizosphere and liquid culture acidification, inoculation with the bacteria enhanced it further. On the other hand, nitrate increased pH of the rhizosphere, but combined with the bacterial inoculation, increase in pH was significantly smaller. Bacterial inoculation with ammonium enhanced plant growth.

  19. [Isolation and purification of Mn-peroxidase from Azospirillum brasilense Sp245].

    Science.gov (United States)

    Kupriashina, M A; Selivanov, N Iu; Nikitina, V E

    2012-01-01

    Homogenous Mn-peroxidase of a 26-fold purity grade was isolated from a culture of Azospirillum brasilense Sp245 cultivated on a medium containing 0.1 mM pyrocatechol. The molecular weight of the enzyme is 43 kD as revealed by electrophoresis in SDS-PAAG. It was shown that the use of pyrocatechol and 2,2'-azino-bis(3-ethylbenzotiazoline-6-sulfonate) at concentrations of 0.1 and I mM as inductors increased the Mn-peroxidase activity by a factor of 3.

  20. Optimization of biofloc production in Azospirillum brasilense (MTCC-125) and evaluation of its adherence with the roots of certain crops.

    Science.gov (United States)

    Joe, Melvin; Karthikeyan, M B; Sekar, C; Deiveekasundaram, M

    2010-10-01

    The phenomenon of flocculation in Azospirillum brasilense (MTCC-125) was studied under different combinations of carbon and nitrogen sources. Fructose and Potassium nitrate at a pH of 6.4 in the cultural medium favour a higher bio-floc production. The biofloc was studied for root adhesion and its survival efficiency in the rhizoplane and rhizosphere of certain crops such as sorghum and sunflower under dryland condition. It has been demonstrated that the flocculated cultures of Azospirillum were found to have maximum adhesion to the root surface and higher survival rate in the rhizoplane and rhizosphere under different moisture stressed conditions as compared to the log phase cells of Azospirillum.

  1. Structural and functional peculiarities of the lipopolysaccharide of Azospirillum brasilense SR55, isolated from the roots of Triticum durum.

    Science.gov (United States)

    Boyko, Alevtina S; Konnova, Svetlana A; Fedonenko, Yulia P; Zdorovenko, Evelina L; Smol'kina, Olga N; Kachala, Vadim V; Ignatov, Vladimir V

    2011-10-20

    Azospirillum brasilense SR55, isolated from the rhizosphere of Triticum durum, was classified as serogroup II on the basis of serological tests. Such serogroup affiliation is uncharacteristic of wheat-associated Azospirillum species. The lipid A of A. brasilense SR55 lipopolysaccharide contained 3-hydroxytetradecanoic, 3-hydroxyhexadecanoic, hexadecanoic and octadecenoic fatty acids. The structure of the lipopolysaccharide's O polysaccharide was established, with the branched octasaccharide repeating unit being represented by l-rhamnose, l-3-O-Me-rhamnose, d-galactose and d-glucuronic acid. The SR55 lipopolysaccharide induced deformations of wheat root hairs. The lipopolysaccharide was not involved in bacterial cell aggregation, but its use to pretreat wheat roots was conducive to cell adsorption. This study shows that Azospirillum bacteria can utilise their own lipopolysaccharide as a carbon source, which may give them an advantage in competitive natural environments.

  2. Insights into the 1.59-Mbp largest plasmid of Azospirillum brasilense CBG497.

    Science.gov (United States)

    Acosta-Cruz, Erika; Wisniewski-Dyé, Florence; Rouy, Zoé; Barbe, Valérie; Valdés, María; Mavingui, Patrick

    2012-09-01

    The plant growth-promoting proteobacterium Azospirillum brasilense enhances growth of many economically important crops, such as wheat, maize, and rice. The sequencing and annotation of the 1.59-Mbp replicon of A. brasilense CBG497, a strain isolated from a maize rhizosphere grown on an alkaline soil in the northeast of Mexico, revealed a GC content of 68.7 % and the presence of 1,430 potential protein-encoding genes, 1,147 of them classified into clusters of orthologous groups categories, and 16 tRNA genes representing 11 tRNA species. The presence of sixty-two genes representatives of the minimal gene set and chromid core genes suggests its importance in bacterial survival. The phaAB → G operon, reported as involved in the bacterial adaptation to alkaline pH in the presence of K(+), was also found on this replicon and detected in several Azospirillum strains. Phylogenetic analysis suggests that it was laterally acquired. We were not able to show its inference on the adaptation to basic pH, giving a hint about the presence of an alternative system for adaptation to alkaline pH.

  3. Growth and yield of corn hybrids in response to association with Azospirillum brasilense and nitrogen fertilization

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    Deniele Marini

    2015-02-01

    Full Text Available There is a growing interest in optimizing the positive effects of the association between Azospirillum bacteria and corn crop in order to reduce the use of nitrogen fertilizers. This study aimed to evaluate the inoculation efficiency of an A. brasilense-based commercial product in association with different rates of nitrogen fertilization in two corn genotypes. The experiment was arranged in a 2 x 2 x 5 factorial randomized block design, with four replications. The treatments consisted of two corn hybrids (30F53 and CD386; with and without inoculation with a commercial product based on A. brasilense and five nitrogen rates (0, 40, 80, 120 and 160 kg ha-1. The variables plant height, basal stem diameter, leaf area, shoot dry matter, leaf nitrogen content, length and diameter of the cob, weight of 100 grains and grain yield were evaluated. Inoculation with A. brasilense provided increases of 11 and 12% in leaf area and shoot dry matter, respectively. There were differences in the response of the corn hybrids for most variables and the increase in nitrogen supply provided increments in the growth and yield of corn.

  4. [Localization of denitrification genes in plasmid DNA of bacteria Azospirillum brasilense].

    Science.gov (United States)

    Petrova, L P; Varshalomidze, O É; Shelud'ko, A V; Katsy, E I

    2010-07-01

    In 85-Mda plasmid (p85) of plant-associated bacteria Azospirillum brasilense Sp245 model strain, the genes encoding copper-containing nitrite reductase (nirK); heterodimeric NO-reductase (norCB); NorQ and NorD proteins affecting synthesis and (or) activation of NirK and (or) NO-reductase (norQD); catalytic subunit I ofcytochrom c oxidase (CccoN); presumable NO sensor carrying two hemeerythrine domains (orf181); and an enzyme required for synthesis of presumable NO antagonist, homocystein (metC) were identified. In the same region of p85, orf293 encoding transcriptional regulator of LysR type, orf208 whose protein product carries a formylmethanofuran dehydrogenase subunit E domain, and an orf164-encoding conservative secretory protein with unknown function were also found. Localization of a set of denitrification genes in the plasmid DNA A. brasilense Sp245 adjacent to IS elements ISAzba1 and ISAzba2 indicates potential mobility of these genes and high probability of their horizontal transfer among populations of rhizospheric bacteria. A site homologous to p85 nirK-orf208-orf181 genes was detected in the 115 kb plasmid of A. brasilense Sp7 type strain.

  5. Azospirillum brasilense does not affect population structure of specific rhizobacterial communities of inoculated maize (Zea mays).

    Science.gov (United States)

    Herschkovitz, Yoav; Lerner, Anat; Davidov, Yaacov; Okon, Yaacov; Jurkevitch, Edouard

    2005-11-01

    Positive response of plant species to plant growth-promoting rhizobacteria have led to an increased interest in their use as bacterial inoculants. However, the introduction of exogenous bacteria into natural ecosystems may perturb bacterial populations within the microbial community and lead to the disruption of indigenous populations performing key functional roles. In this study the effect of Azospirillum brasilense inoculation on maize (Zea mays) rhizosphere Actinobacteria, Bacteroidetes, alpha-Proteobacteria, Pseudomonas and Bdellovibrio spp. was assessed using a polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) approach in conjunction with group-specific primers. The DGGE fingerprints analysis revealed that the introduction of A. brasilense did not alter or disrupt the microbial system at the group-specific level. However, some communities such as the alpha-Proteobacteria and Bdellovibrio were influenced by plant age while the other bacterial groups remained unaffected. Based on these as well as previous data, it can be inferred that inoculation with A. brasilense does not perturb the natural bacterial populations investigated.

  6. Metabolic Adaptations of Azospirillum brasilense to Oxygen Stress by Cell-to-Cell Clumping and Flocculation

    Science.gov (United States)

    Bible, Amber N.; Khalsa-Moyers, Gurusahai K.; Mukherjee, Tanmoy; Green, Calvin S.; Mishra, Priyanka; Purcell, Alicia; Aksenova, Anastasia; Hurst, Gregory B.

    2015-01-01

    The ability of bacteria to monitor their metabolism and adjust their behavior accordingly is critical to maintain competitiveness in the environment. The motile microaerophilic bacterium Azospirillum brasilense navigates oxygen gradients by aerotaxis in order to locate low oxygen concentrations that can support metabolism. When cells are exposed to elevated levels of oxygen in their surroundings, motile A. brasilense cells implement an alternative response to aerotaxis and form transient clumps by cell-to-cell interactions. Clumping was suggested to represent a behavior protecting motile cells from transiently elevated levels of aeration. Using the proteomics of wild-type and mutant strains affected in the extent of their clumping abilities, we show that cell-to-cell clumping represents a metabolic scavenging strategy that likely prepares the cells for further metabolic stresses. Analysis of mutants affected in carbon or nitrogen metabolism confirmed this assumption. The metabolic changes experienced as clumping progresses prime cells for flocculation, a morphological and metabolic shift of cells triggered under elevated-aeration conditions and nitrogen limitation. The analysis of various mutants during clumping and flocculation characterized an ordered set of changes in cell envelope properties accompanying the metabolic changes. These data also identify clumping and early flocculation to be behaviors compatible with the expression of nitrogen fixation genes, despite the elevated-aeration conditions. Cell-to-cell clumping may thus license diazotrophy to microaerophilic A. brasilense cells under elevated oxygen conditions and prime them for long-term survival via flocculation if metabolic stress persists. PMID:26407887

  7. Metabolic adaptations of Azospirillum brasilense to oxygen stress by cell-to-cell clumping and flocculation.

    Science.gov (United States)

    Bible, Amber N; Khalsa-Moyers, Gurusahai K; Mukherjee, Tanmoy; Green, Calvin S; Mishra, Priyanka; Purcell, Alicia; Aksenova, Anastasia; Hurst, Gregory B; Alexandre, Gladys

    2015-12-01

    The ability of bacteria to monitor their metabolism and adjust their behavior accordingly is critical to maintain competitiveness in the environment. The motile microaerophilic bacterium Azospirillum brasilense navigates oxygen gradients by aerotaxis in order to locate low oxygen concentrations that can support metabolism. When cells are exposed to elevated levels of oxygen in their surroundings, motile A. brasilense cells implement an alternative response to aerotaxis and form transient clumps by cell-to-cell interactions. Clumping was suggested to represent a behavior protecting motile cells from transiently elevated levels of aeration. Using the proteomics of wild-type and mutant strains affected in the extent of their clumping abilities, we show that cell-to-cell clumping represents a metabolic scavenging strategy that likely prepares the cells for further metabolic stresses. Analysis of mutants affected in carbon or nitrogen metabolism confirmed this assumption. The metabolic changes experienced as clumping progresses prime cells for flocculation, a morphological and metabolic shift of cells triggered under elevated-aeration conditions and nitrogen limitation. The analysis of various mutants during clumping and flocculation characterized an ordered set of changes in cell envelope properties accompanying the metabolic changes. These data also identify clumping and early flocculation to be behaviors compatible with the expression of nitrogen fixation genes, despite the elevated-aeration conditions. Cell-to-cell clumping may thus license diazotrophy to microaerophilic A. brasilense cells under elevated oxygen conditions and prime them for long-term survival via flocculation if metabolic stress persists.

  8. Purification and characterisation of Azospirillum brasilense N-truncated NtrX protein.

    Science.gov (United States)

    Assumpção, Marcelo Constantino; de Souza, Emanuel Maltempi; Yates, M Geoffrey; de Oliveira Pedrosa, Fábio; Benelli, Elaine Machado

    2007-06-01

    The NtrX protein has been identified as a transcriptional activator of genes involved in the metabolic control of alternative nitrogen sources, acting as a member of a two-component regulatory system. The in silico analysis of the NtrX amino acid sequence shows that this protein contains an N-terminal receiver domain, a central AAA+ superfamily domain and a C-terminal DNA binding domain. To over-express and purify this protein, the ntrX gene of Azospirillum brasilense lacking the first eight codons was cloned into the vector pET29a+. The NtrX protein was over-expressed as an S.Tag fusion protein induced by l-arabinose in the Escherichia coli strain BL21AI and purified by ion exchange and affinity chromatography. The ATPase activity of NtrX was measured by coupling the ATP conversion to ADP with NADH oxidation. The ATPase activity of NtrX was stimulated in the presence of A. brasilense sigma(54)/NtrC-dependent promoter of the glnBA gene. Phosphorylation by carbamyl-phosphate also stimulated ATPase, in a manner similar to the NtrC protein. Together our results suggest that NtrX is active in the phosphorylated form and that there may be a cross-talk between the NtrYX and NtrBC regulatory systems in A. brasilense.

  9. Components of corn crop yield under inoculation with Azospirillum brasilense using integrated crop-livestock system

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    Marcos da Silva Brum

    2016-09-01

    Full Text Available The objective of this study was to evaluate the agronomic characteristics of corn seed inoculated with Azospirillum brasilense, grown on black oat and ryegrass straw, and managed under different grazing strategies and doses of nitrogen. The experiment was conducted in Santa Maria, Rio Grande do Sul State, Brazil, during two agricultural seasons (2012/2013 and 2013/2014 in a randomized, complete block design with three replications. In the winter period, black oat and ryegrass straw were managed at different grazing heights by sheep (0.30, 0.20, 0.10 m, conventional grazing, and no grazing with three doses of nitrogen (0, 50, and 100 kg ha-1, with or without inoculation by A. brasilense. We used the hybrid Pioneer (P1630H® in 2012 and the hybrid Agroeste (AS 1551® in 2013. The height of corn plants was greater when they were grown on black oat and ryegrass straw, and the absence of grazing favored productivity. Under drought conditions, the application of nitrogen to the pasture favored corn development, increasing plant height, ear height, and stem diameter. Inoculation with A. brasilense had a positive effect on the characteristics of yield and productivity of corn, independent of growing season and hybrid used.

  10. Cloning, sequencing, and characterization of the Azospirillum brasilense fhuE gene.

    Science.gov (United States)

    Cui, Yanhua; Tu, Ran; Guan, Yue; Ma, Luyan; Chen, Sanfeng

    2006-03-01

    The fhuE gene of Escherichia coli encodes the FhuE protein, which is a receptor protein in the coprogen-mediated siderophore iron-transport system. A fhuE gene homologue from Azospirillum brasilense, a nitrogen-fixing soil bacterium that lives in association with the roots of cereal grasses, was cloned, sequenced, and characterized. The A. brasilense fhuE encodes a protein of 802 amino acids with a predicted molecular weight of approximately 87 kDa. The deduced amino-acid sequence showed a high level of homology to the sequences of all the known fhuE gene products. The fhuE mutant was sensitive to iron starvation and defective in coprogen-mediated iron uptake. The mutant failed to express one membrane protein of approximately 78 kDa that was induced by iron starvation in the wild type. Complementation studies showed that the A. brasilense fhuE gene, when present on a low-copy number plasmid, could restore the functions of the mutant. Mutation in fhuE gene did not affect nitrogen fixation.

  11. Cellular responses during morphological transformation in Azospirillum brasilense and Its flcA knockout mutant.

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    Xingsheng Hou

    Full Text Available FlcA is a response regulator controlling flocculation and the morphological transformation of Azospirillum cells from vegetative to cyst-like forms. To understand the cellular responses of Azospirillum to conditions that cause morphological transformation, proteins differentially expressed under flocculation conditions in A. brasilense Sp7 and its flcA knockout mutant were investigated. Comparison of 2-DE protein profiles of wild-type (Sp7 and a flcA deletion mutant (Sp7-flcAΔ revealed a total of 33 differentially expressed 2-DE gel spots, with 22 of these spots confidently separated to allow protein identification. Analysis of these spots by liquid chromatography-tandem mass spectrometry (LC-MS/MS and MASCOT database searching identified 48 proteins (≥10% emPAI in each spot. The functional characteristics of these proteins included carbon metabolism (beta-ketothiolase and citrate synthase, nitrogen metabolism (Glutamine synthetase and nitric oxide synthase, stress tolerance (superoxide dismutase, Alkyl hydroperoxidase and ATP-dependent Clp protease proteolytic subunit and morphological transformation (transducer coupling protein. The observed differences between Sp7 wild-type and flcA- strains enhance our understanding of the morphological transformation process and help to explain previous phenotypical observations. This work is a step forward in connecting the Azospirillum phenome and genome.

  12. Cellular responses during morphological transformation in Azospirillum brasilense and Its flcA knockout mutant.

    Science.gov (United States)

    Hou, Xingsheng; McMillan, Mary; Coumans, Joëlle V F; Poljak, Anne; Raftery, Mark J; Pereg, Lily

    2014-01-01

    FlcA is a response regulator controlling flocculation and the morphological transformation of Azospirillum cells from vegetative to cyst-like forms. To understand the cellular responses of Azospirillum to conditions that cause morphological transformation, proteins differentially expressed under flocculation conditions in A. brasilense Sp7 and its flcA knockout mutant were investigated. Comparison of 2-DE protein profiles of wild-type (Sp7) and a flcA deletion mutant (Sp7-flcAΔ) revealed a total of 33 differentially expressed 2-DE gel spots, with 22 of these spots confidently separated to allow protein identification. Analysis of these spots by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and MASCOT database searching identified 48 proteins (≥10% emPAI in each spot). The functional characteristics of these proteins included carbon metabolism (beta-ketothiolase and citrate synthase), nitrogen metabolism (Glutamine synthetase and nitric oxide synthase), stress tolerance (superoxide dismutase, Alkyl hydroperoxidase and ATP-dependent Clp protease proteolytic subunit) and morphological transformation (transducer coupling protein). The observed differences between Sp7 wild-type and flcA- strains enhance our understanding of the morphological transformation process and help to explain previous phenotypical observations. This work is a step forward in connecting the Azospirillum phenome and genome.

  13. Mutational analysis of GlnB residues critical for NifA activation in Azospirillum brasilense.

    Science.gov (United States)

    Inaba, Juliana; Thornton, Jeremy; Huergo, Luciano Fernandes; Monteiro, Rose Adele; Klassen, Giseli; Pedrosa, Fábio de Oliveira; Merrick, Mike; de Souza, Emanuel Maltempi

    2015-02-01

    PII proteins are signal transduction that sense cellular nitrogen status and relay this signals to other targets. Azospirillum brasilense is a nitrogen fixing bacterium, which associates with grasses and cereals promoting beneficial effects on plant growth and crop yields. A. brasilense contains two PII encoding genes, named glnB and glnZ. In this paper, glnB was mutagenised in order to identify amino acid residues involved in GlnB signaling. Two variants were obtained by random mutagenesis, GlnBL13P and GlnBV100A and a site directed mutant, GlnBY51F, was obtained. Their ability to complement nitrogenase activity of glnB mutant strains of A. brasilense were determined. The variant proteins were also overexpressed in Escherichia coli, purified and characterized biochemically. None of the GlnB variant forms was able to restore nitrogenase activity in glnB mutant strains of A. brasilense LFH3 and 7628. The purified GlnBY51F and GlnBL13P proteins could not be uridylylated by GlnD, whereas GlnBV100A was uridylylated but at only 20% of the rate for wild type GlnB. Biochemical and computational analyses suggest that residue Leu13, located in the α helix 1 of GlnB, is important to maintain GlnB trimeric structure and function. The substitution V100A led to a lower affinity for ATP binding. Together the results suggest that NifA activation requires uridylylated GlnB bound to ATP.

  14. Identification and characterization of the two-component NtrY/NtrX regulatory system in Azospirillum brasilense

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    M.L. Ishida

    2002-06-01

    Full Text Available Two Azospirillum brasilense open reading frames (ORFs exhibited homology with the two-component NtrY/NtrX regulatory system from Azorhizobium caulinodans. These A. brasilense ORFs, located downstream to the nifR3ntrBC operon, were isolated, sequenced and characterized. The present study suggests that ORF1 and ORF2 correspond to the A. brasilense ntrY and ntrX genes, respectively. The amino acid sequences of A. brasilense NtrY and NtrX proteins showed high similarity to sensor/kinase and regulatory proteins, respectively. Analysis of lacZ transcriptional fusions by the ß-galactosidase assay in Escherichia coli ntrC mutants showed that the NtrY/NtrX proteins failed to activate transcription of the nifA promoter of A. brasilense. The ntrYX operon complemented a nifR3ntrBC deletion mutant of A. brasilense for nitrate-dependent growth, suggesting a possible cross-talk between the NtrY/X and NtrB/C sensor/regulator pairs. Our data support the existence of another two-component regulatory system in A. brasilense, the NtrY/NtrX system, probably involved in the regulation of nitrate assimilation.

  15. The role of the antimicrobial compound 2,4-diacetylphloroglucinol in the impact of biocontrol Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators.

    Science.gov (United States)

    Couillerot, Olivier; Combes-Meynet, Emeline; Pothier, Joël F; Bellvert, Floriant; Challita, Elita; Poirier, Marie-Andrée; Rohr, René; Comte, Gilles; Moënne-Loccoz, Yvan; Prigent-Combaret, Claire

    2011-06-01

    Pseudomonads producing the antimicrobial metabolite 2,4-diacetylphloroglucinol (Phl) can control soil-borne phytopathogens, but their impact on other plant-beneficial bacteria remains poorly documented. Here, the effects of synthetic Phl and Phl(+) Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators were investigated. Most A. brasilense strains were moderately sensitive to Phl. In vitro, Phl induced accumulation of carotenoids and poly-β-hydroxybutyrate-like granules, cytoplasmic membrane damage and growth inhibition in A. brasilense Cd. Experiments with P. fluorescens F113 and a Phl(-) mutant indicated that Phl production ability contributed to in vitro growth inhibition of A. brasilense Cd and Sp245. Under gnotobiotic conditions, each of the three strains, P. fluorescens F113 and A. brasilense Cd and Sp245, stimulated wheat growth. Co-inoculation of A. brasilense Sp245 and Pseudomonas resulted in the same level of phytostimulation as in single inoculations, whereas it abolished phytostimulation when A. brasilense Cd was used. Pseudomonas Phl production ability resulted in lower Azospirillum cell numbers per root system (based on colony counts) and restricted microscale root colonization of neighbouring Azospirillum cells (based on confocal microscopy), regardless of the A. brasilense strain used. Therefore, this work establishes that Phl(+) pseudomonads have the potential to interfere with A. brasilense phytostimulators on roots and with their plant growth promotion capacity.

  16. Structural studies of the polysaccharides from the lipopolysaccharides of Azospirillum brasilense Sp246 and SpBr14.

    Science.gov (United States)

    Sigida, Elena N; Fedonenko, Yuliya P; Shashkov, Alexander S; Grinev, Vyacheslav S; Zdorovenko, Evelina L; Konnova, Svetlana A; Ignatov, Vladimir V; Knirel, Yuriy A

    2014-10-29

    Lipopolysaccharides from closely related Azospirillum brasilense strains, Sp246 and SpBr14, were obtained by phenol-water extraction. Mild acid hydrolysis of the lipopolysaccharides followed by GPC on Sephadex G-50 resulted in polysaccharide mixtures. On the basis of sugar and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy data, it was concluded that both bacteria possess the same two distinct polysaccharides having structures 1 and 2: [structure: see text]. Structure 1 has been reported earlier for a polysaccharide of A. brasilense 54 [Fedonenko et al., 2011] whereas to our knowledge structure 2 has not been hitherto found in bacterial polysaccharides.

  17. Co2 + interaction with Azospirillum brasilense Sp7 cells: a 57Co emission Mössbauer spectroscopic study

    Science.gov (United States)

    Kamnev, Alexander A.; Tugarova, Anna V.; Biró, Borbála; Kovács, Krisztina; Homonnay, Zoltán; Kuzmann, Ernő; Vértes, Attila

    2012-03-01

    Preliminary 57Co emission Mössbauer spectroscopic data were obtained for the soil bacterium Azospirillum brasilense Sp7 ( T = 80 K) in frozen 57Co2 + -containing suspensions and in their dried residues. The Mössbauer parameters were compared with those for A. brasilense strain Sp245 differing from strain Sp7 by ecological behaviour. Live cells of both strains showed metabolic transformations of 57Co2 + within an hour. Differences in the parameters observed for the two strains under similar conditions suggest dissimilarities in their metabolic response to Co2 + .

  18. Co{sup 2 + } interaction with Azospirillum brasilense Sp7 cells: a {sup 57}Co emission Moessbauer spectroscopic study

    Energy Technology Data Exchange (ETDEWEB)

    Kamnev, Alexander A.; Tugarova, Anna V. [Russian Academy of Sciences, Institute of Biochemistry and Physiology of Plants and Microorganisms (Russian Federation); Biro, Borbala [Hungarian Academy of Sciences, Research Institute for Soil Science and Agricultural Chemistry (Hungary); Kovacs, Krisztina, E-mail: kkriszti@chem.elte.hu; Homonnay, Zoltan; Kuzmann, Erno; Vertes, Attila [Eoetvoes Lorand University, Institute of Chemistry (Hungary)

    2012-03-15

    Preliminary {sup 57}Co emission Moessbauer spectroscopic data were obtained for the soil bacterium Azospirillum brasilense Sp7 (T = 80 K) in frozen {sup 57}Co{sup 2 + }-containing suspensions and in their dried residues. The Moessbauer parameters were compared with those for A. brasilense strain Sp245 differing from strain Sp7 by ecological behaviour. Live cells of both strains showed metabolic transformations of {sup 57}Co{sup 2 + } within an hour. Differences in the parameters observed for the two strains under similar conditions suggest dissimilarities in their metabolic response to Co{sup 2 + }.

  19. Genome Sequence of Azospirillum brasilense CBG497 and Comparative Analyses of Azospirillum Core and Accessory Genomes provide Insight into Niche Adaptation

    Directory of Open Access Journals (Sweden)

    Victor González

    2012-09-01

    Full Text Available Bacteria of the genus Azospirillum colonize roots of important cereals and grasses, and promote plant growth by several mechanisms, notably phytohormone synthesis. The genomes of several Azospirillum strains belonging to different species, isolated from various host plants and locations, were recently sequenced and published. In this study, an additional genome of an A. brasilense strain, isolated from maize grown on an alkaline soil in the northeast of Mexico, strain CBG497, was obtained. Comparative genomic analyses were performed on this new genome and three other genomes (A. brasilense Sp245, A. lipoferum 4B and Azospirillum sp. B510. The Azospirillum core genome was established and consists of 2,328 proteins, representing between 30% to 38% of the total encoded proteins within a genome. It is mainly chromosomally-encoded and contains 74% of genes of ancestral origin shared with some aquatic relatives. The non-ancestral part of the core genome is enriched in genes involved in signal transduction, in transport and in metabolism of carbohydrates and amino-acids, and in surface properties features linked to adaptation in fluctuating environments, such as soil and rhizosphere. Many genes involved in colonization of plant roots, plant-growth promotion (such as those involved in phytohormone biosynthesis, and properties involved in rhizosphere adaptation (such as catabolism of phenolic compounds, uptake of iron are restricted to a particular strain and/or species, strongly suggesting niche-specific adaptation.

  20. Genome Sequence of Azospirillum brasilense CBG497 and Comparative Analyses of Azospirillum Core and Accessory Genomes provide Insight into Niche Adaptation

    Science.gov (United States)

    Wisniewski-Dyé, Florence; Lozano, Luis; Acosta-Cruz, Erika; Borland, Stéphanie; Drogue, Benoît; Prigent-Combaret, Claire; Rouy, Zoé; Barbe, Valérie; Mendoza Herrera, Alberto; González, Victor; Mavingui, Patrick

    2012-01-01

    Bacteria of the genus Azospirillum colonize roots of important cereals and grasses, and promote plant growth by several mechanisms, notably phytohormone synthesis. The genomes of several Azospirillum strains belonging to different species, isolated from various host plants and locations, were recently sequenced and published. In this study, an additional genome of an A. brasilense strain, isolated from maize grown on an alkaline soil in the northeast of Mexico, strain CBG497, was obtained. Comparative genomic analyses were performed on this new genome and three other genomes (A. brasilense Sp245, A. lipoferum 4B and Azospirillum sp. B510). The Azospirillum core genome was established and consists of 2,328 proteins, representing between 30% to 38% of the total encoded proteins within a genome. It is mainly chromosomally-encoded and contains 74% of genes of ancestral origin shared with some aquatic relatives. The non-ancestral part of the core genome is enriched in genes involved in signal transduction, in transport and in metabolism of carbohydrates and amino-acids, and in surface properties features linked to adaptation in fluctuating environments, such as soil and rhizosphere. Many genes involved in colonization of plant roots, plant-growth promotion (such as those involved in phytohormone biosynthesis), and properties involved in rhizosphere adaptation (such as catabolism of phenolic compounds, uptake of iron) are restricted to a particular strain and/or species, strongly suggesting niche-specific adaptation. PMID:24705077

  1. Crecimiento de Azospirillum brasilense en presencia de disacáridos: sacarosa y lactosa

    OpenAIRE

    Yudyt Díaz-Saez; Manuel Díaz-de los Ríos; Mario Alberto-Casas; Arianna Nuñez-Caraballo; Marlene Martínez-Mora

    2013-01-01

    La obtención de biopolímeros a partir de fermentaciones bacterianas requiere de la selección adecuada de las condiciones del proceso. Se evaluó el crecimiento de la cepa Azospirillum brasilense 8-INICA en medio líquido suplementado con disacáridos. Las mediciones se realizaron a las 23 h de incubación con la determinación de la densidad óptica, la concentración de azúcares reductores y concentración de carbohidratos totales, así como la determinación del pH del sobrenadante. Se confirmó el má...

  2. Co-inoculation of Azospirillum brasilense and Herbaspirillum seropedicae in maize

    Directory of Open Access Journals (Sweden)

    Janaína Dartora

    2016-06-01

    Full Text Available ABSTRACT Bacteria from the genera Azospirillum and Herbaspirillum have been associated with increments in maize yield. The aim of this study was to evaluate the maize yield and nutritional content in response to inoculation with A. brasilense and H. seropediceae in association with nitrogen (N fertilization. The experimental design was randomized blocks in a 4 x 5 factorial scheme, with four replicates. The treatments consisted of seed inoculation (control without N and inoculation, A. brasilense strain - AbV5, H. seropediceae strain - SMR1 and co-inoculation AbV5 + SMR1 and N doses (0, 40, 80, 120 and 160 kg ha-1. The following variables were evaluated: ear insertion height, ear length, ear diameter, number of rows per ear, number of grains per row, ear weight, yield and NPK contents in leaves and grains. There was no interaction between the factors studied. Co-inoculation with the strains promoted increments of 12% in leaf P content, compared with control, and N fertilization promoted increase in yield and leaf P content up to the maximum dose studied.

  3. Nitric oxide metabolism and indole acetic acid biosynthesis cross-talk in Azospirillum brasilense SM.

    Science.gov (United States)

    Koul, Vatsala; Tripathi, Chandrakant; Adholeya, Alok; Kochar, Mandira

    2015-04-01

    Production of nitric oxide (NO) and the presence of NO metabolism genes, nitrous oxide reductase (nosZ), nitrous oxide reductase regulator (nosR) and nitric oxide reductase (norB) were identified in the plant-associated bacterium (PAB) Azospirillum brasilense SM. NO presence was confirmed in all overexpressing strains, while improvement in the plant growth response of these strains was mediated by increased NO and indole-3-acetic acid (IAA) levels in the strains. Electron microscopy showed random distribution to biofilm, with surface colonization of pleiomorphic Azospirilla. Quantitative IAA estimation highlighted a crucial role of nosR and norBC in regulating IAA biosynthesis. The NO quencher and donor reduced/blocked IAA biosynthesis by all strains, indicating their common regulatory role in IAA biosynthesis. Tryptophan (Trp) and l-Arginine (Arg) showed higher expression of NO genes tested, while in the case of ipdC, only Trp and IAA increased expression, while Arg had no significant effect. The highest nosR expression in SMnosR in the presence of IAA and Trp, along with its 2-fold IAA level, confirmed the relationship of nosR overexpression with Trp in increasing IAA. These results indicate a strong correlation between IAA and NO in A. brasilense SM and suggest the existence of cross-talk or shared signaling mechanisms in these two growth regulators.

  4. Electro-optical study of the exposure of Azospirillum brasilense carbohydrate epitopes.

    Science.gov (United States)

    Guliy, Olga I; Matora, Larisa Yu; Dykman, Lev A; Staroverov, Sergey A; Burygin, Gennady L; Bunin, Viktor D; Burov, Andrei M; Ignatov, Oleg V

    2015-01-01

    The exposure of Azospirillum brasilense carbohydrate epitopes was investigated by electro-optical analysis of bacterial cell suspensions. To study changes in the electro-optical (EO) properties of the suspensions, we used antibodies generated to the complete lipopolysaccharide of A. brasilense type strain Sp7 and also antibodies to the smooth and rough O polysaccharides of Sp7. After 18 hr of culture growth, the EO signal of the suspension treated with antibodies to smooth O polysaccharide was approximately 20% lower than that of the suspension treated with antibodies to complete lipopolysaccharide (control). After 72 hr of culture growth, the strongest EO signal was observed for the cells treated with antibodies to rough O polysaccharide (approximately 46% greater than the control), whereas for the cells treated with antibodies to smooth O polysaccharide, it was much lower (approximately 23% of the control). These data were confirmed by electron microscopy. The results of the study may have importance for the rapid evaluation of changes in lipopolysaccharide form in microbial biotechnology, when the antigenic composition of the bacterial surface requires close control.

  5. Physiological and biochemical characterization of Azospirillum brasilense strains commonly used as plant growth-promoting rhizobacteria.

    Science.gov (United States)

    Di Salvo, Luciana P; Silva, Esdras; Teixeira, Kátia R S; Cote, Rosalba Esquivel; Pereyra, M Alejandra; García de Salamone, Inés E

    2014-12-01

    Azospirillum is a plant growth-promoting rhizobacteria (PGPR) genus vastly studied and utilized as agriculture inoculants. Isolation of new strains under different environmental conditions allows the access to the genetic diversity and improves the success of inoculation procedures. Historically, the isolation of this genus has been performed by the use of some traditional culture media. In this work we characterized the physiology and biochemistry of five different A. brasilense strains, commonly used as cereal inoculants. The aim of this work is to contribute to pose into revision some concepts concerning the most used protocols to isolate and characterize this bacterium. We characterized their growth in different traditional and non-traditional culture media, evaluated some PGPR mechanisms and characterized their profiles of fatty acid methyl esters and carbon-source utilization. This work shows, for the first time, differences in both profiles, and ACC deaminase activity of A. brasilense strains. Also, we show unexpected results obtained in some of the evaluated culture media. Results obtained here and an exhaustive knowledge revision revealed that it is not appropriate to conclude about bacterial species without analyzing several strains. Also, it is necessary to continue developing studies and laboratory techniques to improve the isolation and characterization protocols.

  6. A hybrid two-component system protein from Azospirillum brasilense Sp7 was involved in chemotaxis.

    Science.gov (United States)

    Cui, Yanhua; Tu, Ran; Wu, Lixian; Hong, Yuanyuan; Chen, Sanfeng

    2011-09-20

    We here report the sequence and functional analysis of org35 of Azospirillum brasilense Sp7, which was originally identified to be able to interact with NifA in yeast-two-hybrid system. The org35 encodes a hybrid two-component system protein, including N-terminal PAS domains, a histidine kinase (HPK) domain and a response regulator (RR) domain in C-terminal. To determine the function of the Org35, a deletion-insertion mutant in PAS domain [named Sp7353] and a complemental strain Sp7353C were constructed. The mutant had reduced chemotaxis ability compared to that of wild-type, and the complemental strain was similar to the wild-type strain. These data suggested that the A. brasilense org35 played a key role in chemotaxis. Variants containing different domains of the org35 were expressed, and the functions of these domains were studied in vitro. Phosphorylation assays in vitro demonstrated that the HPK domain of Org35 possessed the autokinase activity and that the phosphorylated HPK was able to transfer phosphate groups to the RR domain. The result indicated Org35 was a phosphorylation-communicating protein.

  7. Glycogen phosphorylase is involved in stress endurance and biofilm formation in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Lerner, Anat; Castro-Sowinski, Susana; Lerner, Hadas; Okon, Yaacov; Burdman, Saul

    2009-11-01

    Here we report the identification of a glycogen phosphorylase (glgP) gene in the plant growth-promoting rhizobacterium Azospirillum brasilense, Sp7, and the characterization of a glgP marker exchange mutant of this strain. The glgP mutant showed a twofold reduction of glycogen phosphorylase activity and an increased glycogen accumulation as compared with wild-type Sp7, indicating that the identified gene indeed encodes a protein with glycogen phosphorylase activity. Interestingly, the glgP mutant had higher survival rates than the wild type after exposure to starvation, desiccation and osmotic pressure. The mutant was shown to be compromised in its biofilm formation ability. Analysis of the exopolysaccharide sugar composition of the glgP mutant revealed a decrease in the amount of glucose, accompanied by increases in rhamnose, fucose and ribose, as compared with the Sp7 exopolysaccharide. To the best of our knowledge, this is the first study that demonstrates GlgP activity in A. brasilense, and shows that glycogen accumulation may play an important role in the stress endurance of this bacterium.

  8. Extracellular polysaccharide composition of Azospirillum brasilense and its relation with cell aggregation.

    Science.gov (United States)

    Burdman, S; Jurkevitch, E; Soria-Díaz, M E; Serrano, A M; Okon, Y

    2000-08-15

    The exopolysaccharide (EPS) and capsular polysaccharide (CPS) composition of four Azospirillum brasilense strains differing in their aggregation capacity was analyzed by high performance anion exchange chromatography. When growing the different strains in an aggregation inducing medium containing a high carbon:nitrogen (C:N) ratio, both EPS and CPS showed a positive correlation between aggregation and the relative amount of arabinose. Arabinose was not detected in polysaccharides from Sp72002, a pleiotrophic Tn5 mutant strain impaired in aggregation. Arabinose was also not detected in extracellular polysaccharides of bacteria grown in a low C:N ratio, non-inducing aggregation medium, with exception for a relatively small amount found in the CPS of FAJ0204, a super-aggregating mutant strain. The only monosaccharides able to significantly inhibit aggregation at low sugar concentration when tested in a bioassay were arabinose (at a higher extent) and galactose. The possibility that residues of arabinose present in the extracellular polysaccharides are involved in the aggregation of A. brasilense is discussed.

  9. In vitro uridylylation of the Azospirillum brasilense N-signal transducing GlnZ protein.

    Science.gov (United States)

    Araujo, Mariana S; Baura, Valter A; Souza, Emanuel M; Benelli, Elaine M; Rigo, Liu U; Steffens, M Berenice R; Pedrosa, Fabio O; Chubatsu, Leda S

    2004-01-01

    Azospirillum brasilense is a diazotroph which associates with important agricultural crops. The nitrogen fixation process in this organism is highly regulated by ammonium and oxygen, and involves several proteins including the two PII-like proteins, GlnB and GlnZ. Although these proteins are structurally very similar, they play different roles in the control of nitrogen fixation. In this work, we describe the expression, purification, and uridylylation of the GlnZ protein of A. brasilense strain FP2. The amplified glnZ gene was sub-cloned and expressed as a His-tagged fusion protein. After purification, we obtained 30-40 mg of purified GlnZ per liter of culture. This protein was purified to 99% purity and assayed for in vitro uridylylation using a partially purified Escherichia coli GlnD as a source of uridylylyl-transferase activity. Analyses of the uridylylation reactions in non-denaturing and denaturing polyacrylamide gel electrophoresis showed that up to 74% of GlnZ monomers were modified after 30 min reaction. This covalent modification is strictly dependent on ATP and 2-ketoglutarate, while glutamine acts as an inhibitor and promotes deuridylylation.

  10. Structural studies of the O-specific polysaccharide(s) from the lipopolysaccharide of Azospirillum brasilense type strain Sp7.

    Science.gov (United States)

    Sigida, Elena N; Fedonenko, Yuliya P; Shashkov, Alexander S; Zdorovenko, Evelina L; Konnova, Svetlana A; Ignatov, Vladimir V; Knirel, Yuriy A

    2013-10-18

    Lipopolysaccharide was obtained by phenol-water extraction from dried bacterial cells of Azospirillum brasilense type strain Sp7. Mild acid hydrolysis of the lipopolysaccharide followed by GPC on Sephadex G-50 resulted in a polysaccharide mixture, which was studied by composition and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy. The following polysaccharide structures were established, where italics indicate a non-stoichiometric (∼40%) 2-O-methylation of l-rhamnose.

  11. [INFLUENCE OF AZOSPIRILLUM BRASILENSE 10/1 ON ASSOCIATIVE NITROGEN FIXATION AND INTRAVARIETAL POLYMORPHISM OF SPRING TRITICALE].

    Science.gov (United States)

    Patika, V P; Nadkernichna, O V; Shahovnina, O O

    2015-01-01

    It is shown, that the perspective Ukrainian sorts of spring triticale characterizes by considerable polymorphism by associative N2-fixing ability in root zone of plants. Application of active strain Azospirillum brasilense 10/1 promotes the decline of variability of this sign within the limits of sort, increase potential nitrogen activity is on the average in 3,2-4,7 times and also distributing normalizations in the selections of the inoculated plants.

  12. Efficiency of inoculant with Azospirillum brasilense on the growth and yield of second-harvest maize1

    OpenAIRE

    Costa, Raoni Ribeiro Guedes Fonseca; Quirino,Gerciene da Silva Ferreira; Naves,Daniela Cristina de Freitas; Santos,Charles Barbosa; Rocha,Ana Flávia de Souza

    2015-01-01

    ABSTRACTThe misuse of fertilizers has become a growing concern in terms of agricultural and environmental sustainability. One alternative to reduce the impact of chemical fertilizers is the use of inoculants containing nitrogen-fixing and plant growth promoting bacteria. This study aimed at evaluating the effects of an inoculant with Azospirillum brasilense on the growth and yield of secondharvest maize. The experimental design was randomized blocks with three replications, in a 3 × 5 factori...

  13. Comparison of two Cellulomonas strains and their interaction with Azospirillum brasilense in degradation of wheat straw and associated nitrogen fixation

    Energy Technology Data Exchange (ETDEWEB)

    Halsall, D.M.; Gibson, A.H.

    1986-04-01

    A mutant strain of Cellulomonas sp. CS1-17 was compared with Cellulomonas gelida 2480 as the cellulolytic component of a mixed culture which was responsible for the breakdown of wheat straw to support asymbiotic nitrogen fixation by Azospirillum brasilense Sp7 (ATCC 29145). Cellulomonas sp. strain CS1-17 was more efficient than was C. gelida in cellulose breakdown at lower oxygen concentrations and, in mixed culture with A. brasilense, it supported higher nitrogenase activity(C/sub 2/H/sub 2/ reduction) and nitrogen fixation with straw as the carbon source. Based on gravimetric determinations of straw breakdown and total N determinations, the efficiency of nitrogen fixation was 72 and 63 mg of N per g of straw utilized for the mixtures containing Cellulomonas sp. and C. gelida, respectively. Both Cellulomonas spp. and Azospirillum spp. exhibited a wide range of pH tolerance. When introduced into sterilized soil, the Cellulomonas sp.-Azospirillum brasilense association was more effective in nitrogen fixation at a pH of 7.0 than at the native soil pH (5.6). This was also true of the indigenous diazotrophic microflora of this soil. The potential implications of this work to the field situation are discussed. 16 references.

  14. [Effect of the fungicide captan on Azospirillum brasilense Cd in pure culture and associated with Setaria italica].

    Science.gov (United States)

    Di Ciocco, C A; Rodríguez Cáceres, E

    1997-01-01

    The effect of fungicide captan on growth and nitrogenase activity of Azospirillum brasilense Cd was studied in pure cultures and in association with foxtail millet (Setaria italica) cultivar Carapé under laboratory conditions. The 8 h growth in rotary shaker of A. brasilense was inhibited with 1 mg/l pure captan; however, after 4 days the differences diminished compared with the control without captan. Nitrogenase activity was affected with 10 mg/l but the differences were negligible after 48 h of growth. Root dry weight of inoculated plants was diminished by the treatment of foxtail millet cv. Carapé with captan. Inoculation with A. brasilense Cd increased shoot dry weight, but differences were significant only with respect to the control but not in relation to captan treatments.

  15. Regulation of glnB gene promoter expression in Azospirillum brasilense by the NtrC protein.

    Science.gov (United States)

    Huergo, Luciano F; Souza, Emanuel M; Steffens, M Berenice R; Yates, M Geoffrey; Pedrosa, Fabio O; Chubatsu, Leda S

    2003-06-01

    In Azospirillum brasilense the glnB and glnA genes are clustered in an operon regulated by three different promoters: two located upstream of glnB (glnBp1-sigma(70), and glnBp2-sigma(N)) and one as yet unidentified promoter, in the glnBA intergenic region. We have investigated the expression of the glnB gene promoter using glnB-lacZ gene fusions, mutation analysis, heterologous expression and DNA band-shift assays. Deletion of the glnB promoter region showed that NtrC-binding sequences were essential for glnB expression under nitrogen limitation. The A. brasilense NtrC protein activated transcription of glnB-lacZ fusions in the heterologous genetic background of Escherichia coli. Expression of glnB-lacZ fusions in two A. brasilense ntrC mutants differed from that in the wild-type strain. In vitro studies also indicated that the purified NtrC protein from E. coli was able to bind to the glnB promoter region of A. brasilense. Our results show that the NtrC protein activates glnBglnA expression under nitrogen limitation in A. brasilense.

  16. An extra-cytoplasmic function sigma factor and anti-sigma factor control carotenoid biosynthesis in Azospirillum brasilense.

    Science.gov (United States)

    Thirunavukkarasu, Nagarajan; Mishra, Mukti Nath; Spaepen, Stijn; Vanderleyden, Jos; Gross, Carol A; Tripathi, Anil K

    2008-07-01

    Strains Sp7 and Cd of Azospirillum brasilense, a plant growth-promoting rhizobacterium, differ in synthesis of carotenoids. While colonies of strain Sp7 have a white-cream colour on plates, colonies of strain Cd are orange-pink coloured because of the synthesis of carotenoids. Screening of a mini-Tn5 mutant library of A. brasilense Sp7 revealed two orange-pink-coloured mutants that produced carotenoids. Cloning and sequencing of the Tn5 flanking region in both the carotenoid-producing mutants of Sp7 revealed insertion of Tn5 in an ORF encoding anti-sigma factor, a ChrR-like protein. The upstream region of the Tn5-mutated ORF contained another ORF that encoded an extra-cytoplasmic function (ECF)-class sigma factor (sigma(E), RpoE). When the nucleotide sequences of the corresponding ORFs from the carotenoid-producing strain Cd were analysed, the sequence of the Cd sigma(E) was identical to that of the carotenoid non-producing strain Sp7, but the Cd anti-sigma(E) ORF had a deletion that caused frame shifting and creation of a stop codon. This resulted in the premature termination of the protein, which was about 7 kDa smaller than the Sp7 anti-sigma(E). Cloning of Sp7 anti-sigma(E) in a broad-host-range expression vector and expression in A. brasilense Cd and in the anti-sigma(E) knockout mutant of A. brasilense Sp7 resulted in the inhibition of carotenoid synthesis. Similarly, cloning and overexpression of A. brasilense Sp7 sigma(E) in A. brasilense Sp7 resulted in the production of carotenoids. These observations clearly indicate that carotenoid synthesis in A. brasilense is controlled by sigma(E) with its cognate anti-sigma(E).

  17. Tryptophan, thiamine and indole-3-acetic acid exchange between Chlorella sorokiniana and the plant growth-promoting bacterium Azospirillum brasilense.

    Science.gov (United States)

    Palacios, Oskar A; Gomez-Anduro, Gracia; Bashan, Yoav; de-Bashan, Luz E

    2016-06-01

    During synthetic mutualistic interactions between the microalga Chlorella sorokiniana and the plant growth-promoting bacterium (PGPB) Azospirillum brasilense, mutual exchange of resources involved in producing and releasing the phytohormone indole-3-acetic acid (IAA) by the bacterium, using tryptophan and thiamine released by the microalga, were measured. Although increased activities of tryptophan synthase in C. sorokiniana and indole pyruvate decarboxylase (IPDC) in A. brasilense were observed, we could not detect tryptophan or IAA in the culture medium when both organisms were co-immobilized. This indicates that no extra tryptophan or IAA is produced, apart from the quantities required to sustain the interaction. Over-expression of the ipdC gene occurs at different incubation times: after 48 h, when A. brasilense was immobilized alone and grown in exudates of C. sorokiniana and at 96 h, when A. brasilense was co-immobilized with the microalga. When A. brasilense was cultured in exudates of C. sorokiniana, increased expression of the ipdC gene, corresponding increase in activity of IPDC encoded by the ipdC gene, and increase in IAA production were measured during the first 48 h of incubation. IAA production and release by A. brasilense was found only when tryptophan and thiamine were present in a synthetic growth medium (SGM). The absence of thiamine in SGM yielded no detectable IAA. In summary, this study demonstrates that C. sorokiniana can exude sufficient tryptophan and thiamine to allow IAA production by a PGPB during their interaction. Thiamine is essential for IAA production by A. brasilense and these three metabolites are part of a communication between the two microorganisms.

  18. Hydrogenase activity in Azospirillum brasilense is inhibited by nitrite, nitric oxide, carbon monoxide, and acetylene

    Energy Technology Data Exchange (ETDEWEB)

    Tibelius, K.H.; Knowles, R.

    1984-10-01

    Nitrite, NO, CO, and C/sub 2/H/sub 2/ inhibited O/sub 2/-dependent H/sub 2/ uptake (H/sup 3/H oxidation) in denitrifying Azospirillum brasilense Sp7 grown anaerobically on N/sub 2/O or NO/sub 3//sup -/. The apparent K/sub i/ values for inhibition of O/sub 2/-dependent H/sub 2/ uptake were 20 ..mu..M for NO/sub 2//sup -/, 0.4 ..mu..M for NO, 28 ..mu..M for CO, and 88 ..mu..M for C/sub 2/H/sub 2/. These inhibitors also affected methylene blue-dependent H/sub 2/ uptake, presumably by acting directly on the hydrogenase. Nitrite and NO inhibited H/sub 2/ uptake irreversibly, whereas inhibition due to CO was easily reversed by repeatedly evacuating and backfilling with N/sub 2/. The C/sub 2/H/sub 2/ inhibition was not readily reversed, partly due to difficulty in removing the last traces of this gas from solution. The NO/sub 2//sup -/ inhibition of malate-dependent respiration was readily reversed by repeatedly washing the cells, in contrast to the effect of NO/sub 2//sup -/ on H/sub 2/-dependent respiration. These results suggest that the low hydrogenase activities observed in NO/sub 3//sup -/-grown cultures of A. brasilense may be due to the irreversible inhibition of hydrogenase by NO/sub 2//sup -/ and NO produced by NO/sub 3//sup -/ reduction.

  19. Mutants with Enhanced Nitrogenase Activity in Hydroponic Azospirillum brasilense-Wheat Associations

    Science.gov (United States)

    Pereg Gerk, Lily; Gilchrist, Kate; Kennedy, Ivan R.

    2000-01-01

    The effect of a mutation affecting flocculation, differentiation into cyst-like forms, and root colonization on nitrogenase expression by Azospirillum brasilense is described. The gene flcA of strain Sp7 restored these phenotypes in spontaneous mutants of both strains Sp7 and Sp245. Employing both constitutive pLA-lacZ and nifH-lacZ reporter fusions expressed in situ, the colony morphology, colonization pattern, and potential for nitrogenase activity of spontaneous mutants and flcA Tn5-induced mutants were established. The results of this study show that the ability of Sp7 and Sp245 mutant strains to remain in a vegetative form improved their ability to express nitrogenase activity in association with wheat in a hydroponic system. Restoring the cyst formation and colonization pattern to the spontaneous mutant Sp7-S reduced nitrogenase activity rates in association with plants to that of the wild-type Sp7. Although Tn5-induced flcA mutants showed higher potentials for nitrogenase expression than Sp7, their potentials were lower than that of Sp7-S, indicating that other factors in this strain contribute to its exceptional nitrogenase activity rates on plants. The lack of lateral flagella is not one of these factors, as Sp7-PM23, a spontaneous mutant impaired in swarming and lateral-flagellum production but not in flocculation, showed wild-type nitrogenase activity and expression. The results also suggest factors of importance in evolving an effective symbiosis between Azospirillum and wheat, such as increasing the availability of microaerobic niches along the root, increased supply of carbon sources by the plant, and the retention of the bacterial cells in vegetative form for faster metabolism. PMID:10788397

  20. [A STUDY OF THE ISOLATED BACTERIOPHAGE ΦAB-SP7 ADSORPTION ON THE CELL SURFACE OF THE AZOSPIRILLUM BRASILENSE SP7].

    Science.gov (United States)

    Guliy, O I; Karavaeva, O A; Velikov, V A; Sokolov, O I; Pavily, S A; Larionova, O S; Burov, A M; Ignatov, O V

    2016-01-01

    The bacteriophage ΦAb-Sp7 was isolated from the cells of the Azospirillum brasilense Sp7. The morphology, size of the gram-negative colonies, and range of lytic activity against other strains and species of the genus Azospirillum was tested. The isolated phage DNA was examined using electrophoretic and restriction analysis, and the size of the genome were established. The electron microscopy. resuIts show that the phage (capsid) has a strand-like form. The electron microscopy study of the bacteriophage ΦAb-Sp7 adsorption on the A. brasilense Sp7 bacterial surface was performed.

  1. The ipdC, hisC1 and hisC2 genes involved in indole-3-acetic production used as alternative phylogenetic markers in Azospirillum brasilense.

    Science.gov (United States)

    Jijón-Moreno, Saúl; Marcos-Jiménez, Cynthia; Pedraza, Raúl O; Ramírez-Mata, Alberto; de Salamone, I García; Fernández-Scavino, Ana; Vásquez-Hernández, Claudia A; Soto-Urzúa, Lucia; Baca, Beatriz E

    2015-06-01

    Plant growth-promoting bacteria of the genus Azospirillum are present in the rhizosphere and as endophytes of many crops. In this research we studied 40 Azospirillum strains isolated from different plants and geographic regions. They were first characterized by 16S rDNA restriction analysis, and their phylogenetic position was established by sequencing the genes 16S rDNA, ipdC, hisC1, and hisC2. The latter three genes are involved in the indole-3-pyruvic acid (IPyA) biosynthesis pathway of indole-3-acetic acid (IAA). Furthermore, the suitability of the 16S-23S rDNA intergenic spacer sequence (IGS) for the differentiation of closely related Azospirillum taxa and development of PCR protocols allows for specific detection of strains. The IGS-RFLP analysis enabled intraspecies differentiation, particularly of Azospirillum brasilense and Azospirillum lipoferum strains. Results demonstrated that the ipdC, hisC1, and hisC2 genes are highly conserved in all the assessed A. brasilense isolates, suggesting that these genes can be used as an alternative phylogenetic marker. In addition, IAA production determined by HPLC ranged from 0.17 to 98.2 μg mg(-1) protein. Southern hybridization with the A. brasilense ipdC gene probe did not show, a hybridization signal with A. lipoferum, Azospirillum amazonense, Azospirillum halopreferans and Azospirillum irakense genomic DNA. This suggests that these species produce IAA by other pathways. Because IAA is mainly synthesized via the IPyA pathway in A. brasilense strains, a species that is used worldwide in agriculture, the identification of ipdC, hisC1, and hisC2 genes by PCR may be suitable for selecting exploitable strains.

  2. Crecimiento de Azospirillum brasilense en presencia de disacáridos: sacarosa y lactosa

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    Yudyt Díaz-Saez

    2013-01-01

    Full Text Available La obtención de biopolímeros a partir de fermentaciones bacterianas requiere de la selección adecuada de las condiciones del proceso. Se evaluó el crecimiento de la cepa Azospirillum brasilense 8-INICA en medio líquido suplementado con disacáridos. Las mediciones se realizaron a las 23 h de incubación con la determinación de la densidad óptica, la concentración de azúcares reductores y concentración de carbohidratos totales, así como la determinación del pH del sobrenadante. Se confirmó el máximo desarrollo de la biomasa para 33 oC y 7,2 g.L-1 de sal con sacarosa, y 40 oC y 3,5 g.L-1 de sal con lactosa. Se estableció que las condiciones más favorables para la producción de exopolisacáridos se encuentran a temperaturas y concentraciones elevadas de sal para un medio con lactosa; y bajas temperaturas a concentraciones elevadas de sal en presencia de sacarosa. Se constató la marcada influencia de la temperatura en el crecimiento, y la acción positiva de la concentración salina en los mecanismos de supervivencia del micro- organismo.

  3. Mode of utilization of amino acids as growth substrates by Azospirillum brasilense.

    Science.gov (United States)

    Bhattacharya, Pradip

    2005-12-01

    The study was undertaken to analyze the rate of uptake and utilization of various amino acids by Azospirillum brasilense Sp81 (RG) in a basal mineral salts solution under non-nitrogen fixing condition. These amino acids including other nitrogenous compounds were tested for both N- and C-sources. The kinetic constants (Km and Vmax) of uptake of some amino acids (e.g. lysine, arginine, proline, glutamine and glutamic acid) were exploited using a Hanes-Woolf plot, and discussed in the context of nitrogen starvation or both carbon and nitrogen starvation. To summarize all the kinetic data for these amino acids strongly suggested that the mode of these amino acids utilization in this bacterium followed the same general pattern, although the quantitative differences were there. A single amino acid was able to satisfy the nitrogen needs of this bacterium in basal mineral salts solution, and this possibility could be considered for the cost-effective growth medium for this bacterium in the biotechnological industry.

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

    Directory of Open Access Journals (Sweden)

    S.O Procópio

    2011-01-01

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

  5. EFFECT OF MOISTURE LEVELS ON THE GROWTH, NODULATION AND NITROGEN FIXATION IN Dalbergia sissoo BY Azospirillum brasilense AND Acaulospora laevis

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    Gaurav Bhushan

    2014-08-01

    Full Text Available In present study, influence of single and combined inoculation of Azospirillum brasilense and Acaulospora laevis on the growth, nodulation and N2 fixation in Dalbergia sissoo was tested at various moisture levels under pot culture conditions. Combined inoculation of A.brasilense and A.laevis was found best in raising maximum growth, nodulation, nitrogen fixation and percentage of AM colonization in roots. These characters were considerably influenced with the increasing moisture stress from -0.3MPa to -1.5MPa. Furthermore, the values of all the above mentioned parameters show decrement with the increasing of moisture stress. Plants with dual inoculation performed better than single inoculated plants. Observation of the present study counted a protective role played by AM in providing resistance to D.sissoo against injurious effects of moisture stress.

  6. The Azospirillum brasilense Sp7 noeJ and noeL genes are involved in extracellular polysaccharide biosynthesis.

    Science.gov (United States)

    Lerner, Anat; Castro-Sowinski, Susana; Valverde, Angel; Lerner, Hadas; Dror, Rachel; Okon, Yaacov; Burdman, Saul

    2009-12-01

    Azospirillum brasilense is a plant root-colonizing bacterium that exerts beneficial effects on the growth of many agricultural crops. Extracellular polysaccharides of the bacterium play an important role in its interactions with plant roots. The pRhico plasmid of A. brasilense Sp7, also named p90, carries several genes involved in synthesis and export of cell surface polysaccharides. We generated two Sp7 mutants impaired in two pRhico-located genes, noeJ and noeL, encoding mannose-6-phosphate isomerase and GDP-mannose 4,6-dehydratase, respectively. Our results demonstrate that in A. brasilense Sp7, noeJ and noeL are involved in lipopolysaccharide and exopolysaccharide synthesis. noeJ and noeL mutant strains were significantly altered in their outer membrane and cytoplasmic/periplasmic protein profiles relative to the wild-type strain. Moreover, both noeJ and noeL mutations significantly affected the bacterial responses to several stresses and antimicrobial compounds. Disruption of noeL, but not noeJ, affected the ability of the A. brasilense Sp7 to form biofilms. The pleiotropic alterations observed in the mutants could be due, at least partially, to their altered lipopolysaccharides and exopolysaccharides relative to the wild-type.

  7. Enhanced activity of ADP glucose pyrophosphorylase and formation of starch induced by Azospirillum brasilense in Chlorella vulgaris.

    Science.gov (United States)

    Choix, Francisco J; Bashan, Yoav; Mendoza, Alberto; de-Bashan, Luz E

    2014-05-10

    ADP-glucose pyrophosphorylase (AGPase) regulates starch biosynthesis in higher plants and microalgae. This study measured the effect of the bacterium Azospirillum brasilense on AGPase activity in the freshwater microalga Chlorella vulgaris and formation of starch. This was done by immobilizing both microorganisms in alginate beads, either replete with or deprived of nitrogen or phosphorus and all under heterotrophic conditions, using d-glucose or Na-acetate as the carbon source. AGPase activity during the first 72h of incubation was higher in C. vulgaris when immobilized with A. brasilense. This happened simultaneously with higher starch accumulation and higher carbon uptake by the microalgae. Either carbon source had similar effects on enzyme activity and starch accumulation. Starvation either by N or P had the same pattern on AGPase activity and starch accumulation. Under replete conditions, the population of C. vulgaris immobilized alone was higher than when immobilized together, but under starvation conditions A. brasilense induced a larger population of C. vulgaris. In summary, adding A. brasilense enhanced AGPase activity, starch formation, and mitigation of stress in C. vulgaris.

  8. The chemotaxis-like Che1 pathway has an indirect role in adhesive cell properties of Azospirillum brasilense.

    Science.gov (United States)

    Siuti, Piro; Green, Calvin; Edwards, Amanda Nicole; Doktycz, Mitchel J; Alexandre, Gladys

    2011-10-01

    The Azospirillum brasilense chemotaxis-like Che1 signal transduction pathway was recently shown to modulate changes in adhesive cell surface properties that, in turn, affect cell-to-cell aggregation and flocculation behaviors rather than flagellar-mediated chemotaxis. Attachment to surfaces and root colonization may be functions related to flocculation. Here, the conditions under which A. brasilense wild-type Sp7 and che1 mutant strains attach to abiotic and biotic surfaces were examined using in vitro attachment and biofilm assays combined with atomic force microscopy and confocal microscopy. The nitrogen source available for growth is found to be a major modulator of surface attachment by A. brasilense and could be promoted in vitro by lectins, suggesting that it depends on interaction with surface-exposed residues within the extracellular matrix of cells. However, Che1-dependent signaling is shown to contribute indirectly to surface attachment, indicating that distinct mechanisms are likely underlying flocculation and attachment to surfaces in A. brasilense.

  9. Chemical synthesis of the tetrasaccharide repeating unit of the O-polysaccharide isolated from Azospirillum brasilense SR80.

    Science.gov (United States)

    Sarkar, Vikramjit; Mukhopadhyay, Balaram

    2015-04-10

    A linear strategy has been developed for the synthesis of the tetrasaccharide repeating unit of the O-polysaccharide from Azospirillum brasilense SR80. Stepwise glycosylation of the rationally protected thioglycoside donors activated by NIS in the presence of La(OTf)3 furnished the target tetrasaccharide. The glycosylation reactions resulted in the formation of the desired linkage with absolute stereoselectivity and afforded the required derivatives in good to excellent yields. The phthalimido group has been used as the precursor of the desired acetamido group to meet the requirement of 1,2-trans glycosidic linkage.

  10. Identification of an O-linked repetitive glycan chain of the polar flagellum flagellin of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Belyakov, Alexei Ye; Burygin, Gennady L; Arbatsky, Nikolai P; Shashkov, Alexander S; Selivanov, Nikolai Yu; Matora, Larisa Yu; Knirel, Yuriy A; Shchyogolev, Sergei Yu

    2012-11-01

    This is the first report to have identified an O-linked repetitive glycan in bacterial flagellin, a structural protein of the flagellum. Studies by sugar analysis, Smith degradation, (1)H and (13)C NMR spectroscopy, and mass spectrometry showed that the glycan chains of the polar flagellum flagellin of the plant-growth-promoting rhizobacterium Azospirillum brasilense Sp7 are represented by a polysaccharide with a molecular mass of 7.7 kDa, which has a branched tetrasaccharide repeating unit of the following structure:

  11. A CheR/CheB fusion protein is involved in cyst cell development and chemotaxis in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Wu, Lixian; Cui, Yanhua; Hong, Yuanyuan; Chen, Sanfeng

    2011-12-20

    We here report the sequence and functional analysis of cstB of Azospirillum brasilense Sp7. The predicted cstB contains C-terminal two PAS domains and N-terminal part which has similarity with CheB-CheR fusion protein. cstB mutants had reduced swarming ability compared to that of A. brasilense wild-type strain, implying that cstB was involved in chemotaxis in A. brasilense. A microscopic analysis revealed that cstB mutants developed mature cyst cells more quickly than wild type, indicating that cstB is involved in cyst formation. cstB mutants were affected in colony morphology and the production of exopolysaccharides (EPS) which are essential for A. brasilense cells to differentiate into cyst-like forms. These observations suggested that cstB was a multi-effector involved in cyst development and chemotaxis in A. brasilense.

  12. RpoH2 sigma factor controls the photooxidative stress response in a non-photosynthetic rhizobacterium, Azospirillum brasilense Sp7.

    Science.gov (United States)

    Kumar, Santosh; Rai, Ashutosh Kumar; Mishra, Mukti Nath; Shukla, Mansi; Singh, Pradhyumna Kumar; Tripathi, Anil Kumar

    2012-12-01

    Bacteria belonging to the Alphaproteobacteria normally harbour multiple copies of the heat shock sigma factor (known as σ(32), σ(H) or RpoH). Azospirillum brasilense, a non-photosynthetic rhizobacterium, harbours five copies of rpoH genes, one of which is an rpoH2 homologue. The genes around the rpoH2 locus in A. brasilense show synteny with that found in rhizobia. The rpoH2 of A. brasilense was able to complement the temperature-sensitive phenotype of the Escherichia coli rpoH mutant. Inactivation of rpoH2 in A. brasilense results in increased sensitivity to methylene blue and to triphenyl tetrazolium chloride (TTC). Exposure of A. brasilense to TTC and the singlet oxygen-generating agent methylene blue induced several-fold higher expression of rpoH2. Comparison of the proteome of A. brasilense with its rpoH2 deletion mutant and with an A. brasilense strain overexpressing rpoH2 revealed chaperone GroEL, elongation factors (Ef-Tu and EF-G), peptidyl prolyl isomerase, and peptide methionine sulfoxide reductase as the major proteins whose expression was controlled by RpoH2. Here, we show that the RpoH2 sigma factor-controlled photooxidative stress response in A. brasilense is similar to that in the photosynthetic bacterium Rhodobacter sphaeroides, but that RpoH2 is not involved in the detoxification of methylglyoxal in A. brasilense.

  13. Gold(III) reduction by the rhizobacterium Azospirillum brasilense with the formation of gold nanoparticles.

    Science.gov (United States)

    Tugarova, Anna V; Burov, Andrei M; Burashnikova, Marina M; Kamnev, Alexander A

    2014-01-01

    For the soil nitrogen-fixing bacterium Azospirillum brasilense, the ability to reduce [AuCl4](-) and to form gold nanoparticles (GNPs) has been demonstrated, with the appearance of a mauve tint of the culture. To validate the shapes and chemical nature of nanoparticles, transmission electron microscopy (TEM) and X-ray fluorescence analysis were used. For the widely studied agriculturally important wild-type strains A. brasilense Sp7 and Sp245, GNPs formed after 10 days of incubation of cell biomass with 0.25 mM [AuCl4](-) were shown (using TEM) to be mainly of spherical form (5 to 20 nm in diameter), with rare occasional triangles. In the course of cultivation with [AuCl4](-), after 5 days, a mauve tint was already visible for cells of strain Sp245.5, after 6 days for Sp245 and after 10 days for Sp7. Thus, for the mutant strain Sp245.5 (which has significant differences in the structure and composition of cell-surface polysaccharides as compared with Sp245), a more rapid formation of GNPs was observed. Moreover, their TEM images (also obtained after 10 days) showed different shapes: nano-sized spheres, triangles, hexagons and rods, as well as larger round-shaped flower-like nanoparticles about 100 nm in size. Since by the time of GNP formation in our experiments the cells were found to be already not viable, this confirms the dominating role of cell surface structure and chemical composition in shaping the GNPs formed in the course of [AuCl4](-) reduction to Au(0). This finding may be useful for understanding the natural biogeochemical mechanisms of gold reduction and formation of GNPs involving microorganisms. The data obtained may also help in developing protocols for environmentally friendly synthesis of nanoparticles and possible use of bacterial cells with modified surface structure and composition for their fabrication.

  14. Convergent synthesis of a tetrasaccharide repeating unit of the O-specific polysaccharide from the cell wall lipopolysaccharide of Azospirillum brasilense strain Sp7

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    Pintu Kumar Mandal

    2014-01-01

    Full Text Available A straightforward convergent synthesis has been carried out for the tetrasaccharide repeating unit of the O-specific cell wall lipopolysaccharide of the strain Sp7 of Azospirillum brasilense. The target tetrasaccharide has been synthesized from suitably protected monosaccharide intermediates in 42% overall yield in seven steps by using a [2 + 2] block glycosylation approach.

  15. INOCULAÇÃO DE Azospirillum brasilense NA QUALIDADE FISIOLÓGICA DE SEMENTES DE MILHO DOCE

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    SOUZA, Epitácio José de

    2014-06-01

    Full Text Available The aimed of this study was to survey the physiological quality of sweet corn seeds submitted to inoculation of Azospirillum brasilense in different periods and dilutions in water. The experiment was conducted on the entirely randomized design with four replications in a factorial (4 x 4 more a witness without inoculation of A. brasilense. Were evaluated four periods between the inoculation and installation of the tests (0, 24, 48 and 72 hours and four dilutions of the inoculant in water (1:0, 1:1, 1:2 and 1:3. Were utilized seeds of sweet corn evaluated Tropical Plus hybrids and commercial strains of A. brasilense Ab-Ab-V5 and V6, in the dose of the commercial product 4 mL kg-1 seed. The parameters evaluated were germination, first count of germination, percentage of abnormal seeds, emergency, emergency speed index, length of epicotyl and of root, mass of the dry matter of shoots and of root. The dilution of A. brasilense in water did not affect no one parameter evaluated on the seeds of sweet corn, and the increase in the time between inoculation and the installation of the tests provide reduction on the first count of germination, speed emergency index and mass of dry matter of roots. It is concluded that, under laboratory conditions, inoculation of A. brasilense not benefits to germination and seed vigor of sweet corn. Objetivou-se avaliar a qualidade fisiológica de sementes de milho doce submetidas à inoculação de Azospirillum brasilense em diferentes períodos e diluições em água. O experimento foi conduzido no delineamento inteiramente casualizado, com quatro repetições em esquema fatorial (4 x 4, mais uma testemunha que não recebeu inoculação de A. brasilense. Foram avaliados quatro períodos entre a inoculação e a instalação dos testes (0, 24, 48 e 72 horas e quatro diluições do inoculante em água (1:0, 1:1, 1:2 e 1:3. Foram utilizadas sementes de milho doce híbrido comercial Tropical Plus e as estirpes de A

  16. Effect of nitrogen supply and Azospirillum brasilense Sp-248 on the response of wheat to seawater irrigation.

    Science.gov (United States)

    Alamri, Saad A; Mostafa, Yasser S

    2009-10-01

    Response of wheat to Azospirillum brasilense Sp-248 inoculation with different N-fertilizer levels using seawater irrigation was investigated. All inoculated treatments increased plant height, shoot and root dry weight, and tiller number in compared with uninoculated treatments. Yield parameters measured were also increased due to the inoculation. In terms of the effect of saline irrigation, there were no significant differences in growth and yield parameters in plants treated with tap water and others irrigated with 8.0% seawater concentration. This would indicate a relatively high tolerance of A. brasilense to saline irrigation and its ability to reduce the deleterious effects of saline on growth by increasing the plant's adaptation. However, increasing the seawater concentration in the irrigation water to 16.0% significantly decreased all tested parameters. Inoculation treatments generally increased NPKCa contents and decreased sodium ratio of the grains in compared with the uninoculated treatments. Overall results clearly revealed that the Azospirillum inoculation saved about 20 units of N-fertilizer and that saving was made economically feasible by decreasing the chemical fertilizers needed, improving the nitrogen content and counteracting the effects of salinity.

  17. Phenotypical and molecular responses of Arabidopsis thaliana roots as a result of inoculation with the auxin-producing bacterium Azospirillum brasilense.

    Science.gov (United States)

    Spaepen, Stijn; Bossuyt, Stijn; Engelen, Kristof; Marchal, Kathleen; Vanderleyden, Jos

    2014-02-01

    The auxin-producing bacterium Azospirillum brasilense Sp245 can promote the growth of several plant species. The model plant Arabidopsis thaliana was chosen as host plant to gain an insight into the molecular mechanisms that govern this interaction. The determination of differential gene expression in Arabidopsis roots after inoculation with either A. brasilense wild-type or an auxin biosynthesis mutant was achieved by microarray analysis. Arabidopsis thaliana inoculation with A. brasilense wild-type increases the number of lateral roots and root hairs, and elevates the internal auxin concentration in the plant. The A. thaliana root transcriptome undergoes extensive changes on A. brasilense inoculation, and the effects are more pronounced at later time points. The wild-type bacterial strain induces changes in hormone- and defense-related genes, as well as in plant cell wall-related genes. The A. brasilense mutant, however, does not elicit these transcriptional changes to the same extent. There are qualitative and quantitative differences between A. thaliana responses to the wild-type A. brasilense strain and the auxin biosynthesis mutant strain, based on both phenotypic and transcriptomic data. This illustrates the major role played by auxin in the Azospirillum-Arabidopsis interaction, and possibly also in other bacterium-plant interactions.

  18. INOCULAÇÃO COM Azospirillum brasilense ASSOCIADA Á ADUBAÇÃO NITROGENADA REDUZIDA EM HIBRIDOS DE MILHO

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    LIBÓRIO, Paloma Helena da Silva1

    2016-11-01

    Full Text Available Objetivou-se avaliar o comportamento de híbridos de milho inoculados com Azospirillum brasilense, estirpes: AbV5 e AbV6, em dose reduzida de N. Dois experimentos em diferentes anos agrícolas foram conduzidos em delineamento experimental de blocos ao acaso, sendo o primeiro em 2007/08: com quatro repetições composto por um híbrido nos seguintes tratamentos: Testemunha sem N e sem Azospirillum (FAzos; N recomendado para a cultura (dose usual; ½ da dose de N usual; ½ da dose de N usual + FAzos turfoso e ½ da dose de N usual + FAzos líquido. O segundo conduzido na safra 2012/13, foi em arranjo fatorial 2 x 5, com oito repetições, composto por dois híbridos testados nos tratamentos já supracitados com alteração apenas da inoculação com Azospirillum nas formulações (FAzos de veículo turfoso e líquido que tramitou da fase de testes para formulação comercial veículo: líquido. Avaliou-se parâmetros agronômicos no início do florescimento e na colheita. De modo geral, a associação de ½ de dose de N usual e FAzos foi equivalente ao tratamento que recebeu o total da dose de nitrogênio recomendada quanto aos parâmetros avaliados no híbrido DKB 390. Houve superioridade do híbrido DKB 390 VT PRO 2 TM em relação ao BM 502, fato atribuído como próprio do genótipo e não da interação com as estirpes da bactéria. Concluiu-se que a prática de inoculação de sementes com Azospirillum brasilense: estirpes AbV5 e AbV6, pode ocorrer de forma associada a adubação nitrogenada reduzida pela metade, parcelada na semeadura e cobertura. This study aimed to evaluate the behavior of maize hybrids inoculated with Azospirillum brasilense, strains: AbV5 and AbV6 in reduced dose of N. Two experiments in different growing seasons were conducted in experimental design was a randomized complete block, with the first in 2007/08: with four replications composed of a hybrid evaluated in the following treatments: Control without N and without

  19. Residual effect of nitrogen fertilization and Azospirillum brasilense inoculation in the maize cultureEfeito residual da adubação nitrogenada e inoculação de Azospirillum brasilense na cultura do milho

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    Jackson Huzar Novakowiski

    2011-12-01

    Full Text Available The diazotrophic bacteria Azospirillum brasilense is an organism able to fix nitrogen of atmosphere and produce plant hormones. Nevertheless, there is lack of information with regard to use in field conditions, especially in production systems that have presence of animals in a determined year period. The objective of paper was to evaluate the association of the nitrogen residual effect of fertilization in pasture winter and the inoculation with A. brasilense in the maize culture. Were carried two experiments in Guarapuava (PR in season 2008/09 with randomized block design with split plots in three replication. The main parcel consisted of application 5 levels of N (0, 75, 150, 225 e 300 kg ha-1 in pasture of black oat and ryegrass occupied by bovines and 4 treatments in the maize (T1 = control; T2 = inoculation of A. brasilense; T3 = 75 kg ha-1 of N; T4 = 150 kg ha-1 of N. Were evaluated: plants population, ears per plant, productivity, mass thousand grains, damage grains, row per ear and grains per row. There was residual effect of nitrogen applied in the pasture on maize culture. The mass thousand grains and the number of rows per ear of maize presented quadratic response for increase of nitrogen level in the pasture. The inoculation of A. brasilense provided higher productivity than control same with increase the nitrogen level applied in the pasture, with quadratic response in the experiment 1 and linear in the experiment 2.A bactéria diazotrófica Azospirillum brasilense é um organismo capaz de fixar nitrogênio da atmosfera e produzir hormônios vegetais. No entanto, há carência de informações a respeito de seu uso em condições de campo, sobretudo em sistemas de produção que tem a presença de animais em um determinado período do ano. O objetivo do trabalho foi avaliar a associação do efeito residual da adubação nitrogenada na pastagem de inverno e a inoculação de A. brasilense na cultura do milho. Foram conduzidos dois

  20. Bacteriophytochrome controls carotenoid-independent response to photodynamic stress in a non-photosynthetic rhizobacterium, Azospirillum brasilense Sp7.

    Science.gov (United States)

    Kumar, Santosh; Kateriya, Suneel; Singh, Vijay Shankar; Tanwar, Meenakshi; Agarwal, Shweta; Singh, Hina; Khurana, Jitendra Paul; Amla, Devinder Vijay; Tripathi, Anil Kumar

    2012-01-01

    Ever since the discovery of the role of bacteriophytochrome (BphP) in inducing carotenoid synthesis in Deinococcus radiodurans in response to light the role of BphPs in other non-photosynthetic bacteria is not clear yet. Azospirillum brasilense, a non-photosynthetic rhizobacterium, harbours a pair of BphPs out of which AbBphP1 is a homolog of AtBphP1 of Agrobacterium tumefaciens. By overexpression, purification, biochemical and spectral characterization we have shown that AbBphP1 is a photochromic bacteriophytochrome. Phenotypic study of the ΔAbBphP1 mutant showed that it is required for the survival of A. brasilense on minimal medium under red light. The mutant also showed reduced chemotaxis towards dicarboxylates and increased sensitivity to the photooxidative stress. Unlike D. radiodurans, AbBphP1 was not involved in controlling carotenoid synthesis. Proteome analysis of the ΔAbBphP1 indicated that AbBphP1 is involved in inducing a cellular response that enables A. brasilense in regenerating proteins that might be damaged due to photodynamic stress.

  1. Interaction between GInB and the N-terminal domain of NifA in Azospirillum brasilense

    Institute of Scientific and Technical Information of China (English)

    ZHOU XiaoYu; ZOU XiaoXiao; LI JiLun

    2008-01-01

    Azospirillum brasilense is a diazotroph associated with many important agricultural crops and shows potential as a biofertilizer. NifA, the transcriptional activator of nitrogen fixation (nif) genes, and GInB, one of P,, signal transduction family protein, are key proteins in the regulation of nitrogen fixation in A. brasilense. It was previously reported that the regulation of NifA activity in A. brasilense depends on GInB. We report here that GInB was found to interact directly with the N-terminal domain of NifA in vivo under nitrogen-free conditions and the N-terminal mutant of NifA in which the Tyr residues at position 18 and 53 were replaced by Phe (NifA-N-Y18/53F) strengthened the interaction with GInB. Moreover, we also found that the amino acid residues 66-88 and 165-176 in N-terminus of NifA are responsible for the interaction with GInB.

  2. Migration of Azospirillum brasilense Yu62 from Root to Stem and Leaves Inside Rice and Tobacco Plants%巴西固氮螺菌Yu62由水稻和烟草根部向茎、叶的迁移运动

    Institute of Scientific and Technical Information of China (English)

    迟峰; 沈世华; 陈三凤; 荆玉祥

    2004-01-01

    Azospirillum brasilense Tarrand, Krieg et Dobereiner is one of the important plant growthpromotion endophytes. A. brasilense Yu62 tagged with gfp gene was inoculated into roots of rice and tobacco seedlings, which were then, cultured in gnotobiotic condition. At a certain days after inoculation the different portions of the seedling were observed under laser confocal microscope, resulting in that A.brasilense Yu62 bacteria were colonized in epidermal and cortical cells, intercellular spaces and vascular system of stem and leaf tissue interiors besides in roots. Higher populations of the bacteria isolated from roots, stems and leaves indicated that A. brasilense Yu62 bacteria could ascend themselves from roots to stems and leaves of rice and tobacco. This observation lays down the foundation for ecology and cell morphology of bacterial migration inside plants, interaction between A. brasilense Yu62 bacteria and host cells as well as the plant-growth promotion, provides scientific basis for further application, and is of importance in science and practice.%巴西固氮螺菌(Azospirillrm brasilence)是重要的植物促生内生菌之一.用gfp基因标记固氮螺菌后接种无菌的水稻和烟草幼苗的根部,限制培养一定时间后,用共聚焦激光显微镜观察,结果表明:除了根部有发荧光的螺菌定殖外,螺菌还分布在茎、叶的表皮细胞,皮层细胞和维管系统组织的细胞间隙.从根、茎、叶器官分离固氮螺菌,都存在有较高的螺菌群体密度.这一结果证明螺菌在植物内存在着从根部向茎、叶顶端的迁移现象.这一发现为研究巴西固氮螺菌在窠主植物体内的迁移运动的机制、与植物细胞间的分子相互作用及其对植物的促生作用奠定了生态学和细胞形态学的基础,也为实际应用提示了进一步的科学依据,具有重要的科学和实践意义.

  3. Involvement of indole-3-acetic acid produced by Azospirillum brasilense in accumulating intracellular ammonium in Chlorella vulgaris.

    Science.gov (United States)

    Meza, Beatriz; de-Bashan, Luz E; Bashan, Yoav

    2015-01-01

    Accumulation of intracellular ammonium and activities of the enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were measured when the microalgae Chlorella vulgaris was immobilized in alginate with either of two wild type strains of Azospirillum brasilense or their corresponding indole-3-acetic acid (IAA)-attenuated mutants. After 48 h of immobilization, both wild types induced higher levels of intracellular ammonium in the microalgae than their respective mutants; the more IAA produced, the higher the intracellular ammonium accumulated. Accumulation of intracellular ammonium in the cells of C. vulgaris followed application of four levels of exogenous IAA reported for A. brasilense and its IAA-attenuated mutants, which had a similar pattern for the first 24 h. This effect was transient and disappeared after 48 h of incubation. Immobilization of C. vulgaris with any bacteria strain induced higher GS activity. The bacterial strains also had GS activity, comparable to the activity detected in C. vulgaris, but weaker than when immobilized with the bacteria. When net activity was calculated, the wild type always induced higher GS activity than IAA-attenuated mutants. GDH activity in most microalgae/bacteria interactions resembled GS activity. When complementing IAA-attenuated mutants with exogenous IAA, GS activity in co-immobilized cultures matched those of the wild type A. brasilense immobilized with the microalga. Similarity occurred when the net GS activity was measured, and was higher with greater quantities of exogenous IAA. It is proposed that IAA produced by A. brasilense is involved in ammonium uptake and later assimilation by C. vulgaris.

  4. [Diverse morphological types of dormant cells and conditions for their formation in Azospirillum brasilense].

    Science.gov (United States)

    Muliukin, A L; Suzina, N E; Pogorelova, A Iu; Antoniuk, L P; Duda, V I; El'-Registan, G I

    2009-01-01

    Differences in generation of dormant forms (DF) were revealed between two strains of non-sporeforming gram-negative bacteria Azospirillum brasilense, Sp7 (non-endophytic) and Sp245 (endophytic strain). In post-stationary ageing bacterial cultures grown in a synthetic medium with a fivefold decreased initial nitrogen content, strain Sp7 formed two types of cyst-like resting cells (CRC). Strain Sp245 did not form such types of DF under the same conditions. CRC of the first type were formed in strain Sp245 only under phosphorus deficiency (C > P). The endophytic strain was also shown to form structurally differentiated cells under complete starvation, i.e. at a transfer of early stationary cultures, grown in the media with C > N unbalance, to saline solution (pH 7.2). These DF had a complex structure similar to that of azotobacter cysts. The CRC, which are generated by both azospirilla strains and belong to distinct morphological types, possessed the following major features: absence of division; specific ultrastructural organization; long-term maintenance of viability (for 4 months and more); higher heat resistance (50-60 degrees C, 10 min) as compared with vegetative cells, i.e. the important criteria for dormant prokaryotic forms. However, CRC of non-endophytic strain Sp7 had higher heat resistance (50, 55, 60 degrees C). The viability maintenance and the portion of heat-resistant cells depended on the conditions of maturation and storage of CRC populations. Long-term storage (for 4 months and more) of azospirilla DF populations at -20 degrees C was optimal for maintenance of their colony-forming ability (57% of the CFU number in stationary cultures), whereas the largest percentage of heat-resistant cells was in CRC suspensions incubated in a spent culture medium (but not in saline solution) at room temperature. The data on the intraspecies diversity of azospirilla DF demonstrate the relation between certain type DF formation to the type of interaction (non

  5. Effect of Azospirillum brasilense and Burkholderia unamae Bacteria on Maize Photosynthetic Activity Evaluated Using the Photoacoustic Technique

    Science.gov (United States)

    Gordillo-Delgado, F.; Marín, E.; Calderón, A.

    2016-09-01

    In this work, the photosynthetic process of maize plants ( Zea mays), which were grown using seeds inoculated with plant growth promoting bacteria Azospirillum brasilense and Burkholderia unamae, was monitored. Photothermal and photobaric signals obtained by a time-resolved photoacoustic measurement configuration were used for measuring the oxygen evolution rate in situ. A frequency-resolved configuration of the method was utilized to determine the oxygen diffusion coefficient and the thermal diffusivity of the maize leaves. The latter parameters, which can be used as indicators of the photosynthetic activity of maize, are found to vary according to the plant-microbe interaction. Treatment with plant growth promoting bacteria induced a decrease in the oxygen diffusion coefficient of about 20 %.

  6. Fourier transform Raman spectroscopic characterisation of cells of the plant-associated soil bacterium Azospirillum brasilense Sp7

    Science.gov (United States)

    Kamnev, A. A.; Tarantilis, P. A.; Antonyuk, L. P.; Bespalova, L. A.; Polissiou, M. G.; Colina, M.; Gardiner, P. H. E.; Ignatov, V. V.

    2001-05-01

    Structural and compositional features of bacterial cell samples and of lipopolysaccharide-protein complex isolated from the cell surface of the plant-growth-promoting rhizobacterium Azospirillum brasilense (wild-type strain Sp7) were characterised using Fourier transform (FT) Raman spectroscopy. The structural spectroscopic information obtained is analysed and considered together with analytical data on the content of metal cations (Co 2+, Cu 2+ and Zn 2+) in the bacterial cells grown in a standard medium as well as in the presence of each of the cations (0.2 mM). The latter, being taken up by bacterial cells from the culture medium in significant amounts, were shown to induce certain metabolic changes in the bacterium revealed in FT-Raman spectra, which is discussed from the viewpoint of bacterial response to environmental stresses.

  7. Spectroscopic characterization of cell membranes and their constituents of the plant-associated soil bacterium Azospirillum brasilense

    Science.gov (United States)

    Kamnev, A. A.; Antonyuk, L. P.; Matora, L. Yu.; Serebrennikova, O. B.; Sumaroka, M. V.; Colina, M.; Renou-Gonnord, M.-F.; Ignatov, V. V.

    1999-05-01

    Structural and compositional features of bacterial membranes and some of their isolated constituents (cell surface lipopolysaccharide, phospholipids) of the plant-growth-promoting diazotrophic rhizobacterium Azospirillum brasilense (wild-type strain Sp245) were characterized using Fourier transform infrared (FTIR) spectroscopy and some other techniques. FTIR spectra of the cell membranes were shown to comprise the main vibration modes of the relevant lipopolysaccharide and protein components which are believed to be involved in associative plant-bacterium interactions, as well as of phospholipid constituents. The role and functions of metal cations in the structural organization and physicochemical properties of bacterial cell membranes are also discussed considering their accumulation in the membranes from the culture medium.

  8. Mössbauer spectroscopic study of 57Fe metabolic transformations in the rhizobacterium Azospirillum brasilense Sp245

    Science.gov (United States)

    Kamnev, Alexander A.; Tugarova, Anna V.; Kovács, Krisztina; Biró, Borbála; Homonnay, Zoltán; Kuzmann, Ernő

    2014-04-01

    Preliminary 57Fe transmission Mössbauer spectroscopic data were obtained for the first time for live cells of the plant-growth-promoting rhizobacterium Azospirillum brasilense (wild-type strain Sp245) grown aerobically with 57FeIII-nitrilotriacetate (NTA) complex as a sole source of iron. The results obtained have shown that live cells actively reduce part of the assimilated iron(III) to iron(II), the latter amounting up to 33 % of total cellular iron after 18 h of growth, and 48 % after additional 3 days of storage of the dense wet cell suspension in nutrient-free saline solution in air at room temperature (measured at 80 K). The cellular iron(II) was found to be represented by two quadrupole doublets of different high-spin forms, while the parameters of the cellular iron(III) were close to those typical for bacterioferritins.

  9. Optimal fed batch experiment design for estimation of monod kinetics of Azospirillum brasilense: from theory to practice.

    Science.gov (United States)

    Cappuyns, Astrid M; Bernaerts, Kristel; Smets, Ilse Y; Ona, Ositadinma; Prinsen, Els; Vanderleyden, Jos; Van Impe, Jan F

    2007-01-01

    In this paper the problem of reliable and accurate parameter estimation for unstructured models is considered. It is illustrated how a theoretically optimal design can be successfully translated into a practically feasible, robust, and informative experiment. The well-known parameter estimation problem of Monod kinetic parameters is used as a vehicle to illustrate our approach. As known for a long time, noisy batch measurements do not allow for unique and accurate estimation of the kinetic parameters of the Monod model. Techniques of optimal experiment design are, therefore, exploited to design informative experiments and to improve the parameter estimation accuracy. During the design process, practical feasibility has to be kept in mind. The designed experiments are easy to implement in practice and do not require additional monitoring equipment. Both design and experimental validation of informative fed batch experiments are illustrated with a case study, namely, the growth of the nitrogen-fixing bacteria Azospirillum brasilense.

  10. Respuesta de tres cultivares de maíz a la inoculación con Azospirillum brasilense bajo cuatro diferentes dosis de nitrógeno

    Directory of Open Access Journals (Sweden)

    Andrés González Huerta

    2011-01-01

    Full Text Available En el municipio de Huamuxtitlán, estado de Guerrero, México, se evaluó la respuesta de los materiales de maíz VS-535, H-507 y criollo veracruzano en las dosis de 0, 30, 60 y 90 kg de nitrógeno ha-1, con y sin Azospirillum brasilense. En el sintético VS- 535 se registró el mayor rendimiento de grano (5.23 t ha-1. En las dosis de 0, 30, 60 y 90 kg de N ha-1 se registraron 4.38, 4.36, 4.81 y 4.93 t ha-1, respectivamente. La diferencia entre aplicar o no Azospirillum brasilense (promedio sobre los factores A, B y repeticiones fue de 1.47 t ha-1 y de dos días en el ciclo biológico del cultivo.

  11. Activity of two catabolic enzymes of the phosphogluconate pathway in mesquite roots inoculated with Azospirillum brasilense Cd.

    Science.gov (United States)

    Leyva, Luis A; Bashan, Yoav

    2008-10-01

    The mesquite amargo (Prosopis articulate), one of the main nurse trees of the Sonoran Desert in Mexico, is responsible for major, natural re-vegetation processes. It exudes gluconic acid in root exudates, a favorite carbon source for the plant growth-promoting bacterium Azospirillum brasilense. Two enzymes, gluconokinase (EC 2.7.1.12) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44), participating in the phosphogluconate pathway, are active in the bacteria. Bacterial 6-phosphogluconate dehydrogenase is a constitutive enzyme, while gluconokinase is induced upon exposure to gluconic acid. Both enzymes are active in young, non-inoculated mesquite seedlings growing under hydroponic conditions. When A. brasilense Cd bacteria are inoculated on the root system, the roots exhibit much higher activity of gluconokinase, but not 6-phosphogluconate dehydrogenase. Mesquite roots exhibit high levels of root colonization by the inoculating bacteria. At the same time, and also for plants growing under sand culture conditions, the seedlings grew taller, greener, had longer leaves, and were heavier.

  12. Enhanced accumulation of starch and total carbohydrates in alginate-immobilized Chlorella spp. induced by Azospirillum brasilense: II. Heterotrophic conditions.

    Science.gov (United States)

    Choix, Francisco J; de-Bashan, Luz E; Bashan, Yoav

    2012-10-10

    The effect of the bacterium Azospirillum brasilense jointly immobilized with Chlorella vulgaris or C. sorokiniana in alginate beads on total carbohydrates and starch was studied under dark and heterotrophic conditions for 144 h in synthetic growth medium supplemented with either d-glucose or Na-acetate as carbon sources. In all treatments, enhanced total carbohydrates and starch content per culture and per cell was obtained after 24h; only jointly immobilized C. vulgaris growing on d-glucose significantly increased total carbohydrates and starch content after 96 h. Enhanced accumulation of carbohydrate and starch under jointly immobilized conditions was variable with time of sampling and substrate used. Similar results occurred when the microalgae was immobilized alone. In both microalgae growing on either carbon sources, the bacterium promoted accumulation of carbohydrates and starch; when the microalgae were immobilized alone, they used the carbon sources for cell multiplication. In jointly immobilized conditions with Chlorella spp., affinity to carbon source and volumetric productivity and yield were higher than when Chlorella spp. were immobilized alone; however, the growth rate was higher in microalgae immobilized alone. This study demonstrates that under heterotrophic conditions, A. brasilense promotes the accumulation of carbohydrates in two strains Chlorella spp. under certain time-substrate combinations, producing mainly starch. As such, this bacterium is a biological factor that can change the composition of compounds in microalgae in dark, heterotrophic conditions.

  13. Enhanced accumulation of starch and total carbohydrates in alginate-immobilized Chlorella spp. induced by Azospirillum brasilense: I. Autotrophic conditions.

    Science.gov (United States)

    Choix, Francisco J; de-Bashan, Luz E; Bashan, Yoav

    2012-10-10

    The effect of the microalgae-growth promoting bacterium Azospirillum brasilense on accumulation of total carbohydrates and starch in two species of Chlorella (Chlorella vulgaris and Chlorella sorokiniana), when the bacterium and each microalga were jointly immobilized in alginate beads was studied under autotrophic conditions for 144 h in synthetic medium. The interaction of the bacterium with the microalgae enhanced accumulation of total carbohydrate and starch. Cells of Chlorella accumulated the highest amounts of carbohydrate after incubation for 24h. Yet, this did not coincide with the highest affinity and volumetric productivity measured in these cultures. However, after incubation for 72 h, mainly in jointly immobilized treatments of both microalgae species, the cultures reached their highest total carbohydrate content (mainly as starch) and also the highest affinity and volumetric productivity. These results demonstrate the potential of A. brasilense to affect carbohydrates and starch accumulation in Chlorella spp. when both microorganisms are co-cultured, which can be an important tool for applications of microalgae.

  14. Biofertilization with Azospirillum brasilense improves in vitro culture of Handroanthus ochraceus, a forestry, ornamental and medicinal plant.

    Science.gov (United States)

    Llorente, Berta E; Alasia, María A; Larraburu, Ezequiel E

    2016-01-25

    Biofertilization with plant growth-promoting rhizobacteria is a potential alternative to plant productivity. Here, in vitro propagation of Handroanthus ochraceus (yellow lapacho), a forest crop with high economic and environmental value, was developed using the Azospirillum brasilense strains Cd and Az39 during rhizogenesis. Epicotiles of in vitro plantlets were multiplied in Woody Plant Medium (WPM). For rooting, elongated shoots were transferred to auxin-free Murashige-Skoog medium with Gamborg's vitamins and WPM, both at half salt concentration (½MSG and ½WPM), and inoculated with Cd or Az39 at the base of each shoot. Anatomical studies were performed using leaves cleared and stained with safranin for optical microscopy and leaves and roots metalized with gold-palladium for scanning electron microscopy (SEM). In ½WPM auxin-free medium, A. brasilense Cd inoculation produced 55% of rooting, increased root fresh and dry weight (45% and 77%, respectively), and led to lower stomata size and density with similar proportion of open and closed stomata. Both strains selectively increased the size or density of glandular trichomes in ½MSG. Moreover, bacteria were detected on the root surface by SEM. In conclusion, the difference in H. ochraceus response to A. brasilense inoculation depends on the strain and the plant culture media. Cd strain enhanced rooting in auxin-free ½WPM and produced plantlets with features similar to those expected in ex vitro plants. This work presents an innovative in vitro approach using beneficial plant-microorganism interaction as an ecologically compatible strategy in plant biotechnology.

  15. Effects of heavy metals on plant-associated rhizobacteria: comparison of endophytic and non-endophytic strains of Azospirillum brasilense.

    Science.gov (United States)

    Kamnev, Alexander A; Tugarova, Anna V; Antonyuk, Lyudmila P; Tarantilis, Petros A; Polissiou, Moschos G; Gardiner, Philip H E

    2005-01-01

    The plant-associated nitrogen-fixing rhizobacterium Azospirillum brasilense attracts world-wide attention owing to its plant growth-promoting activities. Among hundreds of its strains known up to date, wild-type strain Sp245 has been proved to be capable of colonising both the plant-root interior and exterior (i.e. a facultative endophyte), whereas others are non-endophytes colonising the root surface only. Thus, the different ecological niches occupied by these strains in the rhizosphere suggest that their responses to environmental conditions might differ as well. In this study, responses of A. brasilense strains Sp245 and Sp7 to several heavy metals (Co2+, Cu2+, Zn2+), present in the medium at tolerable concentrations (up to 0.2 mmol/l) and taken up by the bacteria, were compared. Fourier transform infrared (FTIR) spectroscopy was used for controlling the compositional features of whole cells. The results obtained show that in strain Sp7 (non-endophyte) the heavy metals induced an enhanced accumulation of polyester compounds (poly-3-hydroxybutyrate; PHB). In contrast, the response of the endophytic strain Sp245 to heavy metal uptake was found to be much less pronounced. These dissimilarities in their behaviour may be caused by different adaptation abilities of these strains to stress conditions owing to their different ecological status. It was also found that adding 0.2 mmol/l Cu2+ or Cd2+ in the culture medium resulted in noticeably reducing the levels of indole-3-acetic acid (IAA, auxin) produced by both the strains of the bacterium. This can directly affect the efficiency of associative plant-bacterial symbioses involving A. brasilense in heavy-metal-contaminated soil.

  16. Responses of Azospirillum brasilense to nitrogen deficiency and to wheat lectin: a diffuse reflectance infrared fourier transform (DRIFT) spectroscopic study.

    Science.gov (United States)

    Kamnev, Alexander A; Sadovnikova, Julia N; Tarantilis, Petros A; Polissiou, Moschos G; Antonyuk, Lyudmila P

    2008-11-01

    For the rhizobacterium Azospirillum brasilense, the optimal nutritional range of C:N ratios corresponds to the presence of malate (ca. 3 to 5 g l(-1) of its sodium salt) and ammonium (ca. 0.5 to 3 g l(-1) of NH4Cl) as preferred carbon and nitrogen sources, respectively. This microaerophilic aerotactic bacterium is known to have a narrow optimal oxygen concentration range of ca. 3 to 5 microM, which is 1.2% to 2% of oxygen solubility in air-saturated water under normal conditions. In this work, the effects of stress conditions (bound-nitrogen deficiency related to a high C:N ratio in the medium; excess of oxygen) on aerobically grown A. brasilense Sp245, a native wheat-associated endophyte, were investigated in the absence and presence of wheat germ agglutinin (WGA, plant stress protein and a molecular host-plant signal for the bacterium) using FTIR spectroscopy of whole cells in the diffuse reflectance mode (DRIFT). The nutritional stress resulted in the appearance of prominent spectroscopic signs of poly-3-hydroxybutyrate (PHB) accumulation in the bacterial cells; in addition, splitting of the amide I band related to bacterial cellular proteins indicated some stress-induced alterations in their secondary structure components. Similar structural changes were observed in the presence of nanomolar WGA both in stressed A. brasilense cells and under normal nutritional conditions. Comparative analysis of the data obtained and the relevant literature data indicated that the stress conditions applied (which resulted in the accumulation of PHB involved in stress tolerance) and/or the presence of nanomolar concentrations of WGA induced synthesis of bacterial cell-surface (glyco)proteins rich in beta-structures, that could be represented by hemagglutinin and/or porin.

  17. Co-inoculation of Rhizobium tropici and Azospirillum brasilense in common beans grown under two irrigation depths

    Directory of Open Access Journals (Sweden)

    Amanda Ribeiro Peres²*

    2016-04-01

    Full Text Available ABSTRACT The alternative technique of co-inoculation or mixed inoculation with symbiotic and non-symbiotic bacteria has been studied in leguminous plants. However, there are few field studies with common beans and under the influence of the amount of irrigated water. Thus, the objective of this study was to evaluate the efficiency of inoculation and co-inoculation of common beans with Rhizobium tropici and Azospirillum brasilense under two irrigation depths. The experiment was carried out in the winter of 2012 and 2013, in Selvíria, state of Mato Grosso do Sul. The experimental design was composed of randomized blocks in split-plot scheme with two irrigation depths in the plots (recommended for common beans and 75% of the recommended and five forms of nitrogen (N supply in the split-plots (control non-inoculated with 40 kg ha- 1 of N in topdressing, 80 kg ha- 1 of N in topdressing, A. brasilense inoculation with 40 kg ha-1 of N in topdressing, R. tropici inoculation with 40 kg ha-1 of N in topdressing, and co-inoculation of A. brasilense and R. tropici with 40 kg ha- 1 of N in topdressing with four repetitions. Co-inoculation increased nodulation in the second year of cultivation. None of the evaluated treatments increased the grain yield in relation to non-inoculated control with 40 kg ha-1 of nitrogen in topdressing, which presented average yield of 2,200 kg ha-1. The use of 75% of the recommended irrigation depth provides similar grain yield to the recommended irrigation depth in common beans cropped in winter.

  18. Effect of the over-expression of PII and PZ proteins on the nitrogenase activity of Azospirillum brasilense.

    Science.gov (United States)

    Huergo, Luciano F; Filipaki, Angela; Chubatsu, Leda S; Yates, M Geoffrey; Steffens, Maria Berenice; Pedrosa, Fabio O; Souza, Emanuel M

    2005-12-01

    The Azospirillum brasilense PII and PZ proteins, encoded by the glnB and glnZ genes respectively, are intracellular transducers of nitrogen levels with distinct functions. The PII protein participates in nif regulation by controlling the activity of the transcriptional regulator NifA. PII is also involved in transducing the prevailing nitrogen levels to the Fe-protein ADP-ribosylation system. PZ regulates negatively ammonium transport and is involved in nitrogenase reactivation. To further investigate the role of PII and PZ in the regulation of nitrogen fixation, broad-host-range plasmids capable of over-expressing the glnB and glnZ genes under control of the ptac promoter were constructed and introduced into A. brasilense. The nitrogenase activity and nitrate-dependent growth was impaired in A. brasilense cells over-expressing the PII protein. Using immunoblot analysis we observed that the reduction of nitrogenase activity in cells over-expressing PII was due to partial ADP-ribosylation of the Fe-protein under derepressing conditions and a reduction in the amount of Fe-protein. These results support the hypothesis that the unmodified PII protein act as a signal to the DraT enzyme to ADP-ribosylate the Fe-protein in response to ammonium shock, and that it also inhibits nif gene expression. In cells over-expressing the PZ protein the nitrogenase reactivation after an ammonium shock was delayed indicating that the PZ protein is involved in regulation of DraG activity.

  19. Physiological, structural and molecular traits activated in strawberry plants after inoculation with the plant growth-promoting bacterium Azospirillum brasilense REC3.

    Science.gov (United States)

    Guerrero-Molina, M F; Lovaisa, N C; Salazar, S M; Martínez-Zamora, M G; Díaz-Ricci, J C; Pedraza, R O

    2015-05-01

    The plant growth-promoting strain REC3 of Azospirillum brasilense, isolated from strawberry roots, prompts growth promotion and systemic protection against anthracnose disease in this crop. Hence, we hypothesised that A. brasilense REC3 can induce different physiological, structural and molecular responses in strawberry plants. Therefore, the aim of this work was to study these traits activated in Azospirillum-colonised strawberry plants, which have not been assessed until now. Healthy, in vitro micropropagated plants were root-inoculated with REC3 under hydroponic conditions; root and leaf tissues were sampled at different times, and oxidative burst, phenolic compound content, malondialdehyde (MDA) concentration, callose deposition, cell wall fortification and gene expression were evaluated. Azospirillum inoculation enhanced levels of soluble phenolic compounds after 12 h post-inoculation (hpi), while amounts of cell wall bound phenolics were similar in inoculated and control plants. Other early responses activated by REC3 (at 24 hpi) were a decline of lipid peroxidation and up-regulation of strawberry genes involved in defence (FaPR1), bacterial recognition (FaFLS2) and H₂O₂ depuration (FaCAT and FaAPXc). The last may explain the apparent absence of oxidative burst in leaves after bacterial inoculation. Also, REC3 inoculation induced delayed structural responses such as callose deposition and cell wall fortification (at 72 hpi). Results showed that A. brasilense REC3 is capable of exerting beneficial effects on strawberry plants, reinforcing their physiological and cellular characteristics, which in turns contribute to improve plant performance.

  20. Identification, cloning and characterization of cysK, the gene encoding O-acetylserine (thiol)-lyase from Azospirillum brasilense, which is involved in tellurite resistance.

    Science.gov (United States)

    Ramírez, Alberto; Castañeda, Miguel; Xiqui, María L; Sosa, Araceli; Baca, Beatriz E

    2006-08-01

    O-Acetylserine (thiol)-lyase (cysteine synthase) was purified from Azospirillum brasilense Sp7. After hydrolysis of the purified protein, amino acid sequences of five peptides were obtained, which permitted the cloning and sequencing of the cysK gene. The deduced amino acid sequence of cysteine synthase exhibited homology with several putative proteins from Alpha- and Gammaproteobacteria. Azospirillum brasilense Sp7 cysK exhibited 58% identity (72% similarity) with Escherichia coli K12 and Salmonella enterica serovar Typhimurium cysteine synthase proteins. An E. coli auxotroph lacking cysteine synthase loci could be complemented with A. brasilense Sp7 cysK. The 3.0-kb HindIII-EcoRI fragment bearing cysK contained two additional ORFs encoding a putative transcriptional regulator and dUTPase. Insertional disruption of the cysK gene did not produce a cysteine auxotroph, indicating that gene redundancy in the cysteine biosynthetic or other biosynthetic pathways exists in Azospirillum, as already described in other bacteria. Nitrogen fixation was not altered in the mutant strain as determined by acetylene reduction. However, this strain showed an eight-fold reduction in tellurite resistance as compared to the wild-type strain, which was only observed during growth in minimal medium. These data confirm earlier observations regarding the importance of cysteine metabolism in tellurite resistance.

  1. Azospirillum IV

    Energy Technology Data Exchange (ETDEWEB)

    Klingmuller, W.

    1988-01-01

    This book's contents include: Advances in the genetics of Azospirillum brasilense Sp7: Use of Tn5 mutagenesis for gene mapping and identification; Characterization of DNA segments adjacent to the nifHDK genes of Azospirillum brasilense by Sp7 Tn5 site-directed mutagenesis; Selection at the chemostat of Azospirillum brasilense Cd N/sub 2/-fixing at high O/sub 2/ pressure. Root hair deformation induced on maize and medicago by an Azospirillum transconjugant containing a Rhizobium meliloti nodulation region. Azospirilla are bacteria that live in association with the roots of many grain crops. Since these bacteria bind molecular nitrogen from the air and excrete plant growth substances, interest has focussed on their potential to increase crop yields.

  2. Accumulation fatty acids of in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoA carboxylase, temperature, and co-immobilization with Azospirillum brasilense

    Science.gov (United States)

    Leyva, Luis A.; Bashan, Yoav; Mendoza, Alberto; de-Bashan, Luz E.

    2014-10-01

    The relation between fatty acid accumulation, activity of acetyl-CoA carboxylase (ACC), and consequently lipid accumulation was studied in the microalgae Chlorella vulgaris co-immobilized with the plant growth-promoting bacterium Azospirillum brasilense under dark heterotrophic conditions with Na acetate as a carbon source. In C. vulgaris immobilized alone, cultivation experiments for 6 days showed that ACC activity is directly related to fatty acid accumulation, especially in the last 3 days. In co-immobilization experiments, A. brasilense exerted a significant positive effect over ACC activity, increased the quantity in all nine main fatty acids, increased total lipid accumulation in C. vulgaris, and mitigated negative effects of nonoptimal temperature for growth. No correlation between ACC activity and lipid accumulation in the cells was established for three different temperatures. This study demonstrated that the interaction between A. brasilense and C. vulgaris has a significant effect on fatty acid and lipid accumulation in the microalgae.

  3. Accumulation of fatty acids in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoAcarboxylase, temperature, and co-immobilization with Azospirillum brasilense [corrected].

    Science.gov (United States)

    Leyva, Luis A; Bashan, Yoav; Mendoza, Alberto; de-Bashan, Luz E

    2014-10-01

    The relation between fatty acid accumulation, activity of acetyl-CoA carboxylase (ACC), and consequently lipid accumulation was studied in the microalgae Chlorella vulgaris co-immobilized with the plant growth-promoting bacterium Azospirillum brasilense under dark heterotrophic conditions with Na acetate as a carbon source. In C. vulgaris immobilized alone, cultivation experiments for 6 days showed that ACC activity is directly related to fatty acid accumulation, especially in the last 3 days. In co-immobilization experiments, A. brasilense exerted a significant positive effect over ACC activity, increased the quantity in all nine main fatty acids, increased total lipid accumulation in C. vulgaris, and mitigated negative effects of nonoptimal temperature for growth. No correlation between ACC activity and lipid accumulation in the cells was established for three different temperatures. This study demonstrated that the interaction between A. brasilense and C. vulgaris has a significant effect on fatty acid and lipid accumulation in the microalgae.

  4. [The structure of the O-specific polysaccharide from a mutant of nitrogen-fixing rhizobacterium Azospirillum brasilense Sp245 with an altered plasmid content].

    Science.gov (United States)

    Fedonenko, Iu P; Katsy, E I; Petrova, L P; Boĭko, A S; Zdorovenko, E L; Kachala, V V; Shashkov, A S; Knirel', Iu A

    2010-01-01

    The rhizobacteria Azospirillum brasilense Sp245 produce antigenically different lipopolysaccharides LPSI and LPSII, both containing identical pentasaccharides built from D-rhamnose residues as the repeated chains of O-specific oligosaccharides (OPS). In this study, we report the structure of the OPS from A. brasilense LPSI(-)LPSII(-)-mutant Sp245.5, which spontaneously lost the p85 and p120 plasmids upon the formation of a new 300-MDa megaplasmid after the long-term storage of the bacteria in a rich medium. The repeating unit of the A. brasilense mutant Sp245.5 appeared to be a disaccharide consisting of residues of N-acetyl-D-galactosamine and N-acetyl-D-mannosaminuronic acid: [Formula: see text].

  5. In Azospirillum brasilense, mutations in flmA or flmB genes affect polar flagellum assembly, surface polysaccharides, and attachment to maize roots.

    Science.gov (United States)

    Rossi, Fernando Ariel; Medeot, Daniela Beatriz; Liaudat, Juan Pablo; Pistorio, Mariano; Jofré, Edgardo

    2016-09-01

    Azospirillum brasilense is a soil bacterium capable of promoting plant growth. Several surface components were previously reported to be involved in the attachment of A. brasilense to root plants. Among these components are the exopolysaccharide (EPS), lipopolysaccharide (LPS) and the polar flagellum. Flagellin from polar flagellum is glycosylated and it was suggested that genes involved in such a posttranslational modification are the same ones involved in the biosynthesis of sugars present in the O-antigen of the LPS. In this work, we report on the characterization of two homologs present in A. brasilense Cd, to the well characterized flagellin modification genes, flmA and flmB, from Aeromonas caviae. We show that mutations in either flmA or flmB genes of A. brasilense resulted in non-motile cells due to alterations in the polar flagellum assembly. Moreover, these mutations also affected the capability of A. brasilense cells to adsorb to maize roots and to produce LPS and EPS. By generating a mutant containing the polar flagellum affected in their rotation, we show the importance of the bacterial motility for the early colonization of maize roots.

  6. [Genome Rearrangements in Azospirillum brasilense Sp7 with the Involvement of the Plasmid pRhico and the Prophage phiAb-Cd].

    Science.gov (United States)

    Katsy, E I; Petrova, L P

    2015-12-01

    Alphaproteobacteria of the species Azospirillum brasilense have a multicomponent genome that undergoes frequent spontaneous rearrangements, yielding changes in the plasmid profiles of strains. Specifically, variants (Cd, Sp7.K2, Sp7.1, Sp7.4, Sp7.8, etc.) of the type strainA. brasilense Sp7 that had lost a 115-MDa plasmid were previously selected. In many of them, the molecular weight of a 90-MDa plasmid (p90 or pRhico), which is a kind of "depot" for glycopolymer biosynthesis genes, increased. In this study, a collection of primers was designed to the plasmid pRhico and to the DNA of prophage phiAb-Cd integrated in it. The use ofthese primers in polymerase chain reactions allowed the detection of the probable excision of phiAb-Cd phage from the DNA of A. brasilense variants Sp7.4 and Sp7.8 and other alterations of the pRhico structure in A. brasilense strains Cd, Sp7.K2, and Sp7.8. The developed primers and PCR conditions may be recoin mended for primary analysis of spontaneous plasmid rearrangements in A. brasilense Sp7 and related strains.

  7. Influence of tryptophan and indole-3-acetic acid on starch accumulation in the synthetic mutualistic Chlorella sorokiniana-Azospirillum brasilense system under heterotrophic conditions.

    Science.gov (United States)

    Palacios, Oskar A; Choix, Francisco J; Bashan, Yoav; de-Bashan, Luz E

    2016-06-01

    This study measured the relations between tryptophan production, the phytohormone indole-3-acetic acid (IAA) and the metabolism and accumulation of starch during synthetic mutualism between the microalgae Chlorella sorokiniana and the microalgae growth-promoting bacteria Azospirillum brasilense, created by co-immobilization in alginate beads. Experiments used two wild-type A. brasilense strains (Cd and Sp6) and an IAA-attenuated mutant (SpM7918) grown under nitrogen-replete and nitrogen-starved conditions tested under dark, heterotrophic and aerobic growth conditions. Under all incubating conditions, C. sorokiniana, but not A. brasilense, produced tryptophan. A significant correlation between IAA-production by A. brasilense and starch accumulation in C. sorokiniana was found, since the IAA-attenuated mutant was not producing increased starch levels. The highest ADP-glucose pyrophosphorylase (AGPase) activity, starch content and glucose uptake were found during the interaction of A. brasilense wild type strains with the microalgae. When the microalgae were grown alone, they produced only small amounts of starch. Supplementation with synthetic IAA to C. sorokiniana grown alone enhanced the above parameters, but only transiently. Activity of α-amylase decreased under nitrogen-replete conditions, but increased under nitrogen-starved conditions. In summary, this study demonstrated that, during synthetic mutualism, the exchange of tryptophan and IAA between the partners is a mechanism that governs several changes in starch metabolism of C. sorokiniana, yielding an increase in starch content.

  8. Regulation of expression and biochemical characterization of a beta-class carbonic anhydrase from the plant growth-promoting rhizobacterium, Azospirillum brasilense Sp7.

    Science.gov (United States)

    Kaur, Simarjot; Mishra, Mukti Nath; Tripathi, Anil K

    2009-10-01

    Carbonic anhydrase (CA; [EC 4.2.1.1]) is a ubiquitous enzyme catalysing the reversible hydration of CO(2) to bicarbonate, a reaction that supports various biochemical and physiological functions. Genome analysis of Azospirillum brasilense, a nonphotosynthetic, nitrogen-fixing, rhizobacterium, revealed an ORF with homology to beta-class carbonic anhydrases (CAs). Biochemical characteristics of the beta-class CA of A. brasilense, analysed after cloning the gene (designated as bca), overexpressing in Escherichia coli and purifying the protein by affinity purification, revealed that the native recombinant enzyme is a homotetramer, inhibited by the known CA inhibitors. CA activity in A. brasilense cell extracts, reverse transcriptase (RT)-PCR and Western blot analyses showed that bca was constitutively expressed under aerobic conditions. Lower beta-galactosidase activity in A. brasilense cells harbouring bca promoter: lacZ fusion during the stationary phase or during growth on 3% CO(2) enriched air or at acidic pH indicated that the transcription of bca was downregulated by the stationary phase, elevated CO(2) levels and acidic pH conditions. These observations were also supported by RT-PCR analysis. Thus, beta-CA in A. brasilense seems to be required for scavenging CO(2) from the ambient air and the requirement of CO(2) hydration seems to be higher for the cultures growing exponentially at neutral to alkaline pH.

  9. Influence of soil saprophyte fungus Chaetomium cochliodes on associative system "Triticum aestivum – Azospirillum brasilense"

    Directory of Open Access Journals (Sweden)

    E. P. Kopylov

    2009-11-01

    Full Text Available In laboratory and vegetative experiments the ability of soil ascomycete C. cochliodes 3250 to promote the penetration of Azospirillum nitrogen-fixing bacteria into roots’ inner tissues was shown. At the same time the endophytic association: spring wheat – Azospirillum nitrogen-fixing bacteria – soil saprophyte ascomycete C. cochliodes 3250 is forming. It allows activating the nitrogen fixation in the spring wheat roots zone and biosynthetic processes in plants, in particular: to raise glutamine synthetase activity, chlorophylls content in leaves and plants’ productivity.

  10. Effect of genomic rearrangement on heavy metal tolerance in the plant-growth-promoting rhizobacterium Azospirillum brasilense Sp245.

    Science.gov (United States)

    Shelud'ko, Andrei V; Varshalomidze, Olga E; Petrova, Lilia P; Katsy, Elena I

    2012-01-01

    A derivative of Azospirillum brasilense Sp245, Sp245.5, which spontaneously lost 85 and 120 MDa replicons upon the formation of a new megaplasmid, has been shown to produce a novel lipopolysaccharide and to lose Calcofluor-binding polysaccharides. As compared to Sp245, the derivative displays notably increased heavy metal tolerance. The phenotypes of Sp245 and Sp245.5 are characterized by the following minimal inhibitory concentrations (MICs) of heavy metals: 0.5 and 0.9 μmol l(-1) of Ag(+), 0.4 and 0.7 mmol l(-1) of Co(2+), 0.9 and 4.7 mmol l(-1) of Cu(2+), and 3.1 and 11.5 mmol l(-1) of Zn(2+), respectively. In Sp245, in the presence of a nonlethal concentration (0.625 μmol l(-1)) of the efflux pump inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP), the MIC of cobalt, copper, and zinc drop 1.3- to 1.6-fold, but the low tolerance to silver is unaffected. In Sp245.5, CCCP does not affect cobalt tolerance, suppresses tolerance to copper and silver to the wild-type levels, and causes a 1.4-fold decrease in resistance to zinc. Therefore, significant elevation of heavy metal tolerance in Sp245.5 seems caused by the induction/overexpression of the proton-dependent efflux of certain metal ions. The novel cell surface and other unknown factors could also be responsible for the increased tolerance of A. brasilense Sp245.5 to heavy metals.

  11. The Azospirillum brasilense Che1 chemotaxis pathway controls swimming velocity, which affects transient cell-to-cell clumping.

    Science.gov (United States)

    Bible, Amber; Russell, Matthew H; Alexandre, Gladys

    2012-07-01

    The Che1 chemotaxis-like pathway of Azospirillum brasilense contributes to chemotaxis and aerotaxis, and it has also been found to contribute to regulating changes in cell surface adhesive properties that affect the propensity of cells to clump and to flocculate. The exact contribution of Che1 to the control of chemotaxis and flocculation in A. brasilense remains poorly understood. Here, we show that Che1 affects reversible cell-to-cell clumping, a cellular behavior in which motile cells transiently interact by adhering to one another at their nonflagellated poles before swimming apart. Clumping precedes and is required for flocculation, and both processes appear to be independently regulated. The phenotypes of a ΔaerC receptor mutant and of mutant strains lacking cheA1, cheY1, cheB1, or cheR1 (alone or in combination) or with che1 deleted show that Che1 directly mediates changes in the flagellar swimming velocity and that this behavior directly modulates the transient nature of clumping. Our results also suggest that an additional receptor(s) and signaling pathway(s) are implicated in mediating other Che1-independent changes in clumping identified in the present study. Transient clumping precedes the transition to stable clump formation, which involves the production of specific extracellular polysaccharides (EPS); however, production of these clumping-specific EPS is not directly controlled by Che1 activity. Che1-dependent clumping may antagonize motility and prevent chemotaxis, thereby maintaining cells in a metabolically favorable niche.

  12. Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    C.Y. Nishikawa

    2012-02-01

    Full Text Available Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ54 co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL. A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH4Cl but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

  13. Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense.

    Science.gov (United States)

    Nishikawa, C Y; Araújo, L M; Kadowaki, M A S; Monteiro, R A; Steffens, M B R; Pedrosa, F O; Souza, E M; Chubatsu, L S

    2012-02-01

    Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ(54) co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH(4)Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

  14. Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense

    Energy Technology Data Exchange (ETDEWEB)

    Nishikawa, C.Y.; Araújo, L.M.; Kadowaki, M.A.S.; Monteiro, R.A.; Steffens, M.B.R.; Pedrosa, F.O.; Souza, E.M.; Chubatsu, L.S. [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR (Brazil)

    2012-01-27

    Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ{sup 54} factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH{sub 4}Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

  15. [Cloning and functional analysis of glnB from Azospirillum brasilense Yu62].

    Science.gov (United States)

    Li, Z H; Chen, S F; Li, J L

    2001-01-01

    The glnB gene of A. brasilense Yu62 was determined in a 3.7 kb EcoRI + PstI fragment. The glnA is located downstream of glnB and an ORF for hypothetical protein is on upstream of glnB. The deduced amino acid sequence of PII encoded by glnB is 71%, 77%, 79% and 69% identical to that of K. pneumoniae, Bradyrhizobium japonicum, Rhizobium leguninosarum and E. coli, respectively. A Km-casette was inserted into BglII site of glnB coding region and GlnB- mutant was obtained by homologous recombination. The GlnB- mutant has lost the nitrogenase activity, i.e.: Nif-. For the functional confirmation of glnB gene, a complementary test was carried out and it was shown that C-glnB(glnB::Km/glnB) can restore the nitrogenase activity. When the recombinant plasmid pVK-II which containined the coding region of glnB was introduced into A. brasilense Yu62 and A. brasilense Yu62 DraT-, respectively, the Yu62-II (containing pVK-II) and draT-II(containing pVK-II) showed higher nitrogenase activity than wild type. These results confirmed that glnB plays an important role in the regulation of nitrogen in A. brasilense.

  16. Colonization and nitrogenase activity of Triticum aestivum (cv. Baccross and Mahdavi) to the dual inoculation with Azospirillum brasilense and Rhizobium meliloti plus 2,4-D.

    Science.gov (United States)

    Mehry, Askary; Akbar, Mostajeran; Giti, Emtiazi

    2008-06-15

    The potential enhancement of root colonization and nitrogenase activity of wheat cultivars (Baccross and Mahdavi) was studied with application of two Azospirillum brasilense strains (native and Sp7) co-inoculated with two Rhizobium meliloti strains (native and DSMZ 30135). The results indicated that the colonization was different due to the strains and cultivars of wheat were used. Native A. brasilense colonized wheat root better than Sp7 strain. However, Baccross cv. reacted better with native Azospirillum compared to Mahdavi cv. which reacted better with Sp7. When plants inoculated with dual inoculants (SP7 with standard Rhizobium), the colonization of Azospirillum were increased significantly (from 1.67 x 10(5) to 22 x 10(5) cfu g(-1) FW for Baccras cv. and 3.67 x 10(5) to 26 x 10(5) cfu g(-1) FW for Mahdavi cultivar). When the standard Rhizobium as co-inoculants changed to the native Rhizobium, the colonization of Azospirillum was higher when compared to the single inoculants but was almost the same when compared to the standard Rhizobium. When the standard or native strains of Rhizobium used as single inoculation of wheat roots, the number of Rhizobium in the wheat roots were not changed significantly. However, when plants co-inoculated with Rhizobium and Azospirillum, the colonization of Rhizobium was increased. Co-inoculation of standard strain of R. melilot with A. brasilense Sp7 showed that the colonization of Rhizobium were increased from 0.67 x 10(5) to 21 x 10(5) cfu g(-1) FW for Baccross cv. and 0.33 x 10(5) to 18 x 10(5) cfu g(-1) FW for Mahdavi cv. This behavior was the same when inoculation of Rhizobium was happened with the native one. In dual inoculation, the highest nitrogenase activity was measured in combination of the local strains (native A. brasilense with the native R. meliloti) and the lower one belongs to the combination of standard strains (Sp7 with standard R. meliloti). The difference in nirtogenase activity for different cultivars of

  17. pVLT-EGFP载体构建及其在巴西固氮螺菌(Azospirillum brasilense)的表达研究%Construction of an Expression Vector pVLT-EGFP and Its Expression in Azospirillum brasilense

    Institute of Scientific and Technical Information of China (English)

    巩涛; 王继雯; 杨文玲; 甄静; 刘莹莹; 李冠杰; 刘莉; 岳丹丹; 陈国参

    2015-01-01

    为了研究巴西固氮螺菌(Azospirillum brasilense)在植物体内的定殖,利用酶切连接的方法,以增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)基因与表达栽体pVLT-33为基本元件,构建了重组表达载体pVLT-EGFP,电转巴西固氮螺菌R7细胞,并利用实时荧光定量PCR(qPCR)研究了不同温度、不同时间EGFP mRNA的表达情况.酶切及测序结果表明,成功构建了pVLT-EGFP载体,并在荧光显微镜下观察到绿色荧光蛋白表达;qPCR结果显示:30℃,诱导9hEGFP基因的表达水平最高.本研究成功构建了重组表达载体pVLT-EGFP,为实现pVLT-EGFP的可控表达及研究固氮菌在植物体内的定殖规律及促生长机理提供了一种有效的途径.

  18. Surface colonization by Azospirillum brasilense SM in the indole-3-acetic acid dependent growth improvement of sorghum.

    Science.gov (United States)

    Kochar, Mandira; Srivastava, Sheela

    2012-04-01

    The key to improving plant productivity is successful bacterial-plant interaction in the rhizosphere that can be maintained in the environment. The results presented here confirm Azospirillum brasilense strain SM as a competent plant growth promoting bacterium over mid- and long-term associations with sorghum. This study establishes that plant growth can be directly correlated with the associated bacterium's indole-3-acetic acid (IAA) production capability as IAA over-expressing variants, SMp30 and SMΔi3-6 fared better than the wild type strain. The auxin antagonist, p-chlorophenoxy isobutyric acid confirmed the role of bacterial IAA in plant growth promotion and verified the presence of larger amount of IAA available to the seeds on inoculation with IAA over-expressing mutants. Microscopic analysis identified the bacterial association at root tips, root-shoot junction and elongation zone and their surface colonizing nature. Scanning electron microscopy identified larger number of root hairs and extensive exopolysaccharide covering in comparison to untreated ones. In addition, vibroid-shaped Azospirilla attached by means of fibrillar material were dispersed along the elongation zone. The notable difference with IAA over-expressing variants was enhanced number of root hairs. Thus, the variant strains may be more efficient surface colonizers of the sorghum root and used as superior bio-inoculants for improving plant productivity.

  19. cDNA-AFLP analysis of differential gene expression related to cell chemotactic and encystment of Azospirillum brasilense.

    Science.gov (United States)

    Li, Huamin; Cui, Yanhua; Wu, Lixian; Tu, Ran; Chen, Sanfeng

    2011-12-20

    Our previous study indicated org35 was involved in chemotaxis and interacted with nitrogen fixation transcriptional activator NifA via PAS domain. In order to reveal the role of org35 in nitrogen regulation, the downstream target genes of org35 were identified. We here report differentially expressed genes in org35 mutants comparing with wild type Sp7 by means of cDNA-AFLP. Four up-regulated transcript-derived fragments (TDFs) homologues of chemotaxis transduction proteins were found, including CheW, methyl-accepting chemotaxis protein and response regulator CheY-like receiver. Three distinct TDFs (AB46, AB58 and AB63) were similar to PHB de-polymerase C-terminus, cell shape-determining protein and flagellin domain protein. And 11 TDFs showed similarities with signal transduction proteins, including homologous protein of the nitrogen regulation protein NtrY and nitrate/nitrite response regulator protein NarL. These data suggested that the Azospirillum brasilense org35 was a multi-effecter and involved in chemotaxis, cyst development and regulation of nitrogen fixation.

  20. Structure and thermodynamics of effector molecule binding to the nitrogen signal transduction PII protein GlnZ from Azospirillum brasilense.

    Science.gov (United States)

    Truan, Daphné; Bjelić, Saša; Li, Xiao-Dan; Winkler, Fritz K

    2014-07-29

    The trimeric PII signal transduction proteins regulate the function of a variety of target proteins predominantly involved in nitrogen metabolism. ATP, ADP and 2-oxoglutarate (2-OG) are key effector molecules influencing PII binding to targets. Studies of PII proteins have established that the 20-residue T-loop plays a central role in effector sensing and target binding. However, the specific effects of effector binding on T-loop conformation have remained poorly documented. We present eight crystal structures of the Azospirillum brasilense PII protein GlnZ, six of which are cocrystallized and liganded with ADP or ATP. We find that interaction with the diphosphate moiety of bound ADP constrains the N-terminal part of the T-loop in a characteristic way that is maintained in ADP-promoted complexes with target proteins. In contrast, the interactions with the triphosphate moiety in ATP complexes are much more variable and no single predominant interaction mode is apparent except for the ternary MgATP/2-OG complex. These conclusions can be extended to most investigated PII proteins of the GlnB/GlnK subfamily. Unlike reported for other PII proteins, microcalorimetry reveals no cooperativity between the three binding sites of GlnZ trimers for any of the three effectors under carefully controlled experimental conditions.

  1. Cloning and characterisation of the Azospirillum brasilense glnD gene and analysis of a glnD mutant.

    Science.gov (United States)

    Van Dommelen, A; Keijers, V; Somers, E; Vanderleyden, J

    2002-01-01

    Nitrogen regulation in bacteria involves the capacity to sense the availability of fixed nitrogen and to translate a signal indicating nitrogen deficiency or nitrogen excess into a cellular response. One of the key enzymes in this complex regulation process, the uridylyltransferase/uridylyl-removing (UTase) enzyme, encoded by the glnD gene, was characterised in the diazotroph Azospirillum brasilense, which promotes plant growth. The glnD gene product is responsible for the uridylylation of both P(II)-like nitrogen regulatory proteins, P(II) and P(Z), depending on the nitrogen status of the cell. The nitrogen-regulated activity of the main ammonium-assimilating enzyme, glutamine synthetase, is not altered in a glnD-Tn 5-B30 insertion mutant. UTase influences processes that are regulated by the NtrB-NtrC two-component histidine protein kinase system, such as ammonium uptake and nitrate assimilation. Moreover, the glnD gene product is indispensable for the activation of nitrogen fixation. Transcription of glnD is up-regulated under nitrogen-fixing conditions. This regulation is only partially dependent on the global nitrogen regulation (Ntr) system.

  2. Influence of the ADP/ATP ratio, 2-oxoglutarate and divalent ions on Azospirillum brasilense PII protein signalling.

    Science.gov (United States)

    Gerhardt, Edileusa C M; Araújo, Luíza M; Ribeiro, Ronny R; Chubatsu, Leda S; Scarduelli, Marcelo; Rodrigues, Thiago E; Monteiro, Rose A; Pedrosa, Fábio O; Souza, Emanuel M; Huergo, Luciano F

    2012-06-01

    Proteins belonging to the P(II) family coordinate cellular nitrogen metabolism by direct interaction with a variety of enzymes, transcriptional regulators and transporters. The sensing function of P(II) relies on its ability to bind the nitrogen/carbon signalling molecule 2-oxoglutarate (2-OG). In Proteobacteria, P(II) is further subject to reversible uridylylation according to the intracellular levels of glutamine, which reflect the cellular nitrogen status. A number of P(II) proteins have been shown to bind ADP and ATP in a competitive manner, suggesting that P(II) might act as an energy sensor. Here, we analyse the influence of the ADP/ATP ratio, 2-OG levels and divalent metal ions on in vitro uridylylation of the Azospirillum brasilense P(II) proteins GlnB and GlnZ, and on interaction with their targets AmtB, DraG and DraT. The results support the notion that the cellular concentration of 2-OG is a key factor governing occupation of the GlnB and GlnZ nucleotide binding sites by ATP or ADP, with high 2-OG levels favouring the occupation of P(II) by ATP. Both P(II) uridylylation and interaction with target proteins responded to the ADP/ATP ratio within the expected physiological range, supporting the concept that P(II) proteins might act as cellular energy sensors.

  3. Study of the rhizobacterium Azospirillum brasilense Sp245 using Mössbauer spectroscopy with a high velocity resolution: Implication for the analysis of ferritin-like iron cores

    Science.gov (United States)

    Alenkina, I. V.; Oshtrakh, M. I.; Tugarova, A. V.; Biró, B.; Semionkin, V. A.; Kamnev, A. A.

    2014-09-01

    The results of a comparative study of two samples of the rhizobacterium Azospirillum brasilense (strain Sp245) prepared in different conditions and of human liver ferritin using Mössbauer spectroscopy with a high velocity resolution demonstrated the presence of ferritin-like iron (i.e. iron similar to that found in ferritin-like proteins) in the bacterium. Mössbauer spectra of these samples were fitted in two ways: as a rough approximation using a one quadrupole doublet fit (the homogeneous iron core model) and using a superposition of quadrupole doublets (the heterogeneous iron core model). Both results demonstrated differences in the Mössbauer parameters for mammalian ferritin and for bacterial ferritin-like iron. Moreover, some differences in the Mössbauer parameters were observed between the two samples of A. brasilense Sp245 related to the differences in their preparation conditions.

  4. Involvement of glnB, glnZ, and glnD genes in the regulation of poly-3-hydroxybutyrate biosynthesis by ammonia in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Sun, Jun; Van Dommelen, Anne; Van Impe, Jan; Vanderleyden, Jozef

    2002-02-01

    The role of three key nitrogen regulatory genes, glnB (encoding the P(II) protein), glnZ (encoding the P(z) protein), and glnD (encoding the GlnD protein), in regulation of poly-3-hydroxybutyrate (PHB) biosynthesis by ammonia in Azospirillum brasilense Sp7 was investigated. It was observed that glnB glnZ and glnD mutants produce substantially higher amounts of PHB than the wild type produces during the active growth phase. glnB and glnZ mutants have PHB production phenotypes similar to that of the wild type. Our results indicate that the P(II)-P(z) system is apparently involved in nitrogen-dependent regulation of PHB biosynthesis in A. brasilense Sp7.

  5. [Mutants of bacterium Azospirillum brasilense Sp245 with Omegon insertion in mmsB or fabG genes of lipid metabolism are defective in motility and flagellation].

    Science.gov (United States)

    Kovtunov, E A; Shelud'ko, A V; Chernyshova, M P; Petrova, L P; Katsy, E I

    2013-11-01

    Bacteria Azospirillum brasilense have mixed flagellation: in addition to the polar flagellum, numerous lateral flagella are formed in their cells on medium with increased density. Flagella determine the active swimming and swarming capacities of azospirilla. Using A. brasilense Sp245 as an example, we showed that the Omegon-Km artificial transposon insertion into the chromosomal gene for 3-hydroxyisobutyrate dehydrogenase (mmsB) was concurrent with the appearance of significant defects in the formation of polar flagella and with the paralysis of lateral flagella. The Sp245 mutant with the Omegon insertion into the plasmid AZOBR_p1-borne gene for 3-oxoacyl-[acyl-carrier protein]-reductase (fabG) showed the complete loss of flagella and the swarming capacity, as well as significant defects in polar flagellar assembly (though some cells are still motile in liquid medium). The viability of the A. brasilense Sp245 mutants with the Omegon insertion into the mmsB or fabG gene was not reduced. No considerable differences in the fatty acid composition of whole cell lipid extracts were found for the A. brasilense Sp245 strain and its mmsB and fabG mutants.

  6. Accumulation of intra-cellular polyphosphate in Chlorella vulgaris cells is related to indole-3-acetic acid produced by Azospirillum brasilense.

    Science.gov (United States)

    Meza, Beatriz; de-Bashan, Luz E; Hernandez, Juan-Pablo; Bashan, Yoav

    2015-06-01

    Accumulation of intra-cellular phosphate, as polyphosphate, was measured when the microalga Chlorella vulgaris was immobilized in alginate with either of two wild-type strains of the microalgae growth-promoting bacterium Azospirillum brasilense or their corresponding IAA-attenuated mutants. Wild type strains of A. brasilense induced higher amounts of intra-cellular phosphate in Chlorella than their respective mutants. Calculations comparing intra-cellular phosphate accumulation by culture or net accumulation by the cell and the amount of IAA that was produced by each of these strains revealed that higher IAA was linked to higher accumulations of intra-cellular phosphate. Application of four levels of exogenous IAA reported for A. brasilense and their IAA-attenuated mutants to cultures of C. vulgaris enhanced accumulation of intra-cellular phosphate; the higher the content of IAA per culture or per single cell, the higher was the amount of accumulated phosphate. When an IAA-attenuated mutant was complemented with exogenous IAA, accumulation of intra-cellular phosphate at the culture level was even higher than phosphate accumulation with the respective wild type strains. When calculating the net accumulation of intra-cellular phosphate in the complementation experiment, net intra-cellular phosphate induced by the IAA-attenuated mutant was completely restored and was similar to the wild strains. We propose that IAA produced by A. brasilense is linked to polyphosphate accumulation in C. vulgaris.

  7. Purification of the major outer membrane protein of Azospirillum brasilense, its affinity to plant roots, and its involvement in cell aggregation.

    Science.gov (United States)

    Burdman, S; Dulguerova, G; Okon, Y; Jurkevitch, E

    2001-04-01

    The major outer membrane protein (MOMP) of the nitrogen-fixing rhizobacterium Azospirillum brasilense strain Cd was purified and isolated by gel filtration, and antiserum against this protein was obtained. A screening of the binding of outer membrane proteins (OMPs) of A. brasilense to membrane-immobilized root extracts of various plant species revealed different affinities for the MOMP, with a stronger adhesion to extracts of cereals in comparison with legumes and tomatoes. Moreover, this protein was shown to bind to roots of different cereal seedlings in an in vitro adhesion assay. Incubation of A. brasilense cells with MOMP-antiserum led to fast agglutination, indicating that the MOMP is a surface-exposed protein. Cells incubated with Fab fragments obtained from purified MOMP-antiserum immunoglobulin G exhibited significant inhibition of bacterial aggregation as compared with controls. Bacteria preincubated with Fab fragments showed weaker adhesion to corn roots in comparison to controls without Fab fragments. These findings suggest that the A. brasilense MOMP acts as an adhesin involved in root adsorption and cell aggregation of this bacterium.

  8. Assessing the efficacy of co-inoculation of wheat seedlings with the associative bacteria Paenibacillus polymyxa 1465 and Azospirillum brasilense Sp245.

    Science.gov (United States)

    Yegorenkova, Irina V; Tregubova, Kristina V; Burygin, Gennady L; Matora, Larisa Y; Ignatov, Vladimir V

    2016-03-01

    Co-inoculation of associative bacteria, which have high nitrogen-fixing activity, tolerance for environmental conditions, and the ability to compete with the natural microflora, is used widely to enhance the growth and yields of agricultural plants. We evaluated the ability of 2 co-inoculated plant-growth-promoting rhizobacteria, Paenibacillus polymyxa 1465 and Azospirillum brasilense Sp245, to colonize roots of wheat (Triticum aestivum L. 'Saratovskaya 29') seedlings, and we assessed the morphometric parameters of wheat early in its development. Analysis by ELISA with polyclonal antibodies raised against the exopolysaccharide of P. polymyxa 1465 and the lipopolysaccharide of A. brasilense Sp245 demonstrated that the root-colonizing activity of A. brasilense was higher when the bacterium was co-inoculated with P. polymyxa than when it was inoculated singly. Immunofluorescence microscopy with Alexa Fluor 532-labeled antibodies revealed sites of attachment of co-inoculated P. polymyxa and A. brasilense and showed that the 2 bacteria colonized similar regions of the roots. Co-inoculation exerted a negative effect on wheat seedling development, inhibiting root length by 17.6%, total root weight by 11%, and total shoot weight by 12%. Under certain conditions, dual inoculation of wheat may prove ineffective, apparently owing to the competition between the rhizobacteria for colonization sites on the plant roots. The findings from this study may aid in developing techniques for mixed bacterial inoculation of cultivated plants.

  9. Structural analysis of the O-polysaccharide of the lipopolysaccharide from Azospirillum brasilense Jm6B2 containing 3-O-methyl-D-rhamnose (D-acofriose).

    Science.gov (United States)

    Boyko, Alevtina S; Dmitrenok, Andrey S; Fedonenko, Yuliya P; Zdorovenko, Evelina L; Konnova, Svetlana A; Knirel, Yuriy A; Ignatov, Vladimir V

    2012-07-01

    Two types of neutral O-polysaccharides were obtained by mild acid degradation of the lipopolysaccharide isolated by phenol-water extraction from the asymbiotic diazotrophic rhizobacterium Azospirillum brasilense Jm6B2. The following structure of the major O-polysaccharide was established by composition and methylation (ethylation) analyses, Smith degradation, and 1D and 2D (1)H and (13)C NMR spectroscopy: [structure: see text] where a non-stoichiometric (~60%) 3-O-methylation of D-rhamnose is indicated by italics.

  10. Synthesis of the tetrasaccharide repeating unit of the O-glycan from the polar flagellum flagellin of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Pal, Kumar Bhaskar; Mukhopadhyay, Balaram

    2014-12-01

    Chemical synthesis of the tetrasaccharide repeating unit of the O-glycan from the polar flagellum flagellin of Azospirillum brasilense Sp7 in the form of its p-methoxyphenyl glycoside is reported. The required glycosidic linkages have been accomplished by activation of thioglycosides with N-iodosuccinimide in the presence of H2SO4-silica. H2SO4-silica was found to be an effective alternative to the classical acid promoters like TfOH or TMSOTf and it can lead to the formation of both 1,2-cis and 1,2-trans glycosidic linkages depending on the protecting group manipulation and control of the reaction condition.

  11. INOCULAÇÃO DE Azospirillum brasilense E DOSES DE NITROGÊNIO EM MILHO PARA PRODUÇÃO DE SILAGEM E GRÃOS

    OpenAIRE

    Fernando Reimann Skonieski

    2015-01-01

    Esta tese foi desenvolvida com o objetivo de avaliar o efeito da inoculação de Azospirillum brasilense e da fertilização nitrogenada sobre o desenvolvimento da cultura do milho para silagem e grãos. Foram conduzidos experimentos nos municípios de Erechim, RS, nas safras 2012/2013 e 2013/2014 empregando os híbridos AS 1572 e Defender e Santa Maria, RS, na safra 2013/2014 com os híbridos AS 1572 e AG 9030. Os tratamentos foram distribuídos em arranjo fatorial composto por dois hí...

  12. Interaction between PII and NifA in Azospirillum brasilense Sp7

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The interaction between PII and NifA in A. brasilense Sp7 was investigated by using the yeast two-hybrid system. Our experimental results showed that PII directly interacted with the entire NifA protein and its N-terminal domain,but did not interact with the central domain and the C-terminal domain of NifA. No interaction happened if glnB coding for PII was frame-shift mutated. Pz, a homolog of PII, had no interation with NifA.

  13. Different responses of the GlnB and GlnZ proteins upon in vitro uridylylation by the Azospirillum brasilense GlnD protein

    Directory of Open Access Journals (Sweden)

    L.M. Araújo

    2008-04-01

    Full Text Available Azospirillum brasilense is a diazotroph found in association with important agricultural crops. In this organism, the regulation of nitrogen fixation by ammonium ions involves several proteins including the uridylyltransferase/uridylyl-removing enzyme, GlnD, which reversibly uridylylates the two PII proteins, GlnB and GlnZ, in response to the concentration of ammonium ions. In the present study, the uridylylation/deuridylylation cycle of A. brasilense GlnB and GlnZ proteins by GlnD was reconstituted in vitro using the purified proteins. The uridylylation assay was analyzed using non-denaturing polyacrylamide gel electrophoresis and fluorescent protein detection. Our results show that the purified A. brasilense GlnB and GlnZ proteins were uridylylated by the purified A. brasilense GlnD protein in a process dependent on ATP and 2-oxoglutarate. The dependence on ATP for uridylylation was similar for both proteins. On the other hand, at micromolar concentration of 2-oxoglutarate (up to 100 µM, GlnB uridylylation was almost twice that of GlnZ, an effect that was not observed at higher concentrations of 2-oxoglutarate (up to 10 mM. Glutamine inhibited uridylylation and stimulated deuridylylation of both GlnB and GlnZ. However, glutamine seemed to inhibit GlnZ uridylylation more efficiently. Our results suggest that the differences in the uridylylation pattern of GlnB and GlnZ might be important for fine-tuning of the signaling pathway of cellular nitrogen status in A. brasilense.

  14. Different responses of the GlnB and GlnZ proteins upon in vitro uridylylation by the Azospirillum brasilense GlnD protein.

    Science.gov (United States)

    Araújo, L M; Huergo, L F; Invitti, A L; Gimenes, C I; Bonatto, A C; Monteiro, R A; Souza, E M; Pedrosa, F O; Chubatsu, L S

    2008-04-01

    Azospirillum brasilense is a diazotroph found in association with important agricultural crops. In this organism, the regulation of nitrogen fixation by ammonium ions involves several proteins including the uridylyltransferase/uridylyl-removing enzyme, GlnD, which reversibly uridylylates the two PII proteins, GlnB and GlnZ, in response to the concentration of ammonium ions. In the present study, the uridylylation/deuridylylation cycle of A. brasilense GlnB and GlnZ proteins by GlnD was reconstituted in vitro using the purified proteins. The uridylylation assay was analyzed using non-denaturing polyacrylamide gel electrophoresis and fluorescent protein detection. Our results show that the purified A. brasilense GlnB and GlnZ proteins were uridylylated by the purified A. brasilense GlnD protein in a process dependent on ATP and 2-oxoglutarate. The dependence on ATP for uridylylation was similar for both proteins. On the other hand, at micromolar concentration of 2-oxoglutarate (up to 100 microM), GlnB uridylylation was almost twice that of GlnZ, an effect that was not observed at higher concentrations of 2-oxoglutarate (up to 10 mM). Glutamine inhibited uridylylation and stimulated deuridylylation of both GlnB and GlnZ. However, glutamine seemed to inhibit GlnZ uridylylation more efficiently. Our results suggest that the differences in the uridylylation pattern of GlnB and GlnZ might be important for fine-tuning of the signaling pathway of cellular nitrogen status in A. brasilense.

  15. The cyclic-di-GMP diguanylate cyclase CdgA has a role in biofilm formation and exopolysaccharide production in Azospirillum brasilense.

    Science.gov (United States)

    Ramírez-Mata, Alberto; López-Lara, Lilia I; Xiqui-Vázquez, Ma Luisa; Jijón-Moreno, Saúl; Romero-Osorio, Angelica; Baca, Beatriz E

    2016-04-01

    In bacteria, proteins containing GGDEF domains are involved in production of the second messenger c-di-GMP. Here we report that the cdgA gene encoding diguanylate cyclase A (CdgA) is involved in biofilm formation and exopolysaccharide (EPS) production in Azospirillum brasilense Sp7. Biofilm quantification using crystal violet staining revealed that inactivation of cdgA decreased biofilm formation. In addition, confocal laser scanning microscopy analysis of green-fluorescent protein-labeled bacteria showed that, during static growth, the biofilms had differential levels of development: bacteria harboring a cdgA mutation exhibited biofilms with considerably reduced thickness compared with those of the wild-type Sp7 strain. Moreover, DNA-specific staining and treatment with DNase I, and epifluorescence studies demonstrated that extracellular DNA and EPS are components of the biofilm matrix in Azospirillum. After expression and purification of the CdgA protein, diguanylate cyclase activity was detected. The enzymatic activity of CdgA-producing cyclic c-di-GMP was determined using GTP as a substrate and flavin adenine dinucleotide (FAD(+)) and Mg(2)(+) as cofactors. Together, our results revealed that A. brasilense possesses a functional c-di-GMP biosynthesis pathway.

  16. Effects of Azospirillum brasilense with genetically modified auxin biosynthesis gene ipdC upon the diversity of the indigenous microbiota of the wheat rhizosphere.

    Science.gov (United States)

    Baudoin, Ezékiel; Lerner, Anat; Mirza, M Sajjad; El Zemrany, Hamdy; Prigent-Combaret, Claire; Jurkevich, Edouard; Spaepen, Stijn; Vanderleyden, Jos; Nazaret, Sylvie; Okon, Yaacov; Moënne-Loccoz, Yvan

    2010-04-01

    The phytostimulatory properties of Azospirillum inoculants, which entail production of the phytohormone indole-3-acetic acid (IAA), can be enhanced by genetic means. However, it is not known whether this could affect their interactions with indigenous soil microbes. Here, wheat seeds were inoculated with the wild-type strain Azospirillum brasilense Sp245 or one of three genetically modified (GM) derivatives and grown for one month. The GM derivatives contained a plasmid vector harboring the indole-3-pyruvate/phenylpyruvate decarboxylase gene ipdC (IAA production) controlled either by the constitutive promoter PnptII or the root exudate-responsive promoter PsbpA, or by an empty vector (GM control). All inoculants displayed equal rhizosphere population densities. Only inoculation with either ipdC construct increased shoot biomass compared with the non-inoculated control. At one month after inoculation, automated ribosomal intergenic spacer analysis (ARISA) revealed that the effect of the PsbpA construct on bacterial community structure differed from that of the GM control, which was confirmed by 16S rDNA-based denaturing gradient gel electrophoresis (DGGE). The fungal community was sensitive to inoculation with the PsbpA construct and especially the GM control, based on ARISA data. Overall, fungal and bacterial communities displayed distinct responses to inoculation of GM A. brasilense phytostimulators, whose effects could differ from those of the wild-type.

  17. Mutation in a D-alanine-D-alanine ligase of Azospirillum brasilense Cd results in an overproduction of exopolysaccharides and a decreased tolerance to saline stress.

    Science.gov (United States)

    Jofré, Edgardo; Fischer, Sonia; Príncipe, Analía; Castro, Marina; Ferrari, Walter; Lagares, Antonio; Mori, Gladys

    2009-01-01

    Bacteria of the genus Azospirillum are free-living nitrogen-fixing, rhizobacteria that are found in close association with plant roots, where they exert beneficial effects on plant growth and yield in many crops of agronomic importance. Unlike other bacteria, little is known about the genetics and biochemistry of exopolysaccharides in Azospirillum brasilense. In an attempt to characterize genes associated with exopolysaccharides production, we generated an A. brasilense Cd Tn5 mutant that showed exopolysaccharides overproduction, decreased tolerance to saline conditions, altered cell morphology, and increased sensitivity to detergents. Genetic characterization showed that the Tn5 was inserted within a ddlB gene encoding for a d-alanine-d-alanine ligase, and located upstream of the ftsQAZ gene cluster responsible for cell division in different bacteria. Heterologous complementation of the ddlB Tn5 mutant restored the exopolysaccharides production to wild-type levels and the ability to grow in the presence of detergents, but not the morphology and growth characteristics of the wild-type bacteria, suggesting a polar effect of Tn5 on the fts genes. This result and the construction of a nonpolar ddlB mutant provide solid evidence of the presence of transcriptional coupling between a gene associated with peptidoglycan biosynthesis and the fts genes required to control cell division.

  18. Characterization of cell surface and extracellular matrix remodeling of Azospirillum brasilense chemotaxis-like 1 signal transduction pathway mutants by atomic force microscopy.

    Science.gov (United States)

    Edwards, Amanda Nicole; Siuti, Piro; Bible, Amber N; Alexandre, Gladys; Retterer, Scott T; Doktycz, Mitchel J; Morrell-Falvey, Jennifer L

    2011-01-01

    To compete in complex microbial communities, bacteria must sense environmental changes and adjust cellular functions for optimal growth. Chemotaxis-like signal transduction pathways are implicated in the regulation of multiple behaviors in response to changes in the environment, including motility patterns, exopolysaccharide production, and cell-to-cell interactions. In Azospirillum brasilense, cell surface properties, including exopolysaccharide production, are thought to play a direct role in promoting flocculation. Recently, the Che1 chemotaxis-like pathway from A. brasilense was shown to modulate flocculation, suggesting an associated modulation of cell surface properties. Using atomic force microscopy, distinct changes in the surface morphology of flocculating A. brasilense Che1 mutant strains were detected. Whereas the wild-type strain produces a smooth mucosal extracellular matrix after 24 h, the flocculating Che1 mutant strains produce distinctive extracellular fibril structures. Further analyses using flocculation inhibition, lectin-binding assays, and comparison of lipopolysaccharides profiles suggest that the extracellular matrix differs between the cheA1 and the cheY1 mutants, despite an apparent similarity in the macroscopic floc structures. Collectively, these data indicate that disruption of the Che1 pathway is correlated with distinctive changes in the extracellular matrix, which likely result from changes in surface polysaccharides structure and/or composition.

  19. Characterization of cell surface and extracellular matrix remodeling of Azospirillum brasilense chemotaxis-like 1 signal transduction pathway mutants by atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Doktycz, Mitchel John [ORNL; Morrell-Falvey, Jennifer L [ORNL

    2011-01-01

    To compete in complex microbial communities, bacteria must sense environmental changes and adjust cellular functions for optimal growth. Chemotaxis-like signal transduction pathways are implicated in the regulation of multiple behaviors in response to changes in the environment, including motility patterns, exopolysaccharide production, and cell-to-cell interactions. In Azospirillum brasilense, cell surface properties, including exopolysaccharide production, are thought to play a direct role in promoting flocculation. Recently, the Che1 chemotaxis-like pathway from A. brasilense was shown to modulate flocculation, suggesting an associated modulation of cell surface properties. Using atomic force microscopy, distinct changes in the surface morphology of flocculating A. brasilense Che1 mutant strains were detected. Whereas the wild-type strain produces a smooth mucosal extracellular matrix after 24 h, the flocculating Che1 mutant strains produce distinctive extracellular fibril structures. Further analyses using flocculation inhibition, lectin-binding assays, and comparison of lipopolysaccharides profiles suggest that the extracellular matrix differs between the cheA1 and the cheY1 mutants, despite an apparent similarity in the macroscopic floc structures. Collectively, these data indicate that disruption of the Che1 pathway is correlated with distinctive changes in the extracellular matrix, which likely result from changes in surface polysaccharides structure and/or composition.

  20. Characterization of Cell Surface and EPS Remodeling of Azospirillum brasilense Chemotaxis-like 1 Signal Transduction Pathway mutants by Atomic Force Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Billings, Amanda N [ORNL; Siuti, Piro [ORNL; Bible, Amber [University of Tennessee, Knoxville (UTK); Alexandre, Gladys [University of Tennessee, Knoxville (UTK); Retterer, Scott T [ORNL; Doktycz, Mitchel John [ORNL; Morrell-Falvey, Jennifer L [ORNL

    2011-01-01

    To compete in complex microbial communities, bacteria must quickly sense environmental changes and adjust cellular functions for optimal growth. Chemotaxis-like signal transduction pathways are implicated in the modulation of multiple cellular responses, including motility, EPS production, and cell-to-cell interactions. Recently, the Che1 chemotaxis-like pathway from Azospirillum brasilense was shown to modulate flocculation. In A. brasilense, cell surface properties, including EPS production, are thought to play a direct role in promoting flocculation. Using atomic force microscopy (AFM), we have detected distinct changes in the surface morphology of flocculating A. brasilense Che1 mutant strains that are absent in the wild type strain. Whereas the wild type strain produces a smooth mucosal extracellular matrix, the flocculating Che1 mutant strains produce distinctive extracellular fibril structures. Further analyses using flocculation inhibition and lectin-binding assays suggest that the composition of EPS components in the extracellular matrix differs between the cheA1 and cheY1 mutants, despite an apparent similarity in the macroscopic floc structures. Collectively, these data indicate that mutations in the Che1 pathway that result in increased flocculation are correlated with distinctive changes in the extracellular matrix structure produced by the mutants, including likely changes in the EPS structure and/or composition.

  1. Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense

    Energy Technology Data Exchange (ETDEWEB)

    Sotomaior, P.; Araújo, L.M.; Nishikawa, C.Y.; Huergo, L.F.; Monteiro, R.A.; Pedrosa, F.O.; Chubatsu, L.S.; Souza, E.M. [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR (Brazil)

    2012-09-21

    Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.

  2. Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense

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    P. Sotomaior

    2012-12-01

    Full Text Available Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.

  3. Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense.

    Science.gov (United States)

    Sotomaior, P; Araújo, L M; Nishikawa, C Y; Huergo, L F; Monteiro, R A; Pedrosa, F O; Chubatsu, L S; Souza, E M

    2012-12-01

    Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.

  4. An extracytoplasmic function sigma factor cotranscribed with its cognate anti-sigma factor confers tolerance to NaCl, ethanol and methylene blue in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Mishra, Mukti Nath; Kumar, Santosh; Gupta, Namrata; Kaur, Simarjot; Gupta, Ankush; Tripathi, Anil K

    2011-04-01

    Azospirillum brasilense, a plant-growth-promoting rhizobacterium, is exposed to changes in its abiotic environment, including fluctuations in temperature, salinity, osmolarity, oxygen concentration and nutrient concentration, in the rhizosphere and in the soil. Since extra-cytoplasmic function (ECF) sigma factors play an important role in stress adaptation, we analysed the role of ECF sigma factor (also known as RpoE or σ(E)) in abiotic stress tolerance in A. brasilense. An in-frame rpoE deletion mutant of A. brasilense Sp7 was carotenoidless and slow-growing, and was sensitive to salt, ethanol and methylene blue stress. Expression of rpoE in the rpoE deletion mutant complemented the defects in growth, carotenoid biosynthesis and sensitivity to different stresses. Based on data from reverse transcriptase-PCR, a two-hybrid assay and a pull-down assay, we present evidence that rpoE is cotranscribed with chrR and the proteins synthesized from these two overlapping genes interact with each other. Identification of the transcription start site by 5' rapid amplification of cDNA ends showed that the rpoE-chrR operon was transcribed by two promoters. The proximal promoter was less active than the distal promoter, whose consensus sequence was characteristic of RpoE-dependent promoters found in alphaproteobacteria. Whereas the proximal promoter was RpoE-independent and constitutively expressed, the distal promoter was RpoE-dependent and strongly induced in response to stationary phase and elevated levels of ethanol, salt, heat and methylene blue. This study shows the involvement of RpoE in controlling carotenoid synthesis as well as in tolerance to some abiotic stresses in A. brasilense, which might be critical in the adaptation, survival and proliferation of this rhizobacterium in the soil and rhizosphere under stressful conditions.

  5. PRODUTIVIDADE DO TRIGO IRRIGADO SUBMETIDO À APLICAÇÃO DE NITROGÊNIO E À INOCULAÇÃO COM AZOSPIRILLUM BRASILENSE

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    Pedro Henrique Marques Paula Nunes

    2015-02-01

    Full Text Available Inocular sementes de trigo com Azospirillum brasilense pode ser benéfico à cultura do trigo, bem como reduzir a necessidade de aplicação de N. No entanto, são escassos trabalhos que mensurem o benefício dessa prática no trigo irrigado com expectativa de alta produtividade e demanda de N. Objetivou-se com este trabalho avaliar a produtividade e o conteúdo de N na planta do trigo irrigado, submetido a doses de N, com ou sem a inoculação com A. brasilense. Foram conduzidos dois experimentos em 2011 e repetidos em 2012. Em cada ano de cultivo, instalou-se um experimento na área considerada de baixa disponibilidade de N no solo (cultivo em sucessão ao milho e outro na com alta disponibilidade (cultivo em sucessão à soja ou cenoura. Os tratamentos consistiram de cinco doses de N e da inoculação ou não com A. brasilense. As doses de N testadas foram 20, 60, 100, 140 e 180 kg ha-1 na área com baixa disponibilidade de N e 20, 50, 80, 110 e 140 kg ha-1 na com alta disponibilidade. O teor de N na folha índice foi aumentado pela inoculação com A. brasilense. Entretanto, o N absorvido não foi influenciado pela inoculação da bactéria. As doses de N aumentaram a produtividade do trigo e esse aumento foi mais evidente no cultivo na área com baixa disponibilidade de N no solo.

  6. ESTUDIO DEL EFECTO QUIMIOTÁCTICO DE LOS EXUDADOS RADICALES DEL ARROZ SOBRE Azospirillum brasilense Sp7 Y A. brasilense R5(15. PRODUCCIÓN DE AUXINAS POR AMBAS CEPAS

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    Mabel Pazos

    2003-01-01

    Full Text Available El género Azospirillum agrupa bacterias diazotróficas de vida libre, ampliamente encontradas en suelos tropicales y subtropicales, en asociación con raíces de cultivos de importancia económica como arroz, trigo y diversas especies vegetales. La gran mayoría de los estudios realizados con este género han demostrado su distribución prácticamente universal y sus numerosos efectos sobre el crecimiento de las plantas que coloniza. En este trabajo se demuestra el efecto quimiotáctico que ejercen los exudados radicales del arroz variedad J-104 sobre dos cepas de A. brasilense (R5(15 y Sp7, así como la capacidad de ambas de producir auxinas 16.60 mg.mL-1 y 11.82 mg.mL-1 respectivamente.

  7. Inoculação com Azospirillum brasilense, fontes e doses de nitrogênio na cultura do milho em duas épocas de semeadura

    OpenAIRE

    Kaneko, Flávio Hiroshi [UNESP

    2013-01-01

    A adubação nitrogenada na cultura do milho vem passando por modificações em função do uso de tecnologias tais como a fixação biológica de N e o revestimento dos fertilizantes com polímeros. Este trabalho teve como objetivo avaliar o efeito da inoculação de sementes de com Azospirillum brasilense (com e sem inoculação), fontes (Ureia e Ureia revestida com Policote®) e doses de N em cobertura (0, 45, 90, 135 e 180 kg ha-1) na cultura do milho, em esquema fatorial 2x2x5, com 4 repetições. Foram ...

  8. Tn5-mutagenesis and identification of atr operon and trpE gene responsible for indole-3-acetic acid synthesis in Azospirillum brasilense Yu62

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To bring more information about synthesis of indole-3-acetic acid (IAA) from Azospirillum brasilense, a Tn5-insertion library of A. brasilense Yu62 was constructed and subjected to screening for IAA producing mutants. Two mutants with decreased IAA levels, named as A3 and A24, were isolated. The sequence analysis of loci tagged showed that the Tn5-1063a was located in the atrA gene encoding GntR family transcriptional regulator and trpE gene encoding component I of anthranilate synthase respectively. At the same time, atrB encoding phosphotransferase and atrC encoding aminotransferase were cloned downstream the atrA gene and atrA,atrB and atrC were clustered in an operon. Mutagenesis and complementation studies showed that atrA and atrC were involved in IAA synthesis. IAA levels of trpE mutant and wild-type strain could be improved by adding anthranilate into the medium.

  9. A SAM-dependent methyltransferase cotranscribed with arsenate reductase alters resistance to peptidyl transferase center-binding antibiotics in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Singh, Sudhir; Singh, Chhaya; Tripathi, Anil Kumar

    2014-05-01

    The genome of Azospirillum brasilense harbors a gene encoding S-adenosylmethionine-dependent methyltransferase, which is located downstream of an arsenate reductase gene. Both genes are cotranscribed and translationally coupled. When they were cloned and expressed individually in an arsenate-sensitive strain of Escherichia coli, arsenate reductase conferred tolerance to arsenate; however, methyltransferase failed to do so. Sequence analysis revealed that methyltransferase was more closely related to a PrmB-type N5-glutamine methyltransferase than to the arsenate detoxifying methyltransferase ArsM. Insertional inactivation of prmB gene in A. brasilense resulted in an increased sensitivity to chloramphenicol and resistance to tiamulin and clindamycin, which are known to bind at the peptidyl transferase center (PTC) in the ribosome. These observations suggested that the inability of prmB:km mutant to methylate L3 protein might alter hydrophobicity in the antibiotic-binding pocket of the PTC, which might affect the binding of chloramphenicol, clindamycin, and tiamulin differentially. This is the first report showing the role of PrmB-type N5-glutamine methyltransferases in conferring resistance to tiamulin and clindamycin in any bacterium.

  10. Alternative mechanism for the evaluation of indole-3-acetic acid (IAA) production by Azospirillum brasilense strains and its effects on the germination and growth of maize seedlings.

    Science.gov (United States)

    Masciarelli, Oscar; Urbani, Lucia; Reinoso, Herminda; Luna, Virginia

    2013-10-01

    We evaluated the production of indole-3-acetic acid (IAA) by Azospirillum brasilense strains in vitro (cell culture supernatants) and in vivo (stems and roots of maize seedlings) to clarify the role of this phytohormone as a signaling and effector molecule in the symbiotic interaction between maize and A. brasilense. The three strains all showed IAA production when cultured in NFb medium supplemented with 100 μg/ml L-tryptophan. The level of IAA production was 41.5 μg/ml for Yu62, 12.9 μg/ml for Az39, and 0.15 μg/ml for ipdC-. The release of IAA into culture medium by the bacteria appeared to be the main activator of the early growth promotion observed in the inoculated maize seedlings. The application of supernatants with different IAA contents caused significant differences in the seedling growth. This observation provides the basis for novel technological tools for effective quality control procedures on inoculants. The approach described can be incorporated into different inoculation methods, including line sowing, downspout, and foliar techniques, and increase the sustainability of symbiotic plant-bacteria systems.

  11. Scale-up from shake flasks to pilot-scale production of the plant growth-promoting bacterium Azospirillum brasilense for preparing a liquid inoculant formulation.

    Science.gov (United States)

    Trujillo-Roldán, Mauricio A; Valdez-Cruz, Norma A; Gonzalez-Monterrubio, César F; Acevedo-Sánchez, Eduardo V; Martínez-Salinas, Carlos; García-Cabrera, Ramsés I; Gamboa-Suasnavart, Ramsés A; Marín-Palacio, Luz D; Villegas, Jesús; Blancas-Cabrera, Abel

    2013-11-01

    Azospirillum brasilense has industrial significance as a growth promoter in plants of commercial interest. However, there is no report in the literature disclosing a liquid product produced in pilot-scale bioreactors and is able to be stored at room temperature for more than 2 years. The aim of this work was to scale up a process from a shake flask to a 10-L lab-scale and 1,000-L pilot-scale bioreactor for the production of plant growth-promoting bacterium A. brasilense for a liquid inoculant formulation. Furthermore, this work aimed to determine the shelf life of the liquid formulation stored at room temperature and to increase maize crops yield in greenhouses. Under a constant oxygen mass transfer coefficient (K L a), a fermentation process was successfully scaled up from shake flasks to 10- and 1,000-L bioreactors. A concentration ranging from 3.5 to 7.5 × 10(8) CFU/mL was obtained in shake flasks and bioreactors, and after 2 years stored at room temperature, the liquid formulation showed one order of magnitude decrease. Applications of the cultured bacteria in maize yields resulted in increases of up to 95 % in corncobs and 70 % in aboveground biomass.

  12. Screening of the interacting proteins with NifA in Azospirillum brasilense Sp7 by the yeast two-hybrid system

    Institute of Scientific and Technical Information of China (English)

    CHEN Sanfeng; GUAN Yu; TU Ran; SUN Wengai; LI Jilun

    2005-01-01

    NifA in Azospirillum brasilense plays a key role in regulating the synthesis and activity of nitrogenase in response to ammonia and oxygen available. In this work we used the yeast two-hybrid system to identify the proteins that interact with NifA. The nifA gene was fused to the yeast two-hybrid vector pGBD-C2, and three A. brasilense Sp7 genomic libraries for use in yeast two-hybrid studies were constructed. Screening of the libraries identified four clones encoding proteins that interact with NifA. The confirmation of the interactions of each gene product of the four clones and NifA were carried out by exchanging the vectors for nifA and the four clones and by mutageneses of the four clones with shift reading frame experiments in yeast two-hybrid studies. DNA sequence analyses showed that two clones encode proteins containing PAS domains that play an important role in signal transduction. One clone has high similarity with the fhuE gene of Escherichia coli, whose gene product is involved in iron uptake and transportation, and the other clone encodes an unknown protein.

  13. Adenosine diphosphate ribosylation of dinitrogenase reductase and adenylylation of glutamine synthetase control ammonia excretion in ethylenediamine-resistant mutants of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Srivastava, A; Tripathi, A K

    2006-10-01

    Azospirillum brasilense is a nitrogen-fixing, root-colonizing bacterium that brings about plant-growth-promoting effects mainly because of its ability to produce phytohormones. Ethylenediamine (EDA)-resistant mutants of A. brasilense were isolated and screened for their higher ability to decrease acetylene and release ammonia in the medium. One of the mutants showed considerably higher levels of acetylene decrease and ammonia excretion. Nitrogenase activity of this mutant was relatively resistant to inhibition by NH(4)Cl. Adenosine triphosphate ribosylation of dinitrogenase reductase in the mutant did not increase even in presence of 10 mM NH(4)Cl. Although the mutant showed decreased glutamine synthetase (GS) activity, neither the levels of GS synthesized by the mutant nor the NH (4) (+) -binding site in the GS differed from those of the parent. The main reason for the release of ammonia by the mutant seems to be the fixation of higher levels of nitrogen than its GS can assimilate, as well as higher levels of adenylylation of GS, which may decrease ammonia assimilation.

  14. Spontaneous Super-Swarming Derivatives of Azospirillum brasilense Sp245 have Different DNA Profiles and Behavior in the Presence of Various Nitrogen Sources.

    Science.gov (United States)

    Varshalomidze, Olga E; Petrova, Lilia P; Shelud'ko, Andrei V; Katsy, Elena I

    2012-12-01

    Azospirillum brasilense swims in liquid environments and swarms in semisolid media. Five variants of A. brasilense Sp245, Sp245.P1-Sp245.P5, which swarmed faster than Sp245 in a semisolid malate-salt medium, have been isolated. In Sp245.P1-Sp245.P4, a new megaplasmid was revealed instead of an indigenous 85-MDa plasmid (p85). By polymerase chain reactions (PCR) with primers to the segments of p85 important for proper bacterial motility/flagellation and for dissimilatory nitrite and NO reduction, that DNA of p85 was found retained by all the variants. In ERIC- and RAPD-PCR, microdiversity between the total DNAs of Sp245 and its variants was detected. Interstrain differences in growth characteristics in liquid peptone-succinate-salt medium with KNO3 or KNO2 and in KNO2 production/consumption were revealed. Although all the variants swam and swarmed faster than Sp245 in the medium supplemented with NH4Cl or KNO3, not all of them could do so in MPSS with KNO2.

  15. Cultivation factors and population size control the uptake of nitrogen by the microalgae Chlorella vulgaris when interacting with the microalgae growth-promoting bacterium Azospirillum brasilense.

    Science.gov (United States)

    de-Bashan, Luz E; Antoun, Hani; Bashan, Yoav

    2005-10-01

    Growth of and the capacity to take up nitrogen in the freshwater microalgae Chlorella vulgaris were studied while varying the concentrations of ammonium and nitrate, the pH and the source of carbon in a synthetic wastewater growth medium when co-immobilized in alginate beads with the microalgae growth-promoting bacterium Azospirillum brasilense. Analyses of 29 independent experiments showed that co-immobilization of the microalgae with A. brasilense could result in two independent phenomena directly affected by cultivation factors, such as nitrogen species, pH and presence of a carbon source. First, growth of the microalgal population increased without an increase in the capacity of the single cells to take up nitrogen, or second, the capacity of cells to take up nitrogen increased without an increase of the total microalgal population. These phenomena were dependent on the population density of the microalgae, which was in turn affected by cultivation factors. This supports the conclusion that the size of the microalgal population controls the uptake of nitrogen in C. vulgaris cells - the higher the population (regardless the experimental parameters), the less nitrogen each cell takes up.

  16. ROLE OF GLUTAMATE DEHYDROGENASE AND GLUTAMINE SYNTHETASE IN CHLORELLA VULGARIS DURING ASSIMILATION OF AMMONIUM WHEN JOINTLY IMMOBILIZED WITH THE MICROALGAE-GROWTH-PROMOTING BACTERIUM AZOSPIRILLUM BRASILENSE(1).

    Science.gov (United States)

    De-Bashan, Luz E; Magallon, Paola; Antoun, Hani; Bashan, Yoav

    2008-10-01

    Enzymatic activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) participating in the nitrogen metabolism and related ammonium absorption were assayed after the microalga Chlorella vulgaris Beij. was jointly immobilized with the microalgae-growth-promoting bacterium Azospirillum brasilense. At initial concentrations of 3, 6, and 10 mg · L(-1)  NH4 (+) , joint immobilization enhances growth of C. vulgaris but does not affect ammonium absorption capacity of the microalga. However, at 8 mg · L(-1)  NH4 (+) , joint immobilization enhanced ammonium absorption by the microalga without affecting the growth of the microalgal population. Correlations between absorption of ammonium per cell and per culture showed direct (negative and positive) linear correlations between these parameters and microalga populations at 3, 6, and 10 mg · L(-1)  NH4 (+) , but not at 8 mg · L(-1)  NH4 (+) , where the highest absorption of ammonium occurred. In all cultures, immobilized and jointly immobilized, having the four initial ammonium concentrations, enzymatic activities of Chlorella are affected by A. brasilense. Regardless of the initial concentration of ammonium, GS activity in C. vulgaris was always higher when jointly immobilized and determined on a per-cell basis. When jointly immobilized, only at an initial concentration of 8 mg · L(-1)  NH4 (+) was GDH activity per cell higher.

  17. A constitutively expressed pair of rpoE2-chrR2 in Azospirillum brasilense Sp7 is required for survival under antibiotic and oxidative stress.

    Science.gov (United States)

    Gupta, Namrata; Kumar, Santosh; Mishra, Mukti Nath; Tripathi, Anil Kumar

    2013-02-01

    Extracytoplasmic function (ECF) sigma factors (σ(E)) are known to bring about changes in gene expression to enable bacteria to adapt to different stresses. The Azospirillum brasilense Sp245 genome harbours nine genes encoding σ(E), of which two are adjacent to the genes encoding ChrR-type zinc-binding anti-sigma (ZAS) factors. We describe here the role and regulation of a new pair of rpoE-chrR, which was found in the genome of A. brasilense Sp7 in addition to the previously described rpoE-chrR pair (designated rpoE1-chrR1). The rpoE2-chrR2 pair is also cotranscribed, and their products show protein-protein interaction. The -10 and -35 promoter elements of rpoE2-chrR2 and rpoE1-chrR1 were similar but not identical. Unlike the promoter of rpoE1-chrR1, the rpoE2-chrR2 promoter was neither autoregulated nor induced by oxidative stress. Inactivation of chrR2 or overexpression of rpoE2 in A. brasilense Sp7 resulted in an overproduction of carotenoids. It also conferred resistance to oxidative stresses and antibiotics. By controlling the synthesis of carotenoids, initiation and elongation of translation, protein folding and purine biosynthesis, RpoE2 seems to play a crucial role in preventing and repairing the cellular damage caused by oxidative stress. Lack of autoregulation and constitutive expression of rpoE2-chrR2 suggest that RpoE2-ChrR2 may provide a rapid mechanism to cope with oxidative stress, wherein singlet oxygen ((1)O(2))-mediated dissociation of the RpoE2-ChrR2 complex might release RpoE2 to drive the expression of its target genes.

  18. Evidence for ferritin as dominant iron-bearing species in the rhizobacterium Azospirillum brasilense Sp7 provided by low-temperature/in-field Mössbauer spectroscopy.

    Science.gov (United States)

    Kovács, Krisztina; Kamnev, Alexander A; Pechoušek, Jiří; Tugarova, Anna V; Kuzmann, Ernő; Machala, Libor; Zbořil, Radek; Homonnay, Zoltán; Lázár, Károly

    2016-02-01

    For the ubiquitous diazotrophic rhizobacterium Azospirillum brasilense, which has been attracting the attention of researchers worldwide for the last 35 years owing to its significant agrobiotechnological and phytostimulating potential, the data on iron acquisition and its chemical speciation in cells are scarce. In this work, for the first time for azospirilla, low-temperature (at 80 K, 5 K, as well as at 2 K without and with an external magnetic field of 5 T) transmission Mössbauer spectroscopic studies were performed for lyophilised biomass of A. brasilense (wild-type strain Sp7 grown with (57)Fe(III) nitrilotriacetate complex as the sole source of iron) to enable quantitative chemical speciation analysis of the intracellular iron. In the Mössbauer spectrum at 80 K, a broadened quadrupole doublet of high-spin iron(III) was observed with a few percent of a high-spin iron(II) contribution. In the spectrum measured at 5 K, a dominant magnetically split component appeared with the parameters typical of ferritin species from other bacteria, together with a quadrupole doublet of a superparamagnetic iron(III) component and a similarly small contribution from the high-spin iron(II) component. The Mössbauer spectra recorded at 2 K (with or without a 5 T external field) confirmed the assignment of ferritin species. About 20% of total Fe in the dry cells of A. brasilense strain Sp7 were present in iron(III) forms superparamagnetic at both 5 and 2 K, i.e. either different from ferritin cores or as ferritin components with very small particle sizes.

  19. Sequencing and promoter analysis of the nifENXorf3orf5fdxAnifQ operon from Azospirillum brasilense Sp7

    Directory of Open Access Journals (Sweden)

    D.P. Potrich

    2001-11-01

    Full Text Available A 40-kb DNA region containing the major cluster of nif genes has been isolated from the Azospirillum brasilense Sp7 genome. In this region three nif operons have been identified: nifHDKorf1Y, nifENXorf3orf5fdxAnifQ and orf2nifUSVorf4. The operons containing nifENX and nifUSV genes are separated from the structural nifHDKorf1Y operon by about 5 kb and 10 kb, respectively. The present study shows the sequence analysis of the 6045-bp DNA region containing the nifENX genes. The deduced amino acid sequences from the open reading frames were compared to the nif gene products of other diazotrophic bacteria and indicate the presence of seven ORFs, all reading in the same direction as that of the nifHDKorf1Y operon. Consensus sigma54 and NifA-binding sites are present only in the promoter region upstream of the nifE gene. This promoter is activated by NifA protein and is approximately two-times less active than the nifH promoter, as indicated by the ß-galactosidase assays. This result suggests the differential expression of the nif genes and their respective products in Azospirillum.

  20. Increased Salinity Tolerance of Cowpea Plants by Dual Inoculation of an Arbuscular Mycorrhizal Fungus Glomus clarum and a Nitrogen-fixer Azospirillum brasilense.

    Science.gov (United States)

    Rabie, G H; Aboul-Nasr, M B; Al-Humiany, A

    2005-03-01

    Pot greenhouse experiments were carried out to attempt to increase the salinity tolerance of one of the most popular legume of the world; cowpea; by using dual inoculation of an Am fungus Glomus clarum and a nitrogen-fixer Azospirillum brasilense. The effect of these beneficial microbes, as single- or dual inoculation-treatments, was assessed in sterilized loamy sand soil at five NaCl levels (0.0~7.2 ds/m) in irrigating water. The results of this study revealed that percentage of mycorrhizal infection, plant height, dry weight, nodule number, protein content, nitrogenase and phosphatase activities, as well as nutrient elements N, P, K, Ca, Mg were significantly decreased by increasing salinity level in non-mycorrhized plants in absence of NFB. Plants inoculated with NFB showed higher nodule numbers, protein content, nitrogen concentration and nitrogenase activities than those of non-inoculated at all salinity levels. Mycorrhized plants exhibited better improvement in all measurements than that of non-mycorrhized ones at all salinity levels, especially, in the presence of NFB. The concentration of Na(+) was significantly accumulated in cowpea plants by rising salinity except in shoots of mycorrhizal plants which had K(+)/Na(+) ratios higher than other treatments. This study indicated that dual inoculation with Am fungi and N-fixer Azospirillum can support both needs for N and P, excess of NaCl and will be useful in terms of soil recovery in saline area.

  1. The electron transfer flavoprotein fixABCX gene products from Azospirillum brasilense show a NifA-dependent promoter regulation.

    Science.gov (United States)

    Sperotto, Raul Antonio; Gross, Jeferson; Vedoy, Cleber; Passaglia, Luciane Maria Pereira; Schrank, Irene Silveira

    2004-10-01

    The complete nucleotide sequence of the A. brasilense fixA, fixB, fixC, and fixX genes is reported here. Sequence similarities between the protein sequences deduced from fixABCX genes and many electron transfer flavoproteins (ETFs) have been noted. Comparison of the amino acid sequences of both subunits of ETF with the A. brasilense fixA and fixB gene products exhibits an identity of 30%. The amino acid sequence of the other two genes, fixC and fixX, revealed similarity with the membrane-bound electron transfer flavoprotein ubiquinone oxidoreductase (ETF-QO). Using site-directed mutagenesis, mutations were introduced in the fixA promoter element of the A. brasilense fixABCX operon and chimeric p fixA-lacZ reporter gene fusions were constructed. The activation of the fixA promoter of A. brasilense is dependent upon the presence of the NifA protein being approximately 7 times less active than the A. brasilense nifH promoter. These results indicate that NifA from Klebsiella pneumoniae activates the fix promoter of A. brasilense and provide further evidence in support of the regulatory model of NifA activation in A. brasilense. Although no specific function has been assigned to the fixABCX gene products they are apparently required for symbiotic nitrogen fixation. An electron-transferring capacity in the nitrogen fixation pathway has been suggested for the fix gene products based on sequence homologies to the ETFs and ETF-QO proteins and by the absence of orthologous electron transfer proteins NifJ and NifF in A. brasilense.

  2. Gene encoding γ-carbonic anhydrase is cotranscribed with argC and induced in response to stationary phase and high CO2 in Azospirillum brasilense Sp7

    Directory of Open Access Journals (Sweden)

    Mishra Mukti N

    2010-07-01

    Full Text Available Abstract Background Carbonic anhydrase (CA is a ubiquitous enzyme catalyzing the reversible hydration of CO2 to bicarbonate, a reaction underlying diverse biochemical and physiological processes. Gamma class carbonic anhydrases (γ-CAs are widespread in prokaryotes but their physiological roles remain elusive. At present, only γ-CA of Methanosarcina thermophila (Cam has been shown to have CA activity. Genome analysis of a rhizobacterium Azospirillum brasilense, revealed occurrence of ORFs encoding one β-CA and two γ-CAs. Results One of the putative γ-CA encoding genes of A. brasilense was cloned and overexpressed in E. coli. Electrometric assays for CA activity of the whole cell extracts overexpressing recombinant GCA1 did not show CO2 hydration activity. Reverse transcription-PCR analysis indicated that gca1 in A. brasilense is co-transcribed with its upstream gene annotated as argC, which encodes a putative N-acetyl-γ-glutamate-phosphate reductase. 5'-RACE also demonstrated that there was no transcription start site between argC and gca1, and the transcription start site located upstream of argC transcribed both the genes (argC-gca1. Using transcriptional fusions of argC-gca1 upstream region with promoterless lacZ, we further demonstrated that gca1 upstream region did not have any promoter and its transcription occurred from a promoter located in the argC upstream region. The transcription of argC-gca1 operon was upregulated in stationary phase and at elevated CO2 atmosphere. Conclusions This study shows lack of CO2 hydration activity in a recombinant protein expressed from a gene predicted to encode a γ-carbonic anhydrase in A. brasilense although it cross reacts with anti-Cam antibody raised against a well characterized γ-CA. The organization and regulation of this gene along with the putative argC gene suggests its involvement in arginine biosynthetic pathway instead of the predicted CO2 hydration.

  3. Adubação nitrogenada associada à inoculação com Azospirillum brasilense e Herbaspirillum seropedicae na cultura do milho

    Directory of Open Access Journals (Sweden)

    Janaína Dartora

    2013-10-01

    Full Text Available Bactérias dos gêneros Azospirillum e Herbaspirillum são fixadoras de nitrogênio (N atmosférico e consideradas promotoras do crescimento vegetal. Desta forma, desenvolveu-se o presente trabalho com o objetivo de avaliar o desenvolvimento e a produtividade do milho em resposta à inoculação de A. brasilense e H. seropedicae associada à adubação nitrogenada. O experimento foi conduzido em delineamento experimental de blocos ao acaso em esquema fatorial 4 x 5, com quatro repetições. Os tratamentos foram constituídos da inoculação de sementes (testemunha, estirpe de A. brasilense - Ab-V5, estirpe de H. seropedicae - SmR1 e combinação Ab-V5 + SmR1 e doses de N (0, 40, 80, 120 e 160 kg ha-1 de N. Foram avaliados: diâmetro basal do colmo, altura de planta, matéria seca de parte aérea e área foliar, nas fases vegetativa e reprodutiva, e produtividade. A adubação nitrogenada favoreceu o desenvolvimento da cultura até a máxima dose estudada. A inoculação combinada das estirpes proporcionou incrementos de 12% na matéria seca de parte aérea e 7% na produtividade em relação à testemunha indicando benefícios da combinação de bactérias diazotróficas que interagem de forma diferenciada com a planta.

  4. Alkyl hydroperoxide reductase has a role in oxidative stress resistance and in modulating changes in cell-surface properties in Azospirillum brasilense Sp245.

    Science.gov (United States)

    Wasim, Mariam; Bible, Amber N; Xie, Zhihong; Alexandre, Gladys

    2009-04-01

    An ahpC mutant derivative of Azospirillum brasilense Sp245 (strain SK586) that encodes an alkyl hydroperoxide reductase was found to be more sensitive to oxidative stress caused by organic hydroperoxides compared with the wild-type. In addition, the ahpC mutant strain had multiple defects in a large array of cellular functions that were consistent with alteration of cell-surface properties, such as cell morphology in stationary phase, Calcofluor White-, Congo Red- and lectin-binding abilities, as well as cell-to-cell aggregation and flocculation. All phenotypes of the ahpC mutant were complemented by in trans expression of AhpC, and overexpression of AhpC in the wild-type strain was found to affect the same set of phenotypes, suggesting that the pleiotropic effects were caused by the ahpC mutation. SK586 was also found to be fully motile, but it lost motility at a higher rate than the wild-type during growth, such that most SK586 cells were non-motile in stationary phase. Despite these defects, the mutant did not differ from the wild-type in short-term colonization of sterile wheat roots when inoculated alone, and in competition with the wild-type strain; this implied that AhpC activity may not endow the cells with a competitive advantage in colonization under these conditions. Although the exact function of AhpC in affecting these phenotypes remains to be determined, changes in cell morphology, surface properties, cell-to-cell aggregation and flocculation are common adaptive responses to various stresses in bacteria, and the data obtained here suggest that AhpC contributes to modulating such stress responses in A. brasilense.

  5. Evaluación de la eficiencia agronómica de Azospirillum brasilense en la cultura del trigo y los efectos en la calidad de la harina Avaliação da eficiência agronômica de Azospirillum brasilense na cultura do trigo e os efeitos na qualidade de farinha

    Directory of Open Access Journals (Sweden)

    Marcelo Cruz Mendes

    2012-06-01

    Full Text Available

    El objetivo de este estudio fue evaluar la eficacia de la inoculación de la bacteria Azospirillum brasilense sobre parámetros agronómicos del cultivo de trigo en los, donde se utilizó la dosis recomendada de bacterias y tres veces esta dosis, en comparación con dos niveles de nitrógeno abonado aplicado en cobertura. El experimento se llevó a cabo en el campo experimental de la Facultad de Agronomía de la Universidade Estadual do Centro Oeste, en Guarapuava-PR/Brasil. El diseño experimental fue bloques al azar con cinco repeticiones, con un total de 35 parcelas (7 tratamientos x 5 repeticiones. Las parcelas se consistieron de nueve filas (5,0 m de largo x 0,17 m entre filas, con una superficie total de 7,65 m2 y área útil compuesta por las dos filas centrales. Los parámetros de la planta obtenidos muestran que el uso de bacterias Azospirillum brasilense no causó ninguna diferencia estadística en la productividad en comparación con los demás tratamientos. Además, los parámetros de calidad de harina evaluados no difirieran estadísticamente, lo que indica que los tratamientos con y sin reducción de nitrógeno y con y sin el uso de Azospirillum brasilense, no fueron capaces de reducir la sedimentación, proteína y falling number. De este modo, se pude inferir que la bacteria tiene una eficiencia agronómica en la aplicación como tratamiento de semillas con el objetivo principalmente a la reducción de la aplicación de nitrógeno como abono de cobertura.

    O objetivo do trabalho foi o de avaliar a eficiência da inoculação da bactéria Azospirillum brasilense sobre os parâmetros agronômicos da cultura do trigo, onde utilizou-se a dosagem recomendada da bactéria e três vezes essa dosagem, frente a dois níveis de adubação nitrogenada em cobertura. O experimento foi conduzido no campo experimental do Departamento de Agronomia da Universidade

  6. Chemical composition and deoxynivalenol in wheat of Central- Southern Paraná: nitrogen fertilization in top dressing associated with Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    Thiago Montagner Souza

    2014-02-01

    Full Text Available The impact of agricultural management practices on the quality of grain was evaluated in wheat (Triticum aestivum L. BRS Tangará from the South Central region of Paraná State (Ponta Grossa in the crop years of 2010 and 2011. The field trial was carried out in succession with soybean (2010 and corn (2011. The treatments included inoculation of seeds with Azospirillum brasilense and increasing levels of nitrogen application in top dressing (0, 30, 60, 90 and 120 kg ha-1. The experimental design was in randomized block, factorial 2 x 5 (inoculation x N levels, with four replications. The parameters evaluated were water activity, moisture, protein, and grain contamination by deoxynivalenol (DON. The data were subjected to analysis of variance, comparison of means by Tukey’s test (p<0.05 and regression for nitrogen levels. The inoculation of seeds with A. brasilense increased the protein content in grain in 2010 (+1.6%; 16.9 g 100g-1 and 2011 (+1.7%, 15.7 g 100g-1, independently of the nitrogen level (p<0.01. Levels of nitrogen in 2010 presented a positive linear response with protein content, increasing by 14.2% using non-inoculated seeds (p<0.01, R2=0.955 and 14.4% for those inoculated with A. brasilense (p<0.01, R2=0.906. However, in 2011 a quadratic response was observed between nitrogen levels and protein content (p<0.01, R2=0.99, with stabilization or reduction in protein content using high levels of nitrogen (?120 kg ha-1. The contamination by DON was greater using high levels of nitrogen (3574 ?g kg-1, 120 kg ha-1; non-inoculated seeds in 2011, with a quadratic response between nitrogen levels and contamination of grains (p<0.05, R2=0.772. Furthermore, 37.5% of the samples presented contamination by DON higher than the maximum tolerated limit established by Brazilian legislation (2000 ?g kg-1; whole-wheat grain. The data demonstrates that proper management of nitrogen enhances intrinsic effects arising from plant breeding.

  7. Elemental composition of strawberry plants inoculated with the plant growth-promoting bacterium Azospirillum brasilense REC3, assessed with scanning electron microscopy and energy dispersive X-ray analysis.

    Science.gov (United States)

    Guerrero-Molina, M F; Lovaisa, N C; Salazar, S M; Díaz-Ricci, J C; Pedraza, R O

    2014-07-01

    The elemental composition of strawberry plants (Fragaria ananassa cv. Macarena) inoculated with the plant growth-promoting bacterium Azospirillum brasilense REC3, and non-inoculated controls, was studied using scanning electron microscopy (SEM) and energy dispersive X-ray (EDS) analysis. This allowed simultaneous semi-quantification of different elements in a small, solid sample. Plants were inoculated and grown hydroponically in 50% or 100% Hoagland solution, corresponding to limited or optimum nutrient medium, respectively. Bacteria-inoculated plants increased the growth index 45% and 80% compared to controls when grown in 100% and 50% Hoagland solution, respectively. Thus, inoculation with A. brasilense REC3 in a nutrient-limited medium had the strongest effect in terms of increasing both shoot and root biomass and growth index, as already described for Azospirillum inoculated into nutrient-poor soils. SEM-EDS spectra and maps showed the elemental composition and relative distribution of nutrients in strawberry tissues. Leaves contained C, O, N, Na, P, K, Ca and Cu, while roots also had Si and Cl. The organic fraction (C, O and N) accounted for over 96.3% of the total chemical composition; of the mineral fraction, Na had higher accumulation in both leaves and roots. Azospirillum-inoculated and control plants had similar elemental quantities; however, in bacteria-inoculated roots, P was significantly increased (34.33%), which constitutes a major benefit for plant nutrition, while Cu content decreased (35.16%).

  8. Positive effect of reduced aeration rate on growth and stereospecificity of DL-malic acid consumption by Azospirillum brasilense: improving the shelf life of a liquid inoculant formulation.

    Science.gov (United States)

    Carrasco-Espinosa, Karen; García-Cabrera, Ramsés I; Bedoya-López, Andrea; Trujillo-Roldán, Mauricio A; Valdez-Cruz, Norma A

    2015-02-10

    Azospirillum brasilense has significance as a growth promoter in plants of commercial interest. Two industrial native strains (Start and Calf), used as a part of an inoculant formulation in Mexico during the last 15 years, were incubated in laboratory-scale pneumatic bioreactors at different aeration rates. In both strains, the positive effect of decreased aeration was observed. At the lowest (0.1 vvm, air volume/liquid volume×minute), the highest biomass were obtained for Calf (7.8 × 10(10)CFU/ml), and Start (2.9 × 10(9)CFU/ml). These were higher in one magnitude order compared to cultures carried out at 0.5 vvm, and two compared to those at 1.0 vvm. At lower aeration, both stereoisomeric forms of malic acid were consumed, but at higher aeration, just L-malate was consumed. A reduction in aeration allows an increase of the shelf life and the microorganism saved higher concentrations of polyhydroxybutyrate. The selected fermentation conditions are closely related to those prevalent in large-scale bioreactors and offer the possibility of achieving high biomass titles with high shelf life at a reduced costs, due to the complete use of a carbon source at low aeration of a low cost raw material as DL-malic acid mixture in comparison with the L-malic acid stereoisomer.

  9. Fourier transform infrared spectroscopic characterisation of heavy metal-induced metabolic changes in the plant-associated soil bacterium Azospirillum brasilense Sp7

    Science.gov (United States)

    Kamnev, A. A.; Antonyuk, L. P.; Tugarova, A. V.; Tarantilis, P. A.; Polissiou, M. G.; Gardiner, P. H. E.

    2002-06-01

    Structural and compositional features of whole cells of the plant-growth-promoting rhizobacterium Azospirillum brasilense Sp7 under standard and heavy metal-stressed conditions are analysed using Fourier transform infrared (FTIR) spectroscopy and compared with the FT-Raman spectroscopic data obtained previously [J. Mol. Struct. 563-564 (2001) 199]. The structural spectroscopic information is considered together with inductively coupled plasma-mass spectrometric (ICP-MS) analytical data on the content of the heavy metal cations (Co 2+, Cu 2+ and Zn 2+) in the bacterial cells. As a bacterial response to heavy metal stress, all the three metals, being taken up by bacterial cells from the culture medium (0.2 mM) in significant amounts (ca. 0.12, 0.48 and 4.2 mg per gram of dry biomass for Co, Cu and Zn, respectively), are shown to induce essential metabolic changes in the bacterium revealed in the spectra, including the accumulation of polyester compounds in bacterial cells and their enhanced hydration affecting certain IR vibrational modes of functional groups involved.

  10. Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus.

    Science.gov (United States)

    Amavizca, Edgar; Bashan, Yoav; Ryu, Choong-Min; Farag, Mohamed A; Bebout, Brad M; de-Bashan, Luz E

    2017-02-01

    Remote effects (occurring without physical contact) of two plant growth-promoting bacteria (PGPB) Azospirillum brasilense Cd and Bacilus pumilus ES4 on growth of the green microalga Chlorella sorokiniana UTEX 2714 were studied. The two PGPB remotely enhanced the growth of the microalga, up to six-fold, and its cell volume by about three-fold. In addition to phenotypic changes, both bacteria remotely induced increases in the amounts of total lipids, total carbohydrates, and chlorophyll a in the cells of the microalga, indicating an alteration of the microalga's physiology. The two bacteria produced large amounts of volatile compounds, including CO2, and the known plant growth-promoting volatile 2,3-butanediol and acetoin. Several other volatiles having biological functions in other organisms, as well as numerous volatile compounds with undefined biological roles, were detected. Together, these bacteria-derived volatiles can positively affect growth and metabolic parameters in green microalgae without physical attachment of the bacteria to the microalgae. This is a new paradigm on how PGPB promote growth of microalgae which may serve to improve performance of Chlorella spp. for biotechnological applications.

  11. Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus

    Science.gov (United States)

    Amavizca, Edgar; Bashan, Yoav; Ryu, Choong-Min; Farag, Mohamed A.; Bebout, Brad M.; de-Bashan, Luz E.

    2017-01-01

    Remote effects (occurring without physical contact) of two plant growth-promoting bacteria (PGPB) Azospirillum brasilense Cd and Bacilus pumilus ES4 on growth of the green microalga Chlorella sorokiniana UTEX 2714 were studied. The two PGPB remotely enhanced the growth of the microalga, up to six-fold, and its cell volume by about three-fold. In addition to phenotypic changes, both bacteria remotely induced increases in the amounts of total lipids, total carbohydrates, and chlorophyll a in the cells of the microalga, indicating an alteration of the microalga’s physiology. The two bacteria produced large amounts of volatile compounds, including CO2, and the known plant growth-promoting volatile 2,3-butanediol and acetoin. Several other volatiles having biological functions in other organisms, as well as numerous volatile compounds with undefined biological roles, were detected. Together, these bacteria-derived volatiles can positively affect growth and metabolic parameters in green microalgae without physical attachment of the bacteria to the microalgae. This is a new paradigm on how PGPB promote growth of microalgae which may serve to improve performance of Chlorella spp. for biotechnological applications. PMID:28145473

  12. ADP-ribosylation of dinitrogenase reductase in Azospirillum brasilense is regulated by AmtB-dependent membrane sequestration of DraG.

    Science.gov (United States)

    Huergo, Luciano F; Souza, Emanuel M; Araujo, Mariana S; Pedrosa, Fábio O; Chubatsu, Leda S; Steffens, Maria B R; Merrick, Mike

    2006-01-01

    Nitrogen fixation in some diazotrophic bacteria is regulated by mono-ADP-ribosylation of dinitrogenase reductase (NifH) that occurs in response to addition of ammonium to the extracellular medium. This process is mediated by dinitrogenase reductase ADP-ribosyltransferase (DraT) and reversed by dinitrogenase reductase glycohydrolase (DraG), but the means by which the activities of these enzymes are regulated are unknown. We have investigated the role of the P(II) proteins (GlnB and GlnZ), the ammonia channel protein AmtB and the cellular localization of DraG in the regulation of the NifH-modification process in Azospirillum brasilense. GlnB, GlnZ and DraG were all membrane-associated after an ammonium shock, and both this membrane sequestration and ADP-ribosylation of NifH were defective in an amtB mutant. We now propose a model in which membrane association of DraG after an ammonium shock creates a physical separation from its cytoplasmic substrate NifH thereby inhibiting ADP-ribosyl-removal. Our observations identify a novel role for an ammonia channel (Amt) protein in the regulation of bacterial nitrogen metabolism by mediating membrane sequestration of a protein other than a P(II) family member. They also suggest a model for control of ADP-ribosylation that is likely to be applicable to all diazotrophs that exhibit such post-translational regulation of nitrogenase.

  13. Relationship between in vitro enhanced nitrogenase activity of an Azospirillum brasilense Sp7 mutant and its growth-promoting activities in situ.

    Science.gov (United States)

    de Campos, Samanta Bolzan; Roesch, Luiz Fernando Wurdig; Zanettini, Maria Helena Bodanese; Passaglia, Luciane Maria Pereira

    2006-07-01

    In this work, we further analyzed an Azospirillum brasilense Sp7 mutant (Sp7::Tn5-33) showing a pleiotrophic phenotype due to a Tn5 insertion into an open reading frame of 840 bp (orf280). The deduced amino acid sequence of this region has high similarity to a family of universal stress proteins. Because the most interesting property exhibited by the Sp7::Tn5-33 mutant was an enhanced in vitro nitrogen fixation activity, we addressed the question of whether it could benefit the host plant. We found that the increased nitrogenase activity at the free-living state of the mutant bacterium was correlated with an increased production of the nitrogenase reductase protein (NifH), in amounts approximately 1.5 times higher than the wild type. The mutant strain exhibited the same level of auxin production and the same colonization pattern of wheat roots as the wild type. We also observed that Sp7::Tn5-33 increased the total plant dry weight, although the N content did not differ significantly between wheat plants inoculated with mutant or wild-type strains.

  14. [Changes in Cell Surface Properties and Biofilm Formation Efficiency in Azospirillum brasilense Sp245 Mutants in the Putative Genes of Lipid Metabolism mmsB1 and fabG1].

    Science.gov (United States)

    Shumilova, E; Shelud'ko, A V; Filip'echeva, Yu A; Evstigneeva, S S; Ponomareva, E G; Petrova, L P; Katsy, E I

    2016-01-01

    The previously obtained insertion mutants ofAzospirillum brasilense Sp245 in the genes mmsBl and fabG1 (strains SK039 and Sp245.1610, respectively) were characterized by impaired flagellation and motility. The putative products of expression of these genes are 3-hydroxyisobutyrate dehydrogenase and 3-oxoacyl-[acyl-carrier protein] reductase, respectively. In the present work, A. brasilense- Sp245 strains SK039 and Sp245.1610 were found to have differences in the content of 3-hydroxyhexadecanoic, hexadecanoic, 3-hydroxytetradecanoic, hexadecenoic, octadecenoic, and nonadecanoic acids in their lipopolysaccharide prepa- rations, as well as in cell hydrophobicity and hemagglutination activity and dynamics of cell aggregation, in biomass amount, and in the relative content of lipopolysaccharide antigens in mature biofilms formed on hydrophilic or hydrophobic surfaces.

  15. Análise da interação de Azospirillum brasilense FP2 com raízes de milho (Zea mays) por qPCR, microscopia eletrônica e proteômica

    OpenAIRE

    Faleiro, Alexandro Cézar

    2014-01-01

    Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-Graduação em Recursos Genéticos Vegetais, Florianópolis, 2014. As rizobactérias promotoras de crescimento vegetal (RPCV) são um grupo de micro-organismos benéficos às plantas devido à sua capacidade de estimular o crescimento vegetal através de vários processos. Uma das mais importantes e estudadas RPCV é a Azospirillum brasilense, uma diazotrófica associada com importantes culturas tais...

  16. Consórcio de milho e Urochloa ruziziensis e inoculação com Azospirillum brasilense e seu efeito residual associado à adubação nitrogenada em feijoeiro de inverno

    OpenAIRE

    Sabundjian,Michelle Traete

    2016-01-01

    O avanço de estudos das interações entre bactérias capazes de fixar o nitrogênio atmosférico em culturas agrícolas pode ajudar a definir estratégias para aumentar a eficiência do uso e diminuir a dependência do nitrogênio mineral. Assim, o objetivo do trabalho foi avaliar o efeito da inoculação de Azospirillum brasilense sobre o desenvolvimento e a produtividade do milho solteiro ou consorciado com Urochloa ruziziensis e o efeito residual deste consórcio, associados ao manejo de nitrogênio no...

  17. Coberturas vegetais, doses de nitrogênio e inoculação de sementes com Azospirillum brasilense em arroz de terras altas no sistema plantio direto

    Directory of Open Access Journals (Sweden)

    Douglas de Castilho Gitti

    2012-01-01

    Full Text Available O arroz é importante fonte de energia e proteínas para a população mundial, principalmente na Ásia e Oceania. No Brasil, juntamente com o feijão, constitui a base da alimentação. Tecnologias sustentáveis que reduzam custos da produção e aumentem a produtividade do arroz podem garantir seu suprimento em períodos de alta demanda. O objetivo deste trabalho foi avaliar diferentes coberturas vegetais antecessoras (milheto [Pennisetum americanum], crotalária [Crotalaria juncea], guandu [Cajanus cajan], braquiária [Brachiaria ruziziensis], milheto + crotalária e milheto + guandu, doses de nitrogênio (N em cobertura (0, 40, 80 e 120 kg ha-1 e o efeito da inoculação de sementes com Azospirillum brasilense no arroz de terras altas em sistema plantio direto no desenvolvimento e na produtividade. Utilizou-se o delineamento experimental em blocos casualizados em esquema fatorial 6x4x2 com quatro repetições. O estudo foi desenvolvido em um Latossolo Vermelho, em Selvíria, Estado do Mato Grosso do Sul, Brasil, em 2011/2012. O cultivo do guandu antecedendo o arroz proporcionou maior produtividade do arroz somente em comparação a B. ruziziensi. A produtividade do arroz em função das doses de N em cobertura se ajustou a uma função quadrática. Não houve influência da inoculação de sementes com A. brasilense sobre a produtividade do arroz, porém houve interação entre a inoculação e as coberturas vegetais sobre o teor de N foliar, número de panículas por m², matéria seca de plantas de arroz e a massa de cem grãos.

  18. 巴西固氮螺菌Yu62在玉米根的定植%Colonization Pattern of Azospirillum brasilense Yu62 on Maize Roots

    Institute of Scientific and Technical Information of China (English)

    刘元; 陈三凤; 李季伦

    2003-01-01

    Plasmid pVK1001 which carried the gfp gene of GFPmut2, a mutant of GFP, was introduced into Azospirillum brasilense Yu62 by electroporation. Maize seedlings were inoculated with the GFP-labelled bacteria and grown gnotobiotically in flask with semi-solid agar medium. Observations were performed with confocal laser scanning microscopy (CLSM) and electron microscopy, respectively, at 8 d and 12 d after inoculation. Confocal laser scanning microscopy showed that A. brasilense Yu62 could penetrate into the cortex tissue, colonizing in the intercellular spaces of the parenchyma cells of the cortex tissue. Transmission and scanning electron microscopy (TEM) showed that the majority of the bacteria colonized on the root surface and only a minority of them resided in the root interior.%将GFPmut2质粒中的gfp基因 (编码绿色荧光蛋白)克隆到载体pVK100中,构建成重组质粒pVK1001.将pVK1001通过电转化方法导入到联合固氮菌巴西固氮螺菌Yu62中,获得GFP标记的巴西固氮螺菌Yu62菌株.用标记菌株接种限菌培养条件下生长的玉米(农大3318)幼苗,在接种后8 d、12 d,用激光共聚焦扫描显微镜进行观测,结果表明巴西固氮螺菌Yu62菌株能定植于玉米根部皮层的薄壁细胞间隙.用扫描电镜和超薄切片电镜观察表明,大多数细菌主要定植于根表,少数菌可进入玉米根组织内.

  19. Coberturas vegetais, doses de nitrogênio e inoculação de sementes com Azospirillum brasilense em arroz de terras altas no sistema plantio direto Cover crops, nitrogen rates and seeds inoculation with Azospirillum brasilense in upland rice under no-tillage

    Directory of Open Access Journals (Sweden)

    Douglas de Castilho Gitti

    2013-01-01

    Full Text Available O arroz é importante fonte de energia e proteínas para a população mundial, principalmente na Ásia e Oceania. No Brasil, juntamente com o feijão, constitui a base da alimentação. Tecnologias sustentáveis que reduzam custos da produção e aumentem a produtividade do arroz podem garantir seu suprimento em períodos de alta demanda. O objetivo deste trabalho foi avaliar diferentes coberturas vegetais antecessoras (milheto [Pennisetum americanum], crotalária [Crotalaria juncea], guandu [Cajanus cajan], braquiária [Brachiaria ruziziensis], milheto + crotalária e milheto + guandu, doses de nitrogênio (N em cobertura (0, 40, 80 e 120 kg ha-1 e o efeito da inoculação de sementes com Azospirillum brasilense no arroz de terras altas em sistema plantio direto no desenvolvimento e na produtividade. Utilizou-se o delineamento experimental em blocos casualizados em esquema fatorial 6x4x2 com quatro repetições. O estudo foi desenvolvido em um Latossolo Vermelho, em Selvíria, Estado do Mato Grosso do Sul, Brasil, em 2011/2012. O cultivo do guandu antecedendo o arroz proporcionou maior produtividade do arroz somente em comparação a B. ruziziensi. A produtividade do arroz em função das doses de N em cobertura se ajustou a uma função quadrática. Não houve influência da inoculação de sementes com A. brasilense sobre a produtividade do arroz, porém houve interação entre a inoculação e as coberturas vegetais sobre o teor de N foliar, número de panículas por m², matéria seca de plantas de arroz e a massa de cem grãos.Rice is an important source of energy and protein for the world population, mainly in Asia and Oceania. In Brazil, together with common beans, it is the population's basic food. Sustainable technologies that reduce rice production costs and increase productivity can warrant its supply on period of high demand. The objective of this study was to evaluate different preceding cover crops (millet [Pennisetum americanum

  20. Coberturas vegetais, doses de nitrogênio e inoculação de sementes com Azospirillum brasilense em arroz de terras altas no sistema plantio direto Cover crops, nitrogen rates and seeds inoculation with Azospirillum brasilense in upland rice under no-tillage

    Directory of Open Access Journals (Sweden)

    Douglas de Castilho Gitti

    2012-01-01

    Full Text Available O arroz é importante fonte de energia e proteínas para a população mundial, principalmente na Ásia e Oceania. No Brasil, juntamente com o feijão, constitui a base da alimentação. Tecnologias sustentáveis que reduzam custos da produção e aumentem a produtividade do arroz podem garantir seu suprimento em períodos de alta demanda. O objetivo deste trabalho foi avaliar diferentes coberturas vegetais antecessoras (milheto [Pennisetum americanum], crotalária [Crotalaria juncea], guandu [Cajanus cajan], braquiária [Brachiaria ruziziensis], milheto + crotalária e milheto + guandu, doses de nitrogênio (N em cobertura (0, 40, 80 e 120 kg ha-1 e o efeito da inoculação de sementes com Azospirillum brasilense no arroz de terras altas em sistema plantio direto no desenvolvimento e na produtividade. Utilizou-se o delineamento experimental em blocos casualizados em esquema fatorial 6x4x2 com quatro repetições. O estudo foi desenvolvido em um Latossolo Vermelho, em Selvíria, Estado do Mato Grosso do Sul, Brasil, em 2011/2012. O cultivo do guandu antecedendo o arroz proporcionou maior produtividade do arroz somente em comparação a B. ruziziensi. A produtividade do arroz em função das doses de N em cobertura se ajustou a uma função quadrática. Não houve influência da inoculação de sementes com A. brasilense sobre a produtividade do arroz, porém houve interação entre a inoculação e as coberturas vegetais sobre o teor de N foliar, número de panículas por m², matéria seca de plantas de arroz e a massa de cem grãos.Rice is an important source of energy and protein for the world population, mainly in Asia and Oceania. In Brazil, together with common beans, it is the population's basic food. Sustainable technologies that reduce rice production costs and increase productivity can warrant its supply on period of high demand. The objective of this study was to evaluate different preceding cover crops (millet [Pennisetum americanum

  1. Growth of quailbush in acidic, metalliferous desert mine tailings: effect of Azospirillum brasilense Sp6 on biomass production and rhizosphere community structure.

    Science.gov (United States)

    de-Bashan, Luz E; Hernandez, Juan-Pablo; Nelson, Karis N; Bashan, Yoav; Maier, Raina M

    2010-11-01

    Mine tailing deposits in semiarid and arid environments frequently remain devoid of vegetation due to the toxicity of the substrate and the absence of a diverse soil microbial community capable of supporting seed germination and plant growth. The contribution of the plant growth promoting bacterium (PGPB) Azospirillum brasilense Sp6 to the growth of quailbush in compost-amended, moderately acidic, high-metal content mine tailings using an irrigation-based reclamation strategy was examined along with its influence on the rhizosphere bacterial community. Sp6 inoculation resulted in a significant (2.2-fold) increase in plant biomass production. The data suggest that the inoculum successfully colonized the root surface and persisted throughout the 60-day experiment in both the rhizosphere, as demonstrated by excision and sequencing of the appropriate denaturing gradient gel electrophoresis (DGGE) band, and the rhizoplane, as indicated by fluorescent in situ hybridization of root surfaces. Changes in rhizosphere community structure in response to Sp6 inoculation were evaluated after 15, 30, and 60 days using DGGE analysis of 16S rRNA polymerase chain reaction amplicons. A comparison of DGGE profiles using canonical correspondence analysis revealed a significant treatment effect (Sp6-inoculated vs. uninoculated plants vs. unplanted) on bacterial community structure at 15, 30, and 60 days (p < 0.05). These data indicate that in an extremely stressed environment such as acid mine tailings, an inoculated plant growth promoting bacterium not only can persist and stimulate plant growth but also can directly or indirectly influence rhizobacterial community development.

  2. Structural homologues P(II) and P(Z) of Azospirillum brasilense provide intracellular signalling for selective regulation of various nitrogen-dependent functions.

    Science.gov (United States)

    de Zamaroczy, M

    1998-07-01

    P(II) (glnB) is a signal transduction protein that in Azospirillum brasilense is specifically required for nitrogen fixation. Little is known about whether and how its homologue P(Z) (glnZ) participates in the regulation of cellular functions. In this study, we have shown the regulatory action of the two proteins by analysing the relevant single and double null-mutant strains. The transcription of glnZ is monocistronic, and it starts mainly from a sigma54-dependent promoter, activated by NtrC. glnZ expression is dependent on the ntr system, even under conditions of nitrogen excess, and is greatly enhanced in the presence of aspartate. P(Z) is uridylylated in response to nitrogen limitation, like P(II), although different amounts of the two proteins are synthesized. P(II) is required for the dephosphorylation of NtrC. Thus, in the absence of P(II), the repression of nitrate assimilation is not promoted, which, in turn, leads to a high rate of ammonium excretion. Unexpectedly, P(II) and P(Z) proteins are not essential for the reversible modification of glutamine synthetase. (Methyl)ammonium transport into the cell is negatively regulated by P(Z). The growth of a double-mutant strain (glnB::kan; glnZ::omega) is drastically disabled, although wild-type growth is restored by complementation with either glnB or glnZ. We conclude that P(II) and P(Z), despite their structural similarity, are involved in different regulatory processes, except for that required for cell growth.

  3. Monitoring of cobalt(II) uptake and transformation in cells of the plant-associated soil bacterium Azospirillum brasilense using emission Mössbauer spectroscopy.

    Science.gov (United States)

    Kamnev, Alexander A; Antonyuk, Lyudmila P; Kulikov, Leonid A; Perfiliev, Yury D

    2004-08-01

    Interaction of cobalt(II) at micromolar concentrations with live cells of the plant-growth-promoting rhizobacterium Azospirillum brasilense (strain Sp245) and further transformations of the metal cation were monitored using 57Co emission Mössbauer spectroscopy (EMS). Cell suspensions of the bacterial culture (2.4 x 10(8) cells ml(-1)) were doped with radioactive 57CoCl2 (1 mCi; final concentration 2 x 10(-6) M 57Co2+), kept under physiological conditions for various periods of time (from 2 min up to 1 hour) and then rapidly frozen in liquid nitrogen. Analysis of emission Mössbauer spectra of the frozen aqueous suspensions of the bacterial cell samples shows that the primary absorption of cobalt(II) at micromolar concentrations by the bacterial cells is rapid and virtually complete, giving at least two major forms of cobalt(II) species bound to the cells. Within an hour, the metal is involved in further metabolic transformations reflected by changes occurring in the spectra. The Mössbauer parameters calculated from the EMS data by statistical treatment were different for suspensions of live and dead (thermally killed) bacterial cells that had been in contact with 57Co2+ for 1 h, as well as for the cell-free culture medium containing the same concentration of 57Co2+. Chemical after-effects of the nuclear transition (57Co --> 5 7Fe), which provide additional information on the chemical environment of metal ions, are also considered. The data presented demonstrate that EMS is a valuable tool for monitoring the chemical state of cobalt species in biological matter providing information at the atomic level in the course of its uptake and/or metabolic transformations.

  4. [Insertional mutation in the AZOBR_p60120 gene is accompanied by defects in the synthesis of lipopolysaccharide and calcofluor-binding polysaccharides in the bacterium Azospirillum brasilense Sp245].

    Science.gov (United States)

    Katsy, E I; Prilipov, A G

    2015-03-01

    In the bacterium Azospirillum brasilense Sp245, extracellular calcofluor-binding polysaccharides (Cal+ phenotype) and two types of lipopolysaccharides, LPSI and LPSII, were previously identified. These lipopolysaccharides share the same repeating O-polysaccharide unit but have different antigenic structures and different charges of their O-polysaccharides and/or core oligosaccharides. Several dozens of predicted genes involved in the biosynthesis of polysaccharides have been localized in the AZOBR_p6 plasmid of strain Sp245 (GenBank accession no. HE577333). In the present work, it was demonstrated that an artificial transposon Omegon-Km had inserted into the central region of the AZOBR_p60120 gene in the A. brasilense Sp245 LPSI- Cal- KM252 mutant. In A. brasilense strain Sp245, this plasmid gene encodes a putative glycosyltransferase containing conserved domains characteristic of the enzymes participating in the synthesis of O-polysaccharides and capsular polysaccharides (accession no. YP004987664). In mutant KM252, a respective predicted protein is expected to be completely inactivated. As a result of the analysis of the EcoRI fragment of the AZOBR_p6 plasmid, encompassing the AZOBR_p60120 gene and a number of other loci, novel data on the structure of AZOBR_p6 were obtained: an approximately 5-kb gap (GenBank accession no. KM189439) was closed in the nucleotide sequence of this plasmid.

  5. Metabolic profiling of two maize (Zea mays L.) inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense

    Science.gov (United States)

    Brusamarello-Santos, Liziane Cristina; Gilard, Françoise; Brulé, Lenaïg; Quilleré, Isabelle; Gourion, Benjamin; Ratet, Pascal; Maltempi de Souza, Emanuel; Lea, Peter J.; Hirel, Bertrand

    2017-01-01

    Maize roots can be colonized by free-living atmospheric nitrogen (N2)-fixing bacteria (diazotrophs). However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2), already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+). The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts. Moreover, a number

  6. Comparación entre coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja Comparison of coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop

    Directory of Open Access Journals (Sweden)

    S. Benintende

    2010-12-01

    Full Text Available El objetivo fue comparar la coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense con relación a la inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja y su rendimiento. Se realizaron dos experimentos en lotes en los que nunca se cultivó soja en años con diferentes condiciones hídricas. Los tratamientos fueron: testigo (sin inoculación, coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum. La nodulación en ambos experimentos fue superior en la coinoculación, aunque hubo diferencias significativas en el experimento con déficit hídrico. Los rendimientos, en ambos experimentos, presentaron valores medios superiores en la coinoculación, aunque sin diferencias significativas. La acumulación de biomasa aérea y N fue mayor en el año húmedo. El porcentaje de N derivado de la fijación biológica de N en el cultivo en el año seco fue de 26% para la inoculación simple y de 30% en la coinoculación, mientras que en el año húmedo este porcentaje fue de 41 y 47% respectivamente. A partir de estos resultados se concluye que existe un efecto de estimulación del crecimiento del cultivo de soja por la coinoculación. Sin embargo, es necesario continuar con estudios sobre este tema.The aim of this study was to compare coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop in field conditions. Two field tests were carried out for two years with different hydric conditions in areas not previously cultivated with soybean. The treatments were: control, coinoculation using Bradyrhizobium japonicum + Azospirillum brasilense and inoculation with Bradyrhizobium japonicum. Nodulation in both experiments was increased by dual inoculation, although significant differences were

  7. CO-INOCULAÇÃO E MODOS DE APLICAÇÃO DE Bradyrhizobium japonicum e Azospirillum brasilense E ADUBAÇÃO NITROGENADA NA NODULAÇÃO DAS PLANTAS E RENDIMENTO DA CULTURA DA SOJA

    Directory of Open Access Journals (Sweden)

    Alessandro Lucca Braccini

    2016-03-01

    Full Text Available As new cultivation techniques arise, crops require the development of new technologies and management practices that aim higher yields with lower production cost. Thus, the objective of this study was to evaluate the efficiency of co-inoculation and different ways of application of Bradyrhizobium japonicum and Azospirillum brasilense and nitrogen fertilization on nodulation and yield of soybean. The experiment was established in November 2013 in an area located in the Iguatemi Experimental Farm, of State University of Maringá (UEM. The design was a randomized block with four replications. The treatments consisted of a control, fertilization with nitrogen (200 kg N ha-1, standard inoculation in seeds (liquid and peat inoculant, co-inoculation on seed and co-inoculation at sowing, using different doses of inoculant B. japonicum and A. brasilense. The following determinations were carried out in the field and in the laboratory: number of nodes at flowering, mass of nodules at flowering, the shoot dry mass, nitrogen content in the shoot, grain nitrogen content, number of pods/plant, thousand grain mass and yield. The results showed that inoculation through seed treatment with liquid inoculant presents an adequate option to farmers, in the conditions of this experiment. Nitrogen fertilization did not increase yield and yield components, being unnecessary.

  8. Características agronômicas do trigo em função de Azospirillum brasilense, ácidos húmicos e nitrogênio em casa de vegetação

    Directory of Open Access Journals (Sweden)

    Luan F. O. S. Rodrigues

    2014-01-01

    Full Text Available Objetivou-se, neste estudo, verificar o efeito do uso combinado de Azospirillum brasilense, ácidos húmicos e diferentes doses de nitrogênio nas características agronômicas do trigo. Os experimentos foram instalados no dia 21 de maio de 2011 e a colheita foi procedida no dia 13 de setembro de 2011. O delineamento experimental adotado foi o de blocos casualizados em esquema fatorial 4 x 4, com quatro repetições. O primeiro fator se refere à combinação da estirpe Ab-V5 de A. brasilense com os ácidos húmicos, sendo utilizados os seguintes tratamentos: controle; Ab-V5; ácido húmico e Ab-V5+ácido húmico e o segundo fator se refere às doses de nitrogênio (0, 30, 60 e 90 kg ha-1 N. A maioria das variáveis em análise apresentou efeito linear positivo dos tratamentos em função da adubação nitrogenada sendo que apenas para a variável massa de cem grãos este efeito foi linear negativo. Não houve diferença significativa entre as variâncias, na produção de grãos. Pôde-se concluir que o uso combinado de Ab-V5 + ácido húmico promove principalmente a produção de matéria seca de folhas e não é responsivo à adubação nitrogenada para os demais componentes produtivos em estudo nem para a produtividade dos grãos.

  9. Aspectos morfofisiológicos de plantas de milho e bioquímico do solo em resposta à adubação nitrogenada e à inoculação com Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    Tâmara Prado de Morais

    2015-12-01

    Full Text Available RESUMO O nitrogênio (N é um macronutriente que aumenta a produção de grãos na cultura do milho e o seu manejo adequado torna-se indispensável como proposta de uma agricultura sustentável, incluindo o uso de fontes alternativas como bactérias fixadoras de N2. Os objetivos deste trabalho foram avaliar a resposta morfofisiológica de plantas de milho e a atividade de enzimas no solo (urease e fosfatase decorrente da adubação N mineral e da inoculação com Azospirillum brasilense. Dois ensaios foram realizados em condições de casa de vegetação com solo de cerrado. O delineamento estatístico adotado foi o de blocos ao acaso com os tratamentos distribuídos em esquema fatorial 3 x 3, sendo o primeiro fator correspondente a doses de N (0; 100 e 200 kg ha-1 e o segundo, a doses de um inoculante líquido à base de A. brasilense aplicado via sementes (0; 100 e 200 mL ha-1. Ao final desse experimento, outro ensaio foi realizado para verificar o possível efeito residual da inoculação e da adubação nitrogenada. A adição de fertilizantes nitrogenados promoveu maior desenvolvimento das plantas de milho, elevou os teores de clorofila e de nutrientes. Houve aumento da atividade das enzimas relacionadas à disponibilização de amônio e fósforo inorgânico na rizosfera. Além disso, o nitrogênio apresentou efeito residual no desenvolvimento das plantas na semeadura subsequente. A dose de 200 mL ha-1 de A. brasilense associada à dose de 200 kg ha-1 de N aumentou a resposta fisiológica da cultura.

  10. Isolation and Identification of Azospirillum brasilense Excreting High Concentration of IAA and Ammonium%高产吲哚乙酸及高泌氨巴西固氨螺菌的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    王继雯; 谢宝恩; 周伏忠; 陈晓飞

    2009-01-01

    目的:从玉米根际和土壤中分离具有高产吲哚乙酸较强的泌氨能力的巴西固氮螺菌.方法:分别通过半固体NFb培养基、CR培养基、LB培养基分离培养固氮菌株,并经过一系列菌落菌体形态特征、生理生化特性和16S rDNA序列测定等试验对其进行鉴定.结果:经分离纯化获得10株固氮菌,并鉴定均为巴西固氮螺菌(Azospirillum brasilense),其中菌株R7在甘油半固体培养基上能分泌约14mmol/L的氨,在添加了色氨酸的培养基中能够合成58.8μg/ml的吲哚-3-乙酸(IAA).结论:成功筛选得到一株既高产吲哚乙酸又有较强的泌氨能力的巴西固氮螺菌.

  11. Effects of over-expression of the regulatory enzymes DraT and DraG on the ammonium-dependent post-translational regulation of nitrogenase reductase in Azospirillum brasilense.

    Science.gov (United States)

    Huergo, Luciano F; Souza, Emanuel M; Steffens, Maria B R; Yates, M Geoffrey; Pedrosa, Fábio O; Chubatsu, Leda S

    2005-03-01

    Nitrogen fixation in Azospirillum brasilense is regulated at transcriptional and post-translational levels. Post-translational control occurs through the reversible ADP-ribosylation of dinitrogenase reductase (Fe Protein), mediated by the dinitrogenase reductase ADP-ribosyltransferase (DraT) and dinitrogenase reductase glycohydrolase (DraG). Although the DraT and DraG activities are regulated in vivo, the molecules responsible for such regulation remain unknown. We have constructed broad-host-range plasmids capable of over-expressing, upon IPTG induction, the regulatory enzymes DraT and DraG as six-histidine-N-terminal fused proteins (His). Both DraT-His and DraG-His are functional in vivo. We have analyzed the effects of DraT-His and DraG-His over-expression on the post-translational modification of Fe Protein. The DraT-His over-expression led to Fe Protein modification in the absence of ammonium addition, while cells over-expressing DraG-His showed only partial ADP-ribosylation of Fe Protein by adding ammonium. These results suggest that both DraT-His and DraG-His lose their regulation upon over-expression, possible by titrating out negative regulators.

  12. Functional analysis of the GAF domain of NifA in Azospirillum brasilense: effects of Tyr-->Phe mutations on NifA and its interaction with GlnB.

    Science.gov (United States)

    Chen, Sanfeng; Liu, Li; Zhou, Xiaoyu; Elmerich, Claudine; Li, Ji-Lun

    2005-06-01

    Regulation of NifA activity in Azospirillum brasilense depends on GlnB (a PII protein), and it was previously reported that the target of GlnB activity is the N-terminal domain of NifA. Furthermore, mutation of the Tyr residue at position 18 in the N-terminal domain resulted in a NifA protein that did not require GlnB for activity under nitrogen fixation conditions. We report here that a NifA double mutant in which the Tyr residues at positions 18 and 53 of NifA N-were simultaneously replaced by Phe (NifA-Y1853F) displays high nitrogenase activity, which is still regulatable by ammonia, but not by GlnB. The yeast two-hybrid technique was used to investigate whether GlnB can physically interact with wild-type and mutant NifA proteins. GlnB was found to interact directly with the N-terminal GAF domain of wild-type NifA, but not with its central or C-terminal domain. GlnB could still bind to the single NifA mutants Y18F and Y53F. In contrast, no interaction was detected between GlnB and the double mutant NifA-Y18/53F or between GlnB and NifA-Y43.

  13. Effects of saline tolerant Azospirillum species on the growth parameters of mangrove seedlings.

    Science.gov (United States)

    Ravikumar, S; Ignatiammal, S Thadedus Maria; Gnanadesigan, M; Kalaiarasi, A

    2012-09-01

    Five species of Azospirillum isolated from Manakkudi mangrove ecosystem were subjected for their efficiency to find out their growth parameters potential for the successful establishment of mangrove seedlings. Of the isolated five Azospirillum species, Azospirillum lipoferum (60%) was found to be the dominant one. But the level of maximum indole acetic acid (IAA) production (19.8 mg.ml(-1)) and nitrogen fixation (5.9 C2H2hr1) was identified with A. brasilense. Further, A. brasilense showed significant (p brasilense also showed significant (p Azospirillum brasilense was found better for the growth of Avicennia officinalis and Ceriops decandra seedlings, but Azospirillum irakense was found better for Rhizophora apiculata seedlings.

  14. 固氮螺菌 (Azosprillum brasilense)NO40在红壤性水稻土上的接种效应%Effects of inoculation with tropical rice associative N2-fixing bacteria Azospirillum brasilense NO40 in the red paddy soil

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Effects of inoculation with Azospirillum brasilense NO40 isolatedfrom Egypt on rice growth and yields under different nitrogen fertilizer levels were investigated at Dongyang county of Zhejiang province. The results showed that the counts of colony grown on nitrogen-free medium isolated from rice rhizosphere and root were increased by inoculating with NO40 in contrast to the control, irrespective of the nitrogen fertilizer levels and rice growing periods. It was found that the plant height, chlorophyll content of the newly developed rice leaves were significantly increased after inoculating with NO40.%裂区设计田间试验结果表明,在3个供氮水平下接种巴西固氮螺菌(Azospirillumbrasilense)NO40,使分蘖期与灌浆期水稻根际及根内的固氮菌数量均比对照明显增加,而土体中的固氮菌数量未见明显变化;同时,接种该菌株可显著提高分蘖期与灌浆期水稻新展开叶的叶绿素含量及水稻株高,在低氮水平下使水稻单株干物重和单株籽粒产量有所提高。

  15. [The different functions of glnB and glnZ from Azospirillum brasilense YU62 in the control of nitrogen fixation].

    Science.gov (United States)

    Chen, S; Guan, Y; Ying, J; Li, Z; Wang, J; Li, J

    2001-10-01

    The glnB and glnZ genes of A. brasilense have 70% homology at nucleotide sequence. glnB is located in a 3.7 kb Eco RI+ PstI fragment and glnZ is located in a 3.7 kb SalI fragment. Both glnB and glnZ genes were mutagenized by Kmr cassette insertions and glnB- and glnZ- mutants were obtained. glnB- mutant did not have any nitrogenase activity, while glnZ- mutant still has nitrogenase activity. The coding regions of glnB and glnZ were cloned into pVK100 vectors and recombinant plasmids pVK-II and pVK-Z were obtained, respectively. The recombinant plasmids pVK-II and pVK-Z were introduced into glnB- and glnZ- to produce C-glnB and C-glnZ, respectively. C-glnB can restore nitrogenase activity and C-glnZ does not have effect on nitrogenase activity. When pVK-II and pVK-Z were introduced into A. brasilense Yu62 and draT-, respectively, the Yu62-II (containing pVK-II) and draT-II (containing pVK-II) have higher nitrogenase activity than that of wild type Yu62. In contrast, Yu62-Z (containing pVK-Z) and draT-Z (containing pVK-Z) has no effect on nitrogenase activity. The nifA(-)-II (containg pVK-II) and nifA(-)-Z (containing pVK-Z) still have no nitrogenase activity.

  16. Implicações da inoculação com Azospirillum brasilense e de níveis de N em milho transgênico no cerrado

    OpenAIRE

    Queiroz, Isabel Dayane de Sousa

    2014-01-01

    O milho é um dos cereais de maior importância econômica e nutricional. Sua produção está diretamente relacionada ao fornecimento adequado de nutrientes, especialmente o nitrogênio, que pode ser fornecido pela adubação ou pela fixação biológica de nitrogênio. Um dos micro-organismos fixadores de nitrogênio mais importantes nas culturas gramíneas é o Azospirillum sp., que além de fornecer nitrogênio, melhora o ambiente radicular, aumentando a absorção de água e nutrientes pela planta. Objetivou...

  17. Efecto de la coinmovilización de la microalga Chlorella sorokiniana con la bacteria promotora de crecimiento vegetal Azospirillum brasilense, en el crecimiento poblacional microalgal y capacidad de remoción de amonio como uso potencial en el tratamiento..

    Directory of Open Access Journals (Sweden)

    Guillot Monroy Gabriel

    2006-06-01

    Full Text Available Se desarrollaron dos experimentos de coinmovilización de la microalga Chlorella sorokiniana con la bacteria promotora
    de crecimiento vegetal (PGPB Azospirillum brasilense en esferas de alginato, para evaluar la influencia de la bacteria en el crecimiento poblacional y en la capacidad de remoción de amonio de la microalga; con el fin de establecer el potencial que pueda tener la coinmovilización de estos microorganismos en el tratamiento terciario de aguas residuales. Los resultados revelan un significativo incremento de la población cuando C.
    sorokiniana es coinmovilizada con A. brasilense, aunque no se observó un efecto significativo en la remoción de amonio. Los resultados obtenidos en el presente trabajo, reflejan la importancia de avanzar en el estudio de un sistema coinmovilizado altamente eficiente para ser utilizado en tratamiento de aguas residuales. La coinmovilización
    de C. sorokiniana con A. brasilense, muestra la habilidad de esta bacteria para influir de manera positiva en el crecimiento y fisiología de la microalga y por lo tanto la remoción de amonio puede ser incrementada.

  18. Wheat root colonization and nitrogenase activity by Azospirillum isolates from crop plants in Korea.

    Science.gov (United States)

    Kim, Chungwoo; Kecskés, Mihály L; Deaker, Rosalind J; Gilchrist, Kate; New, Peter B; Kennedy, Ivan R; Kim, Seunghwan; Sa, Tongmin

    2005-11-01

    Nitrogen-fixing bacteria were isolated from the rhizosphere of different crops of Korea. A total of 16 isolates were selected and characterized. Thirteen of the isolates produced characteristics similar to those of the reference strains of Azospirillum, and the remaining 3 isolates were found to be Enterobacter spp. The isolates could be categorized into 3 groups based on their ARDRA patterns, and the first 2 groups comprised Azospirillum brasilense and Azospirillum lipoferum. The acetylene reduction activity (ARA) of these isolates was determined for free cultures and in association with wheat roots. There was no correlation between pure culture and plant-associated nitrogenase activity of the different strains. The isolates that showed higher nitrogenase activities in association with wheat roots in each group were selected and sequenced. Isolates of Azospirillum brasilense CW301, Azospirillum brasilense CW903, and Azospirillum lipoferum CW1503 were selected to study colonization in association with wheat roots. We observed higher expression of beta-galactosidase activity in A. brasilense strains than in A. lipoferum strains, which could be attributed to their higher population in association with wheat roots. All strains tested colonized and exhibited the strongest beta-galactosidase activity at the sites of lateral roots emergence.

  19. Cloning and Sequencing of glnZ and the other Genes Clustered Around glnZ from Azospirillum brasilense Yu62%巴西固氮螺菌Yu62 glnZ基因及其相邻基因的克隆和序列分析

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 杨红; 李季伦

    2001-01-01

    通过原位杂交从巴西固氮螺菌(Azospirillum brasilense)Yu62的基因组文库中获得glnZ基因的阳性克隆,对该阳性克隆进行亚克隆和序列分析,结果表明glnZ基因位于3.7 kb的SaLI片段上。glnZ基因编码区长336 bp,编码的产物是Pz蛋白,由112个氨基酸组成,分子量为1.12 kD。用Blastax 软件对Pz蛋白的氨基酸序列在GenBank数据库中进行同源比较,结果表明Pz蛋白与其它几种固氮菌及大肠杆菌的GlnK蛋白的同源性(identities)达66%以上;与 PII蛋白的同源性达64%以上。glnZ基因上游是部分ubiH-like 基因,与E.coli ubiH基因 (编码辅酶Q,ubiquinone)N-端有31%的同源性(identity)和50%相似性(similarity);glnZ基因下游是aat-like基因,与E.coli和 Bacillus subtilis aat基因(编码天冬氨酸氨基转移酶, aspartate aminotransferase)有同源性和相似性都为26% 和42%;aat-like基因下游是部分ftsK-like基因,与E.coli ftsK基因 (编码肽聚糖, peptidoglycan)N-端有42%同源性和56%相似性。这几个基因在GenBank中的登录号是AF279917。%The glnZ gene of A.brasilense was determined in a 3.7 kb SalI fragment,and ubiH-like,aat-like and ftsK-like genes were clustered around glnZ.A partial ubiH-like gene(required for ubiquinone synthesis)precedes glnZ and is transcribed from the opposite strand.The ubiH-likegene of A.brasilense has 31% identitiy and 50% similarity with ubiH gene of E.coli.The aat(aspartate aminotransferase)-like and ftsK(involved in peptidoglycan synthesis during cell septation )-like genes were found clustered downstream of glnZ.The aat-like gene of A.brasilense has 26% identity and 42% similarity with aat gene of E.coli. The ftsK-like gene of A.brasilense has 42% identity and 56% similarity with ftsKgene of E.coli.The glnZ gene encodes 112 amino acid-long Pz polypeptide, maping between positions 1057~1395bp.The amino acid sequence of Pz from A.brasilense is more than 66% identical to that

  20. INTERAÇÃO ENTRE BACTÉRIAS Azospirillum brasilense E Herbaspirillum seropedicae COM ADUBAÇÃO NITROGENADA, VISANDO O FORNECIMENTO DE NITROGÊNIO PARA O MILHO

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    Luiz Claudio Offemann

    2015-12-01

    Full Text Available This study aimed to investigate the interaction between the bacteria A. brasilense and H. seropedicae associated with nitrogen fertilization, aiming to supply nitrogen for corn crop. The experimental design was randomized blocks, in 4 x 3 factorial design with four replications. The first factor was formed by the treatments: no inoculation, inoculation H. seropedicae and A. brasilense in isolation and combination of the two strains. The second factor referred to nitrogen levels, 0, 60 and 120 kg ha-1 of N. Each experimental plot consisted of six corn lines of 3.5 meters, where the evaluation of yield components was performed: number of rows grain and number of kernels per row, thousand grain mass, average mass of grains per spike and productivity. Isolated inoculation with the bacteria A. brasilense (AbV5 and H. seropedicae (SMR1 resulted in gains especially for thousand grain weight, which can be observed that for less than the maximum dose of nitrogen doses, remained the same yield. There were no gains in productivity with respect to the use of corn seed inoculation with rhizobacteria of the genus A. brasilense (AbV5 and H. seropedicae (SMR1 in relation to nitrogen fertilization.

  1. 巴西固氮螺菌中杂合双组分蛋白Org35与NifA的相互作用%The Interaction between NifA and Hybrid Two Component Protein Org35 of Azospirillum brasilense

    Institute of Scientific and Technical Information of China (English)

    崔艳华; 伍丽娴; 洪媛媛; 陈三凤

    2010-01-01

    巴西固氮螺菌(Azospirillum brasilense)可以与玉米(Zea mays)、小麦(Triticum aestivum)等重要作物联合固氮,是重要的潜在生物肥料.NifA蛋白是所有固氮基因转录所必需的激活蛋白.本研究利用融合蛋白下拉分析,探讨了Org35蛋白与NifA蛋白的相互作用.研究表明二者存在互作.Org35蛋白由3个独立结构区域组成,即PAS结构域、组氨酸蛋白激酶结构域和应答调节蛋白区域.本研究采用融合蛋白下拉分析了Org35蛋白三个结构域与NifA的互作关系,结果表明Org35蛋白与NifA蛋白之间的相互作用是由PAS结构域介导的.

  2. Partial characterization of nif genes from the bacterium Azospirillum amazonense.

    Science.gov (United States)

    Potrich, D P; Passaglia, L M; Schrank, I S

    2001-09-01

    Azospirillum amazonense revealed genomic organization patterns of the nitrogen fixation genes similar to those of the distantly related species A. brasilense. Our work suggests that A. brasilense nifHDK, nifENX, fixABC operons and nifA and glnB genes may be structurally homologous to the counterpart genes of A. amazonense. This is the first analysis revealing homology between A. brasilense nif genes and the A. amazonense genome. Sequence analysis of PCR amplification products revealed similarities between the amino acid sequences of the highly conserved nifD and glnB genes of A. amazonense and related genes of A. brasilense and other bacteria. However, the A. amazonense non-coding regions (the upstream activator sequence region and the region between the nifH and nifD genes) differed from related regions of A. brasilense even in nitrogenase structural genes which are highly conserved among diazotrophic bacteria. The feasibility of the 16S ribosomal RNA gene-based PCR system for specific detection of A. amazonense was shown. Our results indicate that the PCR primers for 16S rDNA defined in this article are highly specific to A. amazonense and can distinguish this species from A. brasilense.

  3. Partial characterization of nif genes from the bacterium Azospirillum amazonense

    Directory of Open Access Journals (Sweden)

    D.P. Potrich

    2001-09-01

    Full Text Available Azospirillum amazonense revealed genomic organization patterns of the nitrogen fixation genes similar to those of the distantly related species A. brasilense. Our work suggests that A. brasilense nifHDK, nifENX, fixABC operons and nifA and glnB genes may be structurally homologous to the counterpart genes of A. amazonense. This is the first analysis revealing homology between A. brasilense nif genes and the A. amazonense genome. Sequence analysis of PCR amplification products revealed similarities between the amino acid sequences of the highly conserved nifD and glnB genes of A. amazonense and related genes of A. brasilense and other bacteria. However, the A. amazonense non-coding regions (the upstream activator sequence region and the region between the nifH and nifD genes differed from related regions of A. brasilense even in nitrogenase structural genes which are highly conserved among diazotrophic bacteria. The feasibility of the 16S ribosomal RNA gene-based PCR system for specific detection of A. amazonense was shown. Our results indicate that the PCR primers for 16S rDNA defined in this article are highly specific to A. amazonense and can distinguish this species from A. brasilense.

  4. 巴西固氮螺菌中PⅡ和Pz在固氮调节中的不同作用%THE DIFFERENT FUNCTIONS OF glnB AND glnZ FROM AZOSPIRILLUM BRASILENSE YU62 IN THE CONTROL OF NITROGEN FIXATION

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 管乐; 应娇妍; 李周华; 王娟; 李季伦

    2001-01-01

    在巴西固氮螺菌(Azospirillum brasilense)中,glnB和glnZ是两个高度同源基因,分别位于3.7kb/EcoRI+PstI和3.7kb/SalI的两个不同的染色体片段上.用卡那霉素盒(Kmr-cas-sette)插入法,对glnB和glnZ分别进行定位诱变,并获得相应的突变株,即glnB-和glnZ-.研究表明,glnB-突变株丧失固氮酶活性,表现为Nif-,而glnZ-象野生型菌株一样具有固氮酶活性.为了进一步研究这两个基因的功能,将glnB和glnZ分别构建在pVK100载体上形成重组质粒pVK-Ⅱ和pVK-Z,对glnB-和glnZ突变株进行互补实验,进一步证明了glnB与固氮酶活有直接相关性,而glnZ无此作用.同时,通过三亲接合法将pVK-Ⅱ和pVK-Z分别转移到巴西固氮螺菌野生型Yu62和具有一定抗铵能力的draT-突变株中,使glnB和glnZ的拷贝数增加,进一步比较它们的固氮酶活性.结果表明多拷贝的glnB基因,能显著提高固氮酶活性,而多拷贝的glnZ对固氮酶活性无影响.同时,将pVK-Ⅱ和pVK-Z分别转移到nifA-突变株中,结果表明glnB和glnZ均不能恢复nifA-的固氮酶活性.

  5. Cellulose decomposition and associated nitrogen fixation by mixed cultures of Cellulomonas gelida and Azospirillum species or Bacillus macerans

    Energy Technology Data Exchange (ETDEWEB)

    Halsall, D.M.; Gibson, A.H.

    1985-10-01

    Mixed cultures of Cellulomonas gelida plus Azospirillum lipoferum or Azospirillum brasilense and C. gelida plus Bacillus macerans were shown to degrade cellulose and straw and to utilize the energy-yielding products to fix atmospheric nitrogen. This cooperative process was followed over 30 days in sand-based cultures in which the breakdown of 20% of the cellulose and 28 to 30% of the straw resulted in the fixation of 12 to 14.6 mg of N per g of cellulose and 17 to 19 mg of N per g of straw consumed. Cellulomonas species have certain advantages over aerobic cellulose-degrading fungi in being able to degrade cellulose at oxygen concentrations as low as 1% O/sub 2/ (vol/vol) which would allow a close association between cellulose-degrading and microaerobic diazotrophic microorganisms. Cultures inoculated with initially different proportions of A. brasilense and C. gelida all reached a stable ratio of approximately 1 Azospirillum/3 Cellulomonas cells.

  6. Azospirillum lectin – induced changes in the content of nitric oxide in wheat seedling roots

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    Alen’kina S.A.

    2010-11-01

    Full Text Available The lectin of Azospirillum brasilense Sp7 at 40 μg ml-1 elicited two peaks of induction of nitric oxide synthesis in the roots of wheat seedlings after 3 and 26 h of coincubation. The lectin of A. brasilense Sp7.2.3, a mutant defective in lectin activity, produced the same effect, but the activation of nitric oxide synthesis in the roots was less in the case of 26-h incubation. Exposure to the lectins for 3 h increased citrulline synthesis in the plant cell to the same extent. This finding indicated that the Azospirillum lectins activate nitric oxide production through the NO signal system of plants, thereby acting as inducers of adaptation processes in the roots of wheat seedlings.

  7. STUDY OF AZOSPIRILLUM LECTINS INFLUENCE ON HYDROGEN PEROXIDE PRODUCTION IN WHEAT-ROOTS

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    Alen’kina S.A.

    2009-12-01

    Full Text Available It was found that two cell-surface lectins isolated from the nitrogen-fixing soil bacterium Azospirillum brasilense Sp7 and from its mutant defective in lectin activity, A. brasilense Sp7.2.3 can stimulate rapid formation of hydrogen peroxide, associated with an increase in the activities of oxalate oxidase and peroxidase in the roots of wheat seedlings. The most advantageous and most rapidly induced pathway of hydrogen peroxide formation was the oxidation of oxalic acid by oxalate oxidase because in this case, a 10-min treatment of the roots with the lectins at 10 µg ml-1 was sufficient. The data from this study attest that the Azospirillum lectins can act as inducers of adaptation processes in the roots of wheat seedlings.

  8. [Effect of Azospirillum lectins on the Activity of Proteolytic Enzymes and Their Inhibitors in Wheat Seedling Roots].

    Science.gov (United States)

    Alen'kina, S A; Nikitina, V E

    2015-01-01

    The lectins of associative nitrogen-fixing strains Azospirillum brasilense Sp7 and Sp245 were shown to exerte a multidirectional effect on the activity of acidic (pH 3.5), neutral (6.8), and alkaline (pH 7.8) proteinases. The lectin of the epiphytic A. brasilense Sp7 decreased proteolytic activity at all pH values, whereas the lectin of the endophytic A. brasilense Sp245 activated neutral and alkaline proteinases, while not affecting the alkaline ones. Experiments with protease inhibitors made it possible to conclude that the lectins of the studied A. brasilense strains alter the ratio between the activities of different protease types in germinating seeds. The activity of trypsin inhibitors in wheat seedling roots was found to increase in the presence of the lectins. Our results indicate a broader spectrum of effects of azospirilla lectins on the host plant organism.

  9. Azospirillum canadense sp. nov., a nitrogen-fixing bacterium isolated from corn rhizosphere.

    Science.gov (United States)

    Mehnaz, Samina; Weselowski, Brian; Lazarovits, George

    2007-03-01

    A free-living diazotrophic strain, DS2(T), was isolated from corn rhizosphere. Polyphasic taxonomy was performed including morphological characterization, Biolog analysis, and 16S rRNA, cpn60 and nifH gene sequence analyses. 16S rRNA gene sequence analysis indicated that strain DS2(T) was closely related to the genus Azospirillum (96 % similarity). Chemotaxonomic characteristics (DNA G+C content 67.9 mol%; Q-10 quinone system; major fatty acid 18 : 1omega7c) were also similar to those of the genus Azospirillum. In all the analyses, including phenotypic characterization using Biolog analysis and comparison of cellular fatty acids, this isolate was found to be different from the closely related species Azospirillum lipoferum, Azospirillum oryzae and Azospirillum brasilense. On the basis of these results, a novel species is proposed for this nitrogen-fixing strain. The name Azospirillum canadense sp. nov. is suggested with the type strain DS2(T) (=NCCB 100108(T)=LMG 23617(T)).

  10. Population dynamics of free living, nitrogen fixing bacteria Azospirillum in Manakkudi mangrove ecosystem, India.

    Science.gov (United States)

    Ravikumar, S; Gnanadesigan, M; Ignatiammal, S Thadedus Maria; Sumaya, S

    2012-05-01

    Seasonal variations of population dynamics of free living nitrogen fixing bacteria, Azospirillum in relation to chemical parameters in Manakkudi mangrove ecosystem was assessed in root and rhizosphere soil samples of mangroves and mangrove associated plants. In rhizosphere soil and root samples, the counts of Azospirillum were recorded maximum in Acrostichum aureum as 8.63 +/- 0.92 x 10(4) and 115.48 +/- 17.36 x 10(4) CFU g(-1), respectively. The counts of Azospirillum in non-rhizosphere soil varied from 0.01 +/- 0.001 x 10(4) to 5.77 +/- 0.92 x 10(4) CFU g(-1) and found maximum in February and minimum in March and September. Azospirillum counts in water samples were found maximum (2.24 x 10(4) CFU l(-1)) in February. During seasonal variations maximum counts of Azospirillum were recorded during southwest monsoon season in Avicennia officinalis (1.40 x 10(4) CFU g(-1)) followed by Rhizophora mucronata (1.07 x 10(4) CFU g(-1)). The average maximum population density of Azospirillum counts was found during non monsoon season (9.73 x 10(4) CFU g(-1)) and the average maximum population density of Azospirillum counts was found with the mangrove associated root samples (13.73 x 10(4) CFU g(-1)). Of the selected isolates Azospirillum lipoferum (60%) was found to be predominant followed by Azospirillum brasilense (25%), Azospirillum irakense (5%), Azospirillum halopraeferens (5%) and Azospirillum amazonense (5%). Of the isolated species, A. halopraeferens exhibited better growth at 35 g l(-1) NaCl. The level of Fe, Cu, Zn and Mn were varied from 0.91 to 15.93 ppm. The level of Mn (12.13 ppm) was found maximum during non-monsoon of rhizosphere soil sample. Highest rainfall (192.80 mm) and atmospheric temperature (25.10 degrees C) were recorded during south west monsoon and non monsoon seasons. The increased population density was greatly influenced by the pH (r = +0.686). The present finding provides enough information on the nitrogen flow through biological process in

  11. Polyhydroxyalcanoates of strains of Azospirillum spp. isolated of roots of Lycopersicon esculentum Mill. “tomato” and Oryza sativa L. “rice” in Lambayeque

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    Katty Baca

    2010-12-01

    Full Text Available In this work was determined the concentration of polyhydroxyalkanoates (PHAs of Azospirillum strains isolated from roots of Lycopersicon esculentum Mill "tomato" and Oryza sativa L. "rice" as an alternative to accumulation of petroleum-based plastics. Previously disinfected root were plated in Nfb semisolid medium where nitrogen-fixing bacteria were recognized by a whitish film on the surface and turn from green to blue. The genus Azospirillum was identified in Congo red agar medium, obtained 96 isolates of A. lipoferum and A. brasilense on tomato and rice. Batch fermentation was performed with broth Azotobacter modified feeding a saturated solution of malic acid every 12 hours and were stained with Sudan Black B. Strains were selected with the greatest number of PHAs granules (in tomato, 18 of A. lipoferum and 2 of A. brasilense; in rice, 10 of A. lipoferum and 10 of A. brasilense and quantified the biomass and PHAs. PHAs concentration reached 0.661 gL-1 in A. lipoferum KM(T-73 and 0.738 gL-1 in A. brasilense KM(T-19, both isolated from tomato. Strains of A. lipoferum and A. brasilense isolated from tomato reached a higher concentration of biomass and PHAs against the strains of rice.

  12. Plant Cell Protolytic Enzymes Activity under Exposure to Lectins of Endophytic and Epiphytic Azospirillum Strains

    Directory of Open Access Journals (Sweden)

    S.A. Alen’kina

    2016-05-01

    Full Text Available We studied the ability of lectins isolated from the surface of the two strains of nitrogen-fixing soil bacteria of the genus Azospirillum, A. brasilense Sp7 (epiphytic and A. brasilense Sp245 (endophytic, to show have a regulating effect on the activity of pectinolytic enzymes in the roots of wheat seedlings. Research results showed that the lectins under study can cause the induction of the activity of polygalacturonase, pectinesterase, pectatlyase from the plant cell wall, thereby ensuring the bacteria penetration in the plant tissues, as well as the induction of plants responses which, being combined with growth-stimulating effect of bacteria, contributes to the formation of plants stability and productivity.

  13. Azospirillum doebereinerae sp. nov., a nitrogen-fixing bacterium associated with the C4-grass Miscanthus.

    Science.gov (United States)

    Eckert, B; Weber, O B; Kirchhof, G; Halbritter, A; Stoffels, M; Hartmann, A

    2001-01-01

    A new group of nitrogen-fixing Azospirillum sp. bacteria was isolated from the roots of the C4-gramineous plant Miscanthus. Polyphasic taxonomy was performed, including auxanography using API galleries, physiological tests and 16S rRNA sequence comparison. The ability of the isolates to fix dinitrogen was evaluated by amplification of the nifD gene, immunodetection of the dinitrogenase reductase and acetylene-reduction assay. On the basis of these results, the nitrogen-fixing isolates represent a new species within the genus Azospirillum. Its closest phylogenetic neighbours, as deduced by 16S rDNA-based analysis, are Azospirillum lipoferum, Azospirillum largimobile and Azospirillum brasilense with 96.6, 96.6 and 95.9% sequence similarity, respectively. Two 16S rRNA-targeting oligonucleotide probes were developed which differentiate the new species from the other Azospirillum species by whole-cell fluorescence hybridization. Strains of the new species are curved rods or S-shaped, 1.0-1.5 microm in width and 2,0-3.0 microm in length, Gram-negative and motile with a single polar flagellum. Optimum growth occurs at 30 degrees C and at pH values between 6.0 and 7.0. No growth takes place at 37 degrees C. They have a respiratory type of metabolism, grow well on arabinose, D-fructose, gluconate, glucose, glycerol, malate, mannitol and sorbitol. They differ from A. largimobile and A. lipoferum by their inability to use N-acetylglucosamine and D-ribose, from A. lipoferum by their ability to grow without biotin supplementation and from A. brasilense by their growth with D-mannitol and D-sorbitol as sole carbon sources. Nitrogen fixation occurs in microaerobic nitrogen-limited conditions. For this species, the name Azospirillum doebereinerae sp. nov. is suggested, with strain GSF71T as the type strain (= DSM 13131T; reference strain Ma4 = DSM 13400). Its G+C content is 70.7 mol%.

  14. Transfer of a plant chitinase gene into a nitrogen-fixing Azospirillum and study of its expression.

    Science.gov (United States)

    Jayaraj, Jayaraman; Muthukrishnan, Subbaratnam; Liang, George H

    2004-07-01

    Azospirillum is used extensively in rice and other cereal crops as a biofertilizer. There is a substantial opportunity to improve the efficiency of this bacterium through the transfer of genes of agricultural importance from other organisms. Chitinases are antifungal proteins, and expression of chitinase genes in Azospirillum would help to develop strains with potential antifungal activities. So far there are no reports about transfer of plant genes into Azospirillum and their expression. The present study was aimed at expressing an antifungal gene (a rice chitinase) of plant origin in Azospirillum brasilense. A rice chitinase cDNA (RC 7) that codes for a 35 kDa protein was subcloned into a broad host range plasmid pDSK519 under the control of LacZ promoter. The plasmid was mobilized into the nitrogen-fixing bacterium, Azospirillum brasilense strain SP51eFL1, through biparental mating. The conjugation frequency was in the range of 35-40 x 10(-6). The transconjugants grew in nitrogen-free media and fixed gaseous nitrogen in vitro. However, their growth and nitrogen-fixing ability were slightly less than those of the wild-type. Expression of the protein was demonstrated through western blotting of the total cell protein, which detected a 35 kDa band that was immuno-reactive to a barley chitinase antibody. The cell lysates also hydrolyzed various chitin substrates, which resulted in release of free sugars demonstrating the chitinase activity of transconjugants. The expressed protein also had antifungal activity as demonstrated by inhibition of growth of the plant pathogenic fungus, Rhizoctonia solani.

  15. Azospirillum fermentarium sp. nov., a nitrogen-fixing species isolated from a fermenter.

    Science.gov (United States)

    Lin, Shih-Yao; Liu, You-Cheng; Hameed, Asif; Hsu, Yi-Han; Lai, Wei-An; Shen, Fo-Ting; Young, Chiu-Chung

    2013-10-01

    An aerobic, Gram-stain-negative, spiral or rod-shaped, non-spore-forming, diazotrophic bacterium (strain CC-LY743(T)) was isolated from a fermentative tank in Taiwan. Strain CC-LY743(T) was able to grow at 20-37 °C and pH 6.0-8.0 and tolerated up to 3.0 % (w/v) NaCl. It was positive for nitrogen fixation, with activity of 10.6 nmol ethylene h(-1). 16S rRNA gene sequence analysis of strain CC-LY743(T) showed highest similarity to Azospirillum picis DSM 19922(T) (96.1 %), Azospirillum oryzae JCM 21588(T) (96.0 %) and Azospirillum rugosum DSM 19657(T) (96.0 %) and lower similarity (Azospirillum species. Highest nifH gene sequence similarities were obtained with Azospirillum brasilense BCRC 12270(T) (92.0 %), Azospirillum formosense BCRC 80273(T) (92.3 %) and A. rugosum DSM 19657(T) (91.8 %). It was positive in the rapid identification by a genus-specific primer set. The predominant quinone system was ubiquinone 10 (Q-10) and the DNA G+C content was 69.6±0.1 mol%. The major fatty acids found in strain CC-LY743(T) were n-C16 : 0, C19 : 0 cyclo ω8c, C14 : 0 3-OH/C16 : 1 iso I, C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c. Based on its phylogenetic, phenotypic and chemotaxonomic features, strain CC-LY743(T) is considered to represent a novel species within the genus Azospirillum for which the name Azospirillum fermentarium sp. nov. is proposed. The type strain is CC-LY743(T) ( = BCRC 80505(T) = JCM 18688(T) = LMG 27264(T)).

  16. [Comparative assessment of inductive effects of Azospirillum lectins with different antigenic properties on the signal systems of wheat seedling roots].

    Science.gov (United States)

    Alen'kina, S A; Petrova, L P; Sokolova, M K; Chernyshova, M P; Trutneva, K A; Bogatyrev, V A; Nikitina, V E

    2014-01-01

    The lectins of associative nitrogen-fixing bacteria Azospirillum brasilense Sp7 and its mutant A. brasilense Sp7.2.3 were shown to have different effects on the components of the wheat seedling root signal system, namely to regulate the levels of cAMP, nitric oxide, diacylglycerol, and salicylic acid, as well as to induce the activities of superoxide dismutase and lipoxygenase. Our results make it possible to consider azospirilla lectins as inducers of the signal systems in wheat seedling roots, since they cause development of several flows of primary signals. These data are of general biological importance, since lectins are present in all living organisms and most ot the functions of lectins remain insufficiently understood.

  17. Improvement in bioavailability of tricalcium phosphate to Cymbopogon martinii var. motia by rhizobacteria, AMF and Azospirillum inoculation.

    Science.gov (United States)

    Ratti, N; Kumar, S; Verma, H N; Gautam, S P

    2001-01-01

    The interactive effects of phosphate solubilizing bacteria, N2 fixing bacteria and arbuscular mycorrhizal fungi (AMF) were studied in a low phosphate alkaline soil amended with tricalcium insoluble source of inorganic phosphate on the growth of an aromatic grass palmarosa (Cymbopogon martinii). The microbial inocula consisted of the AM fungus Glomus aggregatum, phosphate solubilizing rhizobacteria Bacillus polymyxa and N2 fixing bacteria Azospirillum brasilense. These rhizobacteria behaved as "mycorrhiza helper" and enhanced root colonization by G. aggregatum in presence of tricalcium phosphate at the rate of 200 mg kg(-1) soil (P1 level). Dual inoculation of G. aggregatum and B. polymyxa yielded 21.5 g plant dry weight (biomass), while it was 21.7 g in B. polymyxa and A. brasilense inoculated plants as compared to 14.9 g of control at the same level. Phosphate content was maximum (0.167%) in the combined treatment of G. aggregatum, B. polymyxa and A. brasilense at P1 level, however acid phosphatase activity was recorded to be 4.75 pmol mg(-1) min(-1) in G. aggregatum, B. polymyxa and A. brasilense treatment at P0 level. This study indicates that all microbes inoculated together help in the uptake of tricalcium phosphate which is otherwise not used by the plants and their addition at 200 mg kg(-1) of soil gave higher productivity to palmarosa plants.

  18. 巴西固氮螺菌Yu62nifA基因克隆、测序及功能分析%CLONING SEQUENCING AND EXPRESSION PATTERN,FUNCTIONAL ANALYSIS OF nifA GENE IN AZOSPIRILLUM BRASILENSE Yu62

    Institute of Scientific and Technical Information of China (English)

    王娟; 陈三凤; 马旅雁; 李季伦

    2001-01-01

    用TD-PCR法克隆了巴西固氮螺菌(Azospirillun brasilense)Yu62的nifA基因.序列分析表明它与巴西固氮螺菌Sp7的nifA序列高度同源(96.5%),其编码的产物NifA蛋白与Sp7菌株NifA的氨基酸序列同源性为97.6%.该基因可以完全互补巴西固氮螺菌Sp7 nifA-突变株的Nif-表型.研究了NH4+和O2对Yu62 nifA基因的表达及NifA活性的影响.结果表明:mfA基因在Yu62菌株中是部分组成型表达的,氨和氧不能完全阻遏其表达,在5mmol/LNH4Cl与微氧(0.5%O2)条件下表达最高;NifA蛋白在0.4%~0.5%O2时活性最高,氧分压降低和提高都使NifA活性下降,1mmol/L NH4Cl足以抑制NifA的活性.

  19. AMPLIFICATION OF AZOSPIRILLUM SP. JG3 GLPD GENE FRAGMENT USING DEGENERATE PRIMERS GENERATED BY WEB-BASED TOOLS

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    Stalis Norma Ethica

    2013-12-01

    Full Text Available Primaclade and In Silico web-based tools were used as a strategy to obtain the correct-size PCR amplicon targeting a fragment of gene encoding glycerol-3-phosphate dehydrogenase (glpD of Azospirillum sp. JG3. The bacterial strains are soil, Gram-negative PGPR (Plant-Growth Promoting Rhizobacteria isolated from an agricultural land in Purwokerto, Central Java, Indonesia, which have ability to produce several commercial enzymes. The aim is to obtain a pair of reliable degenerate primers from a limited number of glpD sequences from other Azospirilla retrieved in GenBank using bioinformatics approach. We demonstrated degenerate primer design that led to successful PCR amplification corresponding to the targeted DNA fragment. Homology analysis showed that the obtained DNA fragment is 61% and 99% similar to sn-glycerol-3-phosphate dehydrogenase genes of Azospirillum brasilense and Stenotrophomonas maltophili respectively.

  20. Azospirillum, a free-living nitrogen-fixing bacterium closely associated with grasses: genetic, biochemical and ecological aspects.

    Science.gov (United States)

    Steenhoudt, O; Vanderleyden, J

    2000-10-01

    Azospirillum represents the best characterized genus of plant growth-promoting rhizobacteria. Other free-living diazotrophs repeatedly detected in association with plant roots, include Acetobacter diazotrophicus, Herbaspirillum seropedicae, Azoarcus spp. and Azotobacter. Four aspects of the Azospirillum-plant root interaction are highlighted: natural habitat, plant root interaction, nitrogen fixation and biosynthesis of plant growth hormones. Each of these aspects is dealt with in a comparative way. Azospirilla are predominantly surface-colonizing bacteria, whereas A. diazotrophicus, H. seropedicae and Azoarcus sp. are endophytic diazotrophs. The attachment of Azospirillum cells to plant roots occurs in two steps. The polar flagellum, of which the flagellin was shown to be a glycoprotein, mediates the adsorption step. An as yet unidentified surface polysaccharide is believed to be essential in the subsequent anchoring phase. In Azoarcus sp. the attachment process is mediated by type IV pili. Nitrogen fixation structural genes (nif) are highly conserved among all nitrogen-fixing bacteria, and in all diazotrophic species of the class of proteobacteria examined, the transcriptional activator NifA is required for expression of other nif genes in response to two major environmental signals (oxygen and fixed N). However, the mechanisms involved in this control can vary in different organisms. In Azospirillum brasilense and H. seropedicae (alpha- and beta-subgroup, respectively), NifA is inactive in conditions of excess nitrogen. Activation of NifA upon removal of fixed N seems to involve, either directly or indirectly, the signal transduction protein P(II). The presence of four conserved cysteine residues in the NifA protein might be an indication that NifA is directly sensitive to oxygen. In Azotobacter vinelandii (gamma-subgroup) nifA is cotranscribed with a second gene nifL. The nifL gene product inactivates NifA in response to high oxygen tension and cellular

  1. Change in the Content of Salicylic Acid and in the Activities of Phenylalanine Ammonia-Lyase and Catalase in Wheat Seedling Roots Under the Effect of Azospirillum Lectins

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    Alen'kina S.A.

    2012-05-01

    Full Text Available We investigated the time course of changes in the endogenous content of salicylic acid, the ratio between the acid's free and bound forms, and changes in the activities of phenylalanine ammonia-lyase and catalase in wheat seedling roots under the effect of the lectins of two strains of the associative nitrogen-fixing bacterium Azospirillum: A. brasilense Sp7 and its mutant defective in lectin activity, A. brasilense Sp7.2.3. Differences in plant response to the action of the lectins from these two strains were established. On the basis of the obtained data, a model was proposed for lectin-assisted induction of resistance, according to which the lectin effect on the roots of seedlings results in accumulation of free salicylic acid, which inhibits catalase activity, ultimately leading to accumulation of hydrogen peroxide and to formation of induced resistance.

  2. Componentes de produção e produtividade da cultura da soja submetida à inoculação de Bradyrhizobium e Azospirillum

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    Lucas Guilherme Bulegon

    2016-04-01

    Full Text Available O presente trabalho teve por objetivo avaliar os componentes de produção e produtividade de dois genótipos de soja cultivados em casa de vegetação e submetidos à inoculação com bactérias diazotróficas do gênero Bradyrhizobium e Azospirillum. Adotou-se delineamento em blocos ao acaso com quatro repetições, em esquema fatorial 2 × 4; no fator 1 foram testados: dois genótipos de soja (BMX Turbo e Coodetec 250 e no fator 2 foram avaliadas quatro combinações de inoculação (sementes não inoculadas mas que receberam 200 kg ha-1 de N na semeadura; sementes inoculado com Bradyrhizobium japonicum; sementes inoculadas com Azospirillum brasilense; sementes com inoculação de B. japonicum e A. brasilense. No estádio R8 foi avaliado as variáveis biométricas: diâmetro de coleto (DC; altura de planta (ALP; número de vagens (NVS; número de grãos por vagem (NGV; massa seca de parte aérea (MPA;massa seca de vagens (MSV; produtividade por planta (PPP; teor de N na parte aérea (TNA e teor de N nos grãos (TNG. Foi observado resposta diferenciada dos genótipos de soja BMX Turbo e Coodetec 250 à inoculação de sementes com bactérias diazotróficas do gênero B. japonicum ou A. brasilense na altura de plantas e nitrogênio na parte aérea. A inoculação conjunta de B. japonicum e A. brasilense em sementes intensifica o desempenho produtivo da cultura da soja no genótipo BMX Turbo.

  3. Polihidroxialcanoatos de cepas de Azospirillum spp. aisladas de raíces de Lycopersicon esculentum Mill. “tomate” y Oryza sativa L. “arroz” en Lambayeque

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    Katty Baca

    2010-01-01

    Full Text Available En este trabajo se determinó la concentración de polihidroxialcanoatos (PH As de cepas de Azospirillum aisladas de raíces de Lycopersicon esculentum Mill. “tomate” y Oryza sativa L. “arroz”, como una alternativa ante la acumulación de plásticos derivados del petróleo. R aíces previamente desinfectadas se sembraron en medio Nfb se misólido, donde las bacterias fijadoras de nitrógeno se reconocieron por una película blanquecina bajo la superficie y el viraje del indicador al azul. El género Azospirillum se identificó en medio rojo de Congo, obteniéndose 96 cepas de A. lipoferum y A. brasilense en tomate y arroz. Se realizó una fermentación discontinua con caldo Azotobacter modificado, alimentando con una solución saturada de ácido málico cada 12 horas y se realizaron tinciones con Sudán Negro B. Se seleccionaron las cepas con mayor nú mero de gránulos de PHAs (en tomate , 18 de A. lipoferum y 2 de A. brasilense y en arroz, 10 de A. lipoferum y 10 de A. brasilense y se cuantificó la biomasa y PHAs. La concentración de PHAs alcanzó 0 . 661 gL - 1 en A. lipoferum KM(T - 73 y 0 . 738 gL - 1 en A. br asilense KM(T - 19. Las cepas de A. lipoferum y A. brasilense aisladas de tomate alcanzaron una mayor concentración de biomasa y PHAs frente a las cepas aisladas de arroz.

  4. Azospirillum and arbuscular mycorrhizal colonization enhance rice growth and physiological traits under well-watered and drought conditions.

    Science.gov (United States)

    Ruíz-Sánchez, Michel; Armada, Elisabet; Muñoz, Yaumara; García de Salamone, Inés E; Aroca, Ricardo; Ruíz-Lozano, Juan Manuel; Azcón, Rosario

    2011-07-01

    The response of rice plants to inoculation with an arbuscular mycorrhizal (AM) fungus, Azospirillum brasilense, or combination of both microorganisms, was assayed under well-watered or drought stress conditions. Water deficit treatment was imposed by reducing the amount of water added, but AM plants, with a significantly higher biomass, received the same amount of water as non-AM plants, with a poor biomass. Thus, the water stress treatment was more severe for AM plants than for non-AM plants. The results showed that AM colonization significantly enhanced rice growth under both water conditions, although the greatest rice development was reached in plants dually inoculated under well-watered conditions. Water level did not affect the efficiency of photosystem II, but both AM and A. brasilense inoculations increased this value. AM colonization increased stomatal conductance, particularly when associated with A. brasilense, which enhanced this parameter by 80% under drought conditions and by 35% under well-watered conditions as compared to single AM plants. Exposure of AM rice to drought stress decreased the high levels of glutathione that AM plants exhibited under well-watered conditions, while drought had no effect on the ascorbate content. The decrease of glutathione content in AM plants under drought stress conditions led to enhance lipid peroxidation. On the other hand, inoculation with the AM fungus itself increased ascorbate and proline as protective compounds to cope with the harmful effects of water limitation. Inoculation with A. brasilense also enhanced ascorbate accumulation, reaching a similar level as in AM plants. These results showed that, in spite of the fact that drought stress imposed by AM treatments was considerably more severe than non-AM treatments, rice plants benefited not only from the AM symbiosis but also from A. brasilense root colonization, regardless of the watering level. However, the beneficial effects of A. brasilense on most of the

  5. Nitrogen fixation associated with development and localization of mixed populations of Cellulomonas species and Azospirillium brasilense grown on cellulose or wheat straw

    Energy Technology Data Exchange (ETDEWEB)

    Halsall, D.M.; Goodchild, D.J.

    1986-04-01

    Mixed cultures of Cellulomonas sp. and Azospirillum brasilense were grown with straw or cellulose as the carbon source under conditions favoring the fixation of atmospheric nitrogen. Rapid increases in cell numbers, up to 10/sup 9/ cells per g of substrate, were evident after 4 and 5 days of incubation at 30 degrees C for cellulose and straw, respectively. Nitrogen fixation (detected by acetylene reduction measured on parallel cultures) commenced after 2 and 4 days of incubation for straw and cellulose, respectively, and continued for the duration of the experiment. Pure cultures of Cellulomonas sp. showed an increase in cell numbers, but CO/sub 2/ production was low, and acetylene reduction was not detected on either cellulose or straw. Pure cultures of A. brasilense on cellulose showed an inital increase in cell numbers (10/sup 7/ cells per g of substrate) over 4 days, followed by a decline presumably caused by the exhaustion of available carbon substrate. On straw, A. brasilense increased to 10/sup 9/ cells per g of substrate over 5 days and then declined slowly; this growth was accompanied by acetylene reduction. Scanning electron micrographs of straw incubated with a mixture under the above conditions for 8 days showed cells of both species in close proximity to each other. Evidence was furnished that the close spatial relatioship of cells from the two species facilitated the mutally beneficial association between them and thus increased the efficiency with which the products of straw breakdown were used for nitrogen fixation. 17 references.

  6. Comparison of prominent Azospirillum strains in Azospirillum-Pseudomonas-Glomus consortia for promotion of maize growth.

    Science.gov (United States)

    Couillerot, Olivier; Ramírez-Trujillo, Augusto; Walker, Vincent; von Felten, Andreas; Jansa, Jan; Maurhofer, Monika; Défago, Geneviève; Prigent-Combaret, Claire; Comte, Gilles; Caballero-Mellado, Jesus; Moënne-Loccoz, Yvan

    2013-05-01

    Azospirillum are prominent plant growth-promoting rhizobacteria (PGPR) extensively used as phytostimulatory crop inoculants, but only few studies are dealing with Azospirillum-containing mixed inocula involving more than two microorganisms. We compared here three prominent Azospirillum strains as part of three-component consortia including also the PGPR Pseudomonas fluorescens F113 and a mycorrhizal inoculant mix composed of three Glomus strains. Inoculant colonization of maize was assessed by quantitative PCR, transcription of auxin synthesis gene ipdC (involved in phytostimulation) in Azospirillum by RT-PCR, and effects on maize by secondary metabolic profiling and shoot biomass measurements. Results showed that phytostimulation by all the three-component consortia was comparable, despite contrasted survival of the Azospirillum strains and different secondary metabolic responses of maize to inoculation. Unexpectedly, the presence of Azospirillum in the inoculum resulted in lower phytostimulation in comparison with the Pseudomonas-Glomus two-component consortium, but this effect was transient. Azospirillum's ipdC gene was transcribed in all treatments, especially with three-component consortia, but not with all plants and samplings. Inoculation had no negative impact on the prevalence of mycorrhizal taxa in roots. In conclusion, this study brought new insights in the functioning of microbial consortia and showed that Azospirillum-Pseudomonas-Glomus three-component inoculants may be useful in environmental biotechnology for maize growth promotion.

  7. Curva de sobrevivência à luz ultravioleta do Azospirlllum brasilense (BEIJERIWCK, 1925

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    Olivia Márcia Nagy Arantes

    1982-11-01

    Full Text Available The Poem - Third and last part of a Thesis - METAPOESIA - presented no Pontifícia Universidade Católica de Porto Alegre as part of the requirements in order to obtain the degree of full-professorship. In the present study, founded on field-work" and on the metalanguage of the modern poetry, are discussed the concept, nature and species of poems. Poem, in this research, is understood not only the conventional poem but all poetic expression (mythic of the life and the nature. In this article is discussed particularly the poem. The written poem, verbal discourse ("the poem which Poet wrote"; the poem of the life ("The poem which Somebody wrote", and of the nature ("the poem which Nobody wrote".Estudo do Azospirillum brasilense, bactéria fixadora de nitrogênio em gramíneas, com o objetivo de fornecer subsídios para estudos genéticos sobre o A. brasilense através da determinação da curva de sobrevivência do microrganismo à luz ultravioleta. Estabeleceu-se o tempo de exposição à radiação de 1 minuto e obteve-se uma taxa de 5% de sobreviventes, representando uma dose recomendável para a indução de mutações, desde que se expõe suficientemente o DNA da bactéria ao agente mutagênico, sem contudo reduzir drasticamente a população irradiada, o que tornaria muito baixa a frequência de mutantes após o tratamento.

  8. ESTUDIO DE LA INTERACCIÓN PLANTA-Azospirillum EN EL CULTIVO CAÑA DE AZÚCAR (Saccharum sp.

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    Juana Pérez

    2005-01-01

    Full Text Available Se realizó el aislamiento y la caracterización de cepas nativas de Azospirillum en la rizosfera de las variedades comerciales de caña de azúcar Ja60-5 y C266-70 en los suelos Ferralítico Rojo y Ferralítico Amarillento de las provincias de La Habana y Ciego de Avila. A los aislamientos se les evaluó la actividad nitrogenasa mediante la técnica de reducción del acetileno y la actividad estimuladora del crecimiento en vitroplantas de caña de azúcar. Las cepas fueron caracterizadas mediante pruebas morfológicas y bioquímicas. Se encontraron concentraciones de bacterias del orden de 104 cel. g -1 de raíces, correspondiendo los mayores valores a la variedad Ja60-5 en ambos suelos. Se aislaron cuatro cepas con características propias del género Azospirillum, seleccionándose la cepa 8 INICA de Azospirillum brasilense para estudios posteriores, por sus elevados valores de reducción de acetileno y su mayor efecto sobre el crecimiento de vitroplantas de caña de azúcar.

  9. Combination of inoculation methods of Azospirilum brasilense with broadcasting of nitrogen fertilizer increases corn yield

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    Tânia Maria Müller

    2015-01-01

    Full Text Available Nitrogen (N is the most limiting nutrient for corn production. Thereby, the goal of the paper was to evaluate inoculation methods of Azospirillum brasilense in order to partially supply N required by the crop. The experiment was carried out in Guarapuava, PR, Brasil, in 2011/2012 growing season. Randomized blocks with factorial 3 inoculation methods (seed treatment, planting furrow and non-inoculated control x 5 doses of nitrogen (0, 75, 150, 225 and 300kg ha-1 x 8 replications was used as the experimental design. Leaf are index, foliar nitrogen content, total chlorophyll, grains per ear and yield were evaluated. There was significant interaction between inoculation methods and nitrogen fertilization to leaf area index, but not for yield. Inoculation with the diazotrophic bacteria provided yield increase of 702kg ha-1 for inoculation in seeding furrow and 432kg ha-1 for inoculation in seed treatment compared to the control, but both treatments did not differ between each other. Furthermore, total chlorophyll, grains per ear and yield were positively affected, with quadratic response, by the nitrogen fertilization in broadcasting

  10. Nitrogen translocation in wheat inoculated with Azospirillum and fertilized with nitrogen

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    RODRIGUES OSMAR

    2000-01-01

    Full Text Available The productivity and the translocation of assimilates and nitrogen (N were compared after inoculation of wheat (Triticum aestivum L., cv. BR-23 seeds with two strains of Azospirillum brasilense (strains 245 and JA 04 under field conditions. The inoculation of wheat seeds was done with a peat inoculant at sowing time. Plant material for evaluations were collected at anthesis and maturity. No differences in grain yield and in the translocation of assimilates resulting from inoculation were detected. Differences were observed in relation to N rates (0, 15, and 60 kg ha-1. N content in the grain increased significantly in the bacteria-inoculated treatments in which N was not added. This increase in N content in the grain with inoculation was probably due to higher N uptake after anthesis without any significant contribution on the grain yield. Such increment was of 8.4 kg ha-1 of N representing 66% more N than in no inoculated treatment. Regardless of the inoculation and the rate of N applied, it was observed that about 70% of the N accumulated at anthesis was translocated from vegetative parts to the grain.

  11. APPLICATION TIMES OF AN Azospirillum BIOPRODUCT IN TOMATO GROWTH, DEVELOPMENT AND YIELD

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    Elein Terry

    2000-01-01

    Full Text Available Con el objetivo de estudiar diferentes momentos de aplicación de un bioproducto a base de Azospirillum brasilense Sp-7 en el cultivo del tomate, se desarrolló un experimento de campo durante dos años sobre un suelo Ferralítico Rojo compactado, realizándose el trabajo en dos etapas: fase de semillero y fase de floración-fructificación, teniéndose en cuenta un total de cinco tratamientos distribuidos completamente aleatorizados en la fase de semillero y bloques al azar en la fase de trasplante. Los resultados mostraron una respuesta positiva a la inoculación de esta rizobacteria en combinación con el fertilizante nitrogenado, manifestado a través del vigor y rendimiento alcanzado por las plantas. Desde el punto de vista de la viabilidad en el manejo y la factibilidad económica, la inoculación en el momento de la siembra resultó ser la más ventajosa.

  12. Plant yield and nitrogen content of a digitgrass in response to azospirillum inoculation

    Energy Technology Data Exchange (ETDEWEB)

    Schank, S.C.; Weier, K.L.; MacRae, I.C.

    1981-02-01

    Two Australian soils, a vertisol (pH 6.8, 0.299% N) and a sandy yellow podzol (pH 6.2, 0.042% N), were used with digitgrass, Digitaria sp. X46-2 (PI 421785), in a growth room experiment. Comparisons were made between plants inoculated with live and autoclaved bacterial suspensions of Australian and Brazilian isolates of Azospirillum brasilense. Seedlings were inoculated on days 10 and 35. Acetylene-reducing activity was measured five times during the experiment. Dry matter yields of the digitgrass on the podzol (low N) inoculated with liver bacteria were 23% higher than those of the controls. On the vertisol (high N), yield increases from inoculation with live bacteria were 8.5%. The higher-yielding plants had significantly lower precent nitrogen, but when total nitrogen of the tops was calculated, the inoculated plants had a higher total N than did the controls (P = 0.04). Acetylene-reducing activity was variable in the experiment, ranging from 0.5 to 11.9 mu mol of C2H2 core -1 day -1. Live bacterial treatment induced a proliferation of roots, possible earlier maturity, higher percent dry matter, and a higher total N in the tops. (Refs. 21).

  13. EVALUACIÓN DE CEPAS NATIVAS DEL GÉNERO Azospirillum Y SU INTERACCIÓN CON EL CULTIVO DEL ARROZ

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    Mabel Pazos

    2001-01-01

    Full Text Available Catorce cepas nativas pertenecientes a la especie Azospirillum brasilense se caracterizaron en cuanto a su ca- pacidad de fijar el nitrógeno atmosférico. Se estudió su efecto en el cultivo del arroz (Oryza sativa, mediante el modelo Microcosmos, realizando determinaciones de la altura de las plantas inoculadas, masa fresca de la raíz y el contenido de fenoles libres y la cuantificación de proteínas en las hojas. Se observó en todos los casos un efecto positivo por la inocula- ción con estas rizobacterias, donde se destaca la cepa R5(15 con los mayores incrementos en los indicadores estudiados.

  14. Strain-specific chemotaxis of Azospirillum spp.

    OpenAIRE

    Reinhold, B; Hurek, T; Fendrik, I

    1985-01-01

    Chemotactic responses of three Azospirillum strains originating from different host plants were compared to examine the possible role of chemotaxis in the adaptation of these bacteria to their respective hosts. The chemotaxis to several sugars, amino acids, and organic acids was determined qualitatively by an agar plate assay and quantitatively by a channeled-chamber technique. High chemotactic ratios, up to 40, were obtained with the latter technique. The chemotactic response did not rely up...

  15. Realocação de nitrogênio e de biomassa para os grãos, em trigo submetido a inoculação de Azospirillum Reallocation of nitrogen and biomass to the seeds in wheat inoculated with Azospirillum bacteria

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    AGOSTINHO DIRCEU DIDONET

    2000-02-01

    Full Text Available Estudou-se, em condições de campo, o efeito de inoculante turfoso em pó contendo bactérias do gênero Azospirillum no rendimento de grãos e na remobilização de N e de biomassa para os grãos de trigo (Triticum aestivum L., cultivar Embrapa 16. Usaram-se como inoculantes, a estirpe de Azospirillum brasilense 245 e o isolado 10 de Azospirillum lipoferum. Em cada tratamento de inoculação, e também sem inoculação, aplicaram-se diferentes doses de N em diferentes estádios de desenvolvimento das plantas, distribuídos em blocos ao acaso com parcelas subdivididas. Na antese e na maturação fisiológica, avaliaram-se o acúmulo de massa seca e N total nas diferentes partes da planta. Na colheita, além do rendimento de grãos, avaliaram-se também o índice de colheita para biomassa e para N, os principais componentes do rendimento e o teor de N total de grãos. Os resultados ainda preliminares evidenciaram que, mesmo não havendo efeito da inoculação no rendimento de grãos, houve um melhor alocamento de N e de biomassa para os grãos, resultando em maior massa de mil grãos e em menor quantidade de N restante na palha das plantas na maturação fisiológica. Esses efeitos resultaram de um menor número de espigas m-2, provavelmente devido à morte de afilhos, fator que determinou maior disponibilidade de N e de biomassa às espigas e grãos restantes.The effect of inoculating wheat (Triticum aestivum L. seeds, cultivar Embrapa 16, with powder peat inoculant containing Azospirillum bacteria on yield and remobilization of nitrogen and biomass was studied under field conditions. The strain of Azospirillum brasilense 245 and the isolate 10 of Azospirillum lipoferum were used as inoculants. Different rates of nitrogen were applied at varying stages of plant growth for each inoculated and non-inoculated treatment, distributed in blocks at random with split plots. The accumulation of dry matter and total nitrogen in plant parts was evaluated

  16. Azospirillum Inoculation Alters Nitrate Reductase Activity and Nitrogen Uptake in Wheat Plant under Water Deficit Conditions

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    N. Aliasgharzad N. Aliasgharzad

    2014-01-01

    Full Text Available Water deficit stress usually diminishes nitrogen uptake by plants. There are evidences that some nitrogen fixing bacteria can alleviate this stress by supplying nitrogen and improving its metabolism in plants. Four Azospirillum strains, A. lipoferum AC45-II, A. brasilense AC46-I, A. irakense AC49-VII and A. irakense AC51-VI were tested for nitrate reductase activity (NRA. In a pot culture experiment using a sandy loam soil, wheat plants (Triticum aestivum L. cv. Sardari were inoculated with these bacterial strains and three ranges of soil water potential (W1: -10 to -20, W2: -40 to -50 and W3: -65 to -75 kPa were applied to the pots. All strains were positive in NRA test and the highest (7.63mg NO2-N.L-1.48h-1 was recorded for AC49-VII and the least (0.23mg NO2-N.L-1.48h-1 was belong to AC51-VI. Leaf and root NRA, root and shoot nitrogen concentrations, and dry weights of root and shoot decreased by increasing water deficit stress. All four bacterial strains caused a significant enhancement in root NRA and in each water deficit level, the higher root NRA was recorded in AC46-I and AC49-VII inoculated plants. The highest leaf NRA was achieved by AC49-VII. The mean increment of root NRA by bacterial strains was 171% compared to the non-bacterial plants. Moreover, at the highest level of water deficit stress, the highest dry weight and nitrogen concentration in root and shoot were obtained by AC46-I and AC49-VII treatments.

  17. Computer simulation and experimental study of the polysaccharide-polysaccharide interaction in the bacteria Azospirillum brasilense Sp245

    Science.gov (United States)

    Arefeva, Oksana A.; Kuznetsov, Pavel E.; Tolmachev, Sergey A.; Kupadze, Machammad S.; Khlebtsov, Boris N.; Rogacheva, Svetlana M.

    2003-09-01

    We have studied the conformational properties and molecular dynamics of polysaccharides by using molecular modeling methods. Theoretical and experimental results of polysaccharide-polysaccharide interactions are described.

  18. Desempenho agronômico a campo de híbridos de milho inoculados com Azospirillum

    Directory of Open Access Journals (Sweden)

    Patricia Dörr de Quadros

    2014-04-01

    Full Text Available O uso de inoculantes na cultura do milho tem sido cada vez mais valorizado, em vista dos benefícios que pode trazer à cultura, como a fixação biológica do nitrogênio e o aumento da quantidade de raízes. Isto pode melhorar a absorção de água e nutrientes pela planta, contribuindo para o desenvolvimento do milho, principalmente em períodos de seca. Este estudo foi desenvolvido com o objetivo de avaliar, em condições de campo, as características agronômicas e o rendimento de grãos de híbridos de milho, inoculados, ou não, com uma mistura de três espécies de Azospirillum (A. brasilense, A. lipoferum, A. oryzae. Foram avaliados o teor relativo de clorofila nas folhas, a altura de planta, a senescência foliar, os componentes de rendimento de grãos, o teor de N, a matéria seca da parte aérea das plantas e o número mais provável de bactérias diazotróficas na rizosfera das plantas. A inoculação manteve o teor de clorofila significativamente maior até o estádio R3 das plantas, para os três híbridos testados, aumentou o rendimento da matéria seca da parte aérea, dos híbridos AS 1575 e SHS 5050, o peso de 1000 grãos, para o híbrido P32R48 e altura, para o AS 1575. Houve interação entre os genótipos de milho e as bactérias inoculadas, visto que, cada híbrido testado respondeu de forma diferente à inoculação. A inoculação de Azospirillum em milho demonstrou estimular o desenvolvimento das plantas no período vegetativo, aumentando a probabilidade de obter-se um estande de plantas uniforme, maior resistência ao estresse e maior concentração de clorofila nas folhas.

  19. INOCULAÇÃO COM Azospirillum brasilense, MANEJO DE PASTEJO E NITROGÊNIO EM INTEGRAÇÃO LAVOURA-PECUÁRIA

    OpenAIRE

    Marcos da Silva Brum

    2015-01-01

    Este trabalho apresenta resultados de um experimento com integração lavoura-pecuária em dois anos agrícolas (2012/2013 e 2013/2014). Nas estações frias se utilizou pastagem formada pela mistura de aveia preta e azevém sob pastejo com ovinos, em um experimento trifatorial (manejo do pastejo, doses de nitrogênio e inoculação de sementes), em delineamento experimental de blocos ao acaso. Para o fator manejo do pastejo a pastagem foi manejada da seguinte forma: (I) pastejo convenci...

  20. ADUBAÇÃO NITROGENADA E INOCULAÇÃO DE Azospirillum brasilense PARA A PRODUÇÃO DE MILHO E TRIGO EM SISTEMA PLANTIO DIRETO

    OpenAIRE

    ANGELO RAFAEL BINI

    2015-01-01

    O fornecimento adequado de nitrogênio (N) é primordial para a obtenção de altos rendimentos de grãos de cereais. Entretanto, a utilização de elevadas doses de N pode reduzir a eficiência na utilização do N e causar problemas de contaminação ambiental. O fornecimento de N via fixação biológica nas culturas de milho e trigo pode contribuir para a sustentabilidade da produção agrícola. Este estudo foi realizado com o objetivo de avaliar (i) as alterações químicas do solo após a inoculação das se...

  1. Estandarización de un medio de cultivo a partir de fuentes agroindustriales para la multiplicación de Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    Diego Rivera

    2012-07-01

    Full Text Available The Plackett-Burman statistical design was initially applied for Screening the most important nutritional variables (four carbon sources, two nitrogen sources, K2 HPO4 y MgSO4 and a trace elements solution affecting microorganism biomass production. Subsequently, a optimising the process by applied a BoxBehnken design for the most important factors tree. The results showed that biomass (UFC/mL production was strongly affected by nutritional and physico conditions of culture medium. The highest yield was obtained in the following conditions: 3g/L sucrose, 6,75 mL/L molasses, 0,77 mL/L glycerol, 14,42 mL/L soybean hydrolyzate, 0,848 g/L yeast hidrolyzate, 0,25 g/L K2 HPO4 , ⅗ aeration and shaking at 120 rpm 26 hours total fermentation time during which 1 x 109 UFC/mL were obtained.

  2. Inoculation with Azospirillum, associated with nitrogen fertilization in maize

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    Maria do Carmo Lana

    2012-06-01

    Full Text Available The biological nitrogen fixation is an alternative to supply the nitrogen needed for maize. The objective of this study was to evaluate the development and yield of maize in response to inoculation with Azospirillum associated with nitrogen fertilization. We conducted two field experiments in the summer harvest, the first in the 2000/2001 crop year in the region of Marechal Cândido Rondon, under conventional tillage, and second in the 2002/2003 agricultural year in the region of Cascavel, under no tillage. The experimental design in both experiments was a randomized complete block, with four replications, 2x2x2 factorial, with two levels of nitrogen at sowing (zero and 20 kg ha-1, two levels of inoculum (zero and 200 g ha-1 and two levels of nitrogen in topdressing (zero and 100 kg ha-1. There was evaluated the height of ear insertion, total plant height, leaf N content, shoot dry biomass and grain yield. The height of ear insertion and total plant height were not influenced by the factors under study. Nitrogen fertilization at sowing increased the leaf N content, causing the opposite effect when combined with inoculation. Inoculation with Azospirillum in the absence of nitrogen, provide productivity increases of 15.4% and 7.4% for 2000/2001 and 2002/2003 crops, respectively. The inoculation provided productivity similar to that obtained with 100 kg ha-1 in topdressing in crop 2000/2001, while in association with the topdressing, reduced productivity and shoot dry biomass in crop 2002/2003.

  3. Plant root transcriptome profiling reveals a strain-dependent response during Azospirillum-rice cooperation

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    Benoît eDrogue

    2014-11-01

    Full Text Available Cooperation involving Plant Growth-Promoting Rhizobacteria results in improvements of plant growth and health. While pathogenic and symbiotic interactions are known to induce transcriptional changes for genes related to plant defence and development, little is known about the impact of phytostimulating rhizobacteria on plant gene expression. This study aims at identifying genes significantly regulated in rice roots upon Azospirillum inoculation, considering possible favored interaction between a strain and its original host cultivar. Genome-wide analyses of Oryza sativa japonica cultivars Cigalon and Nipponbare were performed, by using microarrays, seven days post inoculation with A. lipoferum 4B (isolated from Cigalon or Azospirillum sp. B510 (isolated from Nipponbare and compared to the respective non-inoculated condition. A total of 7,384 genes were significantly regulated, which represent about 16 % of total rice genes. A set of 34 genes is regulated by both Azospirillum strains in both cultivars, including a gene orthologous to PR10 of Brachypodium, and these could represent plant markers of Azospirillum-rice interactions. The results highlight a strain-dependent response of rice, with 83 % of the differentially expressed genes being classified as combination-specific. Whatever the combination, most of the differentially expressed genes are involved in primary metabolism, transport, regulation of transcription and protein fate. When considering genes involved in response to stress and plant defence, it appears that strain B510, a strain displaying endophytic properties, leads to the repression of a wider set of genes than strain 4B. Individual genotypic variations could be the most important driving force of rice roots gene expression upon Azospirillum inoculation. Strain-dependent transcriptional changes observed for genes related to auxin and ethylene signalling highlight the complexity of hormone signalling networks in the Azospirillum

  4. Results in the application of biofertilizers using Azospirillum and Mycorrhiza, in associations of horticultural crops under semi-greenhouse conditions

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    Dania Bárbara Núñez Sosa

    2013-01-01

    Full Text Available The current research was carried out in the greenhouse “La Condesa”, which belongs to the Empresa de Cultivos Varios “Lenin”, in Jovellanos, Matanzas province. Its main objective was to evaluate the effect of the application of biofertilizer (Azospirillum sp and Micorrizas in combinations of crops (lettuce and radish, or beet and radish. The research included four different treatments: witness, Micorrizas, Azospirillum, Azospirillum +Micorrizas. We used an at random block design. Data were processed after evaluating yields components by using the statistical software called Statgraphics, version 5.0. The research showed a positive response to the application of biofertilizers, resulting in a considerable increase of profits.

  5. Influence of Azospirillum spp. on the nitrogen supply of a gramineous host.

    NARCIS (Netherlands)

    Christiansen-Weniger, C.

    1991-01-01

    The main objectives of this study were to identify factors that control the behaviour of Azospirillum in the rhizosphere of a gramineous plant in order to be able to optimize the association between the bacteria and the host plants in terms of nitrogen supply to the host.Plant produced growth substa

  6. Gluconic acid production and phosphate solubilization by the plant growth-promoting bacterium Azospirillum spp.

    Science.gov (United States)

    Rodriguez, Hilda; Gonzalez, Tania; Goire, Isabel; Bashan, Yoav

    2004-11-01

    In vitro gluconic acid formation and phosphate solubilization from sparingly soluble phosphorus sources by two strains of the plant growth-promoting bacteria A. brasilense (Cd and 8-I) and one strain of A. lipoferum JA4 were studied. Strains of A. brasilense were capable of producing gluconic acid when grown in sparingly soluble calcium phosphate medium when their usual fructose carbon source is amended with glucose. At the same time, there is a reduction in pH of the medium and release of soluble phosphate. To a greater extent, gluconic acid production and pH reduction were observed for A. lipoferum JA4. For the three strains, clearing halos were detected on solid medium plates with calcium phosphate. This is the first report of in vitro gluconic acid production and direct phosphate solubilization by A. brasilense and the first report of P solubilization by A. lipoferum. This adds to the very broad spectrum of plant growth-promoting abilities of this genus.

  7. Azospirillum spp. from native forage grasses in Brazilian Pantanal floodplain: biodiversity and plant growth promotion potential.

    Science.gov (United States)

    Souza, Mayara S T; de Baura, Valter A; Santos, Sandra A; Fernandes-Júnior, Paulo Ivan; Reis Junior, Fábio B; Marques, Maria Rita; Paggi, Gecele Matos; da Silva Brasil, Marivaine

    2017-04-01

    A sustainable alternative to improve yield and the nutritive value of forage is the use of plant growth-promoting bacteria (PGPB) that release nutrients, synthesize plant hormones and protect against phytopathogens (among other mechanisms). Azospirillum genus is considered an important PGPB, due to the beneficial effects observed when inoculated in several plants. The aim of this study was to evaluate the diversity of new Azospirillum isolates and select bacteria according to the plant growth promotion ability in three forage species from the Brazilian Pantanal floodplain: Axonopus purpusii, Hymenachne amplexicaulis and Mesosetum chaseae. The identification of bacterial isolates was performed using specific primers for Azospirillum in PCR reactions and partial sequencing of the 16S rRNA and nifH genes. The isolates were evaluated in vitro considering biological nitrogen fixation (BNF) and indole-3-acetic acid (IAA) production. Based on the results of BNF and IAA, selected isolates and two reference strains were tested by inoculation. At 31 days after planting the plant height, shoot dry matter, shoot protein content and root volume were evaluated. All isolates were able to fix nitrogen and produce IAA, with values ranging from 25.86 to 51.26 mg N mL(-1) and 107-1038 µmol L(-1), respectively. The inoculation of H. amplexicaulis and A. purpusii increased root volume and shoot dry matter. There were positive effects of Azospirillum inoculation on Mesosetum chaseae regarding plant height, shoot dry matter and root volume. Isolates MAY1, MAY3 and MAY12 were considered promising for subsequent inoculation studies in field conditions.

  8. Proline and Abscisic Acid Content in Droughted Corn Plant Inoculated with Azospirillum sp. and Arbuscular Mycorrhizae Fungi

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    NOVRI YOULA KANDOWANGKO

    2009-03-01

    Full Text Available Plants that undergo drought stress perform a physiological response such as accumulation of proline in the leaves and increased content abscisic acid. A research was conducted to study proline and abscisic acid (ABA content on drought-stressed corn plant with Azospirillum sp. and arbuscular mycorrhizae fungi (AMF inoculated at inceptisol soil from Bogor, West Java. The experiments were carried out in a green house from June up to September 2003, using a factorial randomized block design. In pot experiments, two factors were assigned, i.e. inoculation with Azospirillum (0, 0.50, 1.00, 1.50 ml/pot and inoculation with AMF Glomus manihotis (0, 12.50, 25.00, 37.50 g/pot. The plants were observed during tasseling up to seed filling periods. Results of experiments showed that the interaction between Azospirillum sp. and AMF was synergistically increased proline, however it decreased ABA.

  9. Improvement of endophytic Azospirillum colonization by co-inoculation with Cellulomonas Uda ATCC 491

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    Mohammad Javad Mehdipour Moghaddam

    2014-04-01

    Full Text Available Introduction: Most of the plant growth promoting rhizobacteria (PGPR such as Azopirillum if accompanied with strong cellulase producing bacteria such as Cellulomonas, their colonization may be increased and their host plants growth improved. Materials and methods: Six endophytic Azospirilla which isolated from three rice and three wheat cultivars and also one strain from commercial biofertilizer (Green Biotech Co., identified by biochemical tests and 16S rDNA analysis and were studied on the basis of cellulase, pectinase and auxin production and also their chemotaxis toward rice and wheat cultivars exudates was investigated. Two cellulase positive (A5 and A6 and two negative (A2 and A3 strains were selected and their interaction with C. uda ATCC 491 on auxin production and colonization on roots were compared. Results: This study showed that none of the strains had pectinase activity, but the strain isolated from rice had more Carboxy methyl cellulase (CMCase activity. Selected isolates and C. uda ATCC 491 showed chemotaxis toward roots exudates. In most of the isolates, rate of auxin production increased by coculture with C. uda ATCC 491. Also, it was determined that C. uda ATCC 491 promoted the colonization of Azospirillum without or with cellulase activity on rice and wheat roots, respectively. Discussion and conclusion: Co-inoculation Azospirillum with C. uda ATCC 491 improves plant root system due to stimulation or additive effect of auxin production and cellulase activity, followed by more uptakes of water and minerals by roots. Also, it raises the number of colonization niches for useful bacteria such as Azospirillum and finally quantitative and qualitative plant parameters.

  10. Promoting the Growth of Chlorella vulgaris in Secondary Wastewater Treatment Effluent of Tofu Industry using Azospirillum sp

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    Wahyunanto Agung Nugroho

    2016-05-01

    Full Text Available The objective of this research is to investigate the influence of growth promoting bacteria (GPB Azospirillum sp on the growth of microalgae Chlorella vulgaris in wastewater of tofu industry as a medium. To observe the influence, about 106 cells/mL of Chlorella vulgaris was cultivated in 1 L of wastewater of tofu industry. The wastewater was an effluent of an aerobic treatment. Six different treatments were set regarding to the Azospirillum sp added to the medium. The glass was marked as A0 as no GPB inoculants added to the medium, and A2, A4, A6, A8 and A10 for 2 mL, 4mL, 6mL, 8 mL and 10 mL of GPB added to the medium respectively. The concentration of GPB inoculant was 108 cfu per mL. The result showed that the highest number of Chlorella vulgaris population was achieved by addition of 6 mL GPB Azospirillum sp in the day 10, while the highest maximum growth rate was achieved by addition of 10 mL GPB Azospirillum sp  to the medium.

  11. Penggunaan Azospirillum pada Tanah Masam dengan Aluminium Tinggi Terhadap Produksi dan Serapan Nitrogen Rumput Setaria splendida dan Chloris gayana

    Directory of Open Access Journals (Sweden)

    P.D.M.H. Karti

    2005-04-01

    Full Text Available High content of Al on the soil maybe harmful (toxic for plant. Red and yellow podzolic soil was marginal land that characterized by high Al content. Azospirillum is free living N fixing bacteria that can be associated with grass. This research was conducted to find the best yield of grass planted on the soil inoculated with Azospirillum. The research consisted of some steps; 1 soil sampling 2 laboratory research: bacterial isolation, isolate selection, standardized of population, content of IAA 3 pod experiment. Pod experiment in the glass house was designed in completely randomized design, that consisted of six treatments. The variables observed were dry mass production of shoot and root, nitrogen content of shoot and root, and nitrogen absorption. Four best isolates chosen were; SM Setaria, OBIS/BD, PO2 and PM2. Azospirillum isolates enhanced shoot and root production, nitrogen content and N total absorption of tolerance one (S. splendida. The susceptible (C. gayana, Azospirillum significantly enhanced shoot and root nitrogen content, but did not affect the growth, production and N total absorption. Root growth that was inhibited by Al toxicity, decreased the symbiotic capability of nitrogen fixing bacteria. PM2 isolate showed the best effect on production and quality of S. splendida as well as on C. gayana. This isolate will be used for future research. PM2 produces 6.4 ppm Indole Acetic Acid that promoted root growth.

  12. KARAKTERISASI SIFAT-SIFAT BIOKIMIA EKSTRAK KASAR LIPASE EKSTRASELULER BAKTERI Azospirillum sp.PRD1

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    Santi Nur Handayani

    2011-11-01

    Full Text Available Enzim lipase mempunyai peranan penting dalam katalis berbagai reaksi industri satu diantaranya pembuatan flavor melalui reaksi esterifikasi. Lipase adalah biokatalis yang berperan besar dalam aplikasi bioteknologi, seperti dalam sintesis biopolimer, biodiesel, produksi obat, dan produksi flavor. Peningkatan penggunaan lipase untuk industri mendorong dilakukan penelitian untuk mendapatkan sumber-sumber lipase baru. Sumber lipase yang potensial salah satunya adalah bakteri Azospirillum sp.PRD1 dari isolat lokal Laboratorium Mikrobiologi, Fakultas Biologi Universitas Jenderal Soedirman. Tujuan penelitian adalah untuk mendapatkan ekstrak kasar lipase dan menentukan karakteristik sifat-sifat biokimiawinya. Metode yang digunakan antara lain peremajaan bakteri Azospirillum sp.PRD1, dan produksi inokulum, penentuan waktu produksi optimum dan fase pertumbuhan bakteri, ekstraksi dan produksi ekstrak kasar lipase dan penentuan karakteristik sifat-sifat biokimiawinya. Hasil penelitian diperoleh ekstrak kasar lipase dari inokulum berumur 7 jam dan medium produksi dengan induser minyak zaitun yang diinkubasi selama 3 jam memiliki aktivitas spesifik 7,0547 Unit/mg. Lipase ekstrak kasar optimum pada pH 7, suhu 40 oC dan waktu inkubasi selama 25 menit. Lipase merupakan metaloenzim dengan kofaktor Zn2+ , Mn2+, Hg2+, Ca2+, Co2+ and Mg2+.

  13. Cloning and Sequencing of glnB and glnZ Genes from Azospirillum brasilense%巴西固氮螺菌Yu62 glnB基因和glnZ基因的克隆和序列分析

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 杨红; 王娟; 李季伦

    2000-01-01

    glnB基因编码的PII蛋白在巴西固氮螺菌的固氮过程中起着非常重要的调控作用,而glnZ是与glnB高度同源的基因,其产物 Pz可能也在固氮调控中起作用.本研究用PCR法克隆了巴西固氮螺菌Yu62 glnB基因和glnZ基因.DNA序列分析表明glnB和glnZ这2个基因的编码区的长度都为336 bp,编码112个氨基酸.将巴西固氮螺菌Yu62 菌株与标准菌株Sp7的glnB基因和glnZ基因分别进行比较,结果表明这2个菌株的glnB基因在编码区的核苷酸顺序完全相同,而glnZ基因在长336 bp的编码区内有4个碱基发生变化,但改变后都是同义密码子,即编码的氨基酸并未改变.glnB基因和glnZ基因在核苷酸顺序上的同源性高达73.2%,氨基酸顺序的同源性达66.7%.

  14. Colonization of Azospirillum brasilense Yu62 in Wheat Via EGFP%巴西固氮螺菌Yu62的EGFP标记及其在小麦体内的定殖研究

    Institute of Scientific and Technical Information of China (English)

    刘华伟; 王庆贺; 张宏; 王蕊; 肖红利; 郭蔼光

    2009-01-01

    以质粒pEGFP-C1为模板,采用PCR方法特异性扩增增强型绿色荧光蛋白(EGFP)基因全长序列,将其与原核表达载体pVK-100连接,构建成重组载体pVK-EGFP.利用电转化法将重组载体导入巴西固氮螺菌Yu62中,得到EGFP标记菌株.用EGFP标记菌接种小麦'小偃107'种子,室内限菌条件下培养10 d后,用荧光显微镜观测标记菌在小麦体内的定殖规律并观察接菌植株的田间生长状况.结果显示,巴西固氮螺菌Yu62能定殖于小麦根毛区、茎组织的细胞间隙等部位,而且接菌小麦'小偃107'植株在根系发育、株高、分蘖数等方面比对照有较明显的优势.研究表明,巴西固氮螺菌Yu62能够定殖于小麦根茎内,并具有促进植物生长的作用.

  15. Effect of Increasing Maize Yields and Saving Nitrogen Fertilizer by inoculation of Azospirillum brasilense UB37%玉米联合固氮工程菌的增产和节约氮肥的效果

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 李季伦; 姚腾云; 于锦香

    2002-01-01

    连续两年(1999~2000年)在肥力中等的轻质砂壤土中,对玉米联合固氮工程菌UB37和野生菌UB1进行了玉米小区田间接种试验.试验结果表明:在不同水平氮肥的条件下,接种工程菌比接种野生菌和不接菌的对照都有不同程度的增产.特别是在施N水平为尿素180 kg/hm2的试验田中,接种工程菌比接种野生菌平均增产10%,比不接菌对照平均增产19%,生物统计差异显著(P=0.05).在尿素180 kg/hm2条件下接种工程菌的玉米产量与在尿素225 kg/hm2下接种工程菌的产量相当,既可节省20%氮肥,又可达到增产19%的效果.

  16. Protein quantity and quality of safflower seed improved by NP fertilizer and rhizobacteria (Azospirillum and Azotobacter spp.

    Directory of Open Access Journals (Sweden)

    Asia eNosheen

    2016-02-01

    Full Text Available Protein is an essential part of human diet. The aim of present study was to improve the protein quality of safflower seed by the application of plant growth promoting rhizobacteria (PGPR in combination with conventional nitrogen and phosphate (NP fertilizers. The seeds of two safflower cultivar Thori and Saif-32, were inoculated with Azospirillum and Azotobacter and grown under field conditions. Protein content and quality was assessed by crude protein, amino acid analysis and SDS-PAGE. Seed crude protein and amino acids (metheonine, phenylanine and glutamic acid showed significant improvement (55%–1250% by Azotobacter supplemented with quarter dose of fertilizers (BTQ at P≤0.05. Additional protein bands were induced in Thori and Saif-32 by BTQ and BTH (Azotobacter supplemented with half dose of fertilizers respectively. The Azospirillum in combination with half dose of fertilizers (SPH and BTQ enhanced the indole acetic acid (90% and gibberellic acid (23%–27% contents in safflower leaf. Taken together, these data suggest that Azospirillum and Azotobacter along with significantly reduced (up to 75% use of NP fertilizers improved the quality and quantity of safflower seed protein.

  17. Protein Quantity and Quality of Safflower Seed Improved by NP Fertilizer and Rhizobacteria (Azospirillum and Azotobacter spp.)

    Science.gov (United States)

    Nosheen, Asia; Bano, Asghari; Yasmin, Humaira; Keyani, Rumana; Habib, Rabia; Shah, Syed T. A.; Naz, Rabia

    2016-01-01

    HIGHLIGHTS Rhizobacteria (Azotobacter spp.) have improved the quality and quantity of safflower seed protein.Protein quality was confirmed by SDS-PAGE and new bands were found in response to different combinations of rhizobacteria and lower doses of fertilizers.The PGPR application has reduced the use of fertilizers upto 50%. Protein is an essential part of the human diet. The aim of this present study was to improve the protein quality of safflower seed by the application of plant growth promoting rhizobacteria (PGPR) in combination with conventional nitrogen and phosphate (NP) fertilizers. The seeds of two safflower cultivars Thori and Saif-32, were inoculated with Azospirillum and Azotobacter and grown under field conditions. Protein content and quality was assessed by crude protein, amino acid analysis, and SDS-PAGE. Seed crude protein and amino acids (methionine, phenylalanine, and glutamic acid) showed significant improvements (55–1250%) by Azotobacter supplemented with a quarter dose of fertilizers (BTQ) at P ≤ 0.05. Additional protein bands were induced in Thori and Saif-32 by BTQ and BTH (Azotobacter supplemented with a half dose of fertilizer) respectively. The Azospirillum in combination with half dose of fertilizer (SPH) and BTQ enhanced both indole acetic acid (IAA) (90%) and gibberellic acid (GA) (23–27%) content in safflower leaf. Taken together, these data suggest that Azospirillum and Azotobacter along with significantly reduced (up to 75%) use of NP fertilizers could improve the quality and quantity of safflower seed protein. PMID:26941744

  18. Characterization of native strains of Azospirillum spp. and its effect on growth of Oryza sativa L. “rice” in Lambayeque

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    Franklin García

    2010-06-01

    Full Text Available This research was conducted to characterize and determine the effect of native strains of Azospirillum spp. on growth of Oryza sativa L. "rice" as an alternative for the reduction of chemical fertilizers. The bacteria were isolated from roots of rice in Lambayeque. Fragments of previously disinfected roots were planted in Nfb semisolid medium, were nitrogen-fixing bacteria were recognized by a whitish film on the surface and turn from green to blue. The genus Azospirillum was identified in Congo red agar medium, obtained 96 isolates with production of 2.69 to 38.02 ppm indoleacetic acid and 7.95 to 29.09 ppm of fixed nitrogen. The four strains with the highest values were inoculated independently and in combination with 50 and 100 % urea in Tinajones INIA508 rice. It used a completely randomized design and including witnesses’ 50 and 100 % urea and absolute control. All native strains increased height and dry matter of shoot and roots, with effectiveness index of up to 21.77, 102.06 and 126.11 % compared to untreated control. Azospirillum sp. GM-86 selected by the highest value of indoleacetic acid, reached the highest level of effectiveness in height and root dry biomass. The application of 50 % úrea with Azospirillum spp. increased rates of effectiveness. It demonstrated the potential of native strains of Azospirillum spp. to increase the growth of rice as well as the possibility to use associate with low doses of urea.

  19. PEMBUATAN DEKSTRIN DARI PATI UBI KAYU MENGGUNAKAN KATALIS AMILASE HASIL FRAKSINASI DARI Azospirillum sp. JG3

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    Zusfahair

    2012-05-01

    Full Text Available Amilase merupakan salah satu enzim potensial dalam proses industri. Amilase dapat diperoleh dari bakteri Azopsirillum sp. JG3. Amilase ini dapat dimanfaatkan untuk hidrolisis pati ubi kayu pada pembuatan dekstrin. Dekstrin memiliki peran yang cukup penting dalam industri seperti pembuatan roti, makanan bayi dan bahan penyalut lapis tipis tablet. Penelitian ini bertujuan untuk mengetahui karakterisasi biokimiawi amilase hasil fraksinasi dari Azospirillum sp. JG3 serta potensinya dalam pembuatan dekstrin. Tahap penelitian ini dimulai dengan memproduksi enzim yang dilakukan dengan cara menumbuhkan inokulum ke dalam medium produksi selama 30 jam kemudian diekstraksi dengan sentrifugasi sehingga diperoleh ekstrak kasar. Ekstrak kasar amilase kemudian difraksinasi secara bertahap (15%, 30%, 45% dan 60% dengan menggunakan ammonium sulfat dan didialisis. Fraksi yang diperoleh diukur aktivitas, volume dan kadar proteinnya. Fraksi dengan aktivitas spesifik tertinggi dikarakterisasi dan digunakan untuk membuat dekstrin dari pati ubi kayu. Dekstrin pati ubi kayu yang diperoleh dikarakterisasi berupa rendemen, kadar air, kadar abu dan dextrose equivalen. Hasil penelitian menunjukkan bahwa fraksi hasil dialisis 45% (FHD 45% menghasilkan aktivitas dan aktivitas spesifik tertinggi berturut-turut sebesar 11,659 U/mL dan 7,319 U/mg. Aktivitas amilase optimum pada suhu 30°C, pH 7 dan konsentrasi substrat 4%. Dekstrin yang diperoleh memiliki rendemen 95,14, kadar air 3,411%, kadar abu 0,209% dan dextrose equivalen 13,4.

  20. Influência da inoculação de Azospirillum sp. em trigo, cevada e aveia / Influence of Azospirillum sp. inoculation in wheat, barley and oats

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    Osmar Roberto Dalla Santa

    2008-05-01

    Full Text Available The aim of this study has been to evaluate the infl uence of inoculation with Azospirillum sp. RAM-7 in wheat, barley and oats on yields, density and N total of grains. The experiments were conducted under fi eld conditions. The treatments involved bacterial inoculation with or without different levels of nitrogen fertilization and absolute control. The inoculation was carried out by the seed coating method with bacterial suspension containing 1010 CFU.mL-1. Statistically signifi cant effects on grain yield in wheat were obtained at the inoculated treatment without any nitrogen fertilizer, increasing the yield by 23.9% over de control (p<0.05. In the inoculated treatment associated with 48 kg of N.ha-1, the yield was similar to that obtained at 60 kg of N.ha-1 with or without inoculation. In barley, the grain yields of the inoculated treatment associated with 32 kg of N.ha-1 was statistically equivalent to that obtained with 40 kg de N.ha-1 with or without inoculation (p<0,05. In oats, there was no signifi cant difference between treatments and their respective controls for yields, density and N total of grains. The results of the present investigations showed the possibility of the partial substitution of the inorganic nitrogen fertilizer with the Azospirillum sp. RAM-7 inoculation in wheat, barley and oats.

  1. Complete Genomic Structure of the Cultivated Rice Endophyte Azospirillum sp. B510

    Science.gov (United States)

    Kaneko, Takakazu; Minamisawa, Kiwamu; Isawa, Tsuyoshi; Nakatsukasa, Hiroki; Mitsui, Hisayuki; Kawaharada, Yasuyuki; Nakamura, Yasukazu; Watanabe, Akiko; Kawashima, Kumiko; Ono, Akiko; Shimizu, Yoshimi; Takahashi, Chika; Minami, Chiharu; Fujishiro, Tsunakazu; Kohara, Mitsuyo; Katoh, Midori; Nakazaki, Naomi; Nakayama, Shinobu; Yamada, Manabu; Tabata, Satoshi; Sato, Shusei

    2010-01-01

    We determined the nucleotide sequence of the entire genome of a diazotrophic endophyte, Azospirillum sp. B510. Strain B510 is an endophytic bacterium isolated from stems of rice plants (Oryza sativa cv. Nipponbare). The genome of B510 consisted of a single chromosome (3 311 395 bp) and six plasmids, designated as pAB510a (1 455 109 bp), pAB510b (723 779 bp), pAB510c (681 723 bp), pAB510d (628 837 bp), pAB510e (537 299 bp), and pAB510f (261 596 bp). The chromosome bears 2893 potential protein-encoding genes, two sets of rRNA gene clusters (rrns), and 45 tRNA genes representing 37 tRNA species. The genomes of the six plasmids contained a total of 3416 protein-encoding genes, seven sets of rrns, and 34 tRNAs representing 19 tRNA species. Eight genes for plasmid-specific tRNA species are located on either pAB510a or pAB510d. Two out of eight genomic islands are inserted in the plasmids, pAB510b and pAB510e, and one of the islands is inserted into trnfM-CAU in the rrn located on pAB510e. Genes other than the nif gene cluster that are involved in N2 fixation and are homologues of Bradyrhizobium japonicum USDA110 include fixABCX, fixNOQP, fixHIS, fixG, and fixLJK. Three putative plant hormone-related genes encoding tryptophan 2-monooxytenase (iaaM) and indole-3-acetaldehyde hydrolase (iaaH), which are involved in IAA biosynthesis, and ACC deaminase (acdS), which reduces ethylene levels, were identified. Multiple gene-clusters for tripartite ATP-independent periplasmic-transport systems and a diverse set of malic enzymes were identified, suggesting that B510 utilizes C4-dicarboxylate during its symbiotic relationship with the host plant. PMID:20047946

  2. Produção, caracterização e aplicação de anticorpo policlonal contra Azospirillum amazonense estirpe Am15 Production, characterization and application of polyclonal antibodies against Azospirillum amazonense strain Am15

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    Marinete Flores da Silva

    2009-01-01

    Full Text Available O uso de ferramentas moleculares e imunológicas permite a detecção e o monitoramento específico de microrganismos usados como inoculantes agrícolas. O maior exemplo de sucesso com uso de bactérias diazotróficas na agricultura é o caso da inoculação de soja no Brasil. Entretanto, a inoculação de bactérias diferentes de rizóbio não permite a localização de um sítio específico onde ocorra A redução do nitrogênio atmosférico, pois não há a formação de estruturas nodulares. Este trabalho foi realizado na Embrapa-Agrobiologia, e objetivou produzir e caracterizar um anticorpo policlonal a partir da inoculação de células intactas da estirpe Am15 pertencente à espécie Azospirillum amazonense (Anti-Am15 utilizando o método de ELISA indireto. O anticorpo foi usado em sementes peletizadas com turfa contendo a bactéria-alvo e avaliada sua sobrevivência durante 60 dias. Este anticorpo foi capaz de quantificar populações bacterianas em amostras cujo número mínimo celular foi superior a valores de 100.000 células por mL. De acordo com os resultados, o soro policlonal foi especifico contra o antígeno de interesse, sendo possível sua utilização em estudos de quantificação e monitoramento do Azospirillum sp em plantas de milho que receberam inóculo e no controle da qualidade de inoculante turfoso.Molecular and immunological approaches are useful for monitoring the localization of microrganism used as agricultural inoculants. A successful example is the use of a diazotrophic bacteria, as soybean in Brazil. However, as diazotrophic bacterias other than rizobium do not form nodular structures, the localization of the specific site where nitrogen reduction occurs is impossible. In this study, carried out at Embrapa Agrobiology, a polyclonal antibody against Azospirillum amazonense (As-Am15 was to produced and characterized through an indirect Enzyme Linked Immunosorbent Assay (ELISA. Maize seedscoated with peat containing

  3. Yield and yield components of wheat as affected by salinity and inoculation with Azospirillum strains from saline or non-saline soil

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    Somayeh Haji Nia

    2012-06-01

    Full Text Available In many arid and semi-arid areas of the world where sustainability of agriculture is limited by salinity, use of biological potential may be a key component of sustainable plant production. A greenhouse experiment was used to test the effectiveness of inoculation with Azospirillum strains isolated from saline or non-saline soil in alleviating the salinity stress in wheat plants grown with irrigation water with different electrical conductivities (ECw of 0.7, 4, 8 and 12 dS m−1. Inoculation with the two isolates increased salinity tolerance of wheat plants; the saline-adapted isolate significantly increased shoot dry weight and grain yield under severe water salinity. The component of grain yield most affected by inoculation was grains per plant. Plants inoculated with saline-adapted Azospirillum strains had higher N concentrations at all water salinity levels.

  4. Actividad nitrogenasa, producción de fitohormonas, sideróforos y antibiosis en cepas de Azospirillum y Klebsiella aisladas de maíz y teocintle

    OpenAIRE

    Moisés Graciano Carcaño-Montiel; Ronald Ferrera-Cerrato; Jesús Pérez-Moreno; José D. Molina-Galán; Yoav Bashan

    2006-01-01

    Las bacterias de los géneros Azospirillum y Klebsiella, conocidas como rizobacterias promotoras del crecimiento vegetal, son fijadoras de nitrógeno y productoras de sustancias reguladoras del crecimiento, por lo que ejercen un efecto benéfico en las plantas con las cuales interaccionan. Esta asociación microorganismo-planta puede mejorar el crecimiento de los cultivos a través de la combinación de fijación biológica de nitrógeno, producción de sustancias hormonales, incremento de la disponi...

  5. Produtividade do milho em resposta à aplicação de nitrogênio e inoculação das sementes com Azospirillum spp Corn produntivity in response to nitrogen application and seed inoculation with Azospirillum spp

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    Luiz Ermindo Cavallet

    2000-04-01

    Full Text Available Em experimento de campo, nas condições de plantio direto em Latossolo Vermelho eutroférrico de fertilidade média a alta, foram testados para o milho (Zea mays, cv. G-159 S da Ciba Sementes no período de outubro/1997 a março/1998, o aumento de produtividade de grãos, o comprimento de espiga, a altura de plantas e o número de linhas de grão por espiga, por conseqüência da inoculação das sementes com o produto biotecnológico comercial denominado "Graminante", à base de Azospirillum spp. Os tratamentos consistiram em ausência e presença de inoculação de "Graminante", com e sem aplicação de nitrogênio tanto, da semeadura (70 kg ha-1 como em cobertura (70 kg ha-1 consistindo, deste modo, numa distribuição fatorial 2 x 2 x 2, em quatro repetições. A inoculação do "Graminante" aumentou significativamente a produtividade média de grãos, de 5211 para 6067 kg ha-1, ou seja, aumento de 17%, e aumentou o comprimento médio das espigas, de 13,6 para 14,4 cm; a inoculação do "Graminante" não teve efeito sobre o número de linhas de grão por espiga e altura de plantas. A aplicação de nitrogênio na semeadura e/ou em cobertura, não causou efeito sobre a produtividade do milho.An evaluation of corn productivity, cob length, plant height and number of corn lines on the cob as a result of nitrogen application and seed inoculation with Azospirillum spp. ("Graminante" was conducted from October 1997 to March 1998. The experiment was carried out on a high fertility Oxisol using Zea mays L. cv. G-1595 (Ciba under no-tillage in a 2 x 2 x 2 factorial design representing application and non-application of nitrogen at planting (70 kg ha-1, application and non-application of broadcast nitrogen (70 kg ha-1 and presence or absence of seed inoculation with four replications. Results showed that with seed inoculation the corn productivity increased by 17% (from 5211 to 6067 kg ha-1 while corn cob length increased from 13.6 to 14.4 cm. On the

  6. Efeito da inoculação de Azospirillum e Herbaspirillum na produção de compostos indólicos em plântulas de milho e arroz Effect of inoculation with Azospirillum and Herbaspirillum on production of indolic compounds and growth of wheat and rice seedlings

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    Tharwat El-Sayed El-Desouk Radwan

    2004-10-01

    Full Text Available Bactérias fixadoras de nitrogênio podem contribuir para o crescimento vegetal pela produção de auxinas. Os objetivos deste trabalho foram quantificar a produção de hormônios de crescimento por estirpes de bactérias fixadoras de nitrogênio e avaliar o efeito da inoculação destas estirpes em plântulas de milho e trigo. Todas as estirpes avaliadas começaram a produzir indóis no final da fase logarítmica. Houve efeito da adição de diferentes concentrações de triptofano ao meio de cultivo no aumento da produção de indóis até o nível de 200 µM. Nas estirpes de Azospirillum, as formas de N: NH4Cl, (NH42SO4, NH4H2PO4 e NH4NO3 estimularam a produção de indóis em níveis baixos. A adição de KNO3, NaNO3 e KNO2 inibiu a produção de indóis em todas as bactérias testadas e no crescimento de células de Azospirillum. Os efeitos da inoculação foram também comparados com concentrações crescentes do ácido 3-indolacético, KNO3 e triptofano. Em condições axênicas, a elevada produção de indóis reduz o comprimento das raízes e colmos na presença de triptofano, especialmente quando submetidas à inoculação de Azospirillum.Nitrogen fixing bacteria can contribute to plant growth by the production of auxin. The objectives of this work were to quantify the production of plant growth hormones by different nitrogen-fixing bacteria and to evaluate the effect of this inoculation on growth of wheat and rice seedlings. All strains tested produced indoles during the late logarithmic phase. Increasing the amount of tryptophan in the medium enhanced the production of indoles by the bacteria up to the level of 200 µM. For the Azospirillum strains studied, the nitrogen sources: NH4Cl, (NH42SO4, NH4H2PO4 and NH4NO3 stimulated indoles production in low levels. The addition of KNO3, NaNO3 and KNO2 inhibited the production of indole compounds for all bacteria tested as well as the Azospirillum growth. The effects of inoculation were

  7. Detecção de Azospirillum amazonense em raízes e rizosfera de orchidaceae e de outras famílias vegetais

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    A. Lange

    2002-06-01

    Full Text Available Azospirillum amazonense é uma bactéria fixadora de N2 atmosférico de ampla ocorrência, principalmente em associações radiculares com gramíneas e palmeiras. Para verificar sua presença em outras espécies vegetais, ainda não estudadas, e a eficiência de meios para sua detecção, foram testados os meios Fam e LGI para contagens em solo rizosférico ou em culturas de enriquecimento com solo rizosférico, ecto e endorrizosfera. A. amazonense foi detectada no solo rizosférico, ecto e endorrizosfera de várias espécies de monocotiledôneas, incluindo Orchidaceae e dicotiledôneas, sendo o meio Fam mais eficiente para sua detecção

  8. Restrição do 16S-23S DNAr intergênico para avaliação da diversidade de Azospirillum amazonense isolado de Brachiaria spp. Restriction of 16S-23S intergenic rDNA for diversity evaluation of Azospirillum amazonense isolated from different Brachiaria spp.

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    Fábio Bueno dos Reis Junior

    2006-03-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade intra-específica de isolados de Azospirillum amazonense e estabelecer a possível influência de diferentes espécies de Brachiaria ssp. e diferentes condições edafoclimáticas. A caracterização da diversidade desses isolados foi conduzida, utilizando-se a análise de restrição da região intergênica 16S-23S DNAr. As estirpes estudadas separaram-se em dois grupos, definidos a 56% de similaridade. As espécies de Brachiaria ssp. influenciaram a diversidade de estirpes. A maioria dos isolados oriundos de B. decumbens e B. brizantha está inserida no primeiro grupo, enquanto os oriundos de B. humidicola concentram-se no segundo grupo.The aim of this work was to study the intra-specific diversity of Azospirillum amazonense isolates and to establish possible influences of different Brachiaria spp. and edaphoclimatic conditions. The characterization of the diversity among the isolates of A. amazonense studied was conducted using restriction analysis of the 16S-23S rDNA intergenic spacer region. The evaluated strains were separated in two groups, defined at 56% of similarity. Brachiaria spp. showed effects on strain diversity. Most part of the isolates from B. decumbens and B. brizantha are inserted in the first group, while B. humidicola isolates concentrate in the second group.

  9. Study on nodulation and nitrogen fixation of Azospirillum brasilense induced by 2,4- D in the roots of maize%2,4-D诱导固氮螺菌在玉米根部结瘤及固氮效果的研究

    Institute of Scientific and Technical Information of China (English)

    李素翠; 聂延富

    2001-01-01

    采用2,4-D诱瘤法诱导固氮螺菌在玉米根部结人工根瘤并且固氮.最佳的2,4-D诱瘤浓度为0.9mg/kg.用乙炔还原法和15N2示踪法均测到了固氮酶活性;并且在用乙炔测定固氮酶活性时发现了4h延迟期的存在.用2,4-D加固氮螺菌处理的玉米比未加2,4-D只用固氮螺菌处理的玉米可以忍耐更高的氧压,这说明2,4-D诱发的人工根瘤的形成为固氮螺菌提供了一个屏氧场所.

  10. Inoculação de Azospirillum amazonense em dois genótipos de milho sob diferentes regimes de nitrogênio Inoculation of Azospirillum amazonense in two maize genotypes under different N treatments

    Directory of Open Access Journals (Sweden)

    Fábio Bueno dos Reis Junior

    2008-06-01

    Full Text Available A adaptabilidade de genótipos de milho a ambientes onde os nutrientes não estejam prontamente disponíveis pode ser relacionada com a adaptação à predominância, nos solos, das formas de N nítrica e amoniacal e à associação com microrganismos benéficos como as bactérias diazotróficas e, ou, promotoras de crescimento de plantas. O objetivo deste trabalho foi verificar o comportamento de dois híbridos intervarietais de milho sob diferentes doses e formas de N, assim como o efeito da inoculação com Azospirillum amazonense, em um ensaio em casa de vegetação, utilizando vasos preenchidos com vermiculita e solução nutritiva de Hoagland como fonte de nutrientes. O delineamento experimental foi de blocos ao acaso em esquema fatorial, constituindo os tratamentos em: dois híbridos intervarietais, H1 (Carioca x Eldorado e H2 (Palha Roxa ES x Sol da Manhã; plantas inoculadas ou não inoculadas com A. amazonense, e três regimes de N, conforme doses e proporções seguintes: 126 mg semana-1 de N (75 % NH4+ : 25 % NO3-; 126 mg semana-1 de N (25 % NH4+ : 75 % NO3-, e 12,6 mg semana-1 de N (50 % NH4+ : 50 % NO3-. Vinte e cinco dias após o plantio, as plantas foram separadas em raiz e parte aérea para determinação da matéria seca, conteúdos totais de N e P, atividade das enzimas nitrato redutase e glutamina sintetase e açúcares solúveis totais. O híbrido H1, mais eficiente em avaliações preliminares em campo, produziu mais matéria seca e apresentou maior eficiência na utilização de N e P. Plantas que receberam a maior dose de N, independentemente do predomínio da forma nítrica ou amoniacal, apresentaram maior acúmulo de N (nas raízes e parte aérea e P (na parte aérea e maior índice de utilização de P. Com o predomínio da forma amoniacal, observou-se incremento na produção de matéria seca da parte aérea e na atividade da glutamina sintetase, bem como decréscimo na concentração de açúcares solúveis totais

  11. Search for endophytic diazotrophs in barley seeds

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    Myriam S. Zawoznik

    2014-06-01

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

  12. Search for endophytic diazotrophs in barley seeds.

    Science.gov (United States)

    Zawoznik, Myriam S; Vázquez, Susana C; Díaz Herrera, Silvana M; Groppa, María D

    2014-01-01

    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.

  13. Enzymology of biological nitrogen fixation. Annual report

    Energy Technology Data Exchange (ETDEWEB)

    Burris, R.H.

    1992-05-01

    Two genes involved in the regulation of nitrogenase activity, draT and draG, were cloned and found to be contiguous on the Azospirillum brasilense chromosome. The nifH gene, encoding dinitrogenase reductase, is near to draT with an intervening gap of 1.9 kb. The organization of these genes in Azospirillum lipoferum and Rhodosprillum rubrum is similar, but nifH and draT are separated by only 400 bp in the organisms. A. brasilense draTG is very similar to draTG in R. rubrum with 91.8% similarity and 85.3% identity at the amino acid level. Apparently A. brasilense uses the normal ATG initiation codon for draT, and draG. The genes for A. brasilense were able to restore function to appropriate mutants of R. rubrum. The heterologous expression of A. brasilense draTG in R. rubrum was not fully normal, as it responded more slowly to darkness and more quickly to ammonia than wild type cells. Our mutational analysis of the draTG region of A. brasilense confirms the function of these genes in the regulation of nitrogenase activity, but it also revealed minor but demonstrable differences in the control systems of R. rubrum and A. brasilense.

  14. Enzymology of biological nitrogen fixation

    Energy Technology Data Exchange (ETDEWEB)

    Burris, R.H.

    1992-01-01

    Two genes involved in the regulation of nitrogenase activity, draT and draG, were cloned and found to be contiguous on the Azospirillum brasilense chromosome. The nifH gene, encoding dinitrogenase reductase, is near to draT with an intervening gap of 1.9 kb. The organization of these genes in Azospirillum lipoferum and Rhodosprillum rubrum is similar, but nifH and draT are separated by only 400 bp in the organisms. A. brasilense draTG is very similar to draTG in R. rubrum with 91.8% similarity and 85.3% identity at the amino acid level. Apparently A. brasilense uses the normal ATG initiation codon for draT, and draG. The genes for A. brasilense were able to restore function to appropriate mutants of R. rubrum. The heterologous expression of A. brasilense draTG in R. rubrum was not fully normal, as it responded more slowly to darkness and more quickly to ammonia than wild type cells. Our mutational analysis of the draTG region of A. brasilense confirms the function of these genes in the regulation of nitrogenase activity, but it also revealed minor but demonstrable differences in the control systems of R. rubrum and A. brasilense.

  15. Inoculación con Azospirillum spp. en la Región Semiárida-Central de Argentina: factores que afectan la colonización rizosférica Inoculation with Azospirillum spp. in the semiarid-central region of Argentina: factors involved in rhizosphere colonization

    Directory of Open Access Journals (Sweden)

    Adriana Abril

    2006-07-01

    Full Text Available La efectividad de la inoculación con Azospirillum spp. en condiciones de campo a menudo es poco consistente. La colonización de la rizosfera por las bacterias del inoculante es un prerrequisito para el éxito de la inoculación. El objetivo de este trabajo fue evaluar el grado de colonización de raíces en 11 ensayos de campo con diferentes tratamientos (localidades, cepas, cultivos, cultivares, etc. en la Región semiárida central de la Argentina. Todos los ensayos se realizaron con similar diseño experimental y metodología de análisis para permitir su comparación. El grado de colonización mostró alta variabilidad entre los casos analizados (CV 173%, rango: -18 a >100%. Sólo 11 de los 32 casos analizados presentaron diferencias significativas entre el tratamiento inoculado y el control. No se detectó un patrón definido que permita explicar la alta heterogeneidad de los resultados, sin embargo, se podría especular, que los principales factores que pueden haber afectado el grado de colonización son: el estrés hídrico y el origen de las cepas del inoculante. Bajo condiciones de estrés puede existir una fuerte competencia entre las poblaciones rizosféricas, situación en la cual, las cepas nativas tienen ventaja por su mayor adaptación al medio.The effectivity of inoculation with Azospirillum spp. often lacks consistency under field conditions. Microbial colonization of the rhizosphere is considered a prerequisite for successful inoculation. The objective of this study was to determine the degree of rhizosphere colonization in 11 field inoculation assays under different treatments (sites, crops, strains, cultivars, etc. performed in the semiarid central region of Argentina in the last 15 years, conforming a total of 32 study cases. For comparative purposes, all the assays were performed following the same experimental design and inoculation and analytic methods. Variation results relative to control treatment were clustered

  16. Evaluación de la zeolita como soporte sólido para la formulación del biofertilizante Azospirillum

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    Dayamí Borges-Rodríguez

    2012-01-01

    Full Text Available Se realizó una caracterización física de la zeolita proveniente de los yacimientos de San Juan de la Yeras, con el objetivo de evaluar sus posibilidades para ser usada como soporte sólido en la formulación del biofertilizante Azospirillum. Se evaluaron fracciones de tres tamaños de partícula, de las que la fracción Zoac (<0,8 mm fue la más apropiada de acuerdo con sus características y disponibilidad. Se realizaron experimentos mediante un diseño factorial 22 para evaluar la estabilidad del inoculante formulado en soporte sólido, teniendo en cuenta dos niveles de relación de inoculación y concentración celular del producto. De acuerdo con los resultados obtenidos, el producto líquido no puede ser almacenado a temperatura ambiente por un período superior a 14 días, mientras que la zeolita provee al microorganismo de un ambiente adecuado para su mantenimiento por un período de 90 días. La inoculación más efectiva resultó ser la realizada al 21 % con una titulación del producto al salir del fermentador de 1 x 109 UFC/ml para una tasa de extinción celular de 1,4 x 106 cel/ml-1/día-1. La poca cantidad de inóculo líquido que es capaz de admitir este material y su bajo contenido de materia orgánica constituyen desventajas que debe ser consideradas para el establecimiento de una formulación definitiva del biofertilizante.

  17. Survival of endophytic diazotrophic bacteria in soil under different moisture levels Sobrevivência de bactérias diazotróficas endofíticas no solo sob diferentes teores de umidade

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    André L.M. Oliveira

    2004-12-01

    Full Text Available The effects of soil moisture on the survival of three diazotrophic bacteria species (Azospirillum amazonense, Gluconacetobacter diazotrophicus and Azospirillum brasilense were tested. Soil moisture had little influence on the survival of A. brasilense, which is considered a free-living species. On the other hand, increased soil moisture extended the survival of the endophytes A. amazonense and G. diazotrophicus. These results indicate that nitrogen-fixing endophytic species are more affected by soil moisture than associative nitrogen-fixing species.Neste trabalho foi avaliado o efeito da umidade do solo na sobrevivência de três espécies de bactérias diazotróficas (Azospirillum amazonense, Gluconacetobacter diazotrophicus e Azospirillum brasilense. O teor de umidade apresentou pouca influência na sobrevivência de A. brasilense, considerada uma espécie cosmopolita, enquanto A. amazonense e G. diazotrophicus, consideradas endofíticas, aumentaram o período de culturabilidade na presença de umidade no solo. Os resultados demonstram que o teor de umidade do solo possui maior influência nas espécies endofíticas, em comparação às espécies associativas.

  18. PHYSIOLOGICAL QUALITY OF CORN SEEDS REINOCULATED WITH DIAZOTROPHIC BACTERIA

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    Ricardo Felipe Braga de Sousa

    2015-09-01

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

  19. Identificação de isolados de Azospirillum amazonense associados a Brachiaria spp., em diferentes épocas e condições de cultivo e produção de fitormônio pela bactéria Identification of Azospirillum amazonense isolates associated to Brachiaria spp. at different stages and growth conditions, and bacterial plant hormone production

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    F. B. Reis Junior

    2004-02-01

    Full Text Available Existem evidências de que pastagens formadas por algumas espécies do gênero Brachiaria poderiam beneficiar-se com o processo de fixação biológica do nitrogênio atmosférico (FBN, garantindo a estas pastagens maior longevidade. Dentre as bactérias diazotróficas encontradas em associação com estas gramíneas forrageiras, destaca-se a espécie Azospirillum amazonense. Neste trabalho, objetivou-se verificar a influência da espécie de Brachiaria, manejo da pastagem e sazonalidade sobre as populações de A. amazonense associadas às raízes destas plantas. Diferentes pastagens (B. humidicola, B. decumbens e B. brizantha foram introduzidas em regiões do ecossistema Cerrado e de Mata Atlântica. Foram avaliados dois sistemas de manejo com diferentes taxas de lotação, e as coletas foram realizadas em diferentes épocas do ano. As populações de A. amazonense foram quantificadas e a identidade dos isolados confirmada, assim como sua capacidade de produção de fitormônios tipo AIA (ácido 3-indol acético em meio de cultivo. Isolados de A. amazonense foram obtidos a partir de amostras de raízes das três espécies de Brachiaria avaliadas. Estimativas das populações desta bactéria variaram de 10³-10(7 células g-1 de raízes. Em amostras do ecossistema Cerrado, a época de coleta apresentou efeito significativo sobre a população destas bactérias. Os dados da região de Mata Atlântica mostraram que plantas de Brachiaria de diferentes espécies e pastagens sob diferentes taxas de lotação podem apresentar números populacionais distintos associados às suas raízes. A técnica de análise de restrição do DNA ribossomal amplificado (ARDRA confirmou a identidade de todos os isolados avaliados. Estes isolados foram capazes de produzir fitormônios tipo AIA.There is evidence that pastures formed by some species of the genus Brachiaria could be benefited by the biological nitrogen fixation process (BNF, which would give these

  20. Assessing the Effect of Organic Compounds, Biofertilizers and Chemical Fertilizers on Morphological Properties,yield and Yield Components of Forage Sorghum (Sorghum bicolor

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    A.H Saeidnejad

    2012-12-01

    Full Text Available Recently, using the source of organic fertilizers and biofertilizers in sustainable crop production is growing. In order to evaluate the effect of organic compounds, biofertilizers and chemical fertilizer on morphological properties, yield and yield components of forage Sorghum (sorghum bicolor a field experiment was conducted in the Research Farm, College of Agriculture, Ferdowsi University of Mashhad in 2008.The treatments were seed inoculation with the combination of Azotobacter chroococcum and Azospirillum brasilense, Compost (15 t/ha, Vermicompost (10 t/ha, seed inoculation with Azotobacter and Azospirillum and compost (10t/ha, seed inoculation with Azotobacter chroococcum and Azospirillum brasilense and Vermicompost (7t/ha, seed inoculation with Pseudomonas flurescence, seed inoculation with Pseudomonas flurescence and Azotobacter chroococcum and Azospirillum brasilense combination, seed inoculation with Pseudomonas flurescence and compost (15t/ha, chemical fertilizer (80 kg/h urea fertilizer and 50 kg/h super phosphate fertilizer and control. Harvesting was performed in 2 cuts in flowering stage. Plant height, number of tiller per plant and SPAD reading was significantly affected by the treatments. Stem diameter was not affected by any treatments. There was a significant difference among all treatments in terms of fresh and dry forage yield. There were no significant differences among all treatments in terms of stem and leaf dry matter. In general, result of this experiment indicated that organic amendments and biofertilizers could be acceptable alternatives for chemical fertilizers.

  1. Effect of plant growth-promoting bacteria on the growth and fructan production of Agave americana L.

    OpenAIRE

    Neyser De La Torre-Ruiz; Víctor Manuel Ruiz-Valdiviezo; Clara Ivette Rincón-Molina; Martha Rodríguez-Mendiola; Carlos Arias-Castro; Federico Antonio Gutiérrez-Miceli; Héctor Palomeque-Dominguez; Reiner Rincón-Rosales

    2016-01-01

    ABSTRACT The effect of plant growth-promoting bacteria inoculation on plant growth and the sugar content in Agave americana was assessed. The bacterial strains ACO-34A, ACO-40, and ACO-140, isolated from the A. americana rhizosphere, were selected for this study to evaluate their phenotypic and genotypic characteristics. The three bacterial strains were evaluated via plant inoculation assays, and Azospirillum brasilense Cd served as a control strain. Phylogenetic analysis based on the 16S rRN...

  2. Optical methods for creating delivery systems of chemical compounds to plant roots

    Science.gov (United States)

    Kuznetsov, Pavel E.; Rogacheva, Svetlana M.; Arefeva, Oksana A.; Minin, Dmitryi V.; Tolmachev, Sergey A.; Kupadze, Machammad S.

    2004-08-01

    Spectrophotometric and fluorescence methods have been used for creation and investigation of various systems of target delivery of chemical compounds to roots of plants. The possibility of using liposomes, incrusted by polysaccharides of the external surface of nitrogen-fixing rizospheric bacteria Azospirillum brasilense SP 245, and nanoparticles incrusted by polysaccharides of wheat roots, as the named systems has been shown. The important role of polysaccharide-polysaccharide interaction in the adsorption processes of bacteria on wheat roots has been demonstrated.

  3. Microorganisms associated to tomato seedlings growing in saline culture act as osmoprotectant

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    Daniel Cortés-Jiménez

    2014-06-01

    Full Text Available Less than 0.5% of total water in the world is available for human consumption and agriculture. The major part of the world's water is saline and salinity in soils interferes in germination of seeds and the posterior development of the plant. In order to increase the osmotolerance of tomato, seedlings were associated with Azospirillum brasilense Cd, Azospirillum brasilense Cd transformed bacteria with a plasmid harboring a trehalose biosynthesis gene-fusion or Chlorella vulgaris. Two plant culture media: Hydroponic and Murashige and Skoog were tested. In the first set of studies seedlings were associated to single free cells meanwhile in a second set single and combined free cells were studied. A positive interaction between transformed Azospirillum and Chlorella vulagris and tomato plants was observed. Seedlings showed a salt concentration tolerance, as sodium chloride, up to 200 mM. According to our results, the association of plants with A. brasilense Cd-BIF and C. vulgaris is a viable approach to increase their salt tolerance and biomass, as consequence the possible use of sea water to irrigate horticultural plants.

  4. PRODUTIVIDADE DA SOJA EM RESPOSTA Á INOCULAÇÃO PADRÃO E COINOCULAÇÃO

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    Ivana Marino Bárbaro

    2009-06-01

    Full Text Available The objectives of this research were to evaluate the response to a standard inoculation with Bradyrhizobium, as well as the co-inoculation with Azospirillum brasilense and Bradyrhizobium in soybean sown area already established with the culture. The rehearsal was installed in Colina-SP, in november of 2007, with the use of the soybean cultivar MG BR 46 (Conquista. The treatments were: 1 - control without N and without inoculation of Bradyrhizobium, 2-control with 200 kg N / ha split, 3-inoculation with Bradyrhizobium, 4-coinoculation with Bradyrhizobium and Azospirillum brasilense peat and 5 - co-inoculation with Bradyrhizobium and Azospirillum brasilense liquid. The experimental design was randomized blocks, with four replications. The experimental plots were constituted by six rows of four meters of length, with spacing of 0,50 m. In V6, were evaluated the nodulation, root dry mass and shoot dry mass. The crop was accomplished manually, in the useful area of the plot, being dear the yield of grains in kg/ha, after obtaining of the mass of grains for portion, being determined the text of water, calculated in 13% of humid base. Besides, the mass of the thousand seeds/plot was evaluated, in grams. In general, it was found that inoculation as well as the coinoculation did not cause an increase in most parameters assessed and soybean yield when grown in area already established with the culture, in Colina-SP.

  5. LA BIOFERTILIZACIÓN CON RIZOBACTERIAS Y HONGOS MICORRÍZICOS ARBUSCULARES EN LA PRODUCCIÓN DE POSTURAS DE TOMATE (Lycopersicon esculentum Mill. Y CEBOLLA (Allium cepa L.. I. CRECIMIENTO VEGETATIVO

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    L. E. Pulido

    2003-01-01

    Full Text Available En áreas experimentales de la Universidad de Ciego de Ávila, sobre un suelo Ferralítico Rojo compactado eútrico y durante dos campañas hortícolas sucesivas, se estudió el efecto de la inoculación, simple y combinada, mediante recubrimiento de las semillas y prescindiendo de la fertilización mineral, con cuatro y cinco especies, respectivamente, de rizobacterias promotoras del crecimiento vegetal -RPCV- (Azospirillum brasilense, Azotobacter chroococcum, Burkholderia cepacia y Pseudomonas fluorescens y de hongos micorrízicos arbusculares -HMA- (Glomus clarum, G. fasciculatum, G. mosseae, G. agregatum y G. versiculiferum, sobre algunos indicadores del crecimiento de posturas de tomate y cebolla, tomando como criterio de evaluación la altura y la longitud radical de las plántulas. Los resultados mostraron que, para el tomate, la inoculación con Azospirillum brasilense, Azotobacter chroococcum y Burkholderia cepacia permitió obtener posturas de calidad equivalente a la alcanzada con la fertilización mineral, mientras que para la cebolla, solo Azospirillum brasilense y Azotobacter chroococcum lograron que las posturas tuvieran dicha calidad. En relación con la inoculación con HMA, las especies Glomus clarum, G. fasciculatum y G. mosseae, para ambos cultivos, produjeron posturas con valores de altura y longitud radical considerados óptimos. Con las coinoculaciones de RPCV + HMA se lograron posturas de calidad superior a la alcanzada con las mejores variantes de inoculación simple, destacándose las combinaciones de G. clarum y G. fasciculatum con A. brasilense para el tomate y de G. clarum y G. fasciculatum con A. chroococcum para la cebolla.

  6. Maize response to inoculation with strains of plant growth-promoting bactéria

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    Janaína Dartora

    Full Text Available ABSTRACT The aim of this study was to evaluate the response of maize to inoculation with strains of plant growth-promoting bacteria (PGPB in two cultivation years. The experiment was set in a randomized block design with four replicates in two cultivation years (2012/13 and 2013/14. The treatments consisted of PGPB inoculation: control (without N and without inoculation; 30 kg of N ha-1 at sowing (N1; 160 kg of N ha-1 (N1 + 130 kg of N ha-1 as top-dressing; N1 + A. brasilense, Ab-V5; N1 + A. brasilense, HM053; N1 + Azospirillum sp. L26; N1 + Azospirillum sp. L27; N1 + Enhydrobacter sp. 4331; N1 + Rhizobium sp. 8121. Basal stem diameter, plant height, leaf area, shoot dry matter and yield were evaluated. The strain of Rhizobium sp. 8121and the isolate Azospirillum sp. L26 associated with 30 kg of N ha-1 at sowing promoted yields equivalent to that of the N fertilization of 160 kg ha-1, demonstrating the potential to be used in the inoculation of maize seeds.

  7. Efecto de la biofertilización y los biorreguladores en la germinación y el crecimiento de Carica papaya L.

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    Maricela Constantino

    2011-01-01

    Full Text Available Título en inglés: Effect of biofertilization and bioregulators on germination and growth of Carica papaya L. Resumen Con el objetivo de incrementar y acelerar el proceso de germinación de las semillas y obtener una alta producción y homogeneidad de plántulas de Carica papaya variedad Maradol en vivero, se evaluó el efecto de tres biofertilizantes aplicados solos o en combinación (Azotobacter chroococcum, Azospirillum brasilense y Glomus intraradices, y un biorregulador del crecimiento vegetal, el ácido giberélico (AG3, en la germinación y el crecimiento vegetal. Se realizó un experimento bajo un diseño completamente al azar con ocho tratamientos y tres repeticiones. A las semillas se les aplicó un pretratamiento germinativo con alternancia de temperatura para superar la dormancia. Los tratamientos simples con A. chroococcum y A. brasilense, incrementaron el porcentaje de germinación a 90,28 y 88,89% respectivamente. Además, con la aplicación de los biofertilizantes y el AG3, la velocidad de germinación se incrementó y el tiempo medio de germinación se redujo. La doble aplicación en semillas y foliar de los biofertilizantes y el AG3 en plántulas mejoró el crecimiento vegetal. La población de A. chroococcum fue mayor cuando se inoculó en combinación con G. intraradices. La prevalencia de colonización de las plántulas inoculadas con G. intraradices varió de 18,53 a 26,67%, con el mayor valor registrado para el tratamiento combinado con A. brasilense. Finalmente, aplicando esta metodología se logró acelerar la germinación, obteniéndose una mayor homogeneidad en la emergencia de las plántulas, disminuyendo así el tiempo de permanencia en el vivero. Palabras clave: Azospirillum brasilense; Azotobacter chroococcum; Glomus intraradices; ácido giberélico; dormancia. Abstract In order to increase and accelerate the process of seed germination and obtain a high yield and homogeneity of papaya seedlings cv. Maradol in

  8. LAS MICORRIZAS ARBUSCULARES Y LAS BACTERIAS RIZOSFÉRICAS COMO ALTERNATIVA A LA NUTRICIÓN MINERAL DEL TOMATE

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    María I. Hernández

    2004-01-01

    Full Text Available El presente estudio se desarrolló en el Instituto de Investigaciones Hortícolas "Liliana Dimitrova" durante los años 1996-2000 con la variedad de tomate HC 38-80 sembrada en período tardío. En la fase de semillero se realizó un screening, con el objetivo de seleccionar las cepas de micorrizas arbusculares y rizobacterias más eficientes para el cultivo del tomate, así como las mejores combinaciones. Para ello se determinaron la altura de la planta, el diámetro del tallo, la longitud radical y la masa seca total. Posteriormente, se evaluó el efecto de los biofertilizantes seleccionados y la fertilización mineral en el rendimiento del cultivo, sus componentes y el estado nutricional de la planta. El mejor comportamiento en la fase de semillero se obtuvo con la inoculación de las cepas Glomus mosseae, Glomus fasciculatum, Azospirillum brasilense, Azotobacter chroococcum, Glomus mosseae + Pseudomonas fluorescens y Glomus mosseae + Azospirillum brasilense. En la fase de campo se observó que el rendimiento y sus componentes se beneficiaron con la aplicación de niveles óptimos de fertilizantes, mientras que para los tratamientos inoculados los mayores valores correspondieron a Glomus mosseae, Glomus mosseae + Pseudomonas flourescens y Glomus mosseae + Azospirillum brasilense combinadas con el 50 % de la fertilización nitrogenada. Los HFMA y su coinoculación con bacterias rizosféricas influyeron de manera positiva en la absorción de nitrógeno y fósforo.

  9. EFFECT OF NITROGEN-FIXING BACTERIA ON GRAIN YIELD AND DEVELOPMENT OF FLOODED IRRIGATED RICE

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    AMAURI NELSON BEUTLER

    2016-01-01

    Full Text Available This study aimed at evaluating the effect of Azospirillum brasilense , a nitrogen - fixing bacterium, on flooded irrigated rice yield. Evaluations were carried out in a shaded nursery, with seedlings grown on an Alfisol. Were performed two sets of experiments. In the first, were carried out four experiments using the flooded rice cultivars INIA Olimar, Puitá Inta - CL, Br Irga 409 and Irga 424; these trials were set up as completely randomized design in a 5x4 factorial scheme, with four replications. Treatments consisted of five nitrogen rates (0, 40, 80, 120 and 160 kg ha - 1 and four levels of liquid inoculant Ab - V5 and Ab - V6 - A. brasilense (0, 1, 2 and 4 times the manufacturer's recommendation without seed treatment. In second set, were performed two experiments using the cultivars Puitá Inta - CL and Br Irga 409, arranged in the same design, but using a 4x2 factorial. In this set, treatments were composed of four levels of Ab - V5 and Ab - V6 - A. brasilense liquid inoculant (0, 1, 2 and 4 times the recommendation of 100 mL ha - 1 , using rice seeds with and without insecticide and fungicide treatment. Shoot dry matter, number of panicles, and rice grain yield per pot were the assessed variables. The results showed that rice seed inoculation with A. brasilense had no effects on rice grain yield of the cultivars INIA Olimar, Puitá Inta - CL, Br Irga 409 and Irga 424.

  10. Molecular characterization of nitrogen-fixing bacteria isolated from brazilian agricultural plants at São Paulo state

    Science.gov (United States)

    Reinhardt, Érica. L.; Ramos, Patrícia L.; Manfio, Gilson P.; Barbosa, Heloiza R.; Pavan, Crodowaldo; Moreira-Filho, Carlos A.

    2008-01-01

    Fourteen strains of nitrogen-fixing bacteria were isolated from different agricultural plant species, including cassava, maize and sugarcane, using nitrogen-deprived selective isolation conditions. Ability to fix nitrogen was verified by the acetylene reduction assay. All potentially nitrogen-fixing strains tested showed positive hybridization signals with a nifH probe derived from Azospirillum brasilense. The strains were characterized by RAPD, ARDRA and 16S rDNA sequence analysis. RAPD analyses revealed 8 unique genotypes, the remaining 6 strains clustered into 3 RAPD groups, suggesting a clonal origin. ARDRA and 16S rDNA sequence analyses allowed the assignment of 13 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Azospirillum, Herbaspirillum, Pseudomonas and Enterobacteriaceae. Two strains were classified as Stenotrophomonas ssp. Molecular identification results from 16S rDNA analyses were also corroborated by morphological and biochemical data. PMID:24031239

  11. Estudio de algunos géneros bacterianos asociados a la rizosfera de los cultivos de gerbera (Gerbera jamesonii y clavel (Dianthus barbatus, Dianthus caryophyllus

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    Annia Hernández

    2000-01-01

    Full Text Available En este trabajo se realizó la caracterización de algunos géneros bacterianos presentes en la rizosfera de los cultivos de gerbera (Gerbera jamesonii y clavel (Dianthus barbatus y Dianthus caryophyllus, en un suelo Ferralítico Rojo compactado de San José de las Lajas. También se estudió la quimioatracción de los exudados radicales de estos cultivos hacia diferentes rizobacterias (Pseudomonas (Burkholderia cepacia, P. fluorescens y Azospirillum brasilense. En ambos casos se utilizó el Modelo Espermosférico. Los resultados obtenidos demostraron que Pseudomonas constituye una población dominante en relación con Azospirillum, Bacillus y Streptomyces. Pseudomonas (Burkholderia cepacia fue la cepa más fuertemente atraída por los exudados radicales de gerbera y clavel, a los 14 días y 40 minutos de exposición

  12. Molecular responses in root-associative rhizospheric bacteria to variations in plant exudates

    Science.gov (United States)

    Abdoun, Hamid; McMillan, Mary; Pereg, Lily

    2015-04-01

    Plant exudates are a major factor in the interface of plant-soil-microbe interactions and it is well documented that the microbial community structure in the rhizosphere is largely influenced by the particular exudates excreted by various plants. Azospirillum brasilense is a plant growth promoting rhizobacterium that is known to interact with a large number of plants, including important food crops. The regulatory gene flcA has an important role in this interaction as it controls morphological differentiation of the bacterium that is essential for attachment to root surfaces. Being a response regulatory gene, flcA mediates the response of the bacterial cell to signals from the surrounding rhizosphere. This makes this regulatory gene a good candidate for analysis of the response of bacteria to rhizospheric alterations, in this case, variations in root exudates. We will report on our studies on the response of Azospirillum, an ecologically, scientifically and agriculturally important bacterial genus, to variations in the rhizosphere.

  13. Molecular characterization of nitrogen-fixing bacteria isolated from brazilian agricultural plants at São Paulo state.

    Science.gov (United States)

    Reinhardt, Erica L; Ramos, Patrícia L; Manfio, Gilson P; Barbosa, Heloiza R; Pavan, Crodowaldo; Moreira-Filho, Carlos A

    2008-07-01

    Fourteen strains of nitrogen-fixing bacteria were isolated from different agricultural plant species, including cassava, maize and sugarcane, using nitrogen-deprived selective isolation conditions. Ability to fix nitrogen was verified by the acetylene reduction assay. All potentially nitrogen-fixing strains tested showed positive hybridization signals with a nifH probe derived from Azospirillum brasilense. The strains were characterized by RAPD, ARDRA and 16S rDNA sequence analysis. RAPD analyses revealed 8 unique genotypes, the remaining 6 strains clustered into 3 RAPD groups, suggesting a clonal origin. ARDRA and 16S rDNA sequence analyses allowed the assignment of 13 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Azospirillum, Herbaspirillum, Pseudomonas and Enterobacteriaceae. Two strains were classified as Stenotrophomonas ssp. Molecular identification results from 16S rDNA analyses were also corroborated by morphological and biochemical data.

  14. Impact of bacterial priming on some stress tolerance mechanisms and growth of cold stressed wheat seedlings

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    Mohammed E.H. Osman

    2014-01-01

    Full Text Available The potential to enhance growth of cold stressed wheat by seed treatment (priming with the beneficial bacteria Bacillus amyloliquefaciens 5113 and Azospirillum brasilense NO40 were tested. Results showed an improved ability of bacteria-treated seedlings to survive at −5°C up to 12 h. Cold stress increased transcript levels of three stress marker genes and increased activity for the ascorbate-glutathione redox enzymes. However, primed and stressed seedlings generally showed smaller effects on the stress markers correlating with better growth and improved stress tolerance. Bacterial priming to improve crop plant performance at low temperature seems a useful strategy to explore further.

  15. Characterization of a major cluster of nif, fix, and associated genes in a sugarcane endophyte, Acetobacter diazotrophicus.

    Science.gov (United States)

    Lee, S; Reth, A; Meletzus, D; Sevilla, M; Kennedy, C

    2000-12-01

    A major 30.5-kb cluster of nif and associated genes of Acetobacter diazotrophicus (syn. Gluconacetobacter diazotrophicus), a nitrogen-fixing endophyte of sugarcane, was sequenced and analyzed. This cluster represents the largest assembly of contiguous nif-fix and associated genes so far characterized in any diazotrophic bacterial species. Northern blots and promoter sequence analysis indicated that the genes are organized into eight transcriptional units. The overall arrangement of genes is most like that of the nif-fix cluster in Azospirillum brasilense, while the individual gene products are more similar to those in species of Rhizobiaceae or in Rhodobacter capsulatus.

  16. Seeds inoculation with diazotrophic bacteria and nitrogen application in side-dressing and leaf in maize

    OpenAIRE

    Claudinei Kappes; Orivaldo Arf; Marcelo Valentini Arf; João Paulo Ferreira; Edjair Augusto Dal Bem; José Roberto Portugal; Rafael Gonçalves Vilela

    2013-01-01

    Considering the importance of nitrogen management and its biological fixation with diazotrophic bacteria, this study was carried out aiming to evaluate the agronomic performance of maize, in response to seed inoculation with Azospirillum brasilense and nitrogen application in side-dressing and leaf. The experiment was conducted in Selvíria, Mato Grosso of Sul State, Brazil, during the growing season 2010/2011, on a clayey Rhodic Haplustox (20º 20’ S and 51º 24’ W, with altitude of 340 m). Six...

  17. Bioinoculants: A sustainable approach to maximize the yield of Ethiopian mustard (Brassica carinata L.) under low input of chemical fertilizers.

    Science.gov (United States)

    Nosheen, Asia; Bano, Asghari; Ullah, Faizan

    2016-02-01

    This study aimed to find out the effect of plant growth-promoting rhizobacteria (PGPR; Azospirillum brasilense and Azotobacter vinelandii) either alone or in combination with different doses of nitrogen and phosphate fertilizers on growth, seed yield, and oil quality of Brassica carinata (L.) cv. Peela Raya. PGPR were applied as seed inoculation at 10(6) cells/mL(-1) so that the number of bacterial cells per seed was 2.6 × 10(5) cells/seed. The chemical fertilizers, namely, urea and diammonium phosphate (DAP) were applied in different doses (full dose (urea 160 kg ha(-1) + DAP 180 kg ha(-1)), half dose (urea 80 kg ha(-1) + DAP 90 kg ha(-1)), and quarter dose (urea 40 kg ha(-1) + DAP 45 kg ha(-1)). The chemical fertilizers at full and half dose significantly increased the chlorophyll, carotenoids, and protein content of leaves and the seed yield (in kilogram per hectare) but had no effect on the oil content of seed. The erucic acid (C22:1) content present in the seed was increased. Azospirillum performed better than Azotobacter and its effect was at par with full dose of chemical fertilizers (CFF) for pigments and protein content of leaves when inoculated in the presence of half dose of chemical fertilizers (SPH). The seed yield and seed size were greater. Supplementing Azospirillum with SPH assisted Azospirillum to augment the growth and yield, reduced the erucic acid (C22:1) and glucosinolates contents, and increased the unsaturation in seed oil. It is inferred that A. brasilense could be applied as an efficient bioinoculant for enhancing the growth, seed yield, and oil quality of Ethiopian mustard at low fertilizer costs and sustainable ways.

  18. Influence of Organic Manures (Biofertilizers on Soil Microbial Population in the Rhizosphere of Mulberry (Morus Indica L.

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    L. Christilda Louis Mary

    2015-03-01

    Full Text Available The effect of different kinds of organic manures on soil microbial population and mulberry production was assessed. A field experiment wascarried out at Periyar EVR College, Tamil Nadu, India in basic soil to study the influence of organic manures on soil bacterial population andmulberry production. The 4 groups of mulberry plants of MR2 variety were biofertilized with FYM, Azospirillum, Phosphobacteria andVermicompost respectively. The biofertilizers lodged bacteria on the rhizosphere of mulberry plants. When the root microorganism areanalyzed Farm yard manure biofertilized mulberry plant root tips had Gluconacobacter diazotrophicus, Bacillus pumilus, Pseudomonas putida,Bacillus coagulans, Bacillus sonorensis, Azotobacter chrococcum; Azospirillum biofertilized mulberry plants root tips had Bacillus coaculans,Azotobactor chrococcum, Azotobactor vinelandii, Bacillus subtilis and Azospirillum brasilense. Phosphobacteria biofertilized mulberry plantroot tips had Pseudomonas putida, Bacillus stearothermophilus, Brevibacillus borslelansis and Streptomycies thermonitrificans andvermicompost biofertilized mulberry plant root tips had lodged bacterias like Bacillus megaterium, Bacillus subtilis, Gluconacobacterdiazotrophicus, Pseudomonas putida, Azotobacter chrococcum, Azotobacter vinelandi, Bacillus stearothermophilus, Brevibacillus borslelansisand Bacillus sonorensis. Microbiology work reveals luxuriant growth of bacteria in all the biofertizer treated rhizosphere in the order FYM <Azospirillum < Phosphobacteria < Vermicompost. Increased availability of NPK and other micronutrients were noticed in T4 treated plantscompared to other treatments.

  19. The genetic diversity of culturable nitrogen-fixing bacteria in the rhizosphere of wheat.

    Science.gov (United States)

    Venieraki, Anastasia; Dimou, Maria; Pergalis, Panagiotis; Kefalogianni, Io; Chatzipavlidis, Iordanis; Katinakis, Panagiotis

    2011-02-01

    A total of 17 culturable nitrogen-fixing bacterial strains associated with the roots of wheat growing in different regions of Greece were isolated and characterized for plant-growth-promoting traits such as auxin production and phosphate solubilization. The phylogenetic position of the isolates was first assessed by the analysis of the PCR-amplified 16S rRNA gene. The comparative sequence analysis and phylogenetic analysis based on 16S rRNA gene sequences show that the isolates recovered in this study are grouped with Azospirillum brasilense, Azospirillum zeae, and Pseudomonas stutzeri. The diazotrophic nature of all isolates was confirmed by amplification of partial nifH gene sequences. The phylogenetic tree based on nifH gene sequences is consistent with 16S rRNA gene phylogeny. The isolates belonging to Azospirillum species were further characterized by examining the partial dnaK gene phylogenetic tree. Furthermore, it was demonstrated that the ipdC gene was present in all Azospirillum isolates, suggesting that auxin is mainly synthesized via the indole-3-pyruvate pathway. Although members of P. stutzeri and A. zeae are known diazotrophic bacteria, to the best of our knowledge, this is the first report of isolation and characterization of strains belonging to these bacterial genera associated with wheat.

  20. Evaluación de la asociación bacterias fijadoras de nitrógeno - líneas interespecíficas de arroz-nitrógeno, en Typic haplustalf. Ibagué, Colombia Evaluation of the association nitrogen fixing bacterias interspecific - rice lines - nitrogen, in typic haplustalf. Ibagué, Colombia

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    Margarita M Vallejo

    2008-01-01

    Full Text Available El estudio se llevó a cabo en la hacienda Cauchitos, municipio de Ibagué, departamento del Tolima (Norte 4° 23' 51" y Oeste 75° 9' 7", 979 msnm, 24.3°C, bosque seco tropical (bs-T, con el objetivo de evaluar las asociaciones entre bacterias fijadoras de nitrógeno con inóculo y sin él en diez líneas interespecíficas de arroz, con tres dosis de nitrógeno (0%, 50% y 100% de 250 kg/ha-1 y tres repeticiones por tratamiento. La inoculación se realizó con 1 cm³ de unidades formadoras de colonias por 250 g de semilla de cada cultivar. Se aislaron 2.260 bacterias de los géneros Azotobacter spp y Azospirillum spp, se identificaron las especies A. brasilense, A. lipoferum, A. amazonense y del género Azotobacter las especies A. chroococcum, A. vinelandii, A. paspali y A. beijerinckii. Respecto al inóculo no se encontraron diferencias significativas al realizar su aplicación, se determinó que Azotobacter spp y Azospirillum spp fueron géneros típicos de la flora bacteriana en el cultivo del arroz y en condiciones de campo hubo efecto de los tratamientos en la flora bacteriana, y Azotobacter spp fue el predominante en cada uno de los tratamientos.The study was carried out at the Cauchitos farm, Ibague municipality department of Tolima, with bounds: North 4°23'51" and west 75°9'7", 979 ansm, the average temperature is 24,3°C, tropical dry forest (bs-t in the Holdridge classification. The purpose of this study was to evaluate the association between the nitrogen fixation bacteries with and without inoculo in 10 interespecific rice lines with three nitrogen dosis (0, 50 and 100% de 250 kg/ ha-1 and three repetitions. The inoculation was realized with 10(8 former units of colonies per millimeter. 2.260 bacteries of the generums Azotobacter spp y Azospirrillum spp., and identification the species: Azospirillum brasilense, Azospirillum lipoferum, Azospirillum amazonense, were identified and from the genus Azotobacter were identified the

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

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    Tharwat El-Sayed El-Desouk Radwan

    2005-10-01

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

  2. GERMINATION OF GRASSES DUE TO INOCULATION DIAZOTROPHIC BACTERIA

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    C. D. A. Moreira

    2014-07-01

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

  3. A novel interaction between plant-beneficial rhizobacteria and roots: colonization induces corn resistance against the root herbivore Diabrotica speciosa.

    Science.gov (United States)

    Santos, Franciele; Peñaflor, Maria Fernanda G V; Paré, Paul W; Sanches, Patrícia A; Kamiya, Aline C; Tonelli, Mateus; Nardi, Cristiane; Bento, José Mauricio S

    2014-01-01

    A number of soil-borne microorganisms, such as mycorrhizal fungi and rhizobacteria, establish mutualistic interactions with plants, which can indirectly affect other organisms. Knowledge of the plant-mediated effects of mutualistic microorganisms is limited to aboveground insects, whereas there is little understanding of what role beneficial soil bacteria may play in plant defense against root herbivory. Here, we establish that colonization by the beneficial rhizobacterium Azospirillum brasilense affects the host selection and performance of the insect Diabrotica speciosa. Root larvae preferentially orient toward the roots of non-inoculated plants versus inoculated roots and gain less weight when feeding on inoculated plants. As inoculation by A. brasilense induces higher emissions of (E)-β-caryophyllene compared with non-inoculated plants, it is plausible that the non-preference of D. speciosa for inoculated plants is related to this sesquiterpene, which is well known to mediate belowground insect-plant interactions. To the best of our knowledge, this is the first study showing that a beneficial rhizobacterium inoculant indirectly alters belowground plant-insect interactions. The role of A. brasilense as part of an integrative pest management (IPM) program for the protection of corn against the South American corn rootworm, D. speciosa, is considered.

  4. A novel interaction between plant-beneficial rhizobacteria and roots: colonization induces corn resistance against the root herbivore Diabrotica speciosa.

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    Franciele Santos

    Full Text Available A number of soil-borne microorganisms, such as mycorrhizal fungi and rhizobacteria, establish mutualistic interactions with plants, which can indirectly affect other organisms. Knowledge of the plant-mediated effects of mutualistic microorganisms is limited to aboveground insects, whereas there is little understanding of what role beneficial soil bacteria may play in plant defense against root herbivory. Here, we establish that colonization by the beneficial rhizobacterium Azospirillum brasilense affects the host selection and performance of the insect Diabrotica speciosa. Root larvae preferentially orient toward the roots of non-inoculated plants versus inoculated roots and gain less weight when feeding on inoculated plants. As inoculation by A. brasilense induces higher emissions of (E-β-caryophyllene compared with non-inoculated plants, it is plausible that the non-preference of D. speciosa for inoculated plants is related to this sesquiterpene, which is well known to mediate belowground insect-plant interactions. To the best of our knowledge, this is the first study showing that a beneficial rhizobacterium inoculant indirectly alters belowground plant-insect interactions. The role of A. brasilense as part of an integrative pest management (IPM program for the protection of corn against the South American corn rootworm, D. speciosa, is considered.

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

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    VANESSA ZIRONDI LONGHINI

    2016-01-01

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

  6. Identification and characterization of trans-3-hydroxy-l-proline dehydratase and Δ1-pyrroline-2-carboxylate reductase involved in trans-3-hydroxy-l-proline metabolism of bacteria

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    Seiya Watanabe

    2014-01-01

    Full Text Available trans-4-Hydroxy-l-proline (T4LHyp and trans-3-hydroxy-l-proline (T3LHyp occur mainly in collagen. A few bacteria can convert T4LHyp to α-ketoglutarate, and we previously revealed a hypothetical pathway consisting of four enzymes at the molecular level (J Biol Chem (2007 282, 6685–6695; J Biol Chem (2012 287, 32674–32688. Here, we first found that Azospirillum brasilense has the ability to grow not only on T4LHyp but also T3LHyp as a sole carbon source. In A. brasilense cells, T3LHyp dehydratase and NAD(PH-dependent Δ1-pyrroline-2-carboxylate (Pyr2C reductase activities were induced by T3LHyp (and d-proline and d-lysine but not T4LHyp, and no effect of T3LHyp was observed on the expression of T4LHyp metabolizing enzymes: a hypothetical pathway of T3LHyp → Pyr2C → l-proline was proposed. Bacterial T3LHyp dehydratase, encoded to LhpH gene, was homologous with the mammalian enzyme. On the other hand, Pyr2C reductase encoded to LhpI gene was a novel member of ornithine cyclodeaminase/μ-crystallin superfamily, differing from known bacterial protein. Furthermore, the LhpI enzymes of A. brasilense and another bacterium showed several different properties, including substrate and coenzyme specificities. T3LHyp was converted to proline by the purified LhpH and LhpI proteins. Furthermore, disruption of LhpI gene from A. brasilense led to loss of growth on T3LHyp, d-proline and d-lysine, indicating that this gene has dual metabolic functions as a reductase for Pyr2C and Δ1-piperidine-2-carboxylate in these pathways, and that the T3LHyp pathway is not linked to T4LHyp and l-proline metabolism.

  7. Plant-microorganism-soil interactions influence the Fe availability in the rhizosphere of cucumber plants.

    Science.gov (United States)

    Pii, Youry; Penn, Alexander; Terzano, Roberto; Crecchio, Carmine; Mimmo, Tanja; Cesco, Stefano

    2015-02-01

    Iron (Fe) is a very important element for plants, since it is involved in many biochemical processes and, often, for the low solubility of the natural Fe sources in soil, plants suffer from Fe - deficiency, especially when grown on calcareous soils. Among the numerous plant growth-promoting rhizobacteria (PGPR) that colonize the rhizosphere of agronomically important crops, Azospirillum brasilense has been shown to exert strong stimulating activities on plants, by inducing alterations of the root architecture and an improvement of mineral nutrition, which could result from an enhancement of ion uptake mechanisms as well as by increased bioavailability of nutrients. Some studies have also established that A. brasilense can act as biocontrol agent, by preventing the growth and/or virulence of phytopathogens, most likely through the production of microbial siderophores that sequester Fe from the soil. Despite microbial siderophores complexed with Fe could be an easily accessible Fe source for plants, the possible involvement of A. brasilense in improving Fe nutrition in plants suffering from the micronutrient deficiency has not been investigated yet. Within the present research, the characterization of the physiological and biochemical effects induced by Fe starvation and PGPR inoculation in cucumber plants (Cucumis sativus L. cv. Chinese Long) was carried out. The analyses of root exudates released by hydroponically grown plants highlighted that cucumber plants respond differently depending on the nutritional status. In addition, following the cultivation period on calcareous soil, also the root exudates found in the extracts suggested a peculiar behaviour of plants as a function of the treatment. Interestingly, the presence of the inoculum in soil allowed a faster recovery of cucumber plants from Fe-deficiency symptoms, i.e. increase in the chlorophyll content, in the biomass and in the Fe content of leaves. These observations might suggest a feasible application of

  8. HONGO ENDOMICORRÍZICO Y BACTERIA FIJADORA DE NITRÓGENO INOCULADAS A Coffea arabica EN VIVERO

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    Juan Francisco Aguirre-Medina

    2011-01-01

    Full Text Available Con el objetivo de determinar el efecto de la inoculación con Azospirillum brasilense y Glomus intraradice en café, se estableció la presente investigación en un vivero en el Campo Experimental Rosario Izapa, Chiapas, México de septiembre de 2005 a abril de 2006. Se utilizó un suelo Andosol¿mólico típico de la región cafetalera del Soconusco Chiapas, México. El A. brasilense tuvo una concentración de 100 x 106 bacterias por gramo de turba y el hongo micorrízico, cuarenta esporas por gramo de suelo con infección de 95% en el sistema radical de cebolla. Los tratamientos utilizados fueron los microorganismos solos, la combinación de ambos y el testigo sin inocular que se distribuyeron en un diseño de bloques completos al azar. Se evaluaron variables morfológicas y fisiológicas del rendimiento cada treinta días a partir del segundo mes de siembra durante siete meses y se determinó el contenido de N, P y Ca en el tejido vegetal. Los resultados se analizaron estadísticamente y las diferencias entre tratamientos se compararon de acuerdo a Tukey 5%. Los resultados indican una respuesta diferencial entre los microorganismos. A. brasilense indujo mayor desarrollo radical y la simbiosis doble G. intraradices + A. brasilense mejoró el desarrollo del tallo y lámina foliar. El contenido de N, P y Ca se presentó de forma consistente con G. intraradices.

  9. Molecular characterization of nitrogen-fixing bacteria isolated from brazilian agricultural plants at São Paulo state Caracterização molecular de bactérias fixadoras de nitrogênio isoladas de plantas brasileiras no estado de São Paulo

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    Érica. L. Reinhardt

    2008-09-01

    Full Text Available Fourteen strains of nitrogen-fixing bacteria were isolated from different agricultural plant species, including cassava, maize and sugarcane, using nitrogen-deprived selective isolation conditions. Ability to fix nitrogen was verified by the acetylene reduction assay. All potentially nitrogen-fixing strains tested showed positive hybridization signals with a nifH probe derived from Azospirillum brasilense. The strains were characterized by RAPD, ARDRA and 16S rDNA sequence analysis. RAPD analyses revealed 8 unique genotypes, the remaining 6 strains clustered into 3 RAPD groups, suggesting a clonal origin. ARDRA and 16S rDNA sequence analyses allowed the assignment of 13 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Azospirillum, Herbaspirillum, Pseudomonas and Enterobacteriaceae. Two strains were classified as Stenotrophomonas ssp. Molecular identification results from 16S rDNA analyses were also corroborated by morphological and biochemical data.Quatorze linhagens de bactérias fixadoras de nitrogênio foram isoladas de diferentes espécies de plantas, incluindo cassava, milho e cana-de-açúcar, usando condições seletivas desprovidas de nitrogênio. A capacidade de fixar nitrogênio foi verificada por ensaio de redução de acetileno. Todas as linhagens fixadoras de nitrogênio testadas apresentaram hibridização positiva com sonda de gene nifH derivada de Azospirillum brasilense. As linhagens foram caracterizadas por RAPD, ARDRA e sequenciamento do gene 16S rDNA. As análises de RAPD revelaram 8 genótipos, as 6 linhagens restantes foram agrupadas em 3 grupos de RAPD, sugerindo uma origem clonal. ARDRA e seqüências de 16S rDNA foram alocadas em 13 grupos conhecidos de bactérias fixadoras de nitrogênio, incluindo organismos dos gêneros Azospirillum, Herbaspirillum, Pseudomonas e Enterobacteriaceae. Duas linhagens foram classificadas como Stenotrophomonas ssp. Os resultados da identifica

  10. Molecular diversity of diazotrophs in oligotrophic tropical seagrass bed communities.

    Science.gov (United States)

    Bagwell, Christopher E; Rocque, Jeannine R; Smith, Garriett W; Polson, Shawn W; Friez, Michael J; Longshore, John W; Lovell, Charles R

    2002-02-01

    Denaturing gradient gel electrophoresis (DGGE) was employed to resolve PCR-amplified nifH sequences from vegetated and unvegetated sediments from two oligotrophic seagrass bed sites on San Salvador Island, Bahamas, in order to assess diazotroph species composition. All DGGE profiles from these sites showed the same prominent bands. These bands were sequenced, yielding 67 different nifH sequences, which were used in phylogenetic reconstructions. Most sequences were from anaerobes, but some were affiliated with the alpha- and (gamma-+beta-) Proteobacteria. Several NifH sequences were nearly identical to those from Azospirillum brasilense and Vibrio diazotrophicus. These seagrass bed sediments support a diverse diazotroph assemblage that is, at least superficially, similar to that associated with an intertidal grass (Spartina alterniflora).

  11. A high efficient carbendazim-degrading bacterial strain: Its isolation and identification%一株多菌灵高效降解菌的分离鉴定及特性

    Institute of Scientific and Technical Information of China (English)

    林秀; 侯振安; 赵思峰; 谢磊; 李会杰; 李谟志

    2011-01-01

    从长期施用多菌灵农药的葡萄园中分离筛选得到一株多菌灵降解菌XJ-H,经Biolog微生物自动分析系统和16S rDNA序列比对以及系统聚类分析等鉴定菌株XJ-H为巴西固氮螺菌(Azospirillum brasilense).多菌灵降解菌XJ-H可利用多菌灵为唯一碳源、氮源进行生长,将其接种在600 mg·L-1多菌灵的无机盐培养基中,11d时多菌灵的降解率达到95.6%,平均降解能力为52.2 mg·L-1·d-1.

  12. Efecto de diferentes concentraciones de ácido antranílico en el crecimiento del maíz

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    José Luis Hernández-Mendoza

    2010-08-01

    Palabras clave: auxinas, fitohormonas, rizobacterias promotoras del crecimiento vegetal, Zea mays L. Abstract: The plant growth-promoting ability of rhizo-bacterium Azospirillum brasilense is based on production of auxins and other hormones, among them outstanding antranilic acid (AA. In this work we determined the antranilic acid (0, 500, 1000, 3000, and 5000 ppm effects on maize growth and dry biomass accumulation under greenhouse conditions using the hybrid 83G66. From 500 to 3000 ppm of AA significantly increased plant growth (leaf area and plant height while 1000 and 3000 ppm of AA significantly increased dry biomass accumulation in maize compared with control (0 ppm. The 90 % of AA in maize plants was assimilated during the early 10 days after treatment. Key words: Auxins, phyto-hormones, plant growth promoting rhizobacteria, Zea mays L.

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

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    Marivaine da Silva Brasil

    2005-04-01

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

  14. Influence of microbially enriched vermicompost on yield, microbial dynamics and soil nutrients

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    Neeldurai Tensingh Baliah

    2016-12-01

    Full Text Available The present study has been conducted to explore the microbial enrichment of vermicompost with microbial inoculants such as Azospirillum brasilense, Bacillus megaterium and Pseudomonas fluorescens. The enrichment had a positive effect on the crop response, soil biological activity and soil nutrient status. The results indicated that the enriched vermicompost significantly increased crop response of Okra with reference to yield attributes. Further, the microbial enriched vermicompost significantly improved the soil microbial dynamics such as bacteria and fungi and nutrient status such as total N and available P in the amended soil. The enrichment with agronomically important microbes such as nitrogen fixer, phosphate solubilizer and plant growth promoting rhizobacteria had positive response but the response varied among the beneficial microorganisms.

  15. nif HDK基因在W80-2上的初步定位%INITIAL LOCALIZATION OF nif HDK GENE IN W80-2 STRAIN

    Institute of Scientific and Technical Information of China (English)

    罗永华; 郭俊

    2002-01-01

    对几种质粒检测方法进行了比较,发现原位裂解法能比较满意地检测到巴西固氮螺菌(Azospirillum brasilense)的巨大质粒.利用改进后的原位裂解法能比较稳定地检测到W80-2菌株中的巨大质粒.通过Southern-blotting的方法将W80-2菌的染色体及巨大质粒转到尼龙膜上,与用地高辛标记的含nifHDK基因的pSA30质粒杂交,发现W80-2菌株的nifHDK基因定位在染色体上.

  16. Effect of plant growth-promoting bacteria on the growth and fructan production of Agave americana L.

    Science.gov (United States)

    De La Torre-Ruiz, Neyser; Ruiz-Valdiviezo, Víctor Manuel; Rincón-Molina, Clara Ivette; Rodríguez-Mendiola, Martha; Arias-Castro, Carlos; Gutiérrez-Miceli, Federico Antonio; Palomeque-Dominguez, Héctor; Rincón-Rosales, Reiner

    2016-01-01

    The effect of plant growth-promoting bacteria inoculation on plant growth and the sugar content in Agave americana was assessed. The bacterial strains ACO-34A, ACO-40, and ACO-140, isolated from the A. americana rhizosphere, were selected for this study to evaluate their phenotypic and genotypic characteristics. The three bacterial strains were evaluated via plant inoculation assays, and Azospirillum brasilense Cd served as a control strain. Phylogenetic analysis based on the 16S rRNA gene showed that strains ACO-34A, ACO-40 and ACO-140 were Rhizobium daejeonense, Acinetobacter calcoaceticus and Pseudomonas mosselii, respectively. All of the strains were able to synthesize indole-3-acetic acid (IAA), solubilize phosphate, and had nitrogenase activity. Inoculation using the plant growth-promoting bacteria strains had a significant effect (pagave plants with proper biological characteristics for agroindustrial and biotechnological use and to increase the sugar content in this agave species.

  17. Robust biological nitrogen fixation in a model grass-bacterial association.

    Science.gov (United States)

    Pankievicz, Vânia C S; do Amaral, Fernanda P; Santos, Karina F D N; Agtuca, Beverly; Xu, Youwen; Schueller, Michael J; Arisi, Ana Carolina M; Steffens, Maria B R; de Souza, Emanuel M; Pedrosa, Fábio O; Stacey, Gary; Ferrieri, Richard A

    2015-03-01

    Nitrogen-fixing rhizobacteria can promote plant growth; however, it is controversial whether biological nitrogen fixation (BNF) from associative interaction contributes to growth promotion. The roots of Setaria viridis, a model C4 grass, were effectively colonized by bacterial inoculants resulting in a significant enhancement of growth. Nitrogen-13 tracer studies provided direct evidence for tracer uptake by the host plant and incorporation into protein. Indeed, plants showed robust growth under nitrogen-limiting conditions when inoculated with an ammonium-excreting strain of Azospirillum brasilense. (11)C-labeling experiments showed that patterns in central carbon metabolism and resource allocation exhibited by nitrogen-starved plants were largely reversed by bacterial inoculation, such that they resembled plants grown under nitrogen-sufficient conditions. Adoption of S. viridis as a model should promote research into the mechanisms of associative nitrogen fixation with the ultimate goal of greater adoption of BNF for sustainable crop production.

  18. Effect of cover crops and nitrogen rates on sprinkler irrigated wheat in low altitude cerrado

    Directory of Open Access Journals (Sweden)

    Michelle Traete Sabundjian

    2016-06-01

    Full Text Available RESUMO A produção de trigo no cerrado brasileiro enfrenta grandes desafios, principalmente por não ser a região tradicionalmente tritícola e pela ocorrência de inverno quente e seco. Assim, o objetivo deste trabalho foi avaliar o efeito residual das culturas antecessoras, milho e Urochloa ruziziensis (R. Germ & Evrard, em cultivo exclusivo e em consórcio entre ambas, na presença e ausência da inoculação de sementes com Azospirillum brasilense e adubação nitrogenada (N em cobertura, no desenvolvimento e na produtividade do trigo. O experimento foi desenvolvido em Selvíria, MS, em 2011/12. O delineamento experimental foi o de blocos ao acaso, disposto em esquema fatorial 8x4, com quatro repetições, cujos tratamentos foram constituídos pela combinação de restos culturais (associações de milho, Urochloa ruziziensis e Azospirillum brasilense e doses de N (0, 30, 60 e 90 kg ha-1 em cobertura, no trigo. Realizaram-se as seguintes avaliações: cobertura vegetal residual (quantidade, características agronômicas, componentes de produção e produtividade da cultura do trigo. As culturas antecessoras apresentaram efeitos positivos na produtividade do trigo cultivado em sucessão, sendo o consórcio de milho e Urochloa ruziziensis, com ou sem inoculação excelente opção de manejo; o incremento da dose de N, em cobertura, até 90 kg ha- 1 aumenta a produtividade de grãos de trigo irrigado, dependendo da cultura antecessora.

  19. Seeds inoculation with diazotrophic bacteria and nitrogen application in side-dressing and leaf in maizeInoculação de sementes com bactéria diazotrófica e aplicação de nitrogênio em cobertura e foliar em milho

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    Claudinei Kappes

    2013-05-01

    Full Text Available Considering the importance of nitrogen management and its biological fixation with diazotrophic bacteria, this study was carried out aiming to evaluate the agronomic performance of maize, in response to seed inoculation with Azospirillum brasilense and nitrogen application in side-dressing and leaf. The experiment was conducted in Selvíria, Mato Grosso of Sul State, Brazil, during the growing season 2010/2011, on a clayey Rhodic Haplustox (20º 20’ S and 51º 24’ W, with altitude of 340 m. Sixteen treatments were established with four replications, in randomized blocks with the combination of the factors A. brasilense (with and without inoculante, nitrogen rate (0 and 90 kg ha-1, in V5 growth stage and urea leaf application (0, 4, 8 and 12%: application in V5 and V8 growth stage. The maize hybrid used was the DKB 390 YG®, sowed in the row spacing of 0.9 m. Parameters measured were productive and morphological components of culture and crop yield. Increase in maize yield by seed inoculation with A. brasilense was observed. The application of 90 kg ha-1 of nitrogen in side-dressing provided higher chlorophyll leaf index, stalk diameter and prolificacy, however, the yield not was increased. The application of urea leaf did not agronomic efficiency and, therefore, should not be used as the unique form of supply and alternative to nitrogen addition to crop. Considerando a importância do manejo do nitrogênio e da sua fixação biológica através de bactérias diazotróficas, conduziu-se este trabalho com o objetivo de avaliar o desempenho agronômico do milho, em função da inoculação das sementes com Azospirillum brasilense e da aplicação de nitrogênio em cobertura e foliar. O experimento foi conduzido no município de Selvíria – MS, durante o ano agrícola 2010/2011, sob Latossolo Vermelho distrófico típico argiloso (20º 20’ S e 51º 24’ W, com altitude de 340 m. Foram estabelecidos dezesseis tratamentos com quatro repeti

  20. PAS domain of the deduced Org35 protein mediates the interaction with NifA

    Institute of Scientific and Technical Information of China (English)

    TU Ran; CUI Yanhua; CHEN Sanfeng; LI Jilun

    2006-01-01

    NifA in Azospirillum brasilense plays a key role in regulating the synthesis of nitrogenase in response to ammonia and oxygen available. Recently,our laboratory has identified four clones, whose gene prodcuts interact with NifA, from A. brasilense Sp7genomic libraries by using the yeast two-hybrid system with NifA as bait. We are interested in clone S35,one of the four clones, because it contains a PAS-domain coding region. The entire open reading frame (ORF) for the PAS domain-containing protein was isolated and designated as org35 here. org35gene is 2211-bp long and encodes a protein of 736aa with a predicted molecular weight of about 78.4 kD.The predicted amino acid sequence of org35 has similarity to some two-component sensor kinase/response regulator hybrids of bacteria. Structural analyses showed that Org35 comprises at least three discrete conserved domains: the N-terminal PAS, the central histidine protein kinase (HPK) and the C-terminal response regulator (RR). The PAS domain of the deduced Org35 protein was found to interact directly with NifA, but the central HPK and the C-terminal RR domains of Org35 were not. These results indicated that interaction between NifA and Org35 was mediated by PAS domain.

  1. Swimming motility plays a key role in the stochastic dynamics of cell clumping

    Science.gov (United States)

    Qi, Xianghong; Nellas, Ricky B.; Byrn, Matthew W.; Russell, Matthew H.; Bible, Amber N.; Alexandre, Gladys; Shen, Tongye

    2013-04-01

    Dynamic cell-to-cell interactions are a prerequisite to many biological processes, including development and biofilm formation. Flagellum induced motility has been shown to modulate the initial cell-cell or cell-surface interaction and to contribute to the emergence of macroscopic patterns. While the role of swimming motility in surface colonization has been analyzed in some detail, a quantitative physical analysis of transient interactions between motile cells is lacking. We examined the Brownian dynamics of swimming cells in a crowded environment using a model of motorized adhesive tandem particles. Focusing on the motility and geometry of an exemplary motile bacterium Azospirillum brasilense, which is capable of transient cell-cell association (clumping), we constructed a physical model with proper parameters for the computer simulation of the clumping dynamics. By modulating mechanical interaction (‘stickiness’) between cells and swimming speed, we investigated how equilibrium and active features affect the clumping dynamics. We found that the modulation of active motion is required for the initial aggregation of cells to occur at a realistic time scale. Slowing down the rotation of flagellar motors (and thus swimming speeds) is correlated to the degree of clumping, which is consistent with the experimental results obtained for A. brasilense.

  2. Plant growth-promoting bacteria associated with nitrogen fertilization at topdressing in popcorn agronomic performance

    Directory of Open Access Journals (Sweden)

    Leandro Teodoski Spolaor

    2016-03-01

    Full Text Available ABSTRACT The use of plant growth-promoting bacteria is a promising alternative with low environmental impact to increase the efficiency of use of chemical fertilizers, ensuring high yield with better cost-effective ratio. In maize crops, several studies have demonstrated an increased yield when Azospirillum-based inoculants are used. In the case of popcorn, there are no available studies related to use of inoculation and its response on yield parameters. Thus, the aim of this study was to evaluate the field performance of popcorn when inoculated with the commercial product Masterfix L (A. brasilense Ab-V5 and A. brasilense Ab-V6 and the non-commercial inoculant UEL (A. brasilense Ab-V5 + Rhizobium sp. 53GRM1 associated with nitrogen fertilization. The trials were conducted in Londrina and Maringá, Paraná State, Brazil, in a randomized block design with four replications, in a split plot design with the inoculation treatments located in the plots (uninoculated, Masterfix L, and UEL and the different N rates located in the subplots where ammonium sulphate was applied in the topdressing at the V6 stage (0, 50, 100, and 150 kg∙ha–1. The variance analysis showed significant effects (p < 0.05 of inoculation (Londrina environment and N rates (both environments only for grain yield. There was no inoculation effect in the grain yield when inoculants were applied together with N-fertilization at topdressing. In the absence of N-fertilization at topdressing, the inoculants Masterfix L. and UEL promoted higher grain yield as compared to the uninoculated plants, with resulting increases of 13.21 and 26.61% in yield, respectively.

  3. In vitro and in vivo inoculation of four endophytic bacteria on Lycopersicon esculentum.

    Science.gov (United States)

    Botta, Anna Lucia; Santacecilia, Alessandra; Ercole, Claudia; Cacchio, Paola; Del Gallo, Maddalena

    2013-09-25

    Four bacteria selected on the basis of their capability of fixing atmospheric nitrogen, stimulating plant-growth, and protecting the host plant from pathogens - Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, Burkholderia ambifaria - were inoculated on tomato seeds either singularly, in couple and in a four bacteria mixer. Aim of this research was to evaluate: (1) effect of single and mixed cultures on the inoculated plant - plant growth, dry weight, root length and surface, number of leaves, among others; (2) colonization and interactions of the bacteria inside the host plant; (3) localization inside the host of single bacterial strains marked with the gusA reporter gene. The results obtained indicate that all selected microbial strains have colonized Lycopersicon esculentum but in a different way, depending on the single species. A. brasilense, G. diazotrophicus inoculated in vitro singularly and together were the best plant colonizers. In vivo essays, instead, B. ambifaria and the four-bacteria mixer gave the best results. It was possible to localize both A. brasilense and H. seropedicae inside the plant by the gusA reporter gene. The bacterial strains occur along the root axis from the apical zone until to the basal stem, on the shoot from the base up to the leaves. The four bacteria actively colonize tomato seeds and establish an endophytic community inside the plant. This review gives new information about colonization processes, in particular how bacteria interact with plants and whether they are likely to establish themselves in the plant environment after field application as biofertilizers or biocontrol agents.

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

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    Glauber Monçon Fipke

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

  5. Increased root exudation of /sup 14/C-compounds by sorghum seedlings inoculated with nitrogen-fixing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Lee, K.J. (Institute of Forest Genetics, Suweon (Republic of Korea)); Gaskins, M.H. (Florida Univ., Gainesville (USA). Dept. of Agriculture)

    1982-01-01

    Organic components leaked from Sorghum bicolor seedlings ('root exudates') were examined by recovering /sup 14/C labelled compounds from root solutions of seedlings inoculated with Azospirillum brasilense, Azotobacter vinelandii or Klebsiella pneumoniae nif-. Up to 3.5% of the total /sup 14/C recovered from shoots, roots, and nutrient solutions was found in the root solutions. Inoculation with Azospirillum and Azotobacter increased the amounts of /sup 14/C and decreased the amounts of carbohydrates in the root solutions. When sucrose was added as a carbon source for the bacteria, the increase of /sup 14/C in the solutions did not occur. Quantities of /sup 14/C found in the root solutions were proportional to amounts of mineral nitrogen supplied to the plants. Bacterial growth also was proportional to nitrogen levels. When sorghum plants were grown in soil and labelled with /sup 14/CO/sub 2/, about 15% of the total /sup 14/C recovered within 48 hours exposure was found in soil leachates.

  6. Nitrogen-fixing bacteria, arbuscular mycorrhizal fungi, and the productivity and structure of prairie grassland communities.

    Science.gov (United States)

    Bauer, Jonathan T; Kleczewski, Nathan M; Bever, James D; Clay, Keith; Reynolds, Heather L

    2012-12-01

    Due to their complementary roles in meeting plant nutritional needs, arbuscular mycorrhizal fungi (AMF) and nitrogen-fixing bacteria (N(2)-fixers) may have synergistic effects on plant communities. Using greenhouse microcosms, we tested the effects of AMF, N(2)-fixers (symbiotic: rhizobia, and associative: Azospirillum brasilense), and their potential interactions on the productivity, diversity, and species composition of diverse tallgrass prairie communities and on the productivity of Panicum virgatum in monoculture. Our results demonstrate the importance of AMF and N(2)-fixers as drivers of plant community structure and function. In the communities, we found a positive effect of AMF on diversity and productivity, but a negative effect of N(2)-fixers on productivity. Both AMF and N(2)-fixers affected relative abundances of species. AMF shifted the communities from dominance by Elymus canadensis to Sorghastrum nutans, and seven other species increased in abundance with AMF, accounting for the increased diversity. N(2)-fixers led to increases in Astragalus canadensis and Desmanthus illinoense, two legumes that likely benefited from the presence of the appropriate rhizobia symbionts. Sorghastrum nutans declined 44 % in the presence of N(2)-fixers, with the most likely explanation being increased competition from legumes. Panicum monocultures were more productive with AMF, but showed no response to N(2)-fixers, although inference was constrained by low Azospirillum treatment effectivity. We did not find interactions between AMF and N(2)-fixers in communities or Panicum monocultures, indicating that short-term effects of these microbial functional groups are additive.

  7. Application effects of biofertilizers on the growth indices of black cumin (Nigella sativa L.

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    sorur khoram del

    2009-06-01

    Full Text Available Application of biofertilizers, especially plant growth promoting rhizobacteria (PGPR and mycorrhiza fungus is one of the most important strategies for plant nutrition compared to chemical fertilizers, especially in sustainable management of agroecosystems. In order to investigate the effect of Azotobacter and Azospirillum bacteria and Mycorrhiza fungus on the growth indices of black cumin (Nigella sativa L., a field experiment was conducted at the Agricultural Research Station of Ferdowsi University of Mashhad during growing season of 2007. A randomized complete block design with three replications was used. Treatments included: (A Azotobacter paspali, (B Azospirillum brasilense, (C the fungus of Glomus intraradaices, C+A, C+B, A+B, A+B+C, and control without using bio-fertilizers. The Azotobacter and Azospirillum inoculations were applied as liquid and the Mycorrhiza inoculation was applied in solid form on the treated seeds with Arabic resin immediately before planting. The Arabic resin was applied to increase the adherence of Mycorrhiza to seeds. In all treatments except control, the amounts of 15 mg of each bio-fertilizer were applied for 110 g of seeds. The results indicated that the inoculation of black cumin with biological fertilizers significantly increased plant height, leaf area index, dry matter accumulation and crop growth rate compared with control. The maximum plant height was observed in Azospirillum+Mycorrhiza at 89 days after emerging. The highest and lowest leaf area index was observed in B+C (0.37 and control (0.22 treatments, respectively. The fast period of vegetative growth and dry matter accumulation were observed at 40-89 days after emerging with a small decline afterwards until physiological maturity. The maximum and minimum amounts of dry matter accumulation were recorded in the B+C treatment with 66.0 gm-2, and control with 38.3 gm-2, respectively. Crop growth rate reached to its peak in 82 days after emergence

  8. Evaluation of the association nitrogen fixing bacterias interspecific – rice lines – nitrogen, in typic haplustalf. Ibagué, Colombia Evaluación de la asociación bacterias fijadoras de nitrógeno – líneas interespecíficas de arroz–nitrógeno, en Typic haplustalf. Ibagué, Colombia

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    Bonilla Correa Carmen Rosa

    2008-03-01

    Full Text Available The study was carried out at the Cauchitos farm, Ibague municipality department of Tolima, with bounds: North 4°23'51"; and west 75°9'7";, 979 ansm, the average temperature is 24,3°C, tropical dry forest (bs–t in the Holdridge classification. The purpose of this study was to evaluate the association between the nitrogen fixation bacteries with and without inoculo in 10 interespecific rice lines with three nitrogen dosis (0, 50 and 100% de 250 kg/ ha–1 and three repetitions. The inoculation was realized with 108 former units of colonies per millimeter. 2.260 bacteries of the generums Azotobacter spp y Azospirrillum spp., and identification the species: Azospirillum brasilense, Azospirillum lipoferum, Azospirillum amazonense, were identified and from the genus Azotobacter were identified the species: A. choroococcum, A. vinelandii, A. paspali and A. veijerinckii. Weren't found none significative differences after the inoculation. Azotobacter spp and Azospirillum spp. were typical generums of the bacterian flora in the rice plantation and in field conditions were effect of the treatment effects in the bacterian . The Azotobacter spp was the predominant in generum in each one of the treatments.El estudio se llevó a cabo en la hacienda Cauchitos, municipio de Ibagué, departamento del Tolima (Norte 4° 23' 51"; y Oeste 75° 9' 7";, 979 msnm, 24.3°C, bosque seco tropical (bs–T, con el objetivo de evaluar las asociaciones entre bacterias fijadoras de nitrógeno con inóculo y sin él en diez líneas interespecíficas de arroz, con tres dosis de nitrógeno (0%, 50% y 100% de 250 kg/ha–1 y tres repeticiones por tratamiento. La inoculación se

  9. CARACTERIZACIÓN DE BACTERIAS DIAZOTRÓFICAS ASIMBIÓTICAS ASOCIADAS AL EUCALIPTO (Eucalyptussp. EN CODAZZI, CESAR (COLOMBIA Characterization of Diazotrophic Bacteria Non-Symbiotic Associated with Eucalyptus (Eucalyptussp. in Codazzi, Cesar (Colombia

    Directory of Open Access Journals (Sweden)

    DOLLY MELISSA OBANDO CASTELLANOS

    Full Text Available Se evaluó el efecto de las épocas climáticas (lluvia y sequía y del estrato de la muestra (suelo rizosférico, raíces y hojas sobre la población de los géneros Azotobacter, Beijerinckia, Derxia, Azospirillum, Herbaspirillum, Gluconacetobacter y Burkholderia en el Eucalipto (Eucalyptus sp.. Así mismo, se evalúo su capacidad en la producción de compuestos indólicos como promotores del crecimiento vegetal y su actividad de reducción de acetileno como indicador de la fijación biológica de nitrógeno. Los resultados no registraron diferencias estadísticas significativas en el test de Tukey (P ≤ 0.05 en la población con respecto a la época climática. Con respecto al estrato de muestra, los aislamientos tentativos de Herbaspirillum sp. y Azospirillum sp. presentaron diferencias significativas en suelo rizosférico y raíces. Se obtuvieron 44 aislamientos de los cuales se agruparon por caracterización fenotípica como: 14 presuntivos de Beijerinckia sp., 12 de Azotobacter sp., ocho de Derxia sp., cuatro de Herbarpirillum sp., cinco de Azospirillum sp., uno de Gluconacetobacter sp. y uno de Burkholderia sp. Por su alto potencial fueron seleccionados y criopreservados los aislamientos C27, C26 y C25, las cuales presentaron los mejores valores de eficiencia in vitro, superando valores de producción de las cepas de referencia utilizadas (A. chroococcum (AC1 y A. brasilense (SP7.The effect of climatic seasons (rainy and dry and the stratum sample (rhizospheric soil, roots and leaves the population of the genera Azotobacter, Beijerinckia, Derxia, Azospirillum, Herbaspirillum, Gluconacetobacter and Burkholderia in soil rhizosphere, roots and leaves of Eucalyptus (Eucalyptus sp.. It also assesses their ability to produce indoles compounds as plant growth promoters and their acetylene reduction activity as an indicator of biological fixation of nitrogen. The results showed no statistically significant differences in the Duncan test (P ≤ 0

  10. CARACTERIZACIÓN DE BACTERIAS DIAZOTRÓFICAS ASIMBIÓTICAS ASOCIADAS AL EUCALIPTO (Eucalyptus sp. EN CODAZZI, CESAR.

    Directory of Open Access Journals (Sweden)

    Divan Baldani Vera Lúcia

    2010-12-01

    Full Text Available Se evaluó el efecto de las épocas climáticas (lluvia y sequía y del estrato de la muestra (Suelo rizosférico, raíces y hojas sobre la población de los géneros Azotobacter, Beijerinckia, Derxia, Azospirillum, Herbaspirillum, Gluconacetobacter y Burkholderia en el Eucalipto (Eucalyptus sp.. Así mismo, se evalúo su capacidad en la producción de compuestos indólicos como promotores del crecimiento vegetal y su actividad de reducción de acetileno como indicador de la fijación biológica de nitrógeno. Los resultados no registraron diferencias estadísticas significativas en el test de Tukey (P ≤ 0.05 en la población con respecto a la época climática. Con respecto al estrato de muestra, los aislamientos tentativos a Herbaspirillum sp. y Azospirillum sp. presentaron diferencias significativas en suelo rizosférico y raíces. Se obtuvieron 44 aislamientos de los cuales se agruparon por caracterización fenotípica como: 14 presuntivos del género Beijerinckia sp., 12 de Azotobacter sp., 8 de Derxia sp., 4 de Herbarpirillum sp., 5 de Azospirillum sp., 1 de Gluconacetobacter sp. y 1 de Burkholderia sp. Por su alto potencial fueron seleccionados y criopreservados los aislamientos C27, C26, C25 y C45, las cuales presentaron los mejores valores de eficiencia in vitro, superando valores de producción de las cepas de referencia utilizadas (A. chroococcum (AC-01 y A. brasilense (SP7.

  11. BIOFERTILIZATION USING RHIZOBACTERIA AND AMF IN THE PRODUCTION OF TOMATO (Lycopersicon esculentum Mill. AND ONION (Allium cepa L. SEEDLINGS. II. ROOT COLONIZATION AND NUTRITIONAL STATUS

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    L. E. Pulido

    2003-01-01

    Full Text Available Como complemento a estudios precedentes de la biofertilización en posturas de plantas hortícolas sobre suelos Ferralíticos Rojos compactados, eútricos y en áreas experimentales de la Universidad de Ciego de Ávila, se evaluaron los efectos de la inoculación simple y la coinoculación, mediante el recubrimiento de semillas y sin aplicar fertilizantes minerales, con rizobacterias promotoras del crecimiento vegetal -RPCV- (Azospirillum brasilense, Azotobacter chroococcum y Burkholderia cepacia y hongos micorrízicos arbusculares - HMA- (Glomus clarum y G. fasciculatum en algunos indicadores de la colonización radical por los microorganismos y el estado nutricional de plántulas de tomate y cebolla. A partir de los resultados, se evidenció que, para ambos cultivos, las poblaciones de A. chroococcum, B. cepacia y A. brasilense se incrementaron significativamente en aquellos tratamientos inoculados con estas rizobacterias, encontrando, en general, los mayores valores en los tratamientos que fueron coinoculados. Respecto a la micorrización, los mayores porcentajes de colonización micorrízica y masa del endófito en tomate se obtuvieron mediante la coinoculación de A. brasilense con ambas especies de HMA y, para la cebolla, la máxima colonización la realizó G. fasciculatum aplicada de forma independiente, mientras que la masa del endófito fue mayor en la coinoculación de G. clarum + A. chroococcum. En relación con el estado nutricional de las plantas, en tomate, los tratamientos con presencia conjunta de A. brasilense y ambas especies de HMA fueron los que hicieron mayores extracciones de N y estuvieron entre los que realizaron mayores extracciones de P y K. En cebolla, todos los tratamientos inoculados con ambos tipos de microorganismos fueron capaces de extraer mayores cantidades de N, P y K. Todos estos resultados permiten explicar las causas de la obtención de posturas de adecuada calidad mediante la biofertilización sin el uso

  12. Fixação de nitrogênio e produção de ácido indolacético in vitro por bactérias diazotróficas endofíticas Nitrogen fixation and in vitro production of indolacetic acid by endophytic diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Anelise Vicentini Kuss

    2007-10-01

    Full Text Available O objetivo deste trabalho foi isolar e quantificar bactérias diazotróficas associadas a raízes de arroz, e avaliar a produção de ácido indolacético e o potencial de fixação biológica de nitrogênio dessas bactérias, a fim de selecionar isolados promissores para inoculação em plantas. Bactérias fixadoras de nitrogênio, habitantes do interior das raízes de cultivares de arroz do Rio Grande do Sul, foram isoladas e quantificadas em nove cultivares. Raízes de arroz superficialmente esterilizadas foram maceradas e introduzidas em meios de crescimento, elaborados sem fonte de nitrogênio e em condições semi-sólidas. Entre os 58 isolados nos meios NFb, LGI e LGI-P, foram escolhidos UFSM-BD-02-06, UFSM-BD-08-06, UFSM-BD-14-06, UFSM-BD-20-06, UFSM-BD-26-06, UFSM-BD-31-06, UFSM-BD-36-06, UFSM-BD-42-06, UFSM-BD-48-06, UFSM-BD-54-06. Avaliaram-se a fixação biológica de nitrogênio e a produção de ácido indolacético in vitro, pelos métodos Kjeldahl e colorimétrico, respectivamente. Azospirillum brasilense e A. lipoferum apresentam maiores valores para N total, 41,08 e 46,82 µg mL-1, respectivamente. A. brasilense e UFSM-BD-31-06 são os maiores produtores de ácido indolacético, 41,09 mg mL-1 e 13,47 µg mL-1, respectivamente.The aim of this work was to isolate and to quantify diazotrophic bacteria associated with rice root, and evaluate their acid indolacetic production and their potential for biological nitrogen fixation, with the purpose of selecion promissing isolates for plant inoculation. N-fixing bactéria, settlers of the root interior of rice cultivars used in Rio Grande do Sul, Brazil, were isolated and quantified in nine cultivars. Rice root superficialy sterilized were macerated and introduced in specific culture media. Among 58 isolates obtained in the culture media NFB, LGI and LGI-P, the following were selected: UFSM-BD-02-06, UFSM-BD-08-06, UFSM-BD-14-06, UFSM-BD-20-06, UFSM-BD-26-06, UFSM-BD-31-06, UFSM-BD-36

  13. Alternative rooting induction of semi-hardwood olive cuttings by several auxin-producing bacteria for organic agriculture systems

    Directory of Open Access Journals (Sweden)

    M. C. Montero-Calasanz

    2013-01-01

    Full Text Available Southern Spain is the largest olive oil producer region in the world. In recent years organic agriculture systems have grown exponentially so that new alternative systems to produce organic olive cuttings are needed. Several bacterial isolates, namely Pantoea sp. AG9, Chryseobacterium sp. AG13, Chryseobacterium sp. CT348, Pseudomonas sp. CT364 and Azospirillum brasilense Cd (ATCC 29729, have been used to induce rooting in olive semi-hardwood cuttings of Arbequina, Hojiblanca and Picual cultivars of olive (Olea europea L. The first four strains were previously selected as auxin-producing bacteria and by their ability to promote rooting in model plants. They have been classified on the basis of their 16S rDNA gene sequence. The known auxin producer A. brasilense Cd strain has been used as a reference. The inoculation of olive cuttings was performed in two different ways: (i by dipping cuttings in a liquid bacterial culture or (ii by immersing them in a paste made of solid bacterial inoculant and sterile water. Under nursery conditions all of the tested bacterial strains were able to induce the rooting of olive cuttings to a similar or greater extent than the control cuttings treated with indole-3-butyric acid (IBA. The olive cultivars responded differently depending on the bacterial strain and the inoculation method. The strain that consistently gave the best results was Pantoea sp. AG9, the only one of the tested bacterial strains to express the enzyme 1-aminocyclopropane-1-carboxylate (ACC deaminase. The results are also discussed in terms of potential commercial interest and nursery feasibility performance of these strains.

  14. Effect of immobilized rhizobacteria and organic amendment in bulk and rhizospheric soil of Cistus albidus L.

    Science.gov (United States)

    Mengual, Carmen Maria; del Mar Alguacil, Maria; Roldan, Antonio; Schoebitz, Mauricio

    2013-04-01

    A field experiment was carried out to assess the effectiveness of the immobilized microbial inoculant and the addition of organic olive residue. The microbial inoculant contained two rhizobacterial species identified as Azospirillum brasilense and Pantoea dispersa immobilized in a natural inert support. Bacterial population densities were 3.5×109 and 4.1×109 CFU g-1 of A. brasilense M3 and P. dispersa C3, respectively. The amendment used was the organic fraction extracted with KOH from composted "alperujo". The raw material was collected from an olive-mill and mixed with fresh cow bedding as bulking agent for composting. The inoculation of rhizobacteria and the addition of organic residue were employed for plant growth promotion of Cistus albidus L. and enhancement of soil physicochemical, biochemical and biological properties in a degraded semiarid Mediterranean area. One year after planting, the available phosphorus and potassium content in the amended soils was about 100 and 70% respectively higher than in the non-amended soil. Microbial inoculant and their interaction with organic residue increased the aggregate stability of the rhizosphere soil of C. albidus (by 12% with respect to control soil) while the organic residue alone not increased the aggregate stability of the rhizosphere of C. albidus. Microbial biomass C content and enzyme activities (dehydrogenase, urease, protease-BAA and alkaline phosphatase) of the rhizosphere of C. albidus were increased by microbial inoculant and organic residue interaction but not by microbial inoculation alone. The microbial inoculant and organic residue interaction were the most effective treatment for stimulating the roots dry weight of C. albidus (by 133% with respect to control plants) and microbial inoculant was the most effective treatment for increase the shoot dry weigh of plants (by 106% with respect to control plants). The combined treatment, involving microbial inoculant and addition of the organic residue

  15. Identification of nif genes in N2-fixing bacterial strains isolated from rice fields along the Yangtze River Plain.

    Science.gov (United States)

    Xie, Guang Hui; Cui, Zongjun; Yu, Jun; Yan, Jing; Hai, Weili; Steinberger, Yosef

    2006-01-01

    The aim of this research was to identify nifH and nifHDKYE ' genes in twenty strains of N2-fixing heterotrophic bacteria isolated from rice fields in the Yangtze River Plain. Southern hybridization of the total DNA from each strain was performed with the Klebsiella pneumoniae nifHDKYE ' gene probe (6.2 kb Eco RI fragment from pSA30) and the Azospirillum brasilense nifH gene probe (0.6 kb Eco RI-Hin dIII fragment from pHU8). We found that Eco RI fragments of total DNA from Aeromonas hydrophila HY2, Bacillus azotoformans FD, Bacillus licheniformis NCH1, NCH5, WH4, Bacillus brevis NC2, Bacillus pumilus NC12, Bacillus cereus NCH2, Citrobacter freundii HY5, HY9, Derxia gummosa HZ5, Pseudomonas mendocina HZ1 and Pseudomonas pseudoalcaligenes WH3 were positively hybridized with both of the probes. Agrobacterium radiobacter HY17, Corynebacterium sp. HY12, YZ and Pseudomonas sp. HY11 had Eco RI fragments hybridized with the K. pneumoniae nifHDKYE ' gene probe. An Eco RI fragment of total DNA from Bacillus megaterium YY4 was positively hybridized to the A. brasilense nifH gene probe. No hybridization sign was found in the total DNA fragments from Alcaligenes cupidus YY6 and Corynebacterium sp. NC11 hybridized with either of the gene probes. The data provide the number and size of EcoRI fragments of the total DNA hybridized with the nif gene probes for these strains of rarely studied species, suggesting additional evidence for N2 fixing and nif gene diversity of N2-fixing bacteria in rice fields along the Yangtze River Plain.

  16. Use of Phosphate Solubilizing Bacteria to Leach Rare Earth Elements from Monazite-Bearing Ore

    Directory of Open Access Journals (Sweden)

    Doyun Shin

    2015-04-01

    Full Text Available In the present study, the feasibility to use phosphate solubilizing bacteria (PSB to develop a biological leaching process of rare earth elements (REE from monazite-bearing ore was determined. To predict the REE leaching capacity of bacteria, the phosphate solubilizing abilities of 10 species of PSB were determined by halo zone formation on Reyes minimal agar media supplemented with bromo cresol green together with a phosphate solubilization test in Reyes minimal liquid media as the screening studies. Calcium phosphate was used as a model mineral phosphate. Among the test PSB strains, Pseudomonas fluorescens, P. putida, P. rhizosphaerae, Mesorhizobium ciceri, Bacillus megaterium, and Acetobacter aceti formed halo zones, with the zone of A. aceti being the widest. In the phosphate solubilization test in liquid media, Azospirillum lipoferum, P. rhizosphaerae, B. megaterium, and A. aceti caused the leaching of 6.4%, 6.9%, 7.5%, and 32.5% of calcium, respectively. When PSB were used to leach REE from monazite-bearing ore, ~5.7 mg/L of cerium (0.13% of leaching efficiency and ~2.8 mg/L of lanthanum (0.11% were leached by A. aceti, and Azospirillum brasilense, A. lipoferum, P. rhizosphaerae and M. ciceri leached 0.5–1 mg/L of both cerium and lanthanum (0.005%–0.01%, as measured by concentrations in the leaching liquor. These results indicate that determination of halo zone formation was found as a useful method to select high-capacity bacteria in REE leaching. However, as the leaching efficiency determined in our experiments was low, even in the presence of A. aceti, further studies are now underway to enhance leaching efficiency by selecting other microorganisms based on halo zone formation.

  17. Construction of plant expression vectors carrying glnA gene encoding glutamine synthetase and regeneration of transgenic rice plants

    Institute of Scientific and Technical Information of China (English)

    苏金; 张雪琴; 颜秋生; 陈章良; 尤崇杓

    1995-01-01

    The glnA gene encoding glutamine synthetase (GS) was amplified from Azospirillum brasilenseSp7 with PCR technique.The amplified 1.4-kb DNA fragment flanked with a BamH Ⅰ site at each end wascloned into EcoR V site of Bluescript-SK vector.A recombinant plasmid pGSJ1 containing this 1.4-kb DNA frag-ment was selected by restriction digestion analysis.The sequencing data also confirmed that the amplified 1.4-kbDNA fragment was undoubtedly the glnA gene of A.brasilense Sp7.Then the 1.4-kb BamH Ⅰ fragment was ex-cised from pGSJ1.A glnA plant expression vector pAGNB92 with rice actin 1 (Act1) promoter was constructedby using colony in situ hybridization to screen positive clones,and 3 rounds of ligation and transformation wereperformed.Protoplasts isolated from rice (Oryza sativa,L.Japonica) cell suspension line (cv.T986) weretransformed with the glnA plant expression vector pAGNB92 carrying neomycin phosphotransferase Ⅱ (NPT Ⅱ)gene by PEG fusion or electroporation.G418~ calli were used to detect NPT Ⅱ enzyme activity.The resultsshow that G418~ calli possess high positive hybridization signal with the frequency of 37%.The regeneratedG418~NPTII~+ rice plants were used for PCR amplification of glnA gene,and a 1.4-kb DNA fragment was ampli-fied from glnA-transgenic rice plants (R0 generation).The results of Southern blot hybridization prove that the1.4-kb DNA fragment amplified from the total DNA of glnA transgenic rice plants is indeed the glnA gene of A.brasilense Sp7.Northern blot hybridization was carried out using the same glnA gene as probe.The glnAgene was expressed in the transgenic rice plants.Bioassays also confirmed that the glnA transgenic rice plantsgrew much better than that of the control plants under a condition with nitrogen poor source (0.75 mmol/L).

  18. Growth promoting characteristics of rhizobacteria and AM Fungi for biomass amelioration of Zea mays

    Directory of Open Access Journals (Sweden)

    Kumar Manoj

    2015-01-01

    Full Text Available Plant growth promoting rhizobacteria (PGPR and mycorrhiza were evaluated on the growth (biomass and yield of Zea mays. In the present study, selective rhizospheric PGPR (Azotobacter chroococcum, Pseudomonas aeruginosa, Azospirillum brasilense and Streptomyces sp. and a combination of six strains of arbuscular mycorrhizal fungi (AMF (Acaulospora morrowae, Gigaspora margarita, Glomus constrictum, Glomus mossae, Glomus aggregatum and Scutellospora calospora were isolated and identified with standard methods and 16S rRNA sequence analysis. PGPR and AMF were checked for their growth-promoting behavior under specific treatment conditions. The 30-48-day-old treated plants in all combinations showed a significantly higher mass value. The average dry weight from the shoot was in a range from 41-52% as compared to the control. This increase also translated into a higher mass value of the roots. Overall, an 82% growth rate was observed in terms of height as the consequence of biomass production, specifically in the case of AMF + rhizobacteria combination. We report an efficient, sustainable and cost-effective biofertilizer for enhanced biomass of Z. mays, one of the staple food crops worldwide.

  19. Trace cobalt speciation in bacteria and at enzymic active sites using emission Moessbauer spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Kamnev, A.A.; Antonyuk, L.P.; Smirnova, V.E.; Serebrennikova, O.B. [Laboratory of Biochemistry, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov (Russian Federation); Kulikov, L.A.; Perfiliev, Yu.D. [Laboratory of Nuclear Chemistry Techniques, Department of Radiochemistry, Faculty of Chemistry, Moscow State University (Russian Federation)

    2002-02-01

    {sup 57}Co emission Moessbauer spectroscopy (EMS) allows the chemical state of cobalt, as influenced by its coordination environment, to be monitored in biological samples at its physiological (trace) concentrations. To draw attention to EMS as a valuable tool for speciation of cobalt in biocomplexes, the process of cobalt(II) metabolism in cells of the plant growth-promoting rhizobacterium Azospirillum brasilense Sp245 was investigated using EMS of {sup 57}Co{sup II}-doped bacterial cells. EMS measurements also showed {sup 57}Co{sup II}-activated glutamine synthetase (GS, a key enzyme of nitrogen metabolism, isolated from this bacterium) to have two different cobalt(II) forms at its active sites, in agreement with data available on other bacterial GSs. Chemical after-effects following electron capture by the nucleus of the parent {sup 57}Co{sup II} during the {sup 57}Co{yields}{sup 57}Fe transition, which contribute to the formation of a stabilised daughter {sup 57}Fe{sup III} component along with the nucleogenic {sup 57}Fe{sup II} forms, are also briefly considered. (orig.)

  20. Effects of head pruning and different nutritional systems (chemical, biological and integrated on seed yield and oil content in medicinal pumpkin (Cucurbita pepo L.

    Directory of Open Access Journals (Sweden)

    Zarei Dariush

    2016-01-01

    Full Text Available To evaluate the effect of head pruning and different nutritional systems (chemical, biological and integrated on yield and seed oil content in medicinal pumpkin (Cucurbita pepo L., an experiment was conducted in Kermanshah/Iran during the 2013 growing season. The experimental treatments consisted of two levels - no head pruning, control (Co and head pruning (C1 allocated to the main plots. Four levels of different fertilizing systems - control (without fertilizer (T0, chemical (T1, biological (a combination of nitrogen fixing bacteria, Azospirillum brasilense and Glomus mosseae (T2, and integrated fertilizing system (biological fertilizer + 50% chemical fertilizer (T3 were assigned to the sub-plots. The experimental treatments were arranged as a split plot based on a randomized complete block design with three replications. The results showed that the highest percentage of seed oil was obtained (37% in the integrated nutritional system along with the head pruning treatment. The highest grain yields of 53 and 50 g per square meter were obtained in integrated and chemical fertilizing systems, respectively while no pruning was applied. The highest fruit yields of 3,710 and 3,668 kg per hectare were produced by chemical and integrated fertilizing systems, respectively. The biological nutrition system required more time to demonstrate its positive effect on the growth and yield of medicinal pumpkin.

  1. Soil factors exhibit greater influence than bacterial inoculation on alfalfa growth and nitrogen fixation.

    Science.gov (United States)

    Neumann, Ute; Kosier, Bob; Jahnke, Joachim; Priefer, Ursula B; Al-Halbouni, Djamila

    2011-09-01

    In order to study the effects of soil factors and bacterial inoculation on alfalfa (Medicago sativa), plants were inoculated with Ensifer meliloti L33 and Azospirillum brasilense Sp7 in pot experiments using two different soils separately as well as in a mixture. One soil was contaminated with chemical waste products; the other was an arable soil. Soil factors, including the availability of macro- and micronutrients as well as carbon and nitrogen contents, were found to exhibit a much greater influence on the growth of alfalfa than any of the inoculations. In contaminated soil, the shoot and root growth of alfalfa was decreased and nodules were diminished and ineffective. Bacterial inoculations did not significantly improve this hostile growth environment. However, in a mixture (44% arable, 22% contaminated soil, 34% vermiculite), growth conditions for alfalfa were improved so that shoot dry weight and nodule numbers increased up to 100- and 20-fold, respectively, compared with the contaminated soil. For the strain L33, its persistence in the rhizosphere was correlated to the presence of its host plant, but its dynamics were influenced by competition with indigenous rhizobia. The strain Sp7, once provided with a suitable soil, was not dependent on the plant's rhizosphere, but it enhanced the performance of L33 and native rhizobia.

  2. Oviposition Attractancy of Bacterial Culture Filtrates: response of Culex quinquefasciatus

    Directory of Open Access Journals (Sweden)

    S Poonam

    2002-04-01

    Full Text Available Oviposition attractants could be used for monitoring as well as controlling mosquitoes by attracting them to lay eggs at chosen sites. In the present study, culture filtrates of seven bacterial species were tested for their attractancy against gravid females of Culex quinquefasciatus. When their oviposition active indices (OAI were studied, the culture filtrates of Bacillus cereus and Pseudomonas fluorescens exhibited oviposition attractancy (OAI = >0.3 at 100 ppm and the OAI were respectively 0.70 and 0.47. Culture filtrates of B. thuringiensis var. israelensis (wild type, B. t. var. israelensis (mutant and B. sphaericus showed attractancy at 2000 ppm with OAI of respectively 0.71, 0.59 and 0.68. However, the OAI of B. megaterium as well as Azospirillum brasilense was 0.13 (at 2000 ppm, which was less than 0.3 required to be considered them as attractants. When the oviposition attractancy of the bacterial culture filtrates were compared with that of a known oviposition attractant, p-cresol (at 10 ppm, the culture filtrates of B. t. var. israelensis (wild type and B. cereus were found to be more active than p-cresol, respectively with 64.2 and 54.3% oviposition.

  3. Densidade e diversidade fenotípica de bactérias diazotróficas endofíticas em solos de mineração de bauxita, em reabilitação Density and phenotypic diversity of endophytic nitrogen fixing bacteria in soils under rehabilitation after bauxite mining

    Directory of Open Access Journals (Sweden)

    R. Melloni

    2004-02-01

    Full Text Available Bactérias diazotróficas endofíticas contribuem para o desenvolvimento das plantas por meio da fixação biológica do nitrogênio, produção e liberação de substâncias reguladoras do crescimento vegetal, podendo, assim, facilitar a revegetação de solos degradados por atividades antrópicas. No entanto, pouco se conhece sobre as populações destas bactérias em solos ou plantas de áreas de mineração. Objetivando avaliar o efeito de diferentes tipos de vegetação e tempo de reabilitação de áreas degradadas por mineração de bauxita na densidade e diversidade de algumas espécies de bactérias diazotróficas endofíticas, realizaram-se, em duas épocas, amostragens de solo, de dois ambientes distintos, submetidos a diferentes processos de reabilitação. A densidade, avaliada pelo número mais provável, utilizando os meios de cultura: NFb, JNFb e Fam, para Azospirillum brasilense e A. lipoferum, Herbaspirillum spp. e A. amazonense, respectivamente, variou de 0 a 2,0 x 10(4 bactérias por grama de solo e mostrou que o tipo de vegetação influiu nas populações destas bactérias. Foram encontradas densidades maiores em solos revegetados com gramíneas: braquiária (Brachiaria decumbens, capim-azevém (Lolium multiflorum e capim-gordura (Melinis minutiflora. Contudo, estas densidades podem ser consideradas baixas, se comparadas às de solos agrícolas, e não apresentaram relação com o tempo de reabilitação da área. Foram encontrados 36 fenótipos culturais em meio batata, entre os 72 isolados obtidos dos três meios de cultura utilizados. A partir destes, foram formados sete grandes grupos com similaridade superior ou igual a 63 %, dos quais cinco, representando 62,5 % do total de isolados obtidos, continham as estirpes-tipo de Burkholderia brasilensis, Herbaspirillum seropedicae e Azospirillum spp. (A. brasilense, A. amazonense, A. lipoferum, A. irakense. Apesar da baixa densidade, este grupo de bactérias apresentou alta

  4. Effect of plant growth-promoting bacteria on the growth and fructan production of Agave americana L.

    Directory of Open Access Journals (Sweden)

    Neyser De La Torre-Ruiz

    Full Text Available ABSTRACT The effect of plant growth-promoting bacteria inoculation on plant growth and the sugar content in Agave americana was assessed. The bacterial strains ACO-34A, ACO-40, and ACO-140, isolated from the A. americana rhizosphere, were selected for this study to evaluate their phenotypic and genotypic characteristics. The three bacterial strains were evaluated via plant inoculation assays, and Azospirillum brasilense Cd served as a control strain. Phylogenetic analysis based on the 16S rRNA gene showed that strains ACO-34A, ACO-40 and ACO-140 were Rhizobium daejeonense, Acinetobacter calcoaceticus and Pseudomonas mosselii, respectively. All of the strains were able to synthesize indole-3-acetic acid (IAA, solubilize phosphate, and had nitrogenase activity. Inoculation using the plant growth-promoting bacteria strains had a significant effect (p < 0.05 on plant growth and the sugar content of A. americana, showing that these native plant growth-promoting bacteria are a practical, simple, and efficient alternative to promote the growth of agave plants with proper biological characteristics for agroindustrial and biotechnological use and to increase the sugar content in this agave species.

  5. The effect of ectomycorrhizal fungi and bacteria on pine seedlings

    Directory of Open Access Journals (Sweden)

    Hanna Dahm

    2014-08-01

    Full Text Available The effect of ecomycorrhizal fungi (Hebelon crustuliniforme(Bull.: Fr. Quél. 5392 and Pisolithus tinctorius (Pers. Coker et Couch 5335 and bacteria (Bacillus polymyxa and Azospirillum brasilense. associated with mycorrhizas on the growth of pine seedligs was investigated. In addition the influence of bacteria on fungal biomass production and the relationship between ectomycorrhizal fungi and fungi pathogenic to root of pine seedlings were determined. In general, the shoot/root ratio was higher in plants inoculated with Hebeloma crustuliniforme and bacteria than in the control seedlings (grown only under sterile conditions. In non-sterile substrate the root/shoot ratio of the mycorrhizal seedlings was lower as compared to the control. Similar phenomenon was noted in plants inoculated with the mycorrhizal fungus Pisolithus tinetorius. The bacteria used as well as the time of introduction of these organisms into the cultures of mycorrhiza fungi affected the production of fungal biomass. Hebeloma crustuliniforme and Pisolithus tinctorius inhibited the growth of Rizoctonia solani and Fusarium oxysporum fungi pathogenic to pine seedlings.

  6. BIOFERTILIZANTES Y PRODUCTOS BIOACTIVOS, ALTERNATIVAS PARA LA ASOCIACIÓN MAÍZ-TOMATE EN EL PERÍODO TEMPRANO DE SIEMBRA

    Directory of Open Access Journals (Sweden)

    Elein Terry

    2006-01-01

    Full Text Available Con el objetivo de conocer el efecto agronómico de la utilización combinada de biofertilizantes y productos bioactivos, sobre el desarrollo, rendimiento y la calidad interna del tomate (variedad ´Amalia´, cultivado fuera del período óptimo bajo un sistema de asociación de cultivos, se llevó a cabo un experimento de campo en el 2000 y 2001, sobre un suelo Ferralítico Rojo lixiviado típico. Los tratamientos consistieron en plantas coinoculadas (Glomus clarum + Azospirillum brasilense y asperjadas foliarmente con los productos bioactivos Biostan y Biobras-16, aplicados al inicio de la floracióndel cultivo. Los resultados mostraron una relación compatibleentre microorganismos y productos bioactivos, permitieron un adecuado balance nutricional de las plantas, así como un rendimiento entre 26 y 29 t.ha-1, siendo superior en un rango del 12-19 % con respecto al testigo de producción; por otra parte, los frutos obtenidos con estos tratamientos presentaron una calidad bromatológica superior.

  7. 巴西固氮螺菌GlnB与NifA蛋白N端结构域的相互作用

    Institute of Scientific and Technical Information of China (English)

    周小愚; 邹笑笑; 李季伦

    2008-01-01

    巴西固氮螺菌(Azospirillum brasilense)可与多种重要作物联合固氮,具有作为生物肥料的潜能.固氮基因(nif)的转录激活蛋白NifA和PⅡ信号转导蛋白家族成员GlnB是参与该菌固氮调控的两个关键蛋白,且NifA的活性调控依赖于GlnB.研究结果表明,在无氮条件下GlnB与NifA蛋白N端结构域存在相互作用,且N端18和53位酪氨酸突变为苯丙氨酸的NifA与GlnB的互作增强.研究还发现,NifA蛋白N端66~88位和165~176位氨基酸区域对于与GlnB蛋白的相互作用是必需的.

  8. Structure of insoluble immune complexes as studied by spectroturbidimetry and dynamic light scattering

    Science.gov (United States)

    Khlebtsov, Boris N.; Burygin, Gennadii L.; Matora, Larisa Y.; Shchyogolev, Sergei Y.; Khlebtsov, Nikolai G.

    2004-07-01

    We describe two variants of a method for determining the average composition of insoluble immune complex particles (IICP). The first variant is based on measuring the specific turbidity (the turbidity per unit mass concentration of the dispersed substance) and the average size of IICP determined from dynamic light scattering (DLS). In the second variant, the wavelength exponent (i.e., the slope of the logarithmic turbidity spectrum) is used in combination with specific turbidity measurements. Both variants allow the average biopolymer volume fraction to be determined in terms of the average refractive index of IICP. The method is exemplified by two experimental antigen+antibody systems: (i) lipopolysaccharide-protein complex (LPPC) of Azospirillum brasilense Sp245+rabbit anti-LPPC; and (ii) human IgG (hIgG)+sheep anti-hIgG. Our measurements by the two methods for both types of systems gave, on the average, the same result: the volume fraction of the IICP biopolymers is about 30%; accordingly, the volume fraction of buffer solvent is 70%.

  9. Bioremediation of hydrocarbons contaminating sewage effluent using man-made biofilms: effects of some variables.

    Science.gov (United States)

    Al-Mailem, D M; Kansour, M K; Radwan, S S

    2014-11-01

    Biofilm samples were established on glass slides by submerging them in oil-free and oil-containing sewage effluent for a month. In batch cultures, such biofilms were effective in removing crude oil, pure n-hexadecane, and pure phenanthrene contaminating sewage effluent. The amounts of the removed hydrocarbons increased with increasing biofilm surface area exposed to the effluent. On the other hand, addition of the reducing agent thioglycollate dramatically inhibited the hydrocarbon bioremediation potential of the biofilms. The same biofilm samples removed contaminating hydrocarbons effectively in three successive batch bioremediation cycles but started to become less effective in the cycles thereafter, apparently due to mechanical biofilm loss during successive transfers. As major hydrocarbonoclastic bacteria, the biofilms harbored species belonging to the genera Pseudomonas, Microvirga, Zavarzinia, Mycobacterium, Microbacterium, Stenotrophomonas, Gordonia, Bosea, Sphingobium, Brachybacterium, and others. The nitrogen fixer Azospirillum brasilense and the microalga Ochromonas distigma were also present; they seemed to enrich the biofilms, with nitrogenous compounds and molecular oxygen, respectively, which are known to enhance microbiological hydrocarbon degradation. It was concluded that man-made biofilms based upon sewage microflora are promising tools for bioremediation of hydrocarbons contaminating sewage effluent.

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

    Science.gov (United States)

    Baldani, José I; Baldani, Vera L D

    2005-09-01

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

  11. USO COMBINADO DE MICROORGANISMOS BENÉFICOS Y PRODUCTOS BIOACTIVOS COMO ALTERNATIVA PARA LA PRODUCCIÓN DE TOMATE (Lycopersicon esculentum, Mill

    Directory of Open Access Journals (Sweden)

    Elein Terry

    2005-01-01

    Full Text Available Con el objetivo de conocer la efectividad agrobiológica de la combinación de microorganismos benéficos y productos bioactivos, sobre el desarrollo, rendimiento y la calidad interna en el cultivo del tomate (variedad 'Amalia', se llevó a cabo un experimento de campo en el 2000 y 2001, sobre un suelo Ferralítico Rojo lixiviado típico. Los tratamientos consistieron en plantas coinoculadas (Glomus clarum- Azospirillum brasilense y asperjadas foliarmente con el producto bioactivo Biostan en diferentes momentos del desarrollo de las plantas. Los resultados mostraron una relación armónica y ecológicamente compatible entre ambos productos, permitiendo un adecuado estado nutricional de las plantas, así como posibilitó un rendimiento de 35 t.ha-1, siendo superior en un 17 % al testigo de producción; por otra parte, los frutos obtenidos con estos tratamientos tuvieron una mejor calidad bromatológica.

  12. Ternary complex formation between AmtB, GlnZ and the nitrogenase regulatory enzyme DraG reveals a novel facet of nitrogen regulation in bacteria.

    Science.gov (United States)

    Huergo, Luciano F; Merrick, Mike; Pedrosa, Fábio O; Chubatsu, Leda S; Araujo, Luíza M; Souza, Emanuel M

    2007-12-01

    Ammonium movement across biological membranes is facilitated by a class of ubiquitous channel proteins from the Amt/Rh family. Amt proteins have also been implicated in cellular responses to ammonium availability in many organisms. Ammonium sensing by Amt in bacteria is mediated by complex formation with cytosolic proteins of the P(II) family. In this study we have characterized in vitro complex formation between the AmtB and P(II) proteins (GlnB and GlnZ) from the diazotrophic plant-associative bacterium Azospirillum brasilense. AmtB-P(II) complex formation only occurred in the presence of adenine nucleotides and was sensitive to 2-oxoglutarate when Mg(2+) and ATP were present, but not when ATP was substituted by ADP. We have also shown in vitro complex formation between GlnZ and the nitrogenase regulatory enzyme DraG, which was stimulated by ADP. The stoichiometry of this complex was 1:1 (DraG monomer : GlnZ trimer). We have previously reported that in vivo high levels of extracellular ammonium cause DraG to be sequestered to the cell membrane in an AmtB and GlnZ-dependent manner. We now report the reconstitution of a ternary complex involving AmtB, GlnZ and DraG in vitro. Sequestration of a regulatory protein by the membrane-bound AmtB-P(II) complex defines a new regulatory role for Amt proteins in Prokaryotes.

  13. بررسی سودمندی قارچ اندوفیت Piriformospora indica و باکتری Azospirillum Sp. در افزایش تحمل گندم رقم سرداری (Triticum aestivum به تنش شوری

    Directory of Open Access Journals (Sweden)

    سمیه حاجی نیا

    2011-08-01

    Full Text Available شوری آب و خاک یکی از مشکلات جدی و رو به توسعه در سطح جهان است، که سطح وسیعی از اراضی کشور نیز با این مشکل مواجه هستند. کاربرد میکروارگانیسم‌‌های اندوفیت که از مهمترین میکروارگانیسم‌‌های خاک محسوب می‌شوند، در کاهش تنش‌‌‌­های محیطی مانند شوری، به یک راهکار جهانی تبدیل شده است. بنابراین به منظوربررسی اثرات تلقیح قارچ اندوفیت Piriformospora indica و ریزوباکتریهای افزاینده رشد گیاه .Azospirillum sp بر رشد و خصوصیات فیزیولوژیکی گیاه گندم تحت تنش شوری، آزمایشی به صورت فاکتوریل در قالب طرح بلوک‌­های کامل تصادفی با سه تکرار در پاییز سال 1389 در گلخانه تحقیقاتی دانشکده کشاورزی دانشگاه ایلام اجرا گردید. تیمارهای آزمایشی شامل کاربرد پنج سطح میکروارگانیسم‌­های اندوفیت (قارچ پیریفورموسپورا ایندیکا، باکتری آزوسپیریلوم سویه‌های سازگار به مناطق شور و غیر شور، تلقیح توأم قارچ و باکتری و شاهد و چهار سطح شوری آب آبیاری (2/0، 4، 8 و 12 دسی زیمنس بر متر بود. نتایج نشان داد که قارچ پیریفورموسپورا ایندیکا تأثیر مثبت و معنی‌­داری بر رشد، میزان زیتوده تر و خشک اندام‌های هوایی، کلروفیل و محلول‌های اسمولیت در گیاهان گندم تلقیح شده تحت شرایط شور و غیر شور داشت، به طوری که کاربرد قارچ منجر به کاهش اثرات سوء شوری و

  14. Bionota: Bacterias promotoras de crecimiento de microalgas: una nueva aproximación en el tratamiento de aguas residuales Microalgae growth-promoting bacteria: A novel approach in wastewater treatment

    Directory of Open Access Journals (Sweden)

    Bashan Yoav

    2003-12-01

    Full Text Available Las bacterias promotoras de crecimiento en plantas (PGPB del género Azospirillum son conocidas porque mejo­ran el crecimiento de numerosas cosechas agrícolas; sin embargo, el presente trabajo pretende extender el uso de estas bacterias a "bacterias promotoras de crecimiento de microalgas" (MPGB para aumentar la capacidad de las microalgas de eliminar nutrientes de aguas residuales. La inoculación deliberada de las microalgas Chlorella spp. con PGPB de origen terrestre no ha sido reportada con anterioridad, tal vez debido al origen diferente de estos dos microorganismos. Al inmovilizar de manera conjunta Chlorella vulgaris y Azospirillum brasilense Cd en esferas de alginato, se obtuvo como resultado un aumento significativo en varios parámetros de crecimiento de la microalga, como el peso fresco y seco, el número total de células, el tamaño de las colonias de microalgas dentro de la esfera, el número de organismos por colonia y la concentración de pigmentos. Además, aumenta­ron los lípidos y la variedad de ácidos grasos. La microalga combinada con la MGPB tiene una mayor capacidad de eliminar amonio y fósforo tanto en agua residual sintética como en agua residual doméstica. Actualmente se ha estado experimentando con otras PGPB (Flavobacterium sp. Azospirillum sp. y Azotobacter sp. para propósitos acuícolas; por ejemplo aumentar el crecimiento de fitoplancton utilizado en el cultivo de carpas y estabilizar cultivos masivos de microalgas marinas utilizadas como alimento para organismos marinos, todo esto con resul­tados promisorios. Si bien el efecto de las PGPB en microorganismos acuáticos aún no ha sido suficientemente explorado, proponemos que la co-inmovilización de microalgas y bacterias promotoras de crecimiento es un medio efectivo para aumentar la población microalgal y también su capacidad de limpiar aguas residuales. Palabras clave: PGPB; microalgas; biotratamiento de aguas residuales; co

  15. EFECTO DE LA INOCULACIÓN DE Azospirillum halopraeferens Y Bacillus amyloliquefaciens EN LA GERMINACIÓN DE Prosopis chilensis

    OpenAIRE

    2010-01-01

    Las Bacterias Promotoras de Crecimiento de Plantas (BPCP), son microorganismos fijadores de nitrógeno atmosférico y a su vez producen hormonas que aprovechan las plantas para llevar a cabo su desarrollo. Es importante la utilización de microorganismos benéficos para reducir la fertilización química. La investigación científica ha incrementado la busqueda y aislamiento de BPCP de regiones especificas como son las zonas áridas, ya que estos microorganismos estan adaptados a ambientes extremos. ...

  16. Identificação e avaliação de rizobactérias isoladas de raízes de milho

    Directory of Open Access Journals (Sweden)

    Eliamar Aparecida Nascimbém Pedrinho

    2010-12-01

    Full Text Available Estudos sobre a atividade microbiológica que ocorre na rizosfera de diversos vegetais levaram ao descobrimento de grupos de microrganismos importantes para o desenvolvimento vegetal. Dentre eles estão as rizobactérias que são capazes de colonizar as raízes, estimulando-a diretamente ou beneficiando o crescimento e o desenvolvimento de diversas plantas. Estas bactérias são chamadas Rizobactérias Promotoras de Crescimento em Plantas (RPCP. Este trabalho teve o objetivo de isolar, identificar, testar a capacidade da solubilização de fosfato e a produção de ácido indol acético (AIA de bactérias que habitam a rizosfera de plantas de milho. A análise parcial do gene 16S rRNA dos 58 isolados possibilitou a identificação dos gêneros, Bacillus, Burkholderia e Azospirillum, sendo os mais frequentes totalizando 68% dos isolados, seguidos de Sphingomonas, Pseudomonas, Herbaspirillum, Pantoea , Bosea. Desses, 27 apresentaram a capacidade de solubilização do fosfato e 18 foram positivos no teste colorimétrico para detecção de produção do AIA. A partir destes resultados, selecionou-se um organismo pertencente ao gênero Sphingomonas para ser testado em casa de vegetação como promotor de crescimento com as estirpes de Azospirillum brasilense (AbV5 e AbV6. As plantas foram avaliadas quanto à altura aos vinte e setenta dias após a germinação e a massa seca da parte aérea (MSPAe parte radicular (MSPR foi quantificada após setenta dias no encerramento do experimento. Os resultados das análises do isolado pertencente ao gênero Sphingomonas foram estatisticamente semelhantes às estirpes AbV5 e AbV6 na planta de milho indicando que este microrganismo possui potencial para ser utilizado como RPCP

  17. Densidade e diversidade fenotípica de bactérias diazotróficas não simbióticas em solos da Reserva Biológica Serra dos Toledos, Itajubá (MG

    Directory of Open Access Journals (Sweden)

    Talita Filomena Silva

    2011-04-01

    Full Text Available Bactérias diazotróficas não simbióticas (BDNS atuam no desenvolvimento das plantas por meio da fixação biológica de nitrogênio e também pela produção e liberação de substâncias reguladoras do crescimento vegetal. Este estudo objetivou avaliar a densidade e diversidade fenotípica desse grupo de bactérias em fragmentos de solo da Reserva Biológica Serra dos Toledos e entorno, em Itajubá/MG. Essa reserva localiza-se na Área de Proteção Ambiental da Mantiqueira, constituindo-se em uma importante área de recarga e de abrigo à flora e fauna endêmicas. Amostras de solo superficial foram coletadas em áreas com diferentes declividades na reserva, em épocas representativas das estações de inverno (setembro/2006 e verão (abril/2007. A densidade, avaliada pelo número mais provável, utilizando os meios de cultura NFb, JNFb e Fam, para Azospirillum spp., Herbaspirillum spp. e A. amazonense, respectivamente, variou de 0,12 a 75,60 (NMP x 10(5 bactérias g-1 solo seco. Foram obtidos 172 e 174 isolados, respectivamente para as amostras de inverno e verão, dos quais 30 e 55 % apresentaram similaridade igual ou superior a 70 % com as estirpes-tipo Azospirillum brasilense, A. amazonense, A. lipoferum, Herbaspirillum seropedicae e Burkholderia brasilensis. O resultado do comportamento dos isolados com base na tolerância à salinidade nem sempre foi semelhante ao obtido pelas características fenotípicas culturais a 70 % de similaridade, sendo indicado para estudos complementares de diversidade desses organismos. As BDNS apresentam potencial de utilização em estudos de avaliação da qualidade e sustentabilidade de ecossistemas. No entanto, apesar da alta densidade e diversidade fenotípica em solos da reserva, maiores valores foram obtidos no entorno, evidenciando o efeito positivo da cobertura vegetal do tipo gramíneas sobre elas, independentemente da variação climática.

  18. Ecology of nitrogen fixation in soils and rhizospheres. Pt. 5

    Energy Technology Data Exchange (ETDEWEB)

    Werner, D.; Stripf, R.; Abramowski, R.; Fiedler, U.

    1980-12-01

    The effects of reduced oxygen concentration on root growth and activities of enzymes of N-metabolism of wheat (Triticum aestivum var. Kolibri) have been studied, for low O/sub 2/ tensions are required for N/sub 2/ fixation by microaerophilic bacteria (e.g. Azospirillum) associated with root systems of grasses. In hydrocultures with oxygen concentrations in the range of 0.2 to 1 mg O/sub 2/ x 1/sup -1/ compared to aerated cultures (8-9 mg O/sub 2/ x 1/sup -1/) root growth was reduced from 10 mg fresh weight x day/sup -1/ x plant/sup -1/ to one tenth 15 to 30 d after sowing. Specific activity of NADH and NADPH dependent glutamate dehydrogenase (E.C. 1.4.1.2 and 1.4.1.4) is reduced by 50% in the cultures with low oxygen concentrations 20 to 30 days after sowing, whereas specific activity of aspartate aminotransferase (E.C. 2.6.1.1) and alanine amino transferase (E.C. 2.6.1.2) is enhanced by a factor of two to three. Specific activity of glutamine synthetase is almost unaffected. Specific activity of glutamate dehydrogenase is lowest in the root tips, medium in young root hair zone and highest in the old root hair zone, glutamine synthetase activity is reverse in the three zones with differences by a factor of 3-5; aspartate aminotransferase is similarly active in the three zones. Nitrate concentration used (100 ..mu..M) for cultivation of the wheat plants was tested with Azospirillum brasilense in pure culture on agar surfaces exposed to air at the same pH (5.8), used for cultivation of the wheat plants. Activiy after a 14 day period (peak activity 70 mmol C/sub 2/H/sub 4/ x mg protein/sup -1/ x h/sup -1/) was not affected, however 1 mM and 5 mM nitrate added reduced the total activity to 50% and 10% respectively.

  19. CO-INOCULAÇÃO NO DESENVOLVIMENTO VEGETATIVO E NODULAÇÃO DE PLÂNTULAS DE SOJA SUBMETIDAS À CALAGEM, FERTILIZAÇÃO NITROGENADA E APLICAÇÃO DE MICRONUTRIENTES

    Directory of Open Access Journals (Sweden)

    LIBÓRIO, Paloma Helena da Silva

    2015-11-01

    Full Text Available In order to evaluate the co-inoculation of soybeans, or mixed inoculation with bacteria of the genus Bradyrhizobium and Azospirillum brasilense, at the expense of traditional inoculation with Bradyrhizobium, two experiments were installed in october 2014, in a greenhouse. The experimental design was completely randomized with eight treatments and eight replications in pots rotation system. Treatments involving different combinations as commercial fertilizer formulations, with or without inoculant application (s and micronutrients cobalt and molybdenum (Co-Mo in the seeds, being: 0-20-20; 4-20-20; 0-20-20 + inoculant (s; 4-20-20 + inoculant (s; 0-20-20 + inoculant (s + Co-Mo; 4-20-20 + inoculant (s + Co-Mo; 0-20-20 + Co-Mo and 4-20-20 + Co-Mo. Evaluations constituted the% of normal seedlings, number and dry weight of nodules / plant, dry weight of shoot and root / plant. In liming performed with only 30 days before sowing, limestone has not had sufficient time to react with soil, which showed deficiency in calcium, in addition to aluminum and manganese toxicity, reducing the % normal seedlings in the pots, which batch seed was already low in certain physiological quality of germination and emergence tests. Nevertheless, the practice of co-inoculation combined with the formulation (0-20-20 and without Co-Mo in the seeds stood out in relation to the evaluated parameters. Com o objetivo de avaliar a co-inoculação de soja, ou seja, inoculação mista com bactérias do gênero Bradyrhizobium e Azospirillum brasilense, em detrimento da inoculação tradicional com Bradyrhizobium foram instalados dois experimentos em outubro de 2014, em casa de vegetação. O delineamento experimental utilizado foi o inteiramente casualizado com oito tratamentos e oito repetições sob rodízio diário dos vasos. Os tratamentos envolveram diferentes combinações quanto a formulações comerciais de adubo, com ou sem aplicação de inoculante (s e micronutrientes

  20. Inoculation two azotobacter enhancing osmotic stress resistance and growth in wheat seedling%接种两种固氮菌增强小麦幼苗抗渗透胁迫及生长能力

    Institute of Scientific and Technical Information of China (English)

    刘华伟; 林晓军; 孙超; 李强; 杨呼; 郭蔼光

    2013-01-01

    Aims The seedling stage is the key stage of matter and energy accumulation in the wheat life cycle. Therefore, drought during the seedling stage affects population formation in late stages. In this study, wheat seedlings were inoculated with azotobacters Azorhizobium caulinodans 'ORS571' and Azospirillum brasilense 'Yu62'. Methods Wheat seedlings germination was screened in normal conditions and with PEG 6000 osmotic stress using seedlings inoculated with azotobacters. Root volume, relative water content (RWC), proline content and soluble protein content of seedling laminas were determined under PEG drought stress using seedlings inoculated with azotobacters on laminas. Important findings The germination rate of wheat seedlings was significantly increased under drought stress when inoculated with azotobacters. Moreover, wheat seedlings inoculated with mixed azotobacters have more obvious growth promotion than when inoculated with a single azotobacter. The former laminas proline content, relative water content, proline content and soluble protein content had increased. The results showed that drought resistance of wheat seedlings was improved when inoculated with mixed azotobacters, which provided the foundation for further study of azotobacter-wheat interaction under drought stress.%苗期是小麦(Triticum aestivum)物质和能量积累的关键时期,苗期干旱影响小麦的后期群体建成.利用田菁茎瘤固氮根瘤菌(Azorhizobium caulinodans)‘ORS571’与巴西固氮螺菌(Azospirillum brasilense)‘Yu62’浸种侵染小麦和聚乙二醇(PEG)模拟渗透胁迫,研究渗透胁迫下接菌小麦种子的发芽状况;利用固氮菌涂抹小麦幼苗叶部,测定PEG模拟渗透胁迫下小麦幼苗根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量,探究固氮菌增强小麦幼苗抗渗透胁迫的能力.结果表明,接种混合固氮菌后在渗透胁迫下小麦种子的发芽率明显提高;在渗透胁迫下叶部

  1. Influencia de lo abono nitrogenado en la calidad de ensilaje de maíz Influência da adubação nitrogenada sobre a qualidade da silagem de milho

    Directory of Open Access Journals (Sweden)

    Simone Basi

    2012-06-01

    Full Text Available

    La calidad final de ensilaje de maíz está directamente relacionada con el estado nutricional de las plantas. De los nutrientes requeridos por el cultivo, el nitrógeno (N es considerado el más importante. Esta revisión tiene como objetivo discutir temas relacionados con la dinámica de este nutriente en la planta de maíz, con énfasis en su influencia en los diversos eventos fisiológicos en los que participa. La producción de materia seca depende de la fuente y forma de suministro de este elemento a la planta. El uso de Azospirillum brasilense ha tenido prominencia a través de la fijación biológica de nitrógeno mediante la reducción de los costes de producción para la economía en el uso de N mineral, así como con el incremento de producción debido al uso más regular de lo nutriente en relación con los fertilizantes minerales, asegurando así una mayor productividad y calidad del material. Con un entendimiento de las funciones de este importante nutriente a la planta, el suministro alternativo y técnicas de gestión puede obtener un mayor valor nutritivo del ensilaje, debido al aumento de la concentración de proteína en la planta. Además con el aumento de la producción de materia seca por unidad de área, aumento de la producción de grano, mayor valor energético, influye en la concentración de fibras del material ensilado y posiblemente en el rendimiento de los animales.

    A qualidade final da silagem de milho está diretamente relacionada ao estado nutricional das plantas. Dos nutrientes exigidos pela cultura, o nitrogênio (N é considerado o de maior importância. A presente revisão tem como objetivo discutir aspectos relacionados à dinâmica deste nutriente na planta de milho, dando ênfase em sua influência nos diversos eventos fisiológicos dos quais participa. A produção de matéria seca depende da fonte e da forma de fornecimento deste elemento à planta. O uso

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

    Directory of Open Access Journals (Sweden)

    FÁBIO BUENO DOS REIS JUNIOR

    2000-05-01

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

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

    Directory of Open Access Journals (Sweden)

    José I. Baldani

    2005-09-01

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

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

    Directory of Open Access Journals (Sweden)

    Vera Lúcia Divan Baldani

    2010-01-01

    Full Text Available O estudo consistiu em avaliar a sobrevivência e a eficiência das bactérias diazotróficas Herbaspirillum seropedicae, Burkholderia sp. e Azospirillum brasilense, inoculadas em dois tipos de turfas. A sobrevivência das bactérias foi determinada pelo método do NúmeroMais Provável (NMP, durante o armazenamento de até seis meses. A eficiência foi avaliada pela contribuição destas bactérias nos parâmetros agronômicos de acúmulo de massa seca e N-total dos grãos nas variedades de arroz IR42 e IAC4440. O experimento foi conduzido emvasos com solo, em delineamento inteiramente casualizado com três repetições, em arranjo fatorial 2 x 4. Os dados obtidos mostraram que o inoculante contendo a estirpe de Burkholderia sp. manteve o número de células viáveis em torno de 108 cel. g-1 de turfa no período testado, enquanto nos demais inoculantes este número ficou em torno de 106. Os ensaios de inoculação não mostraram diferenças significativas quanto à origem das turfas usadas como veículo sobre os parâmetros agronômicos e sobrevivência das bactérias. O tratamento que continha H. seropedicae manteve o acúmulo de massa seca e N-total dos grãos estatisticamente igual ao tratamento que recebeu 40 kg N ha-1, embora tenham sido observados aumentos de 13 e 19,4% nestes parâmetros na variedade IAC4440. Os resultados mostraram-se promissores quanto à utilização da prática de inoculação na cultura do arroz. The study consisted of a comparison of two-peat materials (Brazil and Canada containing different C-total content. The peats were inoculated with strains of Herbaspirillum seropedicae ZAE 94, Burkholderia sp. M130, and Azospirillum brasilense Sp109, and monitored during a period of six months in relation to variation on humidity and survival of bacteria in the inoculant. The quantification of viable cells in the inoculant was measured by the Most Probable Number (MPN method. The rice seeds were pelleted with the respective

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

    Directory of Open Access Journals (Sweden)

    Valéria Marino Rodrigues Sala

    2007-06-01

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

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

    Directory of Open Access Journals (Sweden)

    Salwani S.

    2012-06-01

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

  7. Manejo agronómico integral de sustratos, métodos de siembra y biofertilización en la producción sostenible de tubérculos-semilla de papa por semilla sexual

    Directory of Open Access Journals (Sweden)

    A. Hernández

    2001-01-01

    Full Text Available Con el objetivo de producir tubérculos-semilla de papa a partir de semilla sexual (SSP de forma ecológica y sostenible, se llevaron a cabo entre los años 1994-1998 tres experimentos en el Instituto Nacional de Ciencias Agrícolas (INCA, en el municipio de San José de las Lajas, La Habana, sobre un suelo Ferrálitico Rojo compactado, donde se estudiaron dos métodos de siembra (directa y transplante, el uso de dos fuentes de materia orgánica; dosis de aplicación de la materia orgánica en la composición del sustrato y la combinación de losmejores resultados de estos experimentos, con la inoculación de las semillas con biofertilizantes (bacterias nitrofijadoras, fosfosolubilizadoras y hongos micorrizógenos, utilizando pulso de nitrógeno a razón de 60 kg de N. ha-1, combinando todos estos en un experimento. Las bacterias empleadas fueron Azospirillum brasilense, Pseudomonas cepacia, Pseudomonas fluorescens y la micorriza empleada fue Glomus fasciculatum.Los resultados finales arrojaron que las plantas donde se había utilizado la coinoculación micorriza + Pseudomonascepacia, tuvieron una mayor producción de tubérculos semilla así como el tratamiento inoculado con Pseudomonas fluorescens, siendo los resultados superiores a los del testigo con fertilización química, consiguiéndose con ello una tecnologíade producción más económica y ecológica en este cultivo.

  8. Instrumental analysis of bacterial cells using vibrational and emission Moessbauer spectroscopic techniques

    Energy Technology Data Exchange (ETDEWEB)

    Kamnev, Alexander A. [Laboratory of Biochemistry of Plant-Bacterial Symbioses, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 410049 Saratov (Russian Federation)]. E-mail: aakamnev@ibppm.sgu.ru; Tugarova, Anna V. [Laboratory of Biochemistry of Plant-Bacterial Symbioses, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 410049 Saratov (Russian Federation); Antonyuk, Lyudmila P. [Laboratory of Biochemistry of Plant-Bacterial Symbioses, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 410049 Saratov (Russian Federation); Tarantilis, Petros A. [Laboratory of Chemistry, Department of Science, Agricultural University of Athens, 11855 Athens (Greece); Kulikov, Leonid A. [Laboratory of Nuclear Chemistry Techniques, Department of Radiochemistry, Faculty of Chemistry, M.V. Lomonosov Moscow State University, 119992 Moscow (Russian Federation); Perfiliev, Yurii D. [Laboratory of Nuclear Chemistry Techniques, Department of Radiochemistry, Faculty of Chemistry, M.V. Lomonosov Moscow State University, 119992 Moscow (Russian Federation); Polissiou, Moschos G. [Laboratory of Chemistry, Department of Science, Agricultural University of Athens, 11855 Athens (Greece); Gardiner, Philip H.E. [Division of Chemistry, School of Science and Mathematics, Sheffield Hallam University, Sheffield S1 1WB (United Kingdom)

    2006-07-28

    In biosciences and biotechnology, the expanding application of physicochemical approaches using modern instrumental techniques is an efficient strategy to obtain valuable and often unique information at the molecular level. In this work, we applied a combination of vibrational (Fourier transform infrared (FTIR), FT-Raman) spectroscopic techniques, useful in overall structural and compositional analysis of bacterial cells of the rhizobacterium Azospirillum brasilense, with {sup 57}Co emission Moessbauer spectroscopy (EMS) used for sensitive monitoring of metal binding and further transformations in live bacterial cells. The information obtained, together with ICP-MS analyses for metals taken up by the bacteria, is useful in analysing the impact of the environmental conditions (heavy metal stress) on the bacterial metabolism and some differences in the heavy metal stress-induced behaviour of non-endophytic (Sp7) and facultatively endophytic (Sp245) strains. The results show that, while both strains Sp7 and Sp245 take up noticeable and comparable amounts of heavy metals from the medium (0.12 and 0.13 mg Co, 0.48 and 0.44 mg Cu or 4.2 and 2.1 mg Zn per gram of dry biomass, respectively, at a metal concentration of 0.2 mM in the medium), their metabolic responses differ essentially. Whereas for strain Sp7 the FTIR measurements showed significant accumulation of polyhydroxyalkanoates as storage materials involved in stress endurance, strain Sp245 did not show any major changes in cellular composition. Nevertheless, EMS measurements showed rapid binding of cobalt(II) by live bacterial cells (chemically similar to metal binding by dead bacteria) and its further transformation in the live cells within an hour.

  9. Microbial consortium role in processing liquid waste of vegetables in Keputran Market Surabaya as organic liquid fertilizer ferti-plus

    Science.gov (United States)

    Rizqi, Fauziah; Supriyanto, Agus; Lestari, Intan; Lita Indri D., L.; Elmi Irmayanti, A.; Rahmaniyah, Fadilatur

    2016-03-01

    Many activities in this market is directly proportional to increase production of vegetables waste, especially surabaya. Therefore, in this study aims to utilize liquid waste of vegetables into liquid organic fertilizer by mixing microbial consorsium. The microbial consorsium consist of Azotobacter chrococcum, Azospirillum brasilense, Rhizobium leguminosarum, Bacillus subtilis, Bacillus megaterium, Pseudomonas putida, and Pseudomonas fluorescens. Ttreatment of microbial concentrations (5%, 10%, 15%) and the length of the incubation period (7 days, 14 days, 21 days) used in this research. The parameters used are: C/N ratio, levels of CNP, and BOD value. This study uses a standard organic fertilizer value according SNI19-7030-2004, The results show the value of C/N ratio comply with the ISO standards. C levels showed an increase during the incubation period but not compare with standards. N levels that compare with standards are microbial treatment in all group concentration except control group with an incubation period of 21 days is > 7. P levels compare with the existing standards in the group of microbe concentration of 10% and 15% during the incubation period. The value of the initial BOD liquid waste of vegetable is 790.25 mg / L, this value indicates that the waste should not go into the water body. Accordingly, the results of this study can not be used as a liquid organic fertilizer, but potentially if it is used as a natural career or build natural soil. The Building natural soil is defined as the natural ingredients that can be used to improve soil properties.

  10. Identification and Quantification of Cause and Effects in Symbiosis of Corn with Arbuscular Mycorrehiza Fungus using Structural Equation Modeling Approach

    Directory of Open Access Journals (Sweden)

    M Jahan

    2015-09-01

    Full Text Available Considering to necessity and importance of determining mycorrhizal symbiosis level and its effects on crop agroecological characteristics as well as other related factors affecting this relationship, beside, the lack of an assured method for this purpose, the present study designed based on new perspectives and insights for determining cause and effects relations, latent variables using structural equation modeling (SEM approach. A field experiment was conducted during 2 years. A split plots arrangement based on randomized complete block design (RCBD with three replications was used. Treatments consisted four cropping systems (high, medium, and low input conventional as well as ecological system and four inoculations (mycorrhizae fungus: Glomus intraradices, plant growth promoting rhizobacteria (PGPR: (Azospirillum brasilense, Azotobacter paspali, dual inoculation: (fungus plus bacteria, and no-inoculation (control, which were allocated to main plots and sub plots, respectively. At the first step, a confirmatory factor analyzing was conducted resulted to two distinguished factors, and then the variables which had the most loads (weight on one of these two factors were determined. At the second step, considering ecophysiological basis of crops growth and development, in order to continue analyzing, the first factor (including: leaf area index (LAI, root length colonization percent (RLCP, dry matter (DM, stem diameter (D, SPAD readings and the second factor (including: maximum photosynthesis (Amax, specific root length (SRL, canopy temperature (CT, plant height (H, soil respiration rate (SRR, variable chlorophyll fluorescence to maximum chlorophyll fluorescence (Fv/Fm, plant tissue phosphorus content (%P determined resource capture latent construct and resource utilization latent construct, respectively. Correlation coefficients, squared multiple correlation coefficients, covariance matrices, direct and indirect path coefficients were calculated

  11. Isolation and identification of phosphate solubilizer Azospirillum, Bacillus and Enterobacter strains by 16SrRNA sequence analysis and their effect on growth of wheat (Triticum aestivum L.)

    NARCIS (Netherlands)

    Tahir, M.; Mirza, M.S.; Zaheer, A.; Rocha Dimitrov, M.; Smidt, H.; Hameed, S.

    2013-01-01

    The aim of the present study was to isolate phosphate solubilizing bacteria from wheat rhizosphere and investigate their potential for plant growth promotion. Three phosphate solubilizing bacterial strains were isolated by serial dilution method from the rhizosphere of wheat grown under wheat-cotton

  12. El papel de Azospirillum y hongos micorrízicos arbusculares en la producción de jitomate y su mecanismo de acción /

    OpenAIRE

    Esquivel Cote, Rosalba

    2011-01-01

     tesis que para obtener el grado de Doctor en Ciencias Biológicas, presenta Rosalba Esquivel Cote ; asesor María del Pilar Huante Pérez, María del Rocío Cruz Ortega, Ronald Ferrera-Cerrato. 162 páginas : diagramas, fotografías. Doctorado en Ciencias Biológicas UNAM, Instituto de Ecología, 2011

  13. 用酵母双杂交系统筛选与NifA相互作用的蛋白质

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 管乐; 涂然; 孙文改; 李季伦

    2005-01-01

    利用酵母双杂合系统从巴西固氮螺菌(Azospirillum brasilense)Sp中筛选与NifA有直接相互作用的蛋白质因子. 首先将nifA基因克隆到pGBD-C2载体上, 构建成诱饵质粒pGBD-nifA, 接着将巴西固氮螺菌的染色体DNA构建在pGAD-C1, pGAD-C2和pGAD-C3质粒载体上, 形成3个质粒文库YSPC1, YSPC2和YSPC3. 然后以pGBD-nifA为诱饵, 对YSPC1, YSPC2和YSPC3这3个文库进行了筛选, 得到11个阳性克隆. DNA序列分析表明, 这11个阳性克隆分为4类, 包含4个1~1.3 kb不同的基因片段. 对这4个基因片段进行同源性比较发现, 其中2个编码的蛋白质含有与信号传递有关的PAS结构域, 另外1个基因片段含有与吸收铁有关的fhuE基因, 另一个是未知蛋白. 将这4个基因片段从原来的pGAD载体转移到pGBD载体, 即互换换载体后, 它们仍然与NifA有相互作用; 对这4个基因片段进行移码突变则不再与NifA有相互作用, 说明了它们与NifA相互作用的特异性.

  14. Effect of azotobacter,slurry and their combination on economic characters and yield of maize%菌剂、沼液及其复配对玉米经济性状和产量的影响

    Institute of Scientific and Technical Information of China (English)

    贺国强; 邓志平; 刘展志; 谢剑波; 李朋飞; 董仁杰; 庞昌乐; 陈三凤

    2011-01-01

    本试验旨在研究菌剂、沼液及其复配对玉米经济性状和产量的影响。用固氮芽孢杆菌W6、巴西固氮螺菌Yu62及固氮巨大芽孢杆菌C4这3种固氮菌分别接种,并与沼液复配后接种玉米,以及这3种固氮菌与解磷假单胞菌S20组成的的4种微生物复合菌剂单独或与沼液复配后接种玉米。结果表明:除百粒重外,单菌剂处理组、沼液处理组及二者复配处理组均较清水组在玉米经济性状及单产方面表现优良。各单菌剂处理组中,固氮巨大芽孢杆菌C4效果较佳,相对于清水处理组,干穗单产增产10.47%。沼液与菌剂配合使用处理玉米的效果要优于相应菌剂单独处理%The aim of this research was to study the effect of microbes,biogas slurry and their combination on the economic characters and yield of maize.The nitrogen-fixing bacteria Bacillus megaterium C4,Bacillus sp.W6,Azospirillum brasilense Yu62,the combination of nitrogen-fixing bacteria with Pseudomonas alealigenes S20 and the combination of bacteria with the biogas slurry were applied to maize in the field.Experimental results showed that the treatments of either single bacterium,biogas slurry or their combination gave a more excellent performance than water treatment in terms of maize yield and economic characteristics except kernel weight,Bacteria treatment groups,especially with C4,gave a 10% higher dry ear yield compared to the water treatment group.The combination of biogas slurry with these bacteria was superior to individual use on maize.In particular,the combined biogas slurry with microbial agent C4 had increases of 2.17% kernel weight,25.15% dry ear yield and 12.47% grain yield comparing to the water treatment.

  15. A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control.

    Science.gov (United States)

    Ouyang, Yiwen; Duarte, Gabriela R M; Poe, Brian L; Riehl, Paul S; dos Santos, Fernando M; Martin-Didonet, Claudia C G; Carrilho, Emanuel; Landers, James P

    2015-12-11

    Infrared (IR)-mediated thermal cycling system, a method proven to be a effective for sub-μL scale polymerase chain reaction (PCR) on microchips, has been integrated with DNA extraction and separation on a glass microchip in a fully integrated micro Total Analysis System by Easley et al., in 2006. IR-PCR has been demonstrated on both glass and PMMA microdevices where the fabrication (bonding) is not trivial. Polyester-toner (PeT) microfluidic devices have significant potential as cost-effective, disposable microdevices as a result of the ease of fabrication (∼$0.25 USD and <10 min per device) and availability of commercial substrates. For the first time, we demonstrate here the thermal cycling in PeT microchips on the IR-PCR system. Undesirable IR absorption by the black-toner bonding layer was eliminated with a spatial filter in the form of an aluminum foil mask. The solution heating rate for a black PeT microchip using a tungsten lamp was 10.1 ± 0.7 °C s(-1) with a cooling rate of roughly -12 ± 0.9 °C s(-1) assisted by forced air cooling. Dynamic surface passivation strategies allowed the successful amplification of a 520 bp fragment of the λ-phage genome (in 11 min) and a 1500 bp region of Azospirillum brasilense. Using a centrosymmetric chamber configuration in a multichamber PeT microchip, homogenous temperature distribution over all chambers was achieved with inter-chamber temperature differences at annealing, extension and denaturing steps of less than ±2 °C. The effectiveness of the multichamber system was demonstrated with the simultaneous amplification of a 390 bp amplicon of human β-globin gene in five PeT PCR microchambers. The relative PCR amplification efficiency with a human β-globin DNA fragment ranged from 70% to 90%, in comparison to conventional thermal cyclers, with an inter-chamber standard deviation of ∼10%. Development of PeT microchips for IR-PCR has the potential to provide rapid, low-volume amplification while

  16. Cloning and Functional Analysis of glnB from Azospirilum brasilense Yu62%巴西固氮螺菌Yu62 glnB基因的克隆及其功能分析

    Institute of Scientific and Technical Information of China (English)

    李周华; 陈三凤; 李季伦

    2001-01-01

    通过原位杂交从巴西固氮螺菌Yu62的基因文库中,筛选到glnB基因的阳性克隆,将3.7kb/EcoRI+PstI的阳性克隆亚克隆到pUC19中,进行了全序列分析,其在GenBank中的登记号是AF323960。DNA序列分析表明该阳性克隆含有完整的glnB基因,glnB基因下游是编码谷氨酰胺合成酶(GS)的glnA基因,glnB基因上游是一个编码未知蛋白的ORF。glnB基因编码区长336bp,编码112个氨基酸,与肺炎克氏杆菌、大豆慢生根瘤菌、豌豆根瘤菌及大肠杆菌在氨基酸顺序的同源性分别高达71%、77%、79%和69%。将卡那霉素抗性片段(Km-cas-sette)插入glnB基因的BglⅡ位点,通过三亲杂交法将其引入到巴西固氮螺菌Yu62中,通过同源重组,获得GlnB-突变株(glnB::Km)。为了进一步分析glnB基因的功能,将glnB基因的编码区(339bp)构建在pVK100中,置于Km启动子下组成型表达,形成重组质粒pVK-Ⅱ。将重组质粒pVK-Ⅱ转入到GlnB-突变株,构建成互补株C-glnB(glnB::Km/glnB)。对GlnB-突变株和互补株的固氮酶活性和生长性能的测定表明,GlnB-突变体无固氮酶活性,即表型 为Nif-;而互补株像野生型菌株一样具有固氮酶活性。突变株、互补株及野生型在菌落生长速度上基本相同。将含有glnB基因的重组质粒pVK-Ⅱ分别转移到野生型Yu62菌株和具有一定抗铵能力的DraT-突变株中,使glnB基因的拷贝数增加,并进一步比较它们的固氮酶活性,结果表明多拷贝的glnB基因能显著提高固氮酶活性。

  17. Modificacion del Valor Nutrimental de Acidos Grasos en Semilla de la Halofita Salicornia bigelovii Torr. con la Interaccion de Bacterias Promotoras del Crecimiento de Plantas (Azospirillum halopraeferens y Klebsiella pneumoniae)

    OpenAIRE

    Edgar Omar Rueda Puente; Enrique Troyo Dieguez; Thelma Castellanos Cervantes; Bernardo Murillo-Amador; Bertha Olivia Arredondo Vega; Jose Luis Díaz de Leon Alvarez; Mario Antonio Tarazon Herrera

    2005-01-01

    Una especie vegetal a gran escala se presenta como una alternativa para la producción de ácidos grasos esenciales en nutrición humana por lo que su composición de ácidos grasos cobra especial importancia. Salicornia bigelovii una halófita que se desarrolla en ambientes áridos y costeros como las del estado de Sonora y de la península de Baja California Sur, México, presenta ser una especie alternativa en la producción de aceites comestibles. La fijación de N2 por bacterias asociadas con raíce...

  18. A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control

    Energy Technology Data Exchange (ETDEWEB)

    Ouyang, Yiwen [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Duarte, Gabriela R.M. [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Universidade Federal de Goiás, Goiânia, GO 74690-900 (Brazil); Poe, Brian L.; Riehl, Paul S. [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Santos, Fernando M. dos; Martin-Didonet, Claudia C.G. [Universidade Estadual de Goiás, Anápolis, GO 75132-400 (Brazil); Carrilho, Emanuel [Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP 13566-590 (Brazil); Instituto Nacional de Ciência e Tecnologia de Bioanalítica, CP 6154, Campinas, SP 13083-970 (Brazil); Landers, James P., E-mail: landers@virginia.edu [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Department of Mechanical Engineering, University of Virginia, Charlottesville, VA 22904 (United States); Department of Pathology, University of Virginia Health Science Center, Charlottesville, VA (United States)

    2015-12-11

    Infrared (IR)-mediated thermal cycling system, a method proven to be a effective for sub-μL scale polymerase chain reaction (PCR) on microchips, has been integrated with DNA extraction and separation on a glass microchip in a fully integrated micro Total Analysis System by Easley et al., in 2006. IR-PCR has been demonstrated on both glass and PMMA microdevices where the fabrication (bonding) is not trivial. Polyester-toner (PeT) microfluidic devices have significant potential as cost-effective, disposable microdevices as a result of the ease of fabrication (∼$0.25 USD and <10 min per device) and availability of commercial substrates. For the first time, we demonstrate here the thermal cycling in PeT microchips on the IR-PCR system. Undesirable IR absorption by the black-toner bonding layer was eliminated with a spatial filter in the form of an aluminum foil mask. The solution heating rate for a black PeT microchip using a tungsten lamp was 10.1 ± 0.7 °C s{sup −1} with a cooling rate of roughly −12 ± 0.9 °C s{sup −1} assisted by forced air cooling. Dynamic surface passivation strategies allowed the successful amplification of a 520 bp fragment of the λ-phage genome (in 11 min) and a 1500 bp region of Azospirillum brasilense. Using a centrosymmetric chamber configuration in a multichamber PeT microchip, homogenous temperature distribution over all chambers was achieved with inter-chamber temperature differences at annealing, extension and denaturing steps of less than ±2 °C. The effectiveness of the multichamber system was demonstrated with the simultaneous amplification of a 390 bp amplicon of human β-globin gene in five PeT PCR microchambers. The relative PCR amplification efficiency with a human β-globin DNA fragment ranged from 70% to 90%, in comparison to conventional thermal cyclers, with an inter-chamber standard deviation of ∼10%. Development of PeT microchips for IR-PCR has the potential to provide rapid, low

  19. Estimation of the efficiency of hydrocarbon mineralization in soil by measuring CO2-emission and variations in the isotope composition of carbon dioxide

    Science.gov (United States)

    Dubrovskaya, Ekaterina; Turkovskaya, Olga

    2010-05-01

    Estimation of the efficiency of hydrocarbon mineralization in soil by measuring CO2-emission and variations in the isotope composition of carbon dioxide E. Dubrovskaya1, O. Turkovskaya1, A. Tiunov2, N. Pozdnyakova1, A. Muratova1 1 - Institute of Biochemistry and Physiology of Plants and Microorganisms, RAS, Saratov, 2 - A.N. Severtsov Institute of Ecology and Evolution, RAS, Moscow, Russian Federation Hydrocarbon mineralization in soil undergoing phytoremediation was investigated in a laboratory experiment by estimating the variation in the 13С/12С ratio in the respired СО2. Hexadecane (HD) was used as a model hydrocarbon pollutant. The polluted soil was planted with winter rye (Secale cereale) inoculated with Azospirillum brasilense strain SR80, which combines the abilities to promote plant growth and to degrade oil hydrocarbon. Each vegetated treatment was accompanied with a corresponding nonvegetated one, and uncontaminated treatments were used as controls. Emission of carbon dioxide, its isotopic composition, and the residual concentration of HD in the soil were examined after two and four weeks. At the beginning of the experiment, the CO2-emission level was higher in the uncontaminated than in the contaminated soil. After two weeks, the quantity of emitted carbon dioxide decreased by about three times and did not change significantly in all uncontaminated treatments. The presence of HD in the soil initially increased CO2 emission, but later the respiration was reduced. During the first two weeks, nonvegetated soil had the highest CO2-emission level. Subsequently, the maximum increase in respiration was recorded in the vegetated contaminated treatments. The isotope composition of plant material determines the isotope composition of soil. The soil used in our experiment had an isotopic signature typical of soils formed by C3 plants (δ13C,-22.4‰). Generally, there was no significant fractionation of the carbon isotopes of the substrates metabolized by the

  20. Alterations in seedling vigour and antioxidant enzyme activities in Catharanthus roseus under seed priming with native diazotrophs

    Institute of Scientific and Technical Information of China (English)

    KARTHIKEYAN B.; JALEEL C.A.; GOPI R.; DEIVEEKASUNDARAM M.

    2007-01-01

    An experiment was conducted on Catharanthus roseus to study the effect of seed treatments with native diazotrophs on its seedling growth and antioxidant enzyme activities. The treatments had significant influence on various seedling parameters.There is no significant influence on dry matter production with the diazotrophs, Azospirillum and Azotobacter. However, the vital seedling parameters such as germination percentage and vigour index were improved. Azotobacter treatment influenced maximum of 50% germination, whereas Azospirillum and Azotobacter were on par with C. roseus with respect to their vigour index. There was significant difference in the population of total diazotrophs. Azospirillum and Azotobacter between rhizosphere and non-rhizosphere soils of C. roseus had the same trend and were observed at various locations of the study. The activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) were increased to a significant extent due to the treatment with diazotrophs.

  1. Effects of Vermi-compost and Two Bacterial Bio-fertilizers on some Quality Parameters of Petunia

    Directory of Open Access Journals (Sweden)

    Mina Zarghami MOGHADAM

    2013-05-01

    Full Text Available The present research was conducted to study the effect of vermi-compost and two bio-fertilizer applications on growth, yield and quality of petunia (Petunia hybrida. The experiment laid out in randomized block design with 3 replications and 9 treatment combinations composing of vermi-compost, bio-fertilizers and NPK fertilizer. The treatment receiving Azospirillum sp. + Phosphate solubilizing bacterium + Vermicompost + NPK (25% of recommended dose recorded the highest plant height, number of branches, plant spread, leaf area index, dry matter accumulation and yield attributes such as number of flowers per plant, number of flowers per plot, flower yield/plant, flower yield/plot. The early flower bud initiation, 50 percent flowering and more flowering duration was achieved in the treatment receiving Azospirillum sp. + Phosphate solubilizing bacterium + Vermicompost + NPK (25% of recommended dose. Application of Azospirillum sp. + Phosphate solubilizing bacterium + Vermicompost + NPK (25% of recommended dose registered significantly higher quality parameters such as flower diameter.

  2. Differential growth stimulation response of potato towards inoculation with taxonomically diverse plant growth promoting rhizobacteria

    Directory of Open Access Journals (Sweden)

    Tahir eNaqqash

    2016-02-01

    Full Text Available Rhizosphere engineering with beneficial plant growth promoting bacteria offers great promise for sustainable crop yield. Potato is an important food commodity that needs large inputs of nitrogen and phosphorus fertilizers. To overcome high fertilizer demand (especially nitrogen, five bacteria, i.e. Azospirillum sp.TN10, Agrobacterium sp.TN14, Pseudomonas sp.TN36, Enterobactersp. TN38 and Rhizobium sp. TN42 were isolated from the potato rhizosphere on nitrogen-free malate medium and identified based on their 16S rRNA gene sequences. Three strains, i.e. TN10, TN38 and TN42, showed nitrogen fixation (92.67-134.54 nmol h-1mg-1protein, while all showed the production of indole acetic acid in the presence and/or absence of L-tryptophan. Azospirillum sp. TN10 produced the highest amount of IAA, as measured by spectrophotometry (312.14 µg mL-1 and HPLC (18.3 µg mL-1. Inoculation with these bacteria under axenic conditions resulted in differential growth responses of potato. Azospirillum sp. TN10 incited the highest increase in potato fresh and dry weight over control plants, along with increased N contents of shoot and roots. All strains were able to colonize and maintain their population densities in the potato rhizosphere for upto 60 days, with Azospirillum sp. and Rhizobium sp. showing the highest survival. Plant root colonization potential was analyzed by transmission electron microscopy of root sections inoculated with Azospirillum sp. TN10. Of the five test strains, Azospirillum sp. TN10 has the greatest potential to increase the growth and nitrogen uptake of potato. Hence, it is suggested as a good candidate for the production of potato biofertilizer for integrated nutrient management with potato.

  3. AcEST: DK949478 [AcEST

    Lifescience Database Archive (English)

    Full Text Available amine/glycine ligase OS=Met... 63 6e-09 tr|Q07HQ2|Q07HQ2_RHOP5 Phosphoribosylamine--glycine ligase OS=Rh... ...ment) OS=Azospirillum brasilens... 64 5e-09 tr|B8IDK0|B8IDK0_METNO Phosphoribosyl

  4. INTEGRATED NUTRIENT MANAGEMENT IN SUNFLOWER +APIARY FARMING SYSTEM FOR SUSTAINABLE PRODUCTIVITY

    Directory of Open Access Journals (Sweden)

    G .SUMALATHA AND T. GEETHA JEBARATHNAM

    2014-10-01

    Full Text Available ABSTRACT: Two sets of field experiments were conducted during  June-August months of 2012-2013 at Annamalai University of Cuddalore district under irrigated conditions to formulate site –specific nutrient management and to assess the yield  performance of sunflower +apiary farming system.The experiment consists of two main treatments; i.e, sunflower alone (M1 ,sunflower  with apiary (M2 and nine sub treatments i.e, control (S1, 100%RDF (S2, 100%RDF + Azospirillum (S3,100% RDF+phosphobacteria (S4,100%RDF+vermicompost (S5, 100%RDF+ Azospirillum+ phosphobacteria (S6, 100% RDF + Vermicompost +Azospirillum (S7, 100%RDF+ vermicompost+phosphobacteria (S8,100%RDF + Vermicompost + Azospirillum+phosphobacteria(S9 and it was laid out in split plot design with three replications .Highest values for seed yield and NPK uptake were observed for the combined application of all three nutrient sources in the plots of the experiment where beehive was installed. Perhaps to ensure better outcome. The apiary should be provided with better feeding for a longer period of time, beside  the natural feeding resources .feed consists of 500ml sugar syrup two times a week. Infact the bee foraging activites, improved seed yield, apart from the income through honey production, thus sustaining the crop + apiary system.

  5. Deep-sea fungi as a source of alkaline and cold-tolerant proteases

    Digital Repository Service at National Institute of Oceanography (India)

    Damare, S.; Raghukumar, C.; Muraleedharan, U.; Raghukumar, S.

    and Chang CS. Bleach resistant alkaline protease produced by a Bacillus sp. Isolated from the Korean polychaete, Periserrula leucophryna. Proc Biochem 2004;39:1441-7. [12] Turkiewicz M, Gromek E, Kalinowska H and Zielinska M, Biosynthesis and properties...-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis. Enzyme Microb Technol 2003;32:294-304. [21] Oh KH, Seong CS, Lee SW, Kwon OS and Park YS. Isolation of a psychrotrophic Azospirillum sp. and characterizaion of its extracellular...

  6. Enriching vermicompost by nitrogen fixing and phosphate solubilizing bacteria.

    Science.gov (United States)

    Kumar, V; Singh, K P

    2001-01-01

    The effect of inoculation of vermicompost with nitrogen-fixing Azotobacter chroococcum strains, Azospirillum lipoferum and the phosphate solubilizing Pseudomonas striata on N and P contents of the vermicompost was assessed. Inoculation of N2 fixing bacteria into vermicompost increased contents of N and P. Enriching vermicompost with rock phosphate improved significantly the available P when inoculated with P. striata. During the incubation period, the inoculated bacterial strains proliferated rapidly, fixed N and solubilized added and native phosphate.

  7. Microorganismos rizosféricos, potenciales antagonistas de Fusarium sp. causante de la pudrición radicular de maracuyá (Passiflora edulis Sims)

    OpenAIRE

    2012-01-01

    El cultivo de maracuyá (Passiflora edulis), de gran importancia económica para Colombia, actualmente es afectado por la enfermedad del marchitamiento vascular causado por Fusarium sp. lo que hace necesario la búsqueda de alternativas que permitan un control eficiente de esta enfermedad. Aislados de las bacterias Azotobacter spp., Azospirillum spp. y el hongo Trichoderma spp., fueron evaluados como potenciales biocontroladores de Fusarium sp. en pruebas in vitro e in vivo. Las pruebas de “test...

  8. The effect of salt tolerant nitrogen fixing bacteria on the growth of paddy rice (Oryza sativa. L

    Directory of Open Access Journals (Sweden)

    Suliasih Suliasih

    2014-06-01

    Full Text Available Nitrogen fixing bacteria (Azospirillum and Azotobacter were isolated from coastal mangrove in Pulau seribu. The aims of the experiment was to find out isolates of nitrogen fixing bacteria which were tolerant to high salinity. The isolates can be used as a biofertilizer to support coastal agriculture.A total of 28 isolates (14 isolates of Azospirillum and 14 isolates of Azotobacter were tested their tolerance to salt by growing them in their respective media containing 1% 2% and 3 % NaCl,. Salt tolerant isolates obtained, then used as inoculants in paddy. The experiment laid out factorial based randomized complete block design which was comprised of 7 fertilizer treatments and 5 watering treatments with 5 replicates for each treatment at green house of Microbiology Division, Research Center of Biology, Indonesian Institute of Sciences. The first factor was fertilizer treatments 1. Compost + mixed bacteria (K1, 2. Control without inoculant , 3. Compost, 4. NPK, 5. Compost + NPK + mixed microbial, 6. Azotobacter isolates; 7. Azospirillum isolates. The second factor was watering treatments, plant was watered by : 1. Freshwater, 2. freshwater + sea water at mixture ratio 1:1, 3. Sea water, 4. sea water + 2% NaCl (20 g NaCl / l , 5. freshwater + 5% NaCl (50 g NaCl / l.The result showed that there were 9 isolates of Azospirillum and 4 isolates of Azotobacter which were tolerant to grow at media with 3% NaCl The green house experimental result revealed that the plants were treated with bacteria can survive up to the level of salinity 12.43 dS-1m.

  9. Cellular and molecular-genetic mechanisms of symbiosis and associative interaction of microorganisms with plants in rhizosphere

    Directory of Open Access Journals (Sweden)

    Lioshina L. G.

    2009-02-01

    Full Text Available The review contains the results of research on symbiotic and associative interaction of microorganisms and plants in rhizosphere. A special attention is given to the process of contact association of microorganisms and plants tissues including the concrete molecular structures and dominant role pertaining to protein-carbohydrate interaction. There are common features and distinctions at formation of arbuscular mycorhiza, rhizobia– legume symbiosis and association of non-leguminous plants with Azospirillum

  10. Identification of novel spp. of rice and wheat endophytic diazotrophs by 16S rDNA gene and FTIR analysis

    Directory of Open Access Journals (Sweden)

    Mohammad Javad Mehdipour Moghaddam

    2012-06-01

    Full Text Available In this research, six isolates, including three from three rice roots (PxR1, PxR2 and StR1 and three from three wheat roots (PxW1, PxW2 and PxW3 were isolated as endophytic bacteria and except for StR1, all the isolates were identified as Pseudoxanthomonas based on phenotypic analysis including FTIR and PCR amplification of 16S rDNA. The results showed that PxR1, PxR2, PxW1 and PxW2 were all similar and belonged to a novel species of Pseudoxanthomonas, but PxW3 was from different species. StR1 belonged to a novel species of Stenotrophomonas. Two strains including Azospirillum brasiliense Sp7 (S1 and Azospirillum lipoferum (S2 were selected as standard strains and compared with those isolates however, phenotypic and genotypic analysis verified that those isolates were not Azospirillum. For the first time, it was indicated that Pseudoxanthomonas existed as an endophytic bacterium in rice root.

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

    Directory of Open Access Journals (Sweden)

    Valeria Marino Rodrigues Sala

    2008-06-01

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

  12. Microorganismos benéficos como biofertilizantes eficientes para el cultivo del tomate (Lycopersicon esculentum, Mill Beneficial microorganisms as efficient biofertilisers for tomato crops (Lycopersicon esculentum, Mill

    Directory of Open Access Journals (Sweden)

    Hernández Annia

    2005-12-01

    Full Text Available En el uso y manejo de biofertilizantes en la agricultura, uno de los principales problemas es el desconocimiento de las especies presentes en los agroecosistemas y en la rizosfera de los cultivos, para su posible utilización eficiente. Desde el punto de vista ecológico, es importante conocer los integrantes de la comunidad bacteriana que favorecen su aplicación como inoculantes y propician un efecto agrobiológico positivo en los cultivos agrícolas. Esta investigación se desarrolló con el objetivo de evaluar la efectividad agrobiológica de Azospirillum sp, en el crecimiento, desarrollo y rendimiento en el cultivo del tomate. Para ello, se partió de seleccionar el género microbiano predominante en la rizosfera del cultivo y posteriormente se evaluó el efecto de su inoculación a partir de la respuesta del cultivo. Los resultados demostraron que los géneros Pseudomonas, Azospirillum, Azotobacter, Bacillus y Streptomyces, forman parte de la comunidad microbiana de la rizosfera del tomate, en las condiciones estudiadas, y que Azospirillum es el género dominante. La inoculación artificial de esta rizobacteria causó un efecto positivo sobre el crecimiento de las plántulas, así como en el estado nutricional de las plantas, con un rendimiento agrícola superior a un 11 % con respecto a las plantas testigo. Se obtuvo un alto nivel poblacional en la rizosfera de las plantas inoculadas. Palabras clave: rizosfera, inoculante, crecimiento, rendimiento.One of the main problems regarding the efficient use and management of biofertilizers in agriculture lies in the unknown species present in agro-ecosystems and crop rhizospheres. From the ecological point of view, it is important to know the members of the bacterial population allowing them to be applied as inoculants and enable a positive agro-biological effect on agricultural crops. This investigation was aimed at evaluating the agro-biological effectiveness of Azospirillum sp. on tomato

  13. Biofertilización de café orgánico en etapa de vivero en Chiapas, México Biofertilizer of organic coffee in stage of seedlings in Chiapas, Mexico

    Directory of Open Access Journals (Sweden)

    María de Lourdes Adriano Anaya

    2011-06-01

    Full Text Available En Chiapas, la producción de plántulas de café, se realiza convencionalmente con la aplicación de fertilizantes químicos. La producción de café orgánico, requiere la nutrición de plántulas con biofertilizantes y por ello el objetivo fue evaluar el efecto de algunos de éstos en el desarrollo de plántulas de café (Coffe arábica variedad Bourbon en vivero. El experimento se realizó durante 2007 y 2008 en Cacahohatan,Chiapas. Los inoculantes fueron una cepa Glomus intraradices Schenck y Smith, cepas PACHAZ08 de Azotobacter y 11B de Azospirillum. Se utilizó el diseño factorial 2³ con ocho tratamientos y 100 repeticiones por tratamiento. En las platulas inoculadas, se efectuaron 4 muestreos con intervalos de 28 días, midiendóse la altura, longitud de hojas, longitud de raíz, peso seco de hojas y raíces, contenido de clorofila y nitrógeno, y colonizacion de raíz por los inoculantes. Los datos se sometieron al análisis de varianza y comparacion de medias por Tukey p≤ 0.05. Las mejores características morfológicas y bioquímicas de las plántulas, se obtuvieron con Azospirillum sóla o coinoculada con Glomus y Azotobacter y estadísticamente fueron los mejores tratamientos. Azospirillum modificó la arquitectura de la raíz y estimuló la micorrización. Los diazotrofos en conjunto fueron antagónicos pero esta fue inhibida por Glomus. La interacción de los tres microorganismos indujo en las plántulas un mejor aprovechamiento de nutrimentos, agua, capacidad fotosintética y mayor acumulación de biomasa carbonada.In Chiapas, c offee seedlings product ion is conventionally done with chemical fertilizers. Organic coffee production requires see dlings nutrition with biofertilizers and therefore the objective was to evaluate the effect of some of these in coffee seedlings development (Coffee arabica Bourbon variety in a nursery. The experiment was conducted during 2007 and 2008 in Cacahohatan, Chiapas. Inoculants were a strain

  14. Role of biofertilizers in increasing tea productivity

    Institute of Scientific and Technical Information of China (English)

    P Nepolean; R Jayanthi; R Vidhya Pallavi; A Balamurugan; T Kuberan; T Beulah; R Premkumar

    2012-01-01

    Objective: To increase the tea productivity using biofertilizers like VAM fungi, phosphobacteria, Azosprillium. Methods: Nitrogen fixing Azospirillum, phosphate solubilising bacteria and VAM fungi were isolated from tea soils of southern India and region specific potential strains were selected based on laboratory, greenhouse, as well as field experiments. A technology for their mass multiplication has been developed. Results: Biofertilizers were showed higher yield at the rate of 5g/seedling and VAM fungi with 15 g of formulation/bush. Conclusions: Multifunctional biofertilizers were used to reduce the chemical fertilizers, diseases and to increase soil fertility in tea.

  15. Aplicação de espectrometria de massa no controle de qualidade de inoculantes comerciais agrícolas contendo bactérias promotoras de crescimento vegetal

    OpenAIRE

    Silva,Leilane Caline

    2014-01-01

    Orientador : Prof. Dr. Leonardo Magalhães Cruz Dissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Ciências : Bioquímica. Defesa: Curitiba, 28/02/2014 Inclui referências : f. 70-79 Resumo: As rizobacterias promotoras do crescimento vegetal (PGPR) sao bactérias do solo benéficas para o crescimento de plantas e para a agricultura pois podem influenciar positivamente no rendimento da safra. Bactérias do gênero Azospirillum ...

  16. Biodiversity of Soil Microbes from Rhizosphere at Wamena Biological Garden (WBiG, Jayawijaya, Papua

    Directory of Open Access Journals (Sweden)

    SRI WIDAWATI

    2005-01-01

    Full Text Available The isolation, identification and population of soil microbes from rizosphere at WBiG had been done in the Soil Laboratories Microbiology, Microbiology Division, Research Center of Biology, Indonesian Institute of Sciences (LIPI, Bogor. The soil was collected randomly from 16 sites in WBiG, and taken from 0-15 cm depth. Isolates of microbes were identified by Bergeys manual method for bacteria; Ellis method for fungi, and the morphology of isolate method for Actinomycetes. The population of microbes was estimated by plate count method. The result of isolation, identification and population soil microbes from 16 samples in WBiG showed that 20 isolates of bacteria (Azotobacter sp., Accinetobacter sp. , Bacillus sp., Citrobacter sp., Flavobacterium sp., Klebsiella sp., Nitrosomonas sp., Pseudomonas sp., Rhizobium sp., Thiobacillus sp., Azospirillum sp., Azotobacter chrococcum, Bacillus panthothenticus, Chromobacterium violaceum, C.lividum, Escherrrichia coli, Flavobacterium breve, Klebsiella aerogenes, Spaerotillus natans, and Staphylococcus epidermidis; nine isolates of fungi (Aspergillus niger, Bisporomyces, Monilia sp., Cephalospharium sp., Verticillum sp., Giocladium sp., Penicillium sp., Nelicocephalum sp., and Cuninghamella sp., and seven isolates of Actinomycetes (Streptomyces, Streptosporangium, Nocardia, Thermomonospora, Thermoactinomyces, Micromonospora, Mycobacterium. The population of Bacillus (108-109, Rhizobium (106-107, Azospirillum (106-107, and Thiobacillus (104 -109 were founded all of soil samples.

  17. Phytoremediation of Polycyclic Aromatic Hydrocarbons in Soils Artificially Polluted Using Plant-Associated-Endophytic Bacteria and Dactylis glomerata as the Bioremediation Plant.

    Science.gov (United States)

    Gałązka, Ann; Gałązka, Rafał

    2015-01-01

    The reaction of soil microorganisms to the contamination of soil artificially polluted with polycyclic aromatic hydrocarbons (PAHs) was evaluated in pot experiments. The plant used in the tests was cock's foot (Dactylis glomerata). Three different soils artificially contaminated with PAHs were applied in the studies. Three selected PAHs (anthracene, phenanthrene, and pyrene) were used at the doses of 100, 500, and 1000 mg/kg d.m. of soil and diesel fuel at the doses of 100, 500, and 1000 mg/kg d.m. of soil. For evaluation of the synergistic effect of nitrogen fixing bacteria, the following strains were selected: associative Azospirillum spp. and Pseudomonas stutzerii. Additionally, in the bioremediation process, the inoculation of plants with a mixture of the bacterial strains in the amount of 1 ml suspension per 500 g of soil was used. Chamber pot-tests were carried out in controlled conditions during four weeks of plant growth period. The basic physical, microbiological and biochemical properties in contaminated soils were determined. The obtained results showed a statistically important increase in the physical properties of soils polluted with PAHs and diesel fuel compared with the control and also an important decrease in the content of PAHs and heavy metals in soils inoculated with Azospirillum spp. and P. stutzeri after cock's foot grass growth. The bioremediation processes were especially intensive in calcareous rendzina soil artificially polluted with PAHs.

  18. Selección y caracterización de rizobacterias promotoras de crecimiento vegetal (RPCV asociadas al cultivo de algodón (Gossypium hirsutum

    Directory of Open Access Journals (Sweden)

    Andrés Guzmán

    2012-04-01

    Full Text Available Normal 0 21 false false false ES-CO X-NONE X-NONE MicrosoftInternetExplorer4 Título en ingles: Selection and characterization of plant growth promoting rhizobacteria (PGPR’s associated with cotton crop (Gossypium hirsutum Resumen: Como parte de las estrategias de una agricultura sostenible, se hace necesario disminuir el uso de fertilizantes nitrogenados de síntesis, mediante la utilización de los biofertilizantes. En particular, los géneros Azotobacter y Azospirillum son utilizados como agentes promotores de crecimiento vegetal debido a su capacidad para fijar nitrógeno atmosférico y producir hormonas de tipo indólico. Por tal razón, en este estudio se aislaron bacterias diazotróficas de los géneros Azotobacter y Azospirillum a partir de la rizósfera de cultivos de algodón en el Espinal (Tolima. Las poblaciones microbianas se caracterizaron fenotípicamente en los medios de cultivo semiespecíficos: Ashby y LG (Azotobacter sp. y NFb, LGI y Batata (Azospirillum sp.. La promoción de crecimiento vegetal se determinó mediante la actividad de la enzima nitrogenasa por medio de la técnica de reducción de acetileno y producción de índoles por el método colorimétrico de Salkowsky. Se obtuvieron 9 aislamientos tentativos de Azotobacter sp. y 4 de Azospirillum sp. Se presentaron diferencias significativas en la prueba de reducción de acetileno con las cepas presuntivas de Azotobacter sp.: NAT 9 (206.43 nmol C2H2 mL-1.h-1, NAT 4, (292.77 nmol C2H2 mL-1.h-1, y NAT 6 (460.60 nmol C2H2 mL-1.h-1 y en la producción de índoles de las cepas NAT 19 (19.87 μg.mL-1 y NAT 13 (20.08 μg.mL-1. Por su eficiencia in vitro en la promoción de crecimiento vegetal se seleccionaron las cepas NAT9, NAT4, NAT6, NAT19 y NAT13 para ser evaluadas como principio activo en futuros inoculantes para el algodón en esta zona del departamento del Tolima. Palabras clave: fijación biológica de nitrógeno; producción de índoles; promoción del crecimiento

  19. Reclamation of abandoned ash pond of Neyveli thermal power plant by enrichment and its effect on growth and yield of gingelly

    Energy Technology Data Exchange (ETDEWEB)

    Sheela, A.M.; Shanmugasundaram, R.; Sundaram, M.D. [Annamalai Nagar University, Annamalai, Nagar (India). Dept. of Microbiology

    2003-07-01

    Ash ponds are the common disposal sites of ash generated from India's thermal power stations. An attempt was made to enrich the ash pond of Neyveli Lignite Corporation Ltd., Neyveli, using organic amendments and microbial inoculants to promote crop growth. The oil seed crop gingelly was grown in two seasons and more growth and yield was recorded in the second crop, which was enriched in the second time also. The inoculation effect of the crop with VAM fungus Glomus mosseae and the acid-tolerant Azospirillum isolate, A. lipoferum PAI-1 was studied. The co-inoculation of Azospirillum with VAM enhanced the rhizosphere dehydrogenase enzyme activity, growth, yield, N, and P content of the plant in the enriched ash pond. Hence, the nutritionally and biologically poor medium like ash ponds could be enriched successively to increase in the nutrient contents and microbial activity. Prolonged enrichment would be providing favourable conditions for the establishment of vegetation and the environmental pollution is greatly reduced by the vegetative cover on barren land. 30 refs., 6 tabs.

  20. INFLUENCE OF ORGANIC MANURES AND AMENDMENTS IN SOIL PHYSIOCHEMICAL PROPERTIES AND THEIR IMPACT ON GROWTH, YIELD AND NUTRIENT UPTAKE OF BANANA

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    K VANILARASU

    2014-06-01

    Full Text Available An investigation was carried out during 2010-2011, to study the effect of organic manures (Farmyard manure, Vermicompost, Neem cake and Wood ash, organic amendments (Arbuscular mycorrhizae, Azospirillum, Phosphate Solubilising Bacteria and Trichoderma harzianum and green manures (Sunhemp and Cowpea in comparison with inorganic fertilizers on leaf nutrient and soil physiochemical properties of banana cv. Grand Naine. The treatment T10 with the combined application of organic manures, amendments and green manures (Farmyard manure @ 10 kg + Neem cake @ 1.25 kg + Vermicompost @ 5 kg and Wood ash @ 1.75 kg /plant + Triple green manuring with Sunhemp + Double intercropping of Cow pea + biofertilizers viz., Arbuscular Mycorrhizae @ 25 g , Azospirillum @ 50 g, Phosphate Solubilizing Bacteria @ 50 g and Trichoderma harzianum @ 50 g/plant registered the maximum growth, yield and yield attributes, leaf nutrient status of N, P and K at 5th and 7th month after planting and soil physiochemical properties at harvesting stage because the role of organic manures and amendments to make the soil has healthy as well as possible and also, the unavailable form of soil nutrients to available form by enhancing mineralization and solubilization process in soil by adding organic manures and microbial agents make easy uptake of nutrients when crop required comparing to chemical fertilizers.

  1. Cultivation-Based and Molecular Assessment of Bacterial Diversity in the Rhizosheath of Wheat under Different Crop Rotations.

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    Muhammad Tahir

    Full Text Available A field study was conducted to compare the formationand bacterial communities of rhizosheaths of wheat grown under wheat-cotton and wheat-rice rotation and to study the effects of bacterial inoculation on plant growth. Inoculation of Azospirillum sp. WS-1 and Bacillus sp. T-34 to wheat plants increased root length, root and shoot dry weight and dry weight of rhizosheathsoil when compared to non-inoculated control plants, and under both crop rotations. Comparing both crop rotations, root length, root and shoot dry weight and dry weight of soil attached with roots were higher under wheat-cotton rotation. Organic acids (citric acid, malic acid, acetic acid and oxalic acid were detected in rhizosheaths from both rotations, with malic acid being most abundant with 24.8±2 and 21.3±1.5 μg g(-1 dry soil in wheat-cotton and wheat-rice rotation, respectively. Two sugars (sucrose, glucose were detected in wheat rhizosheath under both rotations, with highest concentrations of sucrose (4.08±0.5 μg g(-1 and 7.36±1.0 μg g(-1 and glucose (3.12±0.5 μg g(-1 and 3.01± μg g(-1 being detected in rhizosheaths of non-inoculated control plants under both rotations. Diversity of rhizosheath-associated bacteria was evaluated by cultivation, as well as by 454-pyrosequencing of PCR-tagged 16S rRNA gene amplicons. A total of 14 and 12 bacterial isolates predominantly belonging to the genera Arthrobacter, Azospirillum, Bacillus, Enterobacter and Pseudomonaswere obtained from the rhizosheath of wheat grown under wheat-cotton and wheat-rice rotation, respectively. Analysis of pyrosequencing data revealed Proteobacteria, Bacteriodetes and Verrucomicrobia as the most abundant phyla in wheat-rice rotation, whereas Actinobacteria, Firmicutes, Chloroflexi, Acidobacteria, Planctomycetes and Cyanobacteria were predominant in wheat-cotton rotation. From a total of 46,971 sequences, 10.9% showed ≥97% similarity with 16S rRNA genes of 32 genera previously shown to include

  2. Isolation and Identification of Phosphate Solubilizing and Nitrogen Fixing Bacteria from Soil in Wamena Biological Garden, Jayawijaya, Papua

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    SRI WIDAWATI

    2005-07-01

    Full Text Available A study was undertaken to investigate the occurrence of phosphate solubilizing bacteria (PSB and nitrogen-fixing bacteria (NFB from soil samples of Wamena Biological Garden (WbiG. Eleven soil samples were collected randomly to estimate microbial population which used plate count method. The result showed that the microbial population ranged from 5.0x103-7.5x106 cells of bacteria/gram of soil and 5.0x103-1.5x107 cells of bacteria/gram of soil for PSB and NFB respectively. There were 17 isolates which have been identified till genus and species. The isolated microorganism were identified as PSB i.e. Bacillus sp., B. pantothenticus, B. megatherium, Flavobacterium sp., F. breve, Klebsiella sp., K. aerogenes, Chromobacterium lividum, Enterobacter alvei, E. agglomerans, Pseudomonas sp., Proteus sp. and as NFB i.e. Azotobacter sp., A. chroococcum, A. paspalii, Rhizobium sp., and Azospirillum sp.

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

    Science.gov (United States)

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

    2013-05-01

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

  4. Efecto de la inoculación con bacterias rizosféricas en dos variedades de trigo. Fase I: condiciones controladas The effect of innoculation with rhizospheric bacteria on two varities of wheat. Phase1: controlled conditions

    Directory of Open Access Journals (Sweden)

    Carlos José Bécquer Granados

    Full Text Available Se llevó a cabo un experimento bajo condiciones controladas para determinar la respuesta de dos variedades de trigo a la inoculación simple y combinada realizada con Sinorhizobium y Azospirillum. Se utilizó una cepa de A. zeae, y dos cepas pertenecientes a S. meliloti. Los materiales y métodos aplicados correspondieron a lo descrito en las metodologías prestablecidas en este campo de estudio. Diseño experimental: completamente aleatorizado, con 20 tratamientos y 4 réplicas. No se utilizó tratamiento fertilizado. Se evaluaron diferentes variables agronómicas, relacionadas con la arquitectura radical y la biomasa aérea. Análisis de varianza bifactorial. En caso de aparecer diferencias, se aplicó análisis de varianza de un factor a la interacción en primera instancia, o a los factores probados. Diferencias entre medias por LSD de Fisher. Se transformaron datos de conteo de dígitos por √x. Se calculó correlación y regresión múltiple entre variables. Se concluye que la inoculación combinada de Sinorhizobium con Azospirillum, así como la inoculación simple con Sinorhizobium, resultaron de alta importancia en las alternativas de inoculación que se realizaron en el experimento. Existió una alta diferenciación entre las dos variedades de trigo en determinadas variables agronómicas, lo que indica una influencia marcada de las características varietales de las plantas. Se observó una fuerte relación estadística entre las variables peso seco aéreo y variables de la raíz para los tratamientos inoculados con A2 y A2+N7, respectivamente.The experiment was carried out under controlled conditions to determine the response of the two wheat varieties to the simple inoculation and the combined inoculation which was carried out with Sinorhizobium, and Azospirillum. A strain of A. zeae and two strains belonging to S. meliloti were used. The materials and methods that were applied corresponded to what was described in the already

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

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    Fátima M.S. Moreira

    2008-12-01

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

  6. Studies on utilization of nitrogen-fixing bacteria for saving energy; Chisso koteikin no katsuyo ni yoru sho energy no kenkyu

    Energy Technology Data Exchange (ETDEWEB)

    Uozumi, T.; Koyama, R.; Horiuchi, M.; Hidaka, M.; Masaki, H. [The University of Tokyo, Tokyo (Japan); Shigematsu, T.; Inoue, A. [New Energy and Industrial Technology Development Organization, Tokyo, (Japan)

    1997-02-01

    This paper describes analysis and enhancement of nitrogen-fixing gene of rice root bacteria, such as Klebsiella oxytoca, Azospirillum lipoferumn and Sphingomonas paucimobilis, for realizing energy saving through conservation of nitrogenous fertilizers. For K. oxytoca, modified strain R-16 was developed, which can fix nitrogen effectively even in the presence of NH4{sup +}. Nitrogen-fixing ability of A. lipoferumn depends on the activity control by the modification of nitrogen-fixing enzyme as well as on the adjustment of transcription level by the transcription activating gene, nifA. The control gene relating to the above was analyzed by making clones. As a result, a modified strain TAl without the control by NH4{sup +} was developed. The R-16 and TAl strains were inoculated into rice sterile-cultured without nitrogen. Consequently, inoculated strains were settled in the root, which resulted in the increased vegetation weight of plant to two times heavier than that without inoculation. 9 refs.

  7. The influence of Plant Growth Promoting Rhizobacteria (PGPR on the reduction of abiotic stresses in crops

    Directory of Open Access Journals (Sweden)

    Omid Alizadeh

    2011-12-01

    Full Text Available Plants are always subjected to biotic and abiotic stresses in the environment which haveinfluences on the growth and development of the plants. Beneficial free-living soil bacteria are usuallyreferred as Plant-Growth Promoting Rhizobacteria or PGPR. Different plant growth-promotingRhizosphere bacteria, including associative bacteria such as: Azospirillum, Bacillus, Pseudomonas andEnterobacter group have been used for their beneficial influences on plants. Typically, PGPRs areassociated with plants root and augment plant productivity and immunity; however, recent worksshowed that PGPRs not have just induced the systemic tolerance to abiotic stress such as salt anddrought, but also they have increased the nutrient uptake from soils, and as a result the hazardousaccumulation of nitrates and phosphates in the agricultural soils can be reduced by usage of them.

  8. Efecto de la inoculación con bacterias rizosféricas en dos variedades de trigo. Fase II: invernadero Effect of inoculation with rihizospheric bacteria in two varieties of wheat. Phase II: greenhouse

    Directory of Open Access Journals (Sweden)

    Carlos José Bécquer Granados

    Full Text Available Se llevó a cabo un experimento de invernadero para evaluar la influencia de la inoculación simple y combinada, efectuada con las bacterias rizosféricas Sinorhizobium y Azospirillum, en dos variedades de trigo. Materiales y métodos según lo descrito en las metodologías convencionales para este campo de estudio. El diseño experimental fue completamente aleatorizado, con 4 réplicas y 10 tratamientos. Análisis estadístico varianza bifactorial. Se utilizó tratamiento fertilizado con NH4NO3 (150 ppm/kg suelo. Se evaluó contenido de clorofila foliar, peso seco aéreo, peso seco radical, longitud del tallo y germinación. En caso de aparecer diferencias, se determinaron mediante la prueba de Duncan, y las diferencias entre las variedades con t-Student. Se concluye que la inoculación combinada de la cepaA2 (Sinorhizobium meliloti con la cepaN7 (Azospirillum zeae, fue la de mayor influencia positiva en el contenido de clorofila de las plantas. Por otra parte, existió una alta diferenciación entre las dos variedades de trigo en la longitud del tallo, peso seco aéreo y peso seco radical. Los resultados en peso seco aéreo y peso seco radical, al combinarse los dos factores estudiados, dependieron notablemente de las características varietales de la planta y del efecto significativo de la población autóctona de rizobacterias. La germinación de las plantas no estuvo vinculada a ninguno de los factores aplicados en el experimento.The experiment was carried out in a greenhouse so that the inoculation, both simple and combined (with the rhizospheric bacteria- Sinorhizobium andAzospirllium, en two varieties of Wheat. Materials and methodologies were used in accordance with previously prescribed conventional methodologies for this study. The experimental design was completely randomized, with 4 replicas and 10 treatments. A statistical analysis using two-way variance was done. Fertilized treatment was applied with NH4NO3 (150 ppm/kg soil. The

  9. Removal of Ni (Ⅱ) and microbial dynamics in single-chamber microbial electrolysis cell%单室微生物电解池除镍途径分析及微生物群落动态特征

    Institute of Scientific and Technical Information of China (English)

    赵欣; 吴忆宁; 王岭; 李伟明; 靳敏; 李帅

    2016-01-01

    [目的]为探究UASB颗粒污泥启动的单室微生物电解池(Single-chamber microbial electrolysis cell,SMEC)对Ni(Ⅱ)的去除途径和SMEC中微生物群落的动态特征.[方法]以乙酸钠为底物,采用单因子控制方法分析SMEC对Ni(Ⅱ)的去除途径和应用Illumina高通量测序技术解析SMEC启动过程中微生物群落的组成和结构动态学特征.[结果]结果表明,SMEC对重金属的去除主要通过吸附和微生物作用.经培养驯化功能菌群发生变化.成熟单室微生物燃料电池(Single-chamber microbial fuel cell,SMFC)阳极生物膜菌群主要是Proteobacteria(变形菌门,91.42%)中的Geobacter sp.(地杆菌属,76.25%);阴极生物膜菌群主要是Bacteroidetes(拟杆菌门,47.99%)中的Niabella sp.(布鲁氏菌属,33.01%)和Proteobacteria(45.74%)中的Ochrobactrum sp.(苍白杆菌属,10.80%).成熟SMFC改装成的SMEC在12.5 mg-Ni(Ⅱ)/L下,阳极生物膜菌群由单一优势菌Geobacter sp.转变为Geobacter sp.(41.56%)和Proteobacteria中的Azospirillum sp.(固氮螺菌属,5.97%);阴极生物膜菌群由Niabella sp.和Ochrobactrum sp.转变为Firmicutes(厚壁菌门,25.21%)中的Acetoanaerobium sp.(19.28%)、Proteobacteria (51.42%)中的Dokdonella sp.(16.48%)和Azospirillum sp.(9.49%).[结论]本研究表明,污泥微生物经SMFC和SMEC驯化过程及Ni(Ⅱ)的淘汰和选择,在电极上形成了稳定、高效产电与除镍菌群,优势菌群为Proteobacteria.

  10. PRODUCCIÓN DE ANTISUEROS PARA LA DETECCIÓN DE ÁCIDO INDOLACÉTICO EN CULTIVOS DE BACTERIAS PROMOTORAS DEL CRECIMIENTO VEGETAL Antisera Production to Detect Indoleacetic Acid in Cultures of Plant-Growth Promoting Bacteria

    Directory of Open Access Journals (Sweden)

    MARCIA M ROJAS

    2012-05-01

    Full Text Available Se obtuvieron antisueros en conejo utilizando como antígeno el AIA adherido a membranas de nitrocelulosa que mostraron un elevado título y especificidad. Mediante la técnica de inmunoadsorción por manchas marcadas con oro coloidal se detectó la producción de esta auxina por cepas de los géneros Gluconacetobacter, Herbaspirillum, Azospirillum, Pseudomonas, Burkholderia y Bacillus empleando como antígenos los sobrenadantes de los cultivos. Para cuantificar la producción de AIA y corroborar los datos obtenidos se empleó la técnica colorimétrica derivada de Salkowski. Los resultados muestran que todos los géneros bacterianos estudiados tienen la capacidad de producir AIA y se demuestra la factibilidad del uso de este antisuero policlonal para la detección de este metabolito. Teniendo en cuenta las potencialidades de estas bacterias, resulta de gran importancia la utilización de antisueros y técnicas serológicas para la detección rápida y sencilla de este tipo de metabolitos en bacterias asociadas a cultivos de interés económico.Rabbit polyclonal antisera against indoleacetic acid (IAA bound to nitrocellulose membrane were obtained, which exhibited a high titer and specificity. The dot immunobinding technique with colloidal gold was used to detect auxin production by several strains belonging to Gluconacetobacter, Herbaspirillum, Azospirillum, Pseudomonas, Burkholderia and Bacillus genera, using culture supernatants as antigens. Moreover, auxin production was quantified by the Salkowski's method to corroborate the previous results. It was found that that all the studied microorganisms produce IAA and the feasibility of using these antisera to detect the metabolite was confirmed. Taking into account the potentialities of plant growth promoting bacteria as biofertilizers, the use of these antisera for a rapid and easy detection of IAA in bacteria associated with important crops is thus recommended.

  11. Identificación de algunos géneros microbianos asociados al cultivo del maíz (Zea mays L. en diferentes suelos de Cuba Identification of some microbial genera associated to the maize crop (Zea mays L in different Cuban soils

    Directory of Open Access Journals (Sweden)

    Heydrich Mayra

    2003-06-01

    Full Text Available Se ha demostrado que la aplicación de bioproductos a partir de microorganismos rizosféricos en la agricultura, provoca incrementos en la productividad de los cultivos. Si se trabaja con cepas nativas aumenta la factibilidad biológica de los mismos. Esta investigación se realizó con el objetivo de determinar algunos géneros microbianos asociados al cultivo del maíz variedad Francisco mejorado en suelos Nitisol Rhodic, Cambisol Eutric-Humic y Cambisol Eutric procedentes de diferentes localidades cubanas. Para ello se emplearon tres métodos de aislamiento: Método Convencional, Tubos Espermosféricos y Modelo Microcosmos. Los aislados fueron clasificados mediante la utilización de técnicas clásicas. Los resultados obtenidos demostraron que los géneros Pseudomona, Azospirillum, Azotobacter, Bacillus y Streptomyces forman parte de la comunidad microbiana de la rizosfera del cultivo del maíz en las condiciones estudiadas, constituyendo Pseudomonas el género dominante. Los métodos de aislamiento emplea­dos resultaron adecuados para la obtención de representantes típicos de las poblaciones microbianas en estudio, demostrándose la superioridad de los Tubos Espermosféricos y el Modelo Microcosmos para estos fines, ya que los mismos permiten aislar los microorganismos capaces de vivir a expensas de los exudados radicales del cultivo, en la interacción planta-bacteria. Palabras clave: rizosfera; maíz; modelo espermosférico; modelo microcosmos; Pseudomonas.It has been demonstrated that applying bio-products based on native rhizosphere micro-organisms in agriculture increases crop productivity. Working with native strains also improves their biological feasibility. Some microbial genera associated with the improved corn variety Francisco were isolated from Nitisol Rhodic, Cambisol Eutric-Humic and Cambisol Eutric soil from different Cuban regions using three isolation methods: the convencional method, the spermosphere model and the

  12. EFFECT OF INTEGRATED NUTRIENT MANAGEMENT ON VEGETATIVE GROWTH AND YIELD OF TRANSPLANTED HYBRID RICE (ORYZA SATIVA L. CROP

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    RAYEES A. SHAH* SANDEEP KUMAR

    2014-09-01

    Full Text Available The integrated use of organic and inorganic sources of plant nutrients can help in sustainable and environmentally sound nutrient management of soils that are low in organic matter. A study was conducted  during 2009- 10 & 2010-11, on sandy loam soils, low in organic carbon, medium in available phosphorus and potash with pH of 7.5 at Agricultural Research Farm of C.C.R.(P.G. College, Muzaffarnagar U.P. India, to investigate the comparative use of  integrated nutrient management on vegetative growth and yield of hybrid rice. The experiment was laid out with fifteen treatments in a RBD. The treatments were; Control (T1, NPK 100% RDF (T2, NPK 75% RDF (T3, NPK 50% RDF + FYM @ 10 tons  ha-1 (T4, NPK 50% RDF + FYM @15 tons ha-1 (T5, NPK 50% RDF + wheat cut straw @10 tons ha-1 (T6, NPK 50% RDF + wheat cut straw @15 tons ha-1 (T7, NPK 50% RDF + Neem cake @ 2.5 tones ha-1 (T8, NPK 50% RDF + Neem cake @ 5 tones ha-1 (T9, NPK 50% RDF + Vermicompost @ 2.5 tons ha-1 (T10, NPK 50% RDF + Vermicompost @ 2.5 tons ha-1 +Azotobacter (T11, NPK 50% RDF + Vermicompost @ 2.5 tones ha-1 +PSB @ 5Kg ha-1 (T12, NPK 50% RDF + Vermicompost @ 2.5 tones ha-1 + Azospirillum (T13, NPK 50% RDF  + Azotobacter + Azospirillum + PSB @ 5 Kg ha-1(T14, and NPK 50% RDF + Neem cake @ 2.5 tones ha-1 +FYM @5 tons  ha-1 + Azotobacter+PSB @ 5Kg ha-1 (T15. The maximum grain yield (63 and 67 q per hectare during 2009 and 2010, respectively was obtained with the integration of NPK 50% RDF + Neem cake @2.5 tonnes ha-1 + FYM @5 tonnes ha-1+Azotobacter+PSB @ 5kg ha-1.

  13. Cuantificación de bacterias diazótrofas aisladas de suelos cacaoteros (Theobroma cacao L., por la técnica de Número Más Probable (NMP

    Directory of Open Access Journals (Sweden)

    Adriana Zulay Argüello Navarro

    2016-07-01

    Full Text Available Esta investigación tuvo como objetivo cuantificar bacterias diazótrofas y comparar fisicoquímicamente suelos rizosféricos de tres cacaotales (Theobroma cacao L. del Departamento Norte de Santander, Colombia; para lo cual se caracterizaron, diferenciándose en área cultivada, manejo agronómico y edad del cultivo. A partir de diluciones seriadas de las muestras y empleando la técnica de Número Más Probable (NMP, en medios de cultivo semiselectivos (NFb, JMV, LGI, JNFb semisólidos, se cuantificaron las diazótrofas, evaluando como positivo la formación de una película subsuperficial en el medio contenido en viales sellados; muestras pares se enviaron al laboratorio Bioambiental (UNET para los análisis fisicoquímicos. Como resultado, las muestras evaluadas mostraron deficiencias en el porcentaje de materia orgánica y elementos como Potasio, Fósforo y Magnesio. Se reportaron estadísticamente diferencias altamente significativas en NMP. La mayor cuantificación de diazótrofas se reportó en la finca Florilandia, que se caracterizó por tener riego por goteo. La mayor cuantificación de diazotrófas se registró en los medios NFb y JMV, demostrándose una mayor presencia de los presuntos géneros Azospirillum sp. y Burkholderia sp. los cuales son fácilmente aislados de suelos rizosféricos, a diferencia de los géneros Herbaspirillum sp. y Gluconacetobacter sp. que por su carácter endófito suelen ser menos predominantes en este tipo de muestras. Se concluye además, que las características fisicoquímicas del suelo, la humedad y las relaciones climáticas al momento de la toma de las muestras, condicionan la cantidad de exudados de las raíces y por tanto son factores que condicionaron la presencia de diazótrofas en las muestras. Palabras clave: Azospirillum, Burkholderia, cacao, rizósfera

  14. Diversity of 16S rRNA genes from bacteria of sugarcane rhizosphere soil.

    Science.gov (United States)

    Pisa, G; Magnani, G S; Weber, H; Souza, E M; Faoro, H; Monteiro, R A; Daros, E; Baura, V; Bespalhok, J P; Pedrosa, F O; Cruz, L M

    2011-12-01

    Sugarcane is an important agricultural product of Brazil, with a total production of more than 500 million tons. Knowledge of the bacterial community associated with agricultural crops and the soil status is a decisive step towards understanding how microorganisms influence crop productivity. However, most studies aim to isolate endophytic or rhizosphere bacteria associated with the plant by culture-dependent approaches. Culture-independent approaches allow a more comprehensive view of entire bacterial communities in the environment. In the present study, we have used this approach to assess the bacterial community in the rhizosphere soil of sugarcane at different times and under different nitrogen fertilization conditions. At the high taxonomic level, few differences between samples were observed, with the phylum Proteobacteria (29.6%) predominating, followed by Acidobacteria (23.4%), Bacteroidetes (12.1%), Firmicutes (10.2%), and Actinobacteria (5.6%). The exception was the Verrucomicrobia phylum whose prevalence in N-fertilized soils was approximately 0.7% and increased to 5.2% in the non-fertilized soil, suggesting that this group may be an indicator of nitrogen availability in soils. However, at low taxonomic levels a higher diversity was found associated with plants receiving nitrogen fertilizer. Bacillus was the most predominant genus, accounting for 19.7% of all genera observed. Classically reported nitrogen-fixing and/or plant growth-promoting bacterial genera, such as Azospirillum, Rhizobium, Mesorhizobium, Bradyrhizobium, and Burkholderia were also found although at a lower prevalence.

  15. Diversity of 16S rRNA genes from bacteria of sugarcane rhizosphere soil

    Directory of Open Access Journals (Sweden)

    G. Pisa

    2011-12-01

    Full Text Available Sugarcane is an important agricultural product of Brazil, with a total production of more than 500 million tons. Knowledge of the bacterial community associated with agricultural crops and the soil status is a decisive step towards understanding how microorganisms influence crop productivity. However, most studies aim to isolate endophytic or rhizosphere bacteria associated with the plant by culture-dependent approaches. Culture-independent approaches allow a more comprehensive view of entire bacterial communities in the environment. In the present study, we have used this approach to assess the bacterial community in the rhizosphere soil of sugarcane at different times and under different nitrogen fertilization conditions. At the high taxonomic level, few differences between samples were observed, with the phylum Proteobacteria (29.6% predominating, followed by Acidobacteria (23.4%, Bacteroidetes (12.1%, Firmicutes (10.2%, and Actinobacteria (5.6%. The exception was the Verrucomicrobia phylum whose prevalence in N-fertilized soils was approximately 0.7% and increased to 5.2% in the non-fertilized soil, suggesting that this group may be an indicator of nitrogen availability in soils. However, at low taxonomic levels a higher diversity was found associated with plants receiving nitrogen fertilizer. Bacillus was the most predominant genus, accounting for 19.7% of all genera observed. Classically reported nitrogen-fixing and/or plant growth-promoting bacterial genera, such as Azospirillum, Rhizobium, Mesorhizobium, Bradyrhizobium, and Burkholderia were also found although at a lower prevalence.

  16. Cost-effective Chlorella biomass production from dilute wastewater using a novel photosynthetic microbial fuel cell (PMFC).

    Science.gov (United States)

    Ma, Jinxing; Wang, Zhiwei; Zhang, Junyao; Waite, T David; Wu, Zhichao

    2017-01-01

    While microalgae have been suggested as a promising substitute to conventional fossil fuels, their cost effective cultivation and harvesting constitutes a major challenge. In the work described here, a novel photosynthetic microbial fuel cell (PMFC) in which a stainless steel mesh with biofilm formed on it serves as both the cathode and filtration material has been developed. Results of this study reveal that, in addition to inducing oxygen reduction reactions under illumination, the biocathode is effective in preventing the washout of algae during continuous operation, resulting in retained biomass concentrations reaching 3.5-6.5 g L(-1). The maximum output current density reached ∼200 mA m(-2) under irradiation, which is comparable with recent PMFC studies. Microbial diversity analyses targeting 16S and 18S rRNA genes indicated that the eukaryotic species belonging to the genus Chlorella was able to sustain its community dominance (>96%) over other competing species over the course of the studies. In the absence of catalysts such as Pt, a consortium of photosynthetic organisms including plant growth-promoting bacteria such as Azospirillum and Rhizobium were overrepresented in the biofilm, with these organisms most likely contributing to cathodic electron transfer. Energy flow analysis suggested that the PMFC system held the potential to achieve theoretical energy balance in simultaneous algae production and wastewater treatment.

  17. Effects of water stress and inoculation with plant growth promoting rhizobacteria (PGPR on antioxidant status and photosynthetic pigments in basil (Ocimum basilicum L.

    Directory of Open Access Journals (Sweden)

    Mostafa Heidari

    2012-01-01

    Full Text Available Effects of water stress and inoculation with plant growth promoting rhizobacteria (PGPR on antioxidant activity and photosynthetic pigments were studied in basil plants. A field experiment was conducted at the University of Zabol in Iran during 2010 growing season. The experiment laid out as split plot based on randomized complete block design with three replications. Three levels of water stress W1 = 80 (control, W2 = 60 and W3 = 40% of the field capacity (FC as main plots and four levels of bacterial species consisting of S1 = Pseudomonades sp., S2 = Bacillus lentus, S3 = Azospirillum brasilens, S4 = combination of three bacterial species and S5 = control (without use of bacterial as sub plots. The results revealed that water stress caused a significant change in the antioxidant activity. The highest concentration CAT and GPX activity were in W3 treatments. By increasing water stress from control to W3, chlorophyll content in leaves was increased but Fv/Fm and APX activity decreased. Application of rhizobacteria under water stress improved the antioxidant and photosynthetic pigments in basil plants. S1 = Pseudomonades sp. under water stress, significantly increased the CAT enzyme activity, but the highest GPX and APX activity and chlorophyll content in leaves under water stress were in S4 = combination of three bacterial species.

  18. The Effect Different Fertilizers, on Germination, Yield, of Vicia vilosa Roth

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    R Kamaei

    2015-09-01

    Full Text Available In order to study the interaction of germination, yield of Vicia vilosa Roth to use of biological fertilizer, chemical, and manure, an experiment was conducted as a randomized complete block design with three replications at Research greenhouse, Faculty of Agriculture, Ferdowsi University of Mashhad, Iran, in 2013-2014 growing season. The experimental treatments was included three kinds of bio fertilizers and their integration with each other and vermicompost and chemical fertilizer as following : 1- mycorhhizaarbuscular species Glomus mosseae+vermicompost2- mycorhhiza+Nitrocsin (included bacteries Azospirillum sp. and Azotobacter sp. 3- mycorhhiza arbuscular+ Rhizobium (Rhizobium sp. 4-mycorhhiza arbuscular + Chemical fertilizer NPK 5- mycorhhizaarbuscular (Glomus moseae 6-control. The results showed that, although the treatments has not significant effects on height of stem , it has significant effects on characteristics of root length colonization percent, number the root node, Root dry weight, soggy yield, yield dry and protein Percent. The results showed that the highest percent of root length colonization(76 percent, number the root node (20, Root dry weight (.94 g, soggy yield (1894.5 g m-2, yield dry (473.63 g m-2 and protein Percent (27.33 percent was gained in integrated mycorhhiza and nitrocsine treatment. On the basis of results, the integration of mycrhhoriza and biological rhizobium is suggested as the best fertilizer treatment for Vicia vilosa Roth.

  19. Fluctuation of microbial activities after influent load variations in a full-scale SBR. Recovery of the biomass after starvation

    Energy Technology Data Exchange (ETDEWEB)

    Cabezas, Angela; Draper, Patricia; Etchebehere, Claudia [Universidad de la Republica, Montevideo (Uruguay). Catedra de Microbiologia, Facultad de Quimica y Facultad de Ciencias

    2009-10-15

    Due to variations in the production levels, a full-scale sequencing batch reactor (SBR) for post-treatment of tannery wastewater was exposed to low and high ammonia load periods. In order to study how these changes affected the N-removal capacity, the microbiology of the reactor was studied by a diverse set of techniques including molecular tools, activity tests, and microbial counts in samples taken along 3 years. The recover capacity of the biomass was also studied in a lab-scale reactor operated with intermittent aeration without feeding for 36 days. The results showed that changes in the feeding negatively affected the nitrifying community, but the nitrogen removal efficiencies could be restored after the concentration stress. Species substitution was observed within the nitrifying bacteria, Nitrosomonas europaea and Nitrobacter predominated initially, and after an ammonia overload period, Nitrosomonas nitrosa and Nitrospira became dominant. Some denitrifiers, with nirS related to Alicycliphilus, Azospirillum, and Marinobacter nirS, persisted during long-term reactor operation, but the community fluctuated both in composition and in abundance. This fluctuating community may better resist the continuous changes in the feeding regime. Our results showed that a nitrifying-denitrifying SBR could be operated with low loads or even without feeding during production shut down periods. (orig.)

  20. Detection of antibiotic-resistant bacteria endowed with antimicrobial activity from a freshwater lake and their phylogenetic affiliation

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    Zothanpuia

    2016-06-01

    Full Text Available Antimicrobial resistance poses a serious challenge to global public health. In this study, fifty bacterial strains were isolated from the sediments of a freshwater lake and were screened for antibiotic resistance. Out of fifty isolates, thirty-three isolates showed resistance against at least two of the selected antibiotics. Analysis of 16S rDNA sequencing revealed that the isolates belonged to ten different genera, namely Staphylococcus(n = 8, Bacillus(n = 7, Lysinibacillus(n = 4, Achromobacter(n=3, bacterium(n = 3, Methylobacterium(n = 2, Bosea(n = 2, Aneurinibacillus(n = 2, Azospirillum(n = 1, Novosphingobium(n = 1. Enterobacterial repetitive intergenic consensus (ERIC and BOX-PCR markers were used to study the genetic relatedness among the antibiotic resistant isolates. Further, the isolates were screened for their antimicrobial activity against bacterial pathogens viz., Staphylococcus aureus(MTCC-96, Pseudomonas aeruginosa(MTCC-2453 and Escherichia coli(MTCC-739, and pathogenic fungi viz., Fusarium proliferatum (MTCC-286, Fusarium oxysporum (CABI-293942 and Fusarium oxy. ciceri (MTCC-2791. In addition, biosynthetic genes (polyketide synthase II (PKS-II and non-ribosomal peptide synthetase (NRPS were detected in six and seven isolates, respectively. This is the first report for the multifunctional analysis of the bacterial isolates from a wetland with biosynthetic potential, which could serve as potential source of useful biologically active metabolites.

  1. Pyrosequencing reveals a core community of anodic bacterial biofilms in bioelectrochemical systems from China

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    Yong eXiao

    2015-12-01

    Full Text Available Bioelectrochemical systems (BESs are promising technologies for energy and product recovery coupled with wastewater treatment, and the core microbial community in electrochemically active biofilm in BESs remains controversy. In the present study, 7 anodic communities from 6 bioelectrochemical systems in 4 labs in southeast, north and south-central of China are explored by 454 pyrosequencing. A total of 251,225 effective sequences are obtained for 7 electrochemically active biofilm samples at 3% cutoff level. While Alpha-, Beta- and Gamma-proteobacteria are the most abundant classes (averaging 16.0-17.7%, Bacteroidia and Clostridia are the two sub-dominant and commonly shared classes. Six commonly shared genera i.e. Azospira, Azospirillum, Acinetobacter, Bacteroides, Geobacter, Pseudomonas and Rhodopseudomonas dominate the electrochemically active communities and are defined as core genera. A total of 25 OTUs with average relative abundance >0.5% were selected and designated as core OTUs, and some species relating to these OTUs have been reported electrochemically active. Furthermore, cyclic voltammetry and chronoamperometry tests show that two strains from Acinetobacter guillouiae and Stappia indica, bacteria relate to two core OTUs, are electrochemically active. Using randomly selected bioelectrochemical systems, the study presented extremely diverse bacterial communities in anodic biofilms, though, we still can suggest some potential microbes for investigating the electrochemical mechanisms in bioelectrochemical systems.

  2. Endophytes in commercial micropropagation - friend or foe?

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    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.

  3. Effects of rhizodeposition of non-transgenic and transplastomic tobaccos on the soil bacterial community.

    Science.gov (United States)

    Brusetti, Lorenzo; Rizzi, Aurora; Abruzzese, Alessandro; Sacchi, Gian Attilio; Ragg, Enzio; Bazzicalupo, Marco; Sorlini, Claudia; Daffonchio, Daniele

    2008-01-01

    The effect of root-released compounds of transplastomic tobacco (Nicotiana tabacum) on the soil bacterial community structure, and their potential to support horizontal gene transfer (HGT) to bacteria have been studied. Soil microcosms were exposed to root-released compounds collected from transplastomic and non-transgenic tobacco cultivars. Cluster analysis of automated ribosomal intergenic spacer analysis (ARISA) profiles of the soil bacterial community after 48 h incubation grouped the transgenic cultivar apart from the non-transgenic, indicating that it had a rhizodeposition pattern different from the parental plants. However, these differences were less than between the two non-transgenic tobacco cultivars studied. NMR characterization of the root-released compounds showed some differences in chemical fingerprinting pattern between the transplastomic and the parental cultivar. However, the effect on bacterial community structure was transient, and tended to disappear after 96 h of incubation. The potential of root-released compounds as a source of transforming DNA for bacteria was investigated by using four potential recipient species. No transformants were obtained following exposure of all the recipients to the root-released compounds. Root-released compounds amended to transgene donor DNA decreased the transformation frequency of Acinetobacter baylyi strain ADP1200, while Azospirillum, Agrobacterium, and Sinorhizobium strains failed to develop competence also in the presence of an external added transgene source. Detection of plastid sequences by PCR suggested that a very low amount of fragmented plastid donor DNA was present in the root-released compounds.

  4. Efecto del uso del suelo sobre rizobacterias fosfatosolubizadoras y diazotroficas en el distrito de riego del río zulia,norte de santander (colombia

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    Ronal Fernando-Cañon

    2009-07-01

    Full Text Available It was quantified the population of diazotrophic and phosphate solubilizer bacteria with the aim of determine the effect of the use of soils during different periods of intervention with culture of rice in three agro-ecological zones of the District of Irrigation of the Zulia river. The results showed that the management of the culture of rice in the different studied zones significantly influenced the population of microorganisms in all the studied culture media, excepting the semi-solid culture media JMV. The populations of the genera Azospirillum spp., Azotobacter spp., Beijerinckia spp., were influenced by the fertility of the soils in the zones of Buena Esperanza and Restauración compared with Limoncito, where the quantities of organic matter and nutritional escential elements were lower. It were obtained 28 isolations of entophytic, associative, free-living diazotrophic and phosphate solubilizer rhizobacteria according with their macroscopic characteristics in the culture media Batata, JMV, Ashby and nutritive, from the analysis of the population in the zones of Buena Esperanza, Restauración and Limoncito. These isolations were purified and preserved in sterilized saline solution (0.85% NaCl at 4°C in the Laboratory of Microbiology of the Colombian Agricultural Institute, ICA, with the purpose to be used in subsequent studies about their potential as biofertilizers in rice cultured soils of the department.

  5. ESTUDIO DE ALGUNOS GÉNEROS MICROBIANOS ASOCIADOS A DIFERENTES VARIEDADES DE TRIGO (Triticum aestivum L. EN SUELO FERRALÍTICO ROJO

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    Annia Hernández

    2002-01-01

    Full Text Available Se estudiaron los géneros bacterianos predominantes en la rizosfera de cuatro variedades de trigo (Triticum aestivum L. crecido en un suelo Ferralítico Rojo. También se evaluó la quimioatracción de algunas rizobacterias hacia los exudados radicales de este cultivo. Se utilizó el Modelo Espermosférico para el aislamiento y los tubos colectores y la cámara de quimiotaxis modificada para la evaluación del efecto quimiotáctico de los exudados radicales. Los resultados obtenidos demostraron que en las variedades Gang y Caeté, Pseudomonas es la población dominante, siendo Pseudomonas (Burkholderia cepacia la especie más fuertemente atraída por los exudados radicales a los 14 días y 40 minutos de exposición. En las variedades Culiacam y Antisama, es el género Azospirillum el predominante en la rizosfera, siendo esta especie la más fuertemente atraída por los exudados radicales a los 14 días y 60 minutos de exposición. Esta investigación permite la aplicación de biofertilizantes a base de los géneros microbianos predominantes en la rizosfera del trigo, potenciando su actividad sin alterar la biología del suelo.

  6. Microorganismos rizosféricos, potenciales antagonistas de Fusarium sp. causante de la pudrición radicular de maracuyá (Passiflora edulis Sims

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    Luisa Fernanda Quiroga-Rojas

    2012-07-01

    Full Text Available El cultivo de maracuyá (Passiflora edulis, de gran importancia económica para Colombia, actualmente es afectado por la enfermedad del marchitamiento vascular causado por Fusarium sp. lo que hace necesario la búsqueda de alternativas que permitan un control eficiente de esta enfermedad. Aislados de las bacterias Azotobacter spp., Azospirillum spp. y el hongo Trichoderma spp., fueron evaluados como potenciales biocontroladores de Fusarium sp. en pruebas in vitro e in vivo. Las pruebas de “test dual” evidenciaron que un aislado nativo de Trichoderma sp. y un producto comercial (Trichoderma lignorum, provocaron la inhibición del crecimiento micelial de Fusarium sp. entre 94.2% y 93.6%, respectivamente. La evaluación de aislados de Trichoderma sobre plántulas de maracuyá en tres momentos de aplicación indicó que la inoculación previa disminuyó el porcentaje de infección de las plantas entre un 75 y 50%, mientras que con aplicaciones después o simultáneamente con el patógeno, el porcentaje de infección disminuyó en 25%. Estos resultados indican que la aplicación de organismos de biocontrol en semillas pregerminadas mejora la protección de las plantas contra el fitopatógeno estudiado y son un recurso importante en el manejo preventivo de las enfermedades de maracuyá.

  7. Effect of florfenicol on performance and microbial community of a sequencing batch biofilm reactor treating mariculture wastewater.

    Science.gov (United States)

    Gao, Feng; Li, Zhiwei; Chang, Qingbo; Gao, Mengchun; She, Zonglian; Wu, Juan; Jin, Chunji; Zheng, Dong; Guo, Liang; Zhao, Yangguo; Wang, Sen

    2017-03-16

    The effects of florfenicol (FF) on the performance, microbial activity and microbial community of a sequencing batch biofilm reactor (SBBR) were evaluated in treating mariculture wastewater. The chemical oxygen demand (COD) and nitrogen removal were inhibited at high FF concentrations. The specific oxygen utilization rate (SOUR), specific ammonium oxidation rate (SAOR), specific nitrite oxidation rate (SNOR) and specific nitrate reduction rate (SNRR) were decreased with an increase in the FF concentration from 0 to 35 mg/L. The chemical compositions of loosely bound extracellular polymeric substances (LB-EPS) and tightly bound EPS (TB-EPS) could be affected with an increase in the FF concentration. The high-throughput sequencing indicated some obvious variations in the microbial community at different FF concentrations. The relative abundance of Nitrosomonas and Nitrospira showed a decreasing tendency with an increase in the FF concentration, suggesting that FF could affect the nitrification process of SBBR. Some genera capable of reducing nitrate to nitrogen gas could be inhibited by the addition of FF in the influent, such as Azospirillum and Hyphomicrobium.

  8. Bacterial communities associated with the lichen symbiosis.

    Science.gov (United States)

    Bates, Scott T; Cropsey, Garrett W G; Caporaso, J Gregory; Knight, Rob; Fierer, Noah

    2011-02-01

    Lichens are commonly described as a mutualistic symbiosis between fungi and "algae" (Chlorophyta or Cyanobacteria); however, they also have internal bacterial communities. Recent research suggests that lichen-associated microbes are an integral component of lichen thalli and that the classical view of this symbiotic relationship should be expanded to include bacteria. However, we still have a limited understanding of the phylogenetic structure of these communities and their variability across lichen species. To address these knowledge gaps, we used bar-coded pyrosequencing to survey the bacterial communities associated with lichens. Bacterial sequences obtained from four lichen species at multiple locations on rock outcrops suggested that each lichen species harbored a distinct community and that all communities were dominated by Alphaproteobacteria. Across all samples, we recovered numerous bacterial phylotypes that were closely related to sequences isolated from lichens in prior investigations, including those from a lichen-associated Rhizobiales lineage (LAR1; putative N(2) fixers). LAR1-related phylotypes were relatively abundant and were found in all four lichen species, and many sequences closely related to other known N(2) fixers (e.g., Azospirillum, Bradyrhizobium, and Frankia) were recovered. Our findings confirm the presence of highly structured bacterial communities within lichens and provide additional evidence that these bacteria may serve distinct functional roles within lichen symbioses.

  9. Bacterias solubilizadoras de fosfato y sus potencialidades de uso en la promoción del crecimiento de cultivos de importancia económica

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    Gloria María Restrepo-Franco

    2015-01-01

    Full Text Available La mayoría de los suelos tropicales y subtropicales son deficientes en fósforo biodisponible, por lo que el empleo de bacterias promotoras del crecimiento vegetal, principalmente las solubilizadoras de fosfato, pueden reducir el uso de fertilizantes químicos. El objetivo de este trabajo es ofrecer una panorámica actual acerca de los principales géneros y mecanismos de acción de las bacterias solubilizadoras de fosfatos (BSF, así como analizar su posible empleo en cultivos de importancia económica. La utilización de géneros bacterianos con mayores potencialidades de uso como Pseudomonas y Bacillus unido a aislados promisorios de Azospirillum y Herbaspirilum en cultivos como arroz y café permitiría reducir a largo plazo el uso de productos químicos en la agricultura, así como desarrollar estrategias agronómicas que preserven el medio ambiente. El éxito de estos inoculantes bacterianos depende de la selección de cepas autóctonas eficientes por tipo de suelo, su capacidad de colonizar la rizosfera y de mantener la actividad biológica.

  10. Isolation of bacterial strains from bovine fecal microflora capable of degradation of ceftiofur.

    Science.gov (United States)

    Rafii, Fatemeh; Williams, Anna J; Park, Miseon; Sims, Lillie M; Heinze, Thomas M; Cerniglia, Carl E; Sutherland, John B

    2009-10-20

    Ceftiofur, a third-generation cephalosporin used to treat bacterial infections in animals, is degraded in bovine feces but the specific bacteria involved are unknown. To find the bacteria involved in ceftiofur metabolism, the bovine fecal microflora was screened. Twenty-one nonidentical strains of bovine fecal bacteria were isolated on media containing 1-32 microg ml(-1) of ceftiofur. The cultures were incubated with 5 microg ml(-1) ceftiofur for different times, then centrifuged and analyzed by high-performance liquid chromatography. Three strains of Bacillus spp., two strains of Roseomonas spp., and one strain of Azospirillum sp. metabolized 5 microg ml(-1) ceftiofur in broth cultures in less than 24h; ten other strains of Roseomonas and one strain of Bacillus pumilus had metabolized it by 120 h. After the ceftiofur had been metabolized by these bacteria, the filter-sterilized supernatants of centrifuged cultures no longer inhibited the growth of a ceftiofur-sensitive strain of Kocuria rhizophila, which indicated that ceftiofur had been transformed to compounds without bactericidal activity. Each isolate was also found to be able to grow in the presence of other beta-lactams, and a nitrocefin assay showed beta-lactamase activity in the 17 strains that metabolized ceftiofur. The results show that some beta-lactamase-producing bacteria from the bovine fecal microflora are capable of transforming ceftiofur to metabolites lacking bactericidal activity.

  11. A Metagenomic Study Highlights Phylogenetic Proximity of Quorum-Quenching and Xenobiotic-Degrading Amidases of the AS-Family

    Science.gov (United States)

    Tannières, Mélanie; Beury-Cirou, Amélie; Vigouroux, Armelle; Mondy, Samuel; Pellissier, Franck; Dessaux, Yves; Faure, Denis

    2013-01-01

    Quorum-sensing (QS) signals of the N-acylhomoserine lactone (NAHL) class are cleaved by quorum-quenching enzymes, collectively named NAHLases. Here, functional metagenomics allowed the discovery of a novel bacterial NAHLase in a rhizosphere that was treated with γ-caprolactone. As revealed by rrs-DGGE and rrs-pyrosequencing, this treatment increased the percentage of the NAHL-degrading bacteria and strongly biased the structure of the bacterial community, among which Azospirillum dominated. Among the 29 760 fosmids of the metagenomic library, a single one was detected that expressed the qsdB gene conferring NAHL-degradation upon E. coli and decreased QS-regulated virulence in Pectobacterium. Phylogenetic analysis of the 34 orfs of the fosmid suggested that it would belong to an unknown Proteobacterium - probably a γ-proteobacterium. qPCR quantification of the NAHLase-encoding genes attM, qsdA, and qsdB revealed their higher abundance in the γ-caprolactone-treated rhizosphere as compared to an untreated control. The purified QsdB enzyme exhibited amidase activity. QsdB is the first amidase signature (AS) family member exhibiting NAHLase-activity. Point mutations in the AS-family catalytic triad K-S-S abolished the NAHLase activity of QsdB. This study extends the diversity of NAHLases and highlights a common phylogenic origin of AS-family enzymes involved in the degradation of natural compounds, such as NAHLs, and xenobiotics, such as nylon and linuron. PMID:23762380

  12. Aislamiento e identificación de microorganismos con potencial biofertilizante de suelos arroceros del distrito de riego del rio zulia, norte de santander

    Directory of Open Access Journals (Sweden)

    Marilyn Tatiana Santos-Torres

    2006-07-01

    Full Text Available It was evaluated the microbial population of soils cultivated with rice at some zones of the district of irrigation of Zulia River, Norte de Santander state, Colombia, and no cultivated soil with rice. It was found that the seeded soil with rice were significantly low in fungi, bacteria and actinomycete populations compa red with no seeded with rice. This situation was due to possibly excessive use of agricultural machine and excessive application of pesticides and continuous rice monoculture. Therefore, with use the biofertilizer enriched with microorganism isolated from zone with similar agroecological conditions, will improve the microorganism adaptation of the rizosphere and will promote a reestablishment of the biological balance of soils and therefore to increase biotic activity improving the quality and productivity of the cultures. A total of 43 bacteria and 4 fungi were isolated in different soils of the studied zone, and were identified 25 microorganisms. According the isolation methodology, the bacteria were separate in two groups: diazotrophic like the species of the genera Azotobacter sp., Azospirillum sp. and Beijerinckia sp. and the phosphate solubilizate inorganic like Bacillus megaterium, B. cereus, Pseudomonas putida and P. aeruginosa. For the fungi identification were evaluate the micro and macroscopic characteristics in the culture medium Czapeck, following the fungi keys of Samson Hoekstra and Oorsehot (1981, which allowed recognize species like Trichoderma sp., Aspergillus niger, Penicillium purpurogenum, P. pinopilum.

  13. The Effects of Winter Cover Crops and Plant Growth Promoting Rhizobacteria on some Soil Fertility Aspects and Crop Yield in an Organic Production System of Ocimum basilicum L.

    Directory of Open Access Journals (Sweden)

    M Jahan

    2014-03-01

    Full Text Available Sustainable agriculture systems emphasized on the on-farm inputs likes use of biofertilizers, crop rotation and cover crops. This experiment was conducted in a split plots arrangement with two factors based on randomized complete block design with three replications during years 2009-2010, at Research Farm of Ferdowsi University of Mashhad. The main factor consisted of cultivation and no cultivation of cover crops in autumn. The sub factor was biofertilizer application with four levels, included 1-Nitroxin® (containing Azotobacter spp. and Azospirillum spp., 2-Biophosphorous® (Bacillus sp. and Pseudomonas sp., 3-Nitroxin® + Biophosphorous® and 4-Control. Results showed that most characteristics, e.g. seed yield and harvest index were increased with no cover crop cultivation. However in control treatment, the biological yield, seed yield and harvest index were more than biofertilizeres treatments, as Nitroxin® and Biophosphorous® ranked after the control. Amongst the biofertilizers, Biophosphorus® had the most positive effects. The maximum grain weight was obtained from Nitroxin®+ Biophosphorous® treatment. The interaction effects of biofertilizer and cover crops were significant among some characteristics. The results showed that the interaction between biofertilizers and no cover crop cultivation was significant, as use of the biofertilizers especially Nitroxin® and Biophosphorous® in no cover crop condition increased the amounts of biological yield and seed yield.

  14. Interactions between diazotrophic bacteria and mycorrhizal fungus in maize genotypes Interações entre bactérias diazotróficas e fungo micorrízico em genótipos de milho

    Directory of Open Access Journals (Sweden)

    Marina Yumi Horta Miyauchi

    2008-01-01

    Full Text Available Some diazotrophic bacteria can fix nitrogen biologically in gramineous host plants. Generally, gramineous plants are also associated with mycorrhizal fungi, that can improve mainly plant P uptake. Among the factors affecting plant-microbe interactions, the plant genotype plays an important role. This study evaluates the effect of diazotrophic bacteria and an arbuscular mycorrhizal fungus (AMF, on five genotypes of maize (Zea mays L., in relation to plant biomass, shoot N and P concentrations, and fine root morphological traits. The experimental design was entirely randomized in a factorial 5 × 4 × 2 arrangement, i.e., five maize genotypes (hybrids C333B, AS3466, and PREMIUM, and the inbreed lines lg40897-1 and lg40505-1, three diazotrophic bacteria (Azospirillum lipoferum, A. amazonense, and Burkholderia sp. in addition to a control without bacterial inoculation, co-inoculated or not with the AMF Glomus clarum. The non-mycorrhizal plants inoculated with Azospirillum exhibited the highest N concentrations. The lines lg40897-1 and lg40505-1 showed higher P concentrations as compared to the hybrids, mainly when colonized by AMF. The higher levels of mycorrhizal colonization (90% occurred in the C333B and lg40897-1 genotypes, which also exhibited a greater root diameter. Mycorrhiza increased shoot and root biomass, besides root traits as total length, specific length, total surface, and incidence of root hairs in all genotypes. In addition, mycorrhiza also stimulated the root colonization by diazotrophic bacteria. The bacteria did not affect root morphological traits and mycorrhizal colonization.Algumas bactérias diazotróficas podem fixar N biologicamente em gramíneas, as quais se associam a fungos micorrízicos, o que pode levar a um aumento principalmente da absorção de P. Dentre os fatores que afetam as interações planta-microrganismos, o genótipo da planta tem importante papel. Esse trabalho avalia o efeito de bactérias diazotr

  15. Plant-microbe interactions promoting plant growth and health: perspectives for controlled use of microorganisms in agriculture.

    Science.gov (United States)

    Berg, Gabriele

    2009-08-01

    Plant-associated microorganisms fulfill important functions for plant growth and health. Direct plant growth promotion by microbes is based on improved nutrient acquisition and hormonal stimulation. Diverse mechanisms are involved in the suppression of plant pathogens, which is often indirectly connected with plant growth. Whereas members of the bacterial genera Azospirillum and Rhizobium are well-studied examples for plant growth promotion, Bacillus, Pseudomonas, Serratia, Stenotrophomonas, and Streptomyces and the fungal genera Ampelomyces, Coniothyrium, and Trichoderma are model organisms to demonstrate influence on plant health. Based on these beneficial plant-microbe interactions, it is possible to develop microbial inoculants for use in agricultural biotechnology. Dependent on their mode of action and effects, these products can be used as biofertilizers, plant strengtheners, phytostimulators, and biopesticides. There is a strong growing market for microbial inoculants worldwide with an annual growth rate of approximately 10%. The use of genomic technologies leads to products with more predictable and consistent effects. The future success of the biological control industry will benefit from interdisciplinary research, e.g., on mass production, formulation, interactions, and signaling with the environment, as well as on innovative business management, product marketing, and education. Altogether, the use of microorganisms and the exploitation of beneficial plant-microbe interactions offer promising and environmentally friendly strategies for conventional and organic