GuaB Activity Is Required in Rhizobium tropici During the Early Stages of Nodulation of Determinate Nodules but Is Dispensable for the Sinorhizobium meliloti–Alfalfa Symbiotic Interaction

The guaB mutant strain Rhizobium tropici CIAT8999-10T is defective in symbiosis with common bean, forming nodules that lack rhizobial content. In order to investigate the timing of the guaB requirement during the nodule formation on the host common bean by the strain CIAT899-10.T, we constructed gene fusions in which the guaB gene is expressed under the control of the symbiotic promoters nodA, bacA, and nifH. Our data indicated that the guaB is required from the early stages of nodulation because full recovery of the wild-type phenotype was accomplished by the nodA-guaB fusion. In addition, we have constructed a guaB mutant derived from Sinorhizobium meliloti 1021, and shown that, unlike R. tropici, the guaB S. meliloti mutant is auxotrophic for guanine and induces wild-type nodules on alfalfa and Medicago truncatula. The guaB R. tropici mutant also is defective in its symbiosis with Macroptilium atropurpureum and Vigna unguiculata but normal with Leucaena leucocephala. These results show that the requirement of the rhizobial guaB for symbiosis is found to be associated with host plants that form determinate type of nodules.Fil: Collavino, Mónica Mariana. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Riccillo, Pablo M.. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; ArgentinaFil: Grasso, Daniel Horacio. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Crespi, Martín. Centre National de la Recherche Scientifique; FranciaFil: Aguilar, Orlando Mario. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentin

Nodulation competitiveness and diversification of symbiosis genes in common beans from the American centers of domestication

Phaseolus vulgaris (common bean), having a proposed Mexican origin within the Americas, comprises three centers of diversification: Mesoamerica, the southern Andes, and the Amotape-Huancabamba Depression in Peru-Ecuador. Rhizobium etli is the predominant rhizobium found symbiotically associated with beans in the Americasalthough closely related Rhizobium phylotypes have also been detected. To investigate if symbiosis between bean varieties and rhizobia evolved affinity, firstly nodulation competitiveness was studied after inoculation with a mixture of sympatric and allopatric rhizobial strains isolated from the respective geographical regions. Rhizobia strains harboring nodC types α and γ , which were found predominant in Mexico and Ecuador, were comparable in nodule occupancy at 50% of each in beans from the Mesoamerican and Andean gene pools, but it is one of those two nodC types which clearly predominated in Ecuadorian-Peruvian beans as well as in Andean beans nodC type γ predominated the sympatric nodC type δ. The results indicated that those beans from Ecuador-Peru and Andean region, respectively exhibited no affinity for nodulation by the sympatric rhizobial lineages that were found to be predominant in bean nodules formed in those respective areas. Unlike the strains isolated from Ecuador, Rhizobium etli isolated from Mexico as well from the southern Andes was highly competitive for nodulation in beans from Ecuador-Peru, and quite similarly competitive in Mesoamerican and Andean beans. Finally, five gene products associated with symbiosis were examined to analyze variations that could be correlated with nodulation competitiveness. A small GTPase RabA2, transcriptional factors NIN and ASTRAY, and nodulation factor receptors NFR1 and NFR5- indicated high conservation but NIN, NFR1 and NFR5 of beans representative of the Ecuador-Peru genetic pool clustered separated from the Mesoamerican and Andean showing diversification and possible different interaction. These results indicated that both host and bacterial genetics are important for mutual affinity, and that symbiosis is another trait of legumes that could be sensitive to evolutionary influences and local adaptation.Fil: Aguilar, Orlando Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Collavino, Mónica Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Mancini Villagra, Ulises Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentin

Analysis of nifH-RNA reveals phylotypes related to Geobacter and Cyanobacteria as important functional components of the N2-fixing community depending on depth and agricultural use of soil

In this survey, a total of 80 787 reads and 28 171 unique NifH protein sequences were retrieved from soil RNA. This dataset extends our knowledge about the structure and diversity of the functional diazotrophic communities in agricultural soils from Argentinean Pampas. Operational taxonomic unit (OTU)-based analyses showed that nifH phylotypes related to Geobacter and Anaeromyxobacter (44.8%), Rhizobiales (29%), Cyanobacteria (16.7%), and Verrucomicrobiales (8%) are key microbial components of N2 fixation in soils associated with no-till management and soil depth. In addition, quantification of nifH gene copies related to Geobacter and Cyanobacteria revealed that these groups are abundant in soils under maize-soybean rotation and soybean monoculture, respectively. The correlation of physicochemical soil parameters with the diazotrophic diversity and composition showed that soil stability and organic carbon might contribute to the functional signatures of particular nifH phylotypes in fields under no-till management. Because crop production relies on soil-borne microorganism´s activities, such as free N2 fixation, the information provided by our study on the diazotrophic population dynamics, associated with the edaphic properties and land-use practices, represents a major contribution to gain insight into soil biology, in which functionally active components are identified.Fil: Calderoli, Priscila Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Collavino, Mónica Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Behrends Kraemer, Filipe. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Suelos; ArgentinaFil: Morrás, Héctor José María. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Suelos; ArgentinaFil: Aguilar, Orlando Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentin

Composición de las comunidades de rizobios asociados a la especie Prosopis ruscifolia

Las adaptaciones de las plantas con sus microorganismos asociados hacen factible la reutilización y revegetación de tierras salinas. Entre las especies pioneras del parque chaqueño, las pertenecientes al género Prosopis crecen en una gran variabilidad de ambientes. P. ruscifolia es una leguminosa nativa de la región chaqueña semiárida, leñosa, que se destaca por su elevada tolerancia a la salinidad, al estrés hídrico y por su capacidad de adaptarse a suelos degradados. Las plantas leguminosas tienen la capacidad de formar una asociación simbiótica para la fijación de nitrógeno con bacterias de la familia Rhizobiaceae, denominadas comúnmente rizobios. El objetivo de este trabajo fue aislar e identificar los rizobios que están asociados simbióticamente a Prosopis ruscifolia. El muestreo se realizó en suelos de vinales de la localidad de Basail, provincia de Chaco, debajo del dosel de 5 árboles de la especie. Por cada árbol se tomó una muestra compuesta de 4 submuestas. La recuperación de los rizobios se realizó utilizando P. ruscifolia como planta trampa. Para ello, 10 semillas escarificadas y esterilizadas se sembraron en macetas con cada muestra de suelo, las mismas fueron mantenidas con riego con agua esterilizada. A los 120 días se procedió a descalzar las plantas y se colectaron todos los nódulos. Los mismos fueron esterilizados y sembrados en medio de extracto de levadura-manitol (YEM) para el aislamiento de los rizobios. Se clasificaron y seleccionarán las diferentes colonias considerando las características que presentan los rizobios crecidos en este medio, velocidad de crecimiento, la producción de ácido o base, y características macroscópicas de las colonias (color, consistencia, tamaño y forma). Los aislamientos que cumplieron con las características esperadas fueron purificados, multiplicados y conservados a -70 ºC. La diversidad genómica de estos aislamientos fue evaluada a través de la amplificación de las regiones genómicas repetitivas (ERIC-PCR). Un clón de cada perfil ERIC-PCR fue analizado por amplificación de los genes 16S ARNr y de nodulación nodC. En los casos que la amplificación resultó negativa para el gen nodC se ensayó la amplificación del gen nodD. Se encontraron 84 aislamientos con características morfotípicas de rizobios, 33 de ellos de crecimiento lento y 51 de crecimiento rápido. El análisis de ERIC-PCR agrupó los aislamientos rápidos en 23 perfiles, mientras que los aislamientos lentos se agruparon en 10 perfiles diferentes. Encontramos que todos los grupos de crecimiento lento amplificaron el gen nodC, mientras que de los 23 grupos ERIC de crecimiento rápido, 9 amplificaron el gen nodC y 4 el gen nodD. Concluyendo, se obtuvo una colección de 33 aislamientos diferentes colectadas de nódulos de P. rucifolia los cuales presentaron una alta variabilidad fenotípica y genotípica. Se observaron tanto cepas de crecimiento lento como de crecimiento rápido, con mayor abundancia y diversidad de estas últimas. Asimismo, la potencialidad de estas cepas de nodular plantas leguminosas fue confirmada por la amplificación del gen nodC en el 100% de los aislamientos lentos y en el 56% de los aislamientos rápidos.Fil: Sotelo, Cristina Esther. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Collavino, Mónica Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Perticari, Alejandro. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis; ArgentinaXXVII Congreso Argentino de la ciencia del SueloArgentinaAsociación Argentina Ciencia del SueloUniversidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Instituto Nacional de Tecnología Agropecuari

GuaB activity is required in Rhizobium tropici during the early stages of nodulation of determinate nodules but is dispensable for the sinorhizobium meliloti-alfalfa symbiotic interaction

The guaB mutant strain Rhizobium tropici CIAT8999-10T is defective in symbiosis with common bean, forming nodules that lack rhizobial content. In order to investigate the timing of the guaB requirement during the nodule formation on the host common bean by the strain CIAT899-10.T, we constructed gene fusions in which the guaB gene is expressed under the control of the symbiotic promoters nodA, bacA, and nifH. Our data indicated that the guaB is required from the early stages of nodulation because full recovery of the wild-type phenotype was accomplished by the nodA-guaB fusion. In addition, we have constructed a guaB mutant derived from Sinorhizobium meliloti 1021, and shown that, unlike R. tropici, the guaB S. meliloti mutant is auxotrophic for guanine and induces wild-type nodules on alfalfa and Medicago truncatula. The guaB R. tropici mutant also is defective in its symbiosis with Macroptilium atropurpureum and Vigna unguiculata but normal with Leucaena leucocephala. These results show that the requirement of the rhizobial guaB for symbiosis is found to be associated with host plants that form determinate type of nodules.Instituto de Biotecnologia y Biologia Molecula

Analysis of nifH-RNA reveals phylotypes related to Geobacter and Cyanobacteria as important functional components of the N₂-fixing community depending on depth and agricultural use of soil

In this survey, a total of 80 787 reads and 28 171 unique NifH protein sequences were retrieved from soil RNA. This dataset extends our knowledge about the structure and diversity of the functional diazotrophic communities in agricultural soils from Argentinean Pampas. Operational taxonomic unit (OTU)-based analyses showed that nifH phylotypes related to Geobacter and Anaeromyxobacter (44.8%), Rhizobiales (29%), Cyanobacteria (16.7%), and Verrucomicrobiales (8%) are key microbial components of N2 fixation in soils associated with no-till management and soil depth. In addition, quantification of nifH gene copies related to Geobacter and Cyanobacteria revealed that these groups are abundant in soils under maize–soybean rotation and soybean monoculture, respectively. The correlation of physicochemical soil parameters with the diazotrophic diversity and composition showed that soil stability and organic carbon might contribute to the functional signatures of particular nifH phylotypes in fields under no-till management. Because crop production relies on soil-borne microorganism's activities, such as free N2 fixation, the information provided by our study on the diazotrophic population dynamics, associated with the edaphic properties and land-use practices, represents a major contribution to gain insight into soil biology, in which functionally active components are identified.Instituto de Biotecnologia y Biologia Molecula

Comparison of in vitro solubilization activity of diverse phosphate-solubilizing bacteria native to acid soil and their ability to promote Phaseolus vulgaris growth

To identify plant growth promotion ability of phosphorus-solubilizing native bacteria, we have examined a collection of isolates representing the diversity of culturable phosphate-solubilizing bacteria from acid soils of the northeast of Argentina. Assays in growth medium supplemented with tricalcium phosphate revealed different phosphorus solubilization activity and temporal patterns of solubilization. Acidification of the broth medium coincided with phosphorus solubilization. The isolates were grouped according to their Rep fingerprinting profiles and phylogenetically classified by 16S rDNA and biochemical analyses. These isolates were assigned to the genera Enterobacter, Pantoea, Pseudomonas, Acinetobacter, Burkholderia, and Exiguobacterium. Four isolates showing high phosphorus solubilizing activity in in vitro assays were inoculated on common beans (Phaseolus vulgaris); some of them promoted plant growth and increased photosynthesis and the P and N content of leaves. The results indicated that the ability to in vitro solubilize P is not necessarily associated to the promotion of plant growth.Instituto de Biotecnología y Biología Molecula

GuaB activity is required in Rhizobium tropici during the early stages of nodulation of determinate nodules but is dispensable for the sinorhizobium meliloti-alfalfa symbiotic interaction

The guaB mutant strain Rhizobium tropici CIAT8999-10T is defective in symbiosis with common bean, forming nodules that lack rhizobial content. In order to investigate the timing of the guaB requirement during the nodule formation on the host common bean by the strain CIAT899-10.T, we constructed gene fusions in which the guaB gene is expressed under the control of the symbiotic promoters nodA, bacA, and nifH. Our data indicated that the guaB is required from the early stages of nodulation because full recovery of the wild-type phenotype was accomplished by the nodA-guaB fusion. In addition, we have constructed a guaB mutant derived from Sinorhizobium meliloti 1021, and shown that, unlike R. tropici, the guaB S. meliloti mutant is auxotrophic for guanine and induces wild-type nodules on alfalfa and Medicago truncatula. The guaB R. tropici mutant also is defective in its symbiosis with Macroptilium atropurpureum and Vigna unguiculata but normal with Leucaena leucocephala. These results show that the requirement of the rhizobial guaB for symbiosis is found to be associated with host plants that form determinate type of nodules.Instituto de Biotecnologia y Biologia Molecula

Analysis of nifH-RNA reveals phylotypes related to Geobacter and Cyanobacteria as important functional components of the N₂-fixing community depending on depth and agricultural use of soil

In this survey, a total of 80 787 reads and 28 171 unique NifH protein sequences were retrieved from soil RNA. This dataset extends our knowledge about the structure and diversity of the functional diazotrophic communities in agricultural soils from Argentinean Pampas. Operational taxonomic unit (OTU)-based analyses showed that nifH phylotypes related to Geobacter and Anaeromyxobacter (44.8%), Rhizobiales (29%), Cyanobacteria (16.7%), and Verrucomicrobiales (8%) are key microbial components of N2 fixation in soils associated with no-till management and soil depth. In addition, quantification of nifH gene copies related to Geobacter and Cyanobacteria revealed that these groups are abundant in soils under maize–soybean rotation and soybean monoculture, respectively. The correlation of physicochemical soil parameters with the diazotrophic diversity and composition showed that soil stability and organic carbon might contribute to the functional signatures of particular nifH phylotypes in fields under no-till management. Because crop production relies on soil-borne microorganism's activities, such as free N2 fixation, the information provided by our study on the diazotrophic population dynamics, associated with the edaphic properties and land-use practices, represents a major contribution to gain insight into soil biology, in which functionally active components are identified.Instituto de Biotecnologia y Biologia Molecula

nifH pyrosequencing reveals the potential for location-specific soil chemistry to influence N2-fixing community dynamics

A dataset of 87 020 nifH reads and 16 782 unique nifH protein sequences obtained over 2 years from four locations across a gradient of agricultural soil types in Argentina were analysed to provide a detailed and comprehensive picture of the diversity, abundance and responses of the N2-fixing community in relation to differences in soil chemistry and agricultural practices. Phylogenetic analysis revealed an expected high proportion of Alphaproteobacteria, Betaproteobacteria and Deltaproteobacteria, mainly relatives to Bradyrhizobium and Methylosinus/Methylocystis, but a surprising paucity of Gammaproteobacteria. Analysis of variance and stepwise regression modelling suggested location and treatment-specific influences of soil type on diazotrophic community composition and organic carbon concentrations on nifH diversity. nifH gene abundance, determined by quantitative real-time polymerase chain reaction, was higher in agricultural soils than in non-agricultural soils, and was influenced by soil chemistry under intensive crop rotation but not under monoculture. At some locations, sustainable increased crop yields might be possible through the management of soil chemistry to improve the abundance and diversity of N2-fixing bacteriaFil: Collavino, Mónica Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Tripp, H. James. University of California. Department of Ocean Sciences; Estados UnidosFil: Frank, Ildiko E.. University of California. Department of Ocean Sciences; Estados UnidosFil: Vidoz, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); ArgentinaFil: Calderoli, Priscila Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Donato, Mariano Humberto. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Laboratorio de Sistemática y Biología Evolutiva; ArgentinaFil: Zehr, Jonathan P.. University of California. Department of Ocean Sciences; Estados UnidosFil: Aguilar, Orlando Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Biotecnología y Biología Molecular; Argentin