Effects of co-inoculation of native Rhizobium and Pseudomonas strains on growth parameters and yield of two contrasting Phaseolus vulgaris L. genotypes under Cuban soil conditions

Sánchez, Ariany Colás; Gutiérrez, Roldán Torres; Santana, René Cupull; Urrutia, Alianny Rodríguez; Fauvart, Maarten; Michiels, Jan; Vanderleyden, Jos

doi:10.1016/j.ejsobi.2014.03.004

Cited by 81 publications

(45 citation statements)

References 45 publications

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“…Thus, in many cases, it is not possible to quantitatively associate the failure or success of plant growth promotion achieved with the inoculated bacterial population at a strain-specific resolution, leaving the outcome of the inoculation unexplained (17). Furthermore, the crop response to inoculation under field conditions heavily depends on the combination of the plant genotype and the bacterial strain (18)(19)(20), stressing the need for methodologies to evaluate the success of plant colonization accurately at a high resolution. Previously, we used whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) …”

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confidence: 99%

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

Stets

Alquéres

Souza

et al. 2015

Appl Environ Microbiol

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bAzospirillum 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 N 2 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 (ϳ10 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.A zospirillum is one of the most important genera of plant growth-promoting rhizobacteria found worldwide under a variety of environmental and soil conditions (1). The diazotroph Azospirillum brasilense is the best-studied species of the genus, is found in close association with many agriculturally important crops, and exerts beneficial effects on plant growth and productivity (2-4). Nitrogen fixation (5, 6) and the production of the auxin 3-indoleacetic acid (IAA) by many representatives of the genus Azospirillum are related to the growth promotion effects observed in inoculated plants, such as increases in root length and the numbers of root hairs and lateral roots (3).The biotechnological use of A. brasilense inoculants in Latin American and in Brazil, in particular, has increased in recent years (7). Strain FP2 is a spontaneous mutant of A. brasilense Sp7 (8). Strain Sp7 has been shown to be capable of stimulating the growth of several members of the family Poaceae and increasing the productivities of wheat and maize crops (2). Strain FP2 can also promote the growth of wheat (9) and enhance maize and wheat productivity under field conditions (unpublished data). Most of the A. brasilense inoculants in B...

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mentioning

confidence: 99%

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

Stets

Alquéres

Souza

et al. 2015

Appl Environ Microbiol

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“…In a previous study, we demonstrated that plants most susceptible to infection and capable of producing effective nodules should have greater potential to fix more atmospheric nitrogen (Sánchez et al, 2014). At 21 days after inoculation, all isolates were able to nodulate the host plant in the pot experiment under greenhouse conditions.…”

Section: Resultsmentioning

confidence: 99%

“…The isolation methodology proposed by Sánchez et al (2014) was used with minimal modification. Briefly, individual nodules were dissected from the roots using a flame sterilized scalpel and tweezers and were washed thoroughly in distilled water using a sieve to remove all traces of soil.…”

Section: Sample Preparation and Bacterial Isolationmentioning

confidence: 99%

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Genetic and phenotypic diversity of Rhizobium isolates from Southern Ecuador

et al. 2017

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Rhizobium-legume symbioses play relevant roles in agriculture but have not been well studied in Ecuador. The aim of this study was to characterize the genetic and phenotypic diversity of Rhizobium isolates associated with Phaseolus vulgaris from southern Ecuador. Morphocultural characterization, biochemical tests and physiological analyses were conducted to authenticate and determine the diversity of bacteria Rhizobium-like isolates. The genetic diversity of the isolates was determined by molecular techniques, which consisted of bacteria DNA extraction and amplification and sequencing of the 16S rRNA gene. The nodulation parameters and nitrogen fixation for P. vulgaris under greenhouse conditions were also assessed to determine the phenotypic diversity among isolates. Furthermore, bacteria indole-acetic-acid production was evaluated by the colorimetric method. Morpho-cultural and biochemical characteristic assessments demonstrated that Rhizobium-like bacteria was associated with the P. vulgaris nodules. The diversity among the isolates, as determined by physiological analyses, revealed the potential of several isolates to grow at different pH values, salinity conditions and temperatures. Partial sequencing of the 16S rRNA gene identified the Rhizobium genus in every sampling site. From a total of 20 aligned sequences, nine species of Rhizobium were identified. Nodule formation and biomass, as well as nitrogen fixation, showed an increase in plant phenotypic parameters, which could be influenced by IAA production, especially for the strains R. mesoamericanum NAM1 and R. leguminosarum bv. viciae COL6. These results demonstrated the efficiency of native symbiotic diazotrophic strains inoculants for legume production. This work can serve as the basis for additional studies of native Rhizobium strains and to help spread the use of biofertilizers in Ecuadorian fields.Index terms: Phaseolus vulgaris; diazotrophic bacteria; 16S rRNA gene; nodulation; indole acetic acid. RESUMOA simbiose Rhizobium-leguminosa desempenha um relevante papel na agricultura, entretanto não tem recebido suficiente atenção de estudos científicos no Equador. O objetivo deste artigo foi caracterizar a diversidade genética e fenotípica de isolados de Rhizobium associados com Phaseolus vulgaris do sul do Equador. A caracterização morfo-cultural, testes bioquímicos e análises fisiológicas foram realizados para autenticar e determinar a diversidade de isolados de bactérias Rhizobium. A diversidade genética foi determinada por por técnicas moleculares consistindo na extração de DNA genômico bacteriano, amplificação e sequenciamento parcial do gene 16SrRNA; e parâmetros de nodulação e fixação de nitrogênio de P. vulgaris sobre condições de estufa foram testados para determinar a diversidade fenotípica entre os isolados. Além disso, a produção de ácido indolacético foi avaliada por um método colorimétrico. A análise fisiológica da diversidade entre os isolados revelou o potencial de crescimento de diversos isolados em diferentes níveis de pH, sa...

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“…E de fato, a coinoculação ou inoculação mista produzindo efeitos múltiplos, tem superado os resultados obtidos quando organismos são utilizados de forma isolada em uma variedade de leguminosas (Martins et al 2004, Kumar e Chandra 2008, Sánchez et al 2014). …”

Section: Milhounclassified

Promoção de crescimento vegetal por <i>Bacillus</i> sp. RZ2MS9: dos genes ao campo

Batista¹

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A Deus, pelas bênçãos diárias e por estar presente em cada decisão tomada em todos os momentos da minha vida Ao meu querido professor e orientador João Lúcio de Azevedo pelo exemplo como pessoa e profissional, pelos ensinamentos sempre tão valiosos e amizade. À querida Maria Carolina Quecine pelo apoio fundamental, orientação, amizade e pelas sugestões nos momentos decisivos que sempre me ajudam muito! À toda a minha família, mãe, pai, por serem minha inspiração e meu apoio, minhas irmãs, sobrinhos e cunhados por toda alegria que me proporcionam. À Jack e Renata pela amizade, convívio diário no laboratório e conselhos. À Maria Letícia e Samyra pela amizade e companheirismo. A todo o pessoal do laboratório Mari, Isa, Bruno, Tiago, Rafa, Gustavo, Everton, Daniel, Giulio, Jessica, Zezo, Joelma, Bia, pela agradável convivência e auxílio de sempre. Ao Lucas Taniguti pela amizade e ajuda com os dados de bioinformática. À amiga Pórtya pela ajuda com as análises estatísticas. À Renatinha, pelo esforço e dedicação durante o estágio, e Fernando Piotto pela ajuda fundamental com os experimentos dos tomatinhos. Ao professor Paul Dennis, da Universidade de Queensland pelo suporte com os experimentos de quimiotaxia e por me receber em seu laboratório e a todo o pessoal da embaixada da Austrália e ao Governo Australiano pelo financiamento e oportunidade concedidos no intercâmbio. Ao professor Labate pelo acolhimento em seu laboratório por um período do doutorado, à Mônica e todos alunos do Max Feffer pelo suporte. Ao professor Roberto pelo auxílio com análises e infraestrutura disponibilizada, técnicos Miguel e Guilherme e todos alunos do laboratório de alógamas, em especial à Miriam pela prontidão em sempre ajudar. Ao Gustavo -Gu e ao German pela ajuda com o milho no campo e ao Txarli pela disponibilização da área de plantio do milho em Taquarituba. Ao grupo de estágio PACES, em especial Carlo, Beto, Guizo e Nak pelo suporte com os experimentos com a soja. Aos funcionários da ESALQ pela colaboração nos testes realizados em campo e às funcionárias responsáveis pela limpeza do laboratório. À Coordenação do Programa de Pós-Graduação em Genética e Melhoramento de Plantas e aos professores pela contribuição em minha formação. À CAPES e CNPq pelo apoio financeiro. Aos demais que, de alguma forma, colaboraram com o trabalho. Para alimentar a população mundial crescente é necessário um aumento sustentável na produtividade agrícola. Nesse sentido, Rizobactérias Promotoras de Crescimento de Plantas (RPCPs) têm sido continuamente buscadas para formulações inoculantes por sua capacidade de incremento na produção vegetal aliado ao seu potencial de redução e/ou substituição do uso de fertilizantes minerais, insumos que causam grandes impactos ambientais, na saúde humana e econômicos. A RPCP Bacillus sp. RZ2MS9, um representante da biodiversidade amazônica brasileira, é uma forte candidata a bionoculante por seu efeito benéfico, previamente descrito, em uma ampla gama de culturas, incluindo milho e soja. Essas duas culturas representam mais...

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Effects of co-inoculation of native Rhizobium and Pseudomonas strains on growth parameters and yield of two contrasting Phaseolus vulgaris L. genotypes under Cuban soil conditions

Cited by 81 publications

References 45 publications

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

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

Genetic and phenotypic diversity of Rhizobium isolates from Southern Ecuador

Promoção de crescimento vegetal por <i>Bacillus</i> sp. RZ2MS9: dos genes ao campo

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