-The objective of this work was to identify the most competitive and effective Rhizobium strains in order to increase common bean yield by nitrogen fixation as alternative or complementation to the nitrogen fertilization. Competitiveness tests were lead in axenic conditions, in Cerrado soil pots and in three field experiments, with native Rhizobium strains that were previously identified, according to their effectiveness and genetic variability. The identification of strains in nodules was performed using serological tests (axenic conditions) -agglutination and enzyme linked immunosorbent (Elisa) assays -and random amplified polymorfic DNA (RAPD) (Cerrado soil). Plant yield was determined using the dry weight (greenhouse conditions), total N and grain yield (field experiments). Among the analyzed Rhizobium strains, native strain SLA 2.2 and commercial strain CIAT 899 were the dominant nodules in plants of the most productive plots, presenting yield productivity similar or higher to those obtained in treatments where 20 kg ha -1 of N were applied.Index terms: Phaseolus vulgaris, nodule occupancy, Elisa, RAPD. Capacidade competitiva de estirpes de Rhizobium na nodulação do feijoeiro em solos de CerradoResumo -O objetivo deste trabalho foi identificar as estirpes de Rhizobium mais efetivas e competitivas, a fim de maximizar a produtividade do feijoeiro por meio da fixação de nitrogênio, como alternativa à adubação nitrogenada. Foram conduzidos testes de competitividade em condições axênicas, em vasos com solo do Cerrado e em três experimentos de campo, com estirpes de Rhizobium nativas, previamente selecionadas quanto à efetividade e à variabilidade genética. A identificação das estirpes nos nódulos foi efetuada por meio das técnicas de aglutinação e ensaio imunoabsorvente de ligação de enzimas (Elisa), em condições de casa de vegetação, e pela técnica de DNA polimórfico amplificado ao acaso (RAPD), em solo de Cerrado. A produtividade das plantas foi determinada pela produção de matéria seca, teor de N e produção de grãos (condições de campo). A estirpe nativa SLA 2.2 e a estirpe comercial CIAT 899 foram dominantes nos nódulos das plantas das parcelas mais produtivas, com índices de produtividade iguais ou superiores aos obtidos nos tratamentos em que foram aplicados 20 kg ha -1 de N.Termos para indexação: Phaseolus vulgaris, ocupação nodular, Elisa, RAPD.
High temperatures can affect the survival, establishment and symbiotic properties of Rhizobium strains. Bean nodulating Rhizobium strains are considered particularly sensitive because on this strains genetic recombinations and/or deletions occur frequently, thus compromising the use of these bacteria as inoculants. In this study R. tropici and R. leguminosarum bv. phaseoli strains isolated from Cerrado soils were exposed to thermal stress and the strains’ growth, survival and symbiotic relationships as well as alterations in their genotypic and phenotypic characteristics were analyzed. After successive thermal shocks at 45ºC for four hours, survival capacity appeared to be strain-specific, independent of thermo-tolerance and was more apparent in R. tropici strains. Certain R. leguminosarum bv. phaseoli strains had significant alterations in plant dry weight and DNA patterns obtained by AP-PCR method. R. tropici strains (with the exception of FJ2.21) were more stable than R. leguminosarum bv. phaseoli strains because no significant phenotypic alterations were observed following thermal treatments and they maintained their original genotypic pattern after inoculation in plants.
Irregular response to bean plants to Rhizobium inoculation has been attributed to among other factors, low competitive ability, low N2 fixation efficiency and genetic instability of the symbiont. This genetic instability caused by high rates of genomic rearrangements and/or plasmid deletions can be accentuated by high temperatures. This fact may limit the utilization of these strains as inoculants, especially in tropical soils. In this study, the variability of isolated colonies derived from effective R. leguminosarum bv. phaseoli (SLP1.3 and BR 10.026) and R tropici (SLA2.2 and BR322) strains was evaluated before and after exposure to high temperatures (four consecutive thermal shocks at 45ºC). This evaluation involved plant dry matter analysis of inoculated plants and genotypic (plasmid profile and genomic patterns via RAPD) analysis of the Rhizobium strains. The results evidenced that high temperature improve the natural performance variability especially between isolated colonies from R. leguminosarum bv. phaseoli strains. The plasmid profile of isolated colonies from R. tropici strains were identical regardless of temperature treatment whereas isolated colonies from R. leguminosarum bv. phaseoli alterations were detected especially after the thermal treatment. The genomic patterns generated by AP-PCR showed more alterations and genetic variation in isolated colonies from R. leguminosarum bv. phaseoli strains indicating that R. tropici strains are more stable and lower affected by high temperature.
The genetic relationships of 85 Arachis pintoi nodulating Rhizobium strains were determined using the random amplified polymorphic DNA (RAPD) methods. The analysis included 75 strains isolated from Cerrado soils and 10 other ones of different origins. The results indicated that there is a high level of similarity between these strains and that geographic distribution may affect their phylogenetic relationship. In addition, the results allowed the selection of the most suitable primers for characterisation of these Rhizobium strains which will be useful for implementation of competitiveness studies in Cerrado soils.
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