Direct amplification of nodD from community DNA reveals the genetic diversity of Rhizobium leguminosarum in soil

Sequences of nodD, a gene found only in rhizobia, were amplified from total community DNA isolated from a pasture soil. The polymerase chain reaction (PCR) primers used, Y5 and Y6, match nodD from Rhizobium leguminosarum biovar trifolii, R. leguminosarum biovar viciae and Sinorhizobium meliloti. The PCR product was cloned and yielded 68 clones that were identified by restriction pattern as derived from biovar trifolii [11 restriction fragment length polymorphism (RFLP) types] and 15 clones identified as viciae (seven RFLP types). These identifications were confirmed by sequencing. There were no clones related to S. meliloti nodD. For comparison, 122 strains were isolated from nodules of white clover (Trifolium repens) growing at the field site, and 134 from nodules on trap plants of T. repens inoculated with the soil. The nodule isolates were of four nodD RFLP types, with 77% being of a single type. All four of these patterns were also found among the clones from soil DNA, and the same type was the most abundant, although it made up only 34% of the trifolii-like clones. We conclude that clover selects specific genotypes from the available soil population, and that R. leguminosarum biovar trifolii was approximately five times more abundant than biovar viciae in this pasture soil, whereas S. meliloti was rare

Eficiência e competitividade de variantes espontâneos isolados de estirpes de Bradyrhizobium spp recomendadas para a cultura da soja (Glycine max) Effectiveness and competitiveness of spontaneous mutants isolated from Bradyrhizobium spp strains recommended for soybean crop (Glycine max)

O cultivo sucessivo de soja inoculada numa mesma área proporcionou a adaptação de uma população de rizóbios, que podem não ser tão eficientes quanto à capacidade de fixação de N2, mas apresentam alta competitividade, dificultando a introdução de novas estirpes mais eficientes. Com a finalidade de avaliar o desempenho simbiótico (eficiência e competitividade) de variantes espontâneos isolados de estirpes de B. japonicum (SEMIA 5079 e SEMIA 5080) e B. elkanii (SEMIA 587 e SEMIA 5019), realizou-se um experimento em casa de vegetação onde os variantes foram inoculados isoladamente e em diferentes combinações entre os variantes e uma estirpe comprovadamente mais competitiva (SEMIA 587 ou SEMIA 5019) a partir da adição de inóculos mistos (1/1; v/v) no cultivar de soja BR-16. Por meio da avaliação das variáveis analisadas (nodulação, produção de matéria de seca da parte aérea, N total acumulado na parte aérea e ocupação nodular), foi possível constatar que o determinante da maior eficiência em tratamentos co-inoculados não foi a ocupação nodular de determinada estirpe ou variante presente no inóculo, mas, sim, o tipo de interação (sinérgica ou antagônica) predominante no tratamento co-inoculado e que é possível selecionar variantes eficientes e competitivos para a cultura da soja a partir de estirpes parentais que já apresentam características desejáveis para utilização em inoculantes comerciais.<br>The continuous cultivation of inoculated soybean in the same area can determine the soil colonization with a rhizobia population presenting low nitrogen fixation effectiveness. This fact can be a problem for the establishment of a more effective population. A greenhouse experiment was carried out to evaluate the symbiotic effectiveness and competitiveness of spontaneous mutants isolated from B. japonicum (SEMIA 5079 and SEMIA 5080) and B. elkanii (SEMIA 587 and SEMIA 5019) strains. The soybean biovar BR 16 was inoculated with the same proportion of inoculants (1:1; v/v) containing just one of the spontaneous mutants or in different combinations among them in comparison to a competitive strain (SEMIA 587 and 5019). Through the evaluation of nodulation, dry weight yields of plants, total N in plant and nodules occupancy, its was verified that the determinant factor for microorganisms symbiotic effectiveness in co-inoculated treatments was the synergism and the antagonistic interaction and not the nodule occupancy. It was also possible to select effective an competitive spontaneous mutants for soybean from parental strains recommended for commercial inoculants

Symbiotic effectiveness and ecological characterization of indigenous Rhizobium loti populations in Uruguay Eficiência simbiótica e caracterização ecológica de uma população nativa de Rhizobium loti no Uruguai

The objectives of this work were to describe the distribution, density and seasonal variation of the indigenous populations of Rhizobium loti in different Uruguayan soils and to determine the symbiotic effectiveness and stress tolerance factors of different isolates, both with the aim of obtaining selected strains to re-introduce as inoculants in Lotus pastures. R. loti was present in ten soils studied and their densities varied from year to year and within each soil. All the isolates nodulated Lotus corniculatus effectively. The nodules in Lotus pedunculatus and Lotus subbiflorus were small, red on the surface and ineffective in nitrogen fixation. The study of 50 isolates from the ten soils showed high variability in their symbiotic efficiency and tolerance to pH. The indigenous population was acid tolerant in culture medium (pH 4.5), 83% of them could grow at pH 4.5 in 3 days. This work showed that there was a great diversity between the strains of R. loti isolated from Uruguayan soils and supports the importance of selecting among them the most efficient and resistant strains to be included in the inoculants.<br>Os objetivos deste trabalho foram descrever a distribuição, densidade e variação sazonal de populações nativas de Rhizobium loti em diferentes solos uruguaios, e determinar a eficiência simbiótica e os fatores de tolerância a estresse de diversos isolados, tendo em vista a obtenção de cepas a serem reintroduzidas como inoculantes em pastagens de Lotus. Rhizobium loti estava presente em dez solos estudados, e suas densidades variavam de ano para ano e em cada solo. Todos os isolados nodularam efetivamente o Lotus corniculatus. Os nódulos no Lotus pedunculatus e no Lotus subbiflorus eram pequenos, vermelhos na superfície, e ineficazes na fixação de N. O estudo de 50 isolados dos dez solos mostraram alta variabilidade na sua eficiência simbiótica, e alta tolerância ao pH. A população nativa era tolerante à acidez em meio de cultura (pH 4,5); 83% dela pode crescer em pH 4,5 em três dias. O presente trabalho mostrou que há grande diversidade entre as cepas de R. loti isoladas de solos uruguaios, e acentua a importância de se selecionar entre elas as mais eficientes e resistentes, para serem incluídas como inoculantes