No abstract

In the last column of Table 2, the third and fourth entries should be Ͻ0.69 D instead of Ͻ5 C.

The fungal population dynamics in soil and in the rhizospheres of two maize cultivars grown in tropical soils were studied by a cultivation-independent analysis of directly extracted DNA to provide baseline data. Soil and rhizosphere samples were taken from six plots 20, 40, and 90 days after planting in two consecutive years. A 1.65-kb fragment of the 18S ribosomal DNA (rDNA) amplified from the total community DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and by cloning and sequencing. A rhizosphere effect was observed for fungal populations at all stages of plant development. In addition, pronounced changes in the composition of fungal communities during plant growth development were found by DGGE. Similar types of fingerprints were observed in two consecutive growth periods. No major differences were detected in the fungal patterns of the two cultivars. Direct cloning of 18S rDNA fragments amplified from soil or rhizosphere DNA resulted in 75 clones matching 12 dominant DGGE bands. The clones were characterized by their HinfI restriction patterns, and 39 different clones representing each group of restriction patterns were sequenced. The cloning and sequencing approach provided information on the phylogeny of dominant amplifiable fungal populations and allowed us to determine a number of fungal phylotypes that contribute to each of the dominant DGGE bands. Based on the sequence similarity of the 18S rDNA fragment with existing fungal isolates in the database, it was shown that the rhizospheres of young maize plants seemed to select the Ascomycetes order Pleosporales, while different members of the Ascomycetes and basidiomycetic yeast were detected in the rhizospheres of senescent maize plants.

Microvirga vignae sp. nov., a root nodule symbiotic bacterium isolated from cowpea grown in semi-arid Brazil isolated from nodules of cowpea collected from a semi-arid region of Brazil showed 97 % similarity to sequences of recently described rhizobial species of the genus Microvirga. Phylogenetic analyses of four housekeeping genes (gyrB, recA, dnaK and rpoB), DNA-DNA relatedness and AFLP further indicated that these strains belong to a novel species within the genus Microvirga. Our data support the hypothesis that genes related to nitrogen fixation were obtained via horizontal gene transfer, as sequences of nifH genes were very similar to those found in members of the genera Rhizobium and Mesorhizobium, which are not immediate relatives of the genus Microvirga, as shown by 16S rRNA gene sequence analysis. Phenotypic traits, such as host range and carbon utilization, differentiate the novel strains from the most closely related species, Microvirga lotononidis, Microvirga zambiensis and Microvirga lupini. Therefore, these symbiotic nitrogen-fixing bacteria are proposed to be representatives of a novel species, for which the name Microvirga vignae sp. nov. is suggested. The type strain is BR3299 T (5HAMBI 3457 T ).

Resumo -O objetivo deste trabalho foi avaliar as relações filogenéticas de estirpes de Bradyrhizobium e a contribuição destas estirpes para a fixação biológica de nitrogênio em caupi, em solos do Cerrado. Na avaliação da relação filogenética, o gene 16S rDNA de cada uma das estirpes foi amplificado e seqüenciado, e para a análise da eficiência simbiótica, determinou-se: N total, matéria seca das plantas, massa de nódulos e redução de acetileno, em casa de vegetação, e ocupação nodular, em experimento de campo. A maioria das estirpes estudadas pertence a B. elkanii e, pelo menos dez das estirpes, independentemente da espécie, apresentaram bom desempenho quanto à fixação biológica de N 2 . As estirpes BR3262, BR3280 (caracterizadas como B. elkanii) e BR3267, BR3287 e BR3288 (Bradyrhizobium sp.) mostram-se como inoculantes potenciais para o caupi, em razão do bom desempenho tanto na eficiência simbiótica quanto na ocupação nodular.Termos para indexação: fixação biológica de nitrogênio, 16S rDNA, Bradyrhizobium, Bradyrhizobium elkanii. Symbiotic efficiency of cowpea Bradyrhizobium strains in Cerrado soilsAbstract -The obejctive of this study was to evaluate the phylogenetic relationships of Bradyrhizobium strains, and the contribution of these strains to cowpea biological nitrogen fixation in Cerrado soils. To elucidate the phylogenetic relationships among strains, their 16S rDNA gene was extracted, amplified and sequenced. In order to evaluate the symbiotic efficiency, total N, plant dry matter, nodular mass and acetylene reduction in a greenhouse experiment, and nodular occupation, in a field experiment, were determined. Most part of the strains belonge to B. elkanii, and at least ten of the analyzed strains, independently of the species, presented high capacity to fix N 2 . The strains BR3262, BR3280 (characterized as B. elkanii), BR3267, BR3287 and BR3288 (Bradyrhizobium sp.) exhibit a potential as inoculant for cowpea, due to their high biological nitrogen fixation.Index terms: biological nitrogen fixation, 16S rDNA, Bradyrhizobium, Bradyrhizobium elkanii. IntroduçãoNo Brasil, o caupi (Vigna unguiculata (L.) Walp.) é cultivado, basicamente, em regime de subsistência, nas regiões Norte e Nordeste, principalmente por sua adaptação às condições edafoclimáticas. Nessas regiões, esta cultura desempenha um papel importante na alimentação e na geração de empregos para a população de baixa renda. Segundo Freire Filho et al. (1999), a cultura do caupi no Brasil gera, anualmente, cerca de 2,4 milhões de empregos diretos, além de compor a dieta de pelo menos 27,5 milhões de pessoas.O caupi tem sido introduzido, recentemente, também em áreas de grandes produtores agrícolas nos Cerrados dos Estados do Piauí e Maranhão, principalmente por sua compatibilidade com o sistema de rotação de cultura e o regime pluviométrico regional. Nessas áreas, os agricultores introduzem o caupi na "safrinha", após a colheita do arroz, no final da estação das chuvas, tanto em lavouras recém-abertas, como em rotações de cultivos bianuais com so...

Aim: To evaluate the rpoB gene as a biomarker for PCR‐DGGE microbial analyses using soil DNA from the Cerrado, Brazil. 
Methods: DNA extraction from soil was followed by Polymerase Chain Reaction (PCR) amplification of rpoB and 16S rRNA genes. PCR products were compared by Denaturing Gradient Gel Electrophoresis (DGGE) to compare gene/community profiles. 
Results: The rpoB DGGE profiles comprised fewer bands than the 16S rDNA profiles and were easier to delineate and therefore to analyse. Comparison of the community profiles revealed that the methods were complementary. 
Conclusions, Significance and Impact of the Study: The gene for the beta subunit of the RNA polymerase, rpoB, is a single copy gene unlike 16S rDNA. Multiple copies of 16S rRNA genes in bacterial genomes complicate diversity assessments made from DGGE profiles. Using the rpoB gene offers a better alternative to the commonly used 16S rRNA gene for microbial community analyses based on DGGE.

No abstract

RESUMOO estudo objetivou avaliar a contribuição da fixação biológica de nitrogênio (FBN) promovida por estirpes de rizóbio para o desenvolvimento e rendimento de grãos do feijão-caupi em Roraima. Nos anos de 2005 e 2006 foram conduzidos experimentos em área de cerrado e mata alterada, onde foram testadas as estirpes INPA 03-11B, UFLA 3-84, BR3267 (recomendadas à cultura), a estirpe BR3299 e BR3262, duas doses de nitrogênio mineral (50 e 80 kg ha -1 de N) e um controle. As variáveis avaliadas foram: nodulação e produção de massa seca da parte aérea de plantas de feijão-caupi e, o rendimento de grãos na colheita. Na média geral, foi observado que a estirpe BR3262 proporcionou número e massa de nódulos significativamente maiores ao controle, ao passo que entre as estirpes recomendadas, isto só ocorreu de forma esporádica com INPA 03-11B e BR3267. Além disso, também foi observado que a população de rizóbio do solo foi determinante à nodulação das plantas dos experimentos. Comparativamente as demais estirpes, BR3262 juntamente com BR3267, proporcionaram maior efetividade na FBN à produção de massa seca da parte aérea. Em relação à produtividade de grãos, as estirpes BR3267 e INPA 03-11B apresentaram melhores resultadas comparadas a UFLA 3-84, entretanto, apenas a estirpe BR3262 proporcionou rendimento de grãos (na média geral cerca de 1700 kg ha -1 ) igual à dose de 50 kg ha -1 de N e superior ao controle em três dos quatro experimentos conduzidos, mostrando ser a mais indicada para a inoculação do feijão-caupi em Roraima. PALAVRAS-CHAVE:Vigna unguiculata, FBN, cerrado, mata alterada, Amazônia. Contribution of rhizobia strains to cowpea development and grain yield in Roraima -Brazil ABSTRACTThe study aimed to evaluate the contribution of biological nitrogen fixation (BNF), promoted by rhizobium strains, to development and grain yield of cowpea in Roraima. In the years 2005 and 2006 experiments were performed in cerrado and mata alterada areas, where were tested the strains INPA 03-11B, UFLA 3-84, BR3267 (recommended to the cowpea), the strains BR3299 and BR3262, two mineral nitrogen doses (50 and 80 kg ha -1 of N) and a control. The analyzed variables were: nodulation and cowpea plants dry matter production, and the grain yield in the harvest. It was observed, in the mean, that BR3262 strain provided a number and nodule dry mass significantly larger than the control, while among the recommended strains, this only occurred in a sporadic form with INPA 03-11B and BR3267. Besides, it was also observed that soil rhizobium population was determinant to plants nodulation in the experiments. Comparatively to the other strains, BR3262 together with BR3267, provided superior effectiveness in BNF to plant dry mass production. In relation to grain yield, the strain BR3267 and INPA 03-11B presented better resulted compared to UFLA 3-84, however, just the strains BR3262 provided grain yield (in the mean about 1700 kg h -1 ), equal to 50 kg ha -1 N dose and superior to the control in three of the four experiments performed, showin...