Molecular analysis of endophytic bacteria from the genus Bacillus isolated from tropical maize (Zea mays L. ) / Análise molecular de bactérias endofíticas do gênero Bacillus isoladas de milho tropical (Zea mays L. )

Endophytic bacteria play an important role in agriculture by improving plant performance and adaptation against biotic and abiotic stresses. In the present study molecular methods were used for identifying Bacillus endophytic bacteria isolated from Brazilian sweet corn. SDS-PAGE of wholecell protein extract of fortytwo isolates revealed a high number of scrutinable bands. Twenty-four isolates were identified in nine different groups of duplicated bacteria and eighteen were identified as unique. Some high-accumulated polipeptides with variable length were observed in almost isolates. Partial sequencing of 16S ribosomal gene revealed that all isolates are Bacillus sp. and among thirteen isolates with similar protein profiles, two were different strains. Among the forty-two isolates identified by rDNA sequencing, Bacillus subitilis and B. pumilus were the most frequenty species (15 and 12 isolates, respectively) followed by B. licheniformes (7 isolates), B. cereus (5 isolates) and B. amiloliquefascens (3 isolates). According to present results, SDS-PAGE technique could be used as a fast and cheap first tool for identifying interspecific variation in maize endophytic bacterial collections while rDNA sequencing could be applied for analyzing intraspecific variation among isolates with similar protein profile as well as for taxonomic studies.

Bactérias endofíticas desempenham papel importante na agricultura, melhorando a performance e adaptação de plantas contra estresses bióticos e abióticos. No presente estudo, métodos moleculares foram empregados para identificar bactérias endofíticas do gênero Bacillus isoladas de cultivares de milho doce brasileiro. SDS-PAGE de extratos protéicos totais de quarenta e dois isolados revelaram elevado número de bandas escrutináveis. Vinte e quatro isolados formaram nove grupos diferentes de réplicas bactérianas e dezoito foram considerados como únicos. Entre os isolados, alguns polipeptídios, de tamanhos variados, foram altamente acumulados. Seqüenciamento parcial do gene ribosomal 16S revelou que todos os isolados pertencem ao gênero Bacillus e que, entre treze isolados com padrão protéico similar, dois eram linhagens diferentes. Entre os quarenta e dois isolados identificados por seqüenciamento de rDNA, Bacillus subtilis e B. pumilus foram mais frequentes (15 e 12 isolados, respectivamente), seguido por, B. licheniformes (7 isolados), B. cereus (5 isolados) e B. amiloliquefascens (3 isolados). Baseado nos resultados, conclui-se que a técnica de SDS-PAGE poderá ser usada como primeiro procedimento, rápido e barato, para identificar variação inter-específica em coleções de bactérias endofíticas isoladas do milho, enquanto o método de seqüenciamento de rDNA poderá ser aplicado para analisar variações intra-específica entre isolados com padões similares de proteínas e estudos de taxonomia.

Factors affecting "in vitro" plant development and root colonization of sweet potato by Glomus etunicatum Becker & Gerd

Nutrients media (Murashige & Skoog, Hoagland & Arnon and White's media) supplemented or not with sucrose and substrates (vermiculite, agar and natrosol) were tested for their effects on plant development and root colonization of micropropagated sweet potato, cv. White Star, by Glomus etunicatum Becker & Gerdemann (isolate INVAM FL S329). Addition of sucrose (3 per cent) did not affect plant development. However, hyphal growth and root colonization were depressed. Contrasting responses to media nutrient concentration were observed for plant height, root colonization, and hyphal growth. The highest concentration of nutrients in Murashige & Skoog medium improved plant development, but this medium decreased hypha growth and inhibited root colonization. Plants growing in vermiculite substrate had higher (p£0.05) development and mycorrhizal root colonization than those growing in agar or natrosol. The results indicate that colonization of micropropagated sweet potato by arbuscular mycorrhizal fungi is affected by media composition and type of substrate.

The interactive effect of phosphorus and nitrogen on "In vitro" spore germination of glomus etunicatum Becker & Gerdemann, root growth and mycorrhizal colonization

The effects of P and N amendment and its interactions on spore germination, root growth and colonized root length by Glomus etunicatum Becker & Gerdemann (INVAM S329) was studied "in vitro"in RiT-DNA transformed roots of Anthylis vulneraria sub sp. Sampaiana (Kidney vetch). Three N media concentrations (5, 10 and 50 mg/l) at P constant level (2 mg/l) and three P media concentrations (2, 10 and 20 mg/l) at N constant level (5 mg/l) were utilized as a treatment. Bécard & Fortin medium was used as a basal medium for root growth and colonized root length, and water/agar 0.8(per cent) media was the control for spore germination. Spore germination of G. etunicatum at low P level was reduced by N addition in relation to the control media, and at low N level addition of P stimulated spore germination. Total root length was stimulated by N addtion at low P level, but no significant difference (p<0.05) was observed among P concentrations. Colonized root lenght by G. etunicatum increased significantly (pz0,05) with P additions at low N levels. Under low P level no significant differences was found between 10 and 50 mg/l of N. These results demonstrate that the interaction between P and N affect differently spore germination, root growth and colonized root lenght.