RESUMO
Fungi are an inexhaustible source of bioactive metabolites that can be driven to find medicines for chronic diseases, as Alzheimer's disease. In the present work, five species of soil-originated fungi (Aspergillus chevalieri, Clonostachys rogersoniana, Fusarium nygamai, Penicillium sp., and Talaromyces calidicanius) were submitted to mutual biotic stress aiming at activating the expression of metabolites capable of inhibiting the enzyme acetylcholinesterase. HPLC profiles showed that the in vitro biotic stress triggered the biosynthesis of metabolite-mediated defense responses. Five compounds present in the complex co-culture matrix were identified by Paper Spray Mass Spectrometry (PS-MS). The approach enhanced the biosynthesis of acetylcholinesterase inhibitors (up to 99.6% inhibition) in comparison with the individual cultures. The mutual biotic stress between T. calidicanius and A. Chevalieri led to the biosynthesis of a pool of metabolites statistically as efficient as serine (p < 0.05), the positive control used in the experiments (99.6 and 99.1% inhibition, respectively).
Assuntos
Inibidores da Colinesterase , Penicillium , Acetilcolinesterase/metabolismo , Inibidores da Colinesterase/química , Fungos/metabolismo , Penicillium/metabolismo , Estresse FisiológicoRESUMO
Rock phosphate is an alternative form of phosphorus (P) fertilizer; however, there is no information regarding the influence of P fertilizer sources in Brazilian Cerrado soils upon microbial genes coding for phosphohydrolase enzymes in crop rhizospheres. Here, we analyze a field experiment comparing maize and sorghum grown under different P fertilization (rock phosphate and triple superphosphate) upon crop performance, phosphatase activity and rhizosphere microbiomes at three levels of diversity: small subunit rRNA marker genes of bacteria, archaea and fungi; a suite of alkaline and acid phosphatase and phytase genes; and ecotypes of individual genes. We found no significant difference in crop performance between the fertilizer sources, but the accumulation of fertilizer P into pools of organic soil P differed. Phosphatase activity was the only biological parameter influenced by P fertilization. Differences in rhizosphere microbiomes were observed at all levels of biodiversity due to crop type, but not fertilization. Inspection of phosphohydrolase gene ecotypes responsible for differences between the crops suggests a role for lateral genetic transfer in establishing ecotype distributions. Moreover, they were not reflected in microbial community composition, suggesting that they confer competitive advantage to individual cells rather than species in the sorghum rhizosphere.
Assuntos
Fósforo , Rizosfera , Brasil , Fertilização , Monoéster Fosfórico Hidrolases , Filogenia , Solo , Microbiologia do SoloRESUMO
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.