ABSTRACT
Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.
ABSTRACT
Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.
Subject(s)
Bacillus/metabolism , Food/metabolism , Pennisetum/growth & development , Pennisetum/microbiology , Endophytes/metabolism , Indoleacetic Acids/metabolism , Phosphates/analysis , Phosphates/metabolism , Bacillus/genetics , Siderophores/metabolism , Plant Roots/growth & development , Plant Roots/metabolism , Plant Roots/microbiology , Pennisetum/metabolism , Endophytes/genetics , Iron/metabolismABSTRACT
A espécie Rhizoctonia solani é um patógeno importante na cultura do algodão associada a doenças conhecidas como tombamento e mela. A variabilidade entre os isolados são extremamente importantes, porque existem diferenças entre os grupos de anastomose, tomadas como aqueles isolados capazes de trocar informações genéticas entre si. A caracterização morfológica, quando confirmada pela a caracterização genética fornece informações concretas sobre a distribuição do isolados quando agem como um agente patogênico. O objetivo foi identificar e caracterizar os grupos de anastomose no Brasil e confirmá-los pela caracterização genética. Foram consideradas 51 isolados de Rhizoctonia solani com o objetivo de caracterizar os grupos de anastomose e determinar a patogenicidade. A caracterização morfológica foi realizada observando-se o número de núcleos, morfologia da colônia e identificação de grupos de anastomose (AG). Na caracterização genética foram sequenciados e analisados os fragmentos genômicos contendo as regiões 5.8 S, ITS1 e ITS2, comparando-se os isolados listados no GenBank. A patogenicidade foi avaliada utilizando a escala de infecção com graus que variam de 1 a 4. Considerando-se o AG foram identificados 36 de 51 isolados como AG-4 e dois isolados como AG-7, 13 isolados não tiveram classificação. Em análises moleculares foram confirmados os 36 isolados identificados pela caracterização morfológica e mais 10 isolados como AG-4. Todos os isolados AG-7 foram confirmados e encontrado mais um nas análises moleculares. Para a patogenicidade verificou-se que cinco isolados não diferem do controle. A virulência intermediária e alta virulência foram observadas em 16 e 24 isolados, respectivamente, com médias 2,52-3,3 e 3,4-3,9.
The Rhizoctonia solani is an important pathogen in cotton crop associated with damping-off disease. The variability among isolates are extremely important, because differences exist between anastomosis groups, taken as those isolates capable of exchanging genetic information with each other. Morphological characterization, when confirmed by genetic characterization provides concrete information about the isolates distribution when it acts like a pathogen. Our study was to identify and characterize the anastomosis groups in Brazil and confirm them by genetic characterization. We considered 51 Rhizoctonia solani isolates aiming to characterize the anastomose group and determine their pathogenicity. The morphological characterization was done observing the number of cores, colony morphology and identification of anastomosis groups (AG). In genetic characterization were sequenced and analyzed the genomic fragments containing the 5.8 S, ITS1 and ITS2 regions and compared them to Rhizoctonia isolates listed in the GenBank. The pathogenicity was evaluated for the disease severity, using the scale of infection with grades ranging from 1 to 4. Considering the AG we identified 36 of 51 isolates as AG-4 and two isolates as AG-7 and 13 isolates were listed as unrated. In molecular analyses were confirmed those 36 isolates and were identified more 10 isolates as AG-4. All the AG- 7 isolates were confirmed and we found one more considering the molecular analyses. For the pathogenicity was found that five strains did not differ from the control. Intermediate virulence and high virulence were observed in 16 and 24 isolates, respectively with averages from 2.52 to 3.3 and from 3.4 to 3.9.
Subject(s)
Rhizoctonia , Virulence , Gossypium , Databases, Nucleic Acid , Molecular BiologyABSTRACT
Several diseases have affected apple production, among them there is Glomerella leaf spot (GLS) caused by Colletotrichum spp. The first report of this disease in apple was in plants nearby citrus orchards in São Paulo State, Brazil. The origin of this disease is still not clear, and studies based on the molecular phylogeny could relate the organisms evolutionarily and characterize possible mechanisms of divergent evolution. The amplification of 5.8S-ITS (Internal Transcribed Spacer) of rDNA of 51 pathogenic Colletotrichum spp. isolates from apples, pineapple guava and citrus produced one fragment of approximately 600 bases pairs (bp) for all the isolates analyzed. The amplified fragments were cleaved with restriction enzymes, and fragments from 90 to 500bp were obtained. The sequencing of this region allowed the generation of a phylogenetic tree, regardless of their hosts, and 5 isolated groups were obtained. From the "in silico" comparison, it was possible to verify a variation from 93 to 100 percent of similarity between the sequences studied and the Genbank data base. The causal agent of GLS is nearly related (clustered) to isolates of pineapple guava and to the citrus isolates used as control.
A produção de maçã vem sendo comprometida pela ocorrência de muitas doenças, entre as quais se destaca a Mancha Foliar de Glomerella (MFG), causada por Colletotrichum spp. O primeiro relato dessa doença em maçã foi registrado em plantas próximas a pomares de citrus no Estado de São Paulo, Brasil. A origem da MFG ainda não está bem clara, e estudos baseados na filogenia permitirão relacionar o organismo evolutivamente, possibilitando caracterizar possíveis mecanismos divergentes de evolução. A amplificação da região 5.8S-ITS (espaçador interno transcrito) do rDNA de 51 isolados de Colletotrichum patogênicos em de maçã, goiabeira serrana e citrus produziu um fragmento de aproximadamente 600 pares de bases (pb) para todos os isolados analisados. Os fragmentos amplificados foram digeridos por enzimas de restrição, sendo obtidos fragmentos de 90 a 500pb. O sequenciamento dessa região permitiu a construção de uma árvore filogenética com a distribuição dos isolados em cinco grupos, independentemente de seus hospedeiros. A partir da comparação in silico, foi possível verificar uma variação de 93 a 100 por cento de similaridade entre as sequências estudadas e o banco de dados do GenBank. O agente causal da MFG está relacionado (agrupado) a isolados de goiabeira serrana e aos isolados padrões de citrus.
ABSTRACT
We characterized indigenous common bean rhizobia from five districts of the state of Minas Gerais, Brazil. The isolates were trapped by two common bean varieties, the Mineiro Precoce (Andean origin) and Ouro Negro (Mesoamerican origin). Analysis by BOX-PCR of selected isolates detected a high level of genetic diversity.
Subject(s)
Genetic Variation , In Vitro Techniques , Polymerase Chain Reaction , Phaseolus nanus/isolation & purification , Rhizobium/isolation & purification , Methods , Methods , VirulenceABSTRACT
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.