Black leaf streak disease (BLSD; also known as black Sigatoka) caused by Mycosphaerella fijiensis Morelet is one of the main diseases affecting banana trees worldwide. Associated microorganisms in the phylloplane have been previously studied for their control. An exhaustive understanding of biocontrol mechanisms will potentiate the biological control agent. Therefore, strain Bacillus pumilus CCIBP-C5, isolated from banana leaves surface was evaluated for in vitro antagonism against M. fijiensis and the effect of culture filtrate on fungal growth and disease progress was determined. Bacterial cell and its diffused metabolites inhibited M. fijiensis growth. The culture filtrate of B. pumilus affected the mycelial growth, with antifungal effect on hyphal tip, membrane permeability and integrity. Also, a reduction of number of BLSD necrotic lesions and disease progression were found after application of the B. pumilus CCIBP-5 culture filtrate in banana plants. Results indicate that CCIBP-C5 has potential for biological control of BLSD.
Rhizobium-legume symbioses play relevant roles in agriculture but have not been well studied in Ecuador. The aim of this study was to characterize the genetic and phenotypic diversity of Rhizobium isolates associated with Phaseolus vulgaris from southern Ecuador. Morphocultural characterization, biochemical tests and physiological analyses were conducted to authenticate and determine the diversity of bacteria Rhizobium-like isolates. The genetic diversity of the isolates was determined by molecular techniques, which consisted of bacteria DNA extraction and amplification and sequencing of the 16S rRNA gene. The nodulation parameters and nitrogen fixation for P. vulgaris under greenhouse conditions were also assessed to determine the phenotypic diversity among isolates. Furthermore, bacteria indole-acetic-acid production was evaluated by the colorimetric method. Morpho-cultural and biochemical characteristic assessments demonstrated that Rhizobium-like bacteria was associated with the P. vulgaris nodules. The diversity among the isolates, as determined by physiological analyses, revealed the potential of several isolates to grow at different pH values, salinity conditions and temperatures. Partial sequencing of the 16S rRNA gene identified the Rhizobium genus in every sampling site. From a total of 20 aligned sequences, nine species of Rhizobium were identified. Nodule formation and biomass, as well as nitrogen fixation, showed an increase in plant phenotypic parameters, which could be influenced by IAA production, especially for the strains R. mesoamericanum NAM1 and R. leguminosarum bv. viciae COL6. These results demonstrated the efficiency of native symbiotic diazotrophic strains inoculants for legume production. This work can serve as the basis for additional studies of native Rhizobium strains and to help spread the use of biofertilizers in Ecuadorian fields.Index terms: Phaseolus vulgaris; diazotrophic bacteria; 16S rRNA gene; nodulation; indole acetic acid. RESUMOA simbiose Rhizobium-leguminosa desempenha um relevante papel na agricultura, entretanto não tem recebido suficiente atenção de estudos científicos no Equador. O objetivo deste artigo foi caracterizar a diversidade genética e fenotípica de isolados de Rhizobium associados com Phaseolus vulgaris do sul do Equador. A caracterização morfo-cultural, testes bioquímicos e análises fisiológicas foram realizados para autenticar e determinar a diversidade de isolados de bactérias Rhizobium. A diversidade genética foi determinada por por técnicas moleculares consistindo na extração de DNA genômico bacteriano, amplificação e sequenciamento parcial do gene 16SrRNA; e parâmetros de nodulação e fixação de nitrogênio de P. vulgaris sobre condições de estufa foram testados para determinar a diversidade fenotípica entre os isolados. Além disso, a produção de ácido indolacético foi avaliada por um método colorimétrico. A análise fisiológica da diversidade entre os isolados revelou o potencial de crescimento de diversos isolados em diferentes níveis de pH, sa...
The effect of antifungal activity of culture filtrate (CF) of strain CCIBP-C5, an isolate from a phyllosphere of banana () leaves, was determined on challenged banana plants. The CF was shown to decrease the fungal biomass and induce changes in banana plant. In this sense, at 70 days post inoculation (dpi), a lower infection index as well as a decrease in fungal biomass after 6 dpi was obtained in treated plants with respect to control ones. At the same time, changes in the activities of several enzymes related to plant defense responses, such as phenylalanine ammonia lyase, chitinases, β-1,3-glucanases and peroxidases were observed. These results indicate that CCIBP-C5 has a potential role for biological control of possibly due to the production of antifungal metabolites.
Introduction. Due to the absence of totally effective either economically viable chemical agents for the control of Fusarium wilt, the use of antagonistic microorganisms is of great interest since it could represent a more economically and ecologically sustainable alternative. Objective. To analyze the antifungal effect of the Bacillus amyloliquefaciens CCIBP-A5 strain against Fusarium oxysporum. Materials and methods. The work was carried out in the Laboratory of Applied Microbiology of the Instituto de Biotecnología de las Plantas, Cuba, between September 2017 and June, 2018. The in vitro and in vivo antifungal activity of its culture filtrate and cell against F. oxysporum has been assayed. Results. The results indicated that the metabolites present in the culture filtrate of B. amyloliquefaciens CCIBP-A5 significantly influenced the growth and morphology of the mycelium and the conidia. They also caused oxidative damage to the lipid molecules of F. oxysporum. In addition, this strain showed inhibitory effects on the development of the disease under controlled conditions. These aspects are key when selecting a bacterial candidate as a biological control agent. Conclusions. The results showed that the B. amyloliquefaciens CCIBP-A5 strain, isolated from Musa sp., had an in vitro antifungal effect against the vegetative and reproductive structures of Foc race 1 as well as on the Musa spp.-F. oxysporum interaction. This strain is suggested for the development of a bioproduct for Fusarium wilt management.
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