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| Author(s): |
Harold Alexander Vargas Hoyos
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Piracicaba. |
| Institution: | Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC) |
| Defense date: | 2018-02-23 |
| Examining board members: |
Itamar Soares de Melo;
Cleusa Maria Mantovanello Lucon;
Gabriel Padilla Maldonado;
Katia de Lima Nechet
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| Advisor: | Itamar Soares de Melo |
| Abstract | |
In this study, the taxonomic diversity and biological potential of actinobacteria isolated from different Brazilian biomes with antagonistic activity against Sclerotinia sclerotiorum were accessed. In total, 354 isolates were obtained from the Embrapa - Environment Collection of Microorganisms of Agricultural and Environmental Importance (CMAA). The antagonistic activity against S. sclerotiorum was evaluated by in vitro assay. In total, 55 isolates were obtained, all isolates showed some percentage of visible growth inhibition against S. sclerotiorum. The identification and analysis of the diversity of the isolates using the 16S rRNA gene showed that 49 isolates belong to the Streptomyces genus, 4 isolates belonging to the genus Micromonospora, 1 isolated to the genus Kocuria and 1 isolated to the genus Actinomadura. All the actinobacteria showed compatibility with the soil beneficial bacteria evaluated except the isolate Caat P8 35. Three isolates, 1AS2a, Caat P555 and Caat P8 79 inhibited both mycelia and S. sclerotiorum sclerotia. The isolates Bc V1 06 and 3AS4 showed hydrolysis using organic and inorganic phosphorus media. In order to know the possible mechanism of action for solubilization of phosphorus, the aqueous extract of the 3AS4 isolate was submitted to HPLC analysis, the chromatographic profile obtained was similar to gluconic acid. Greenhouse evaluation showed differences in height of soybean plants after 6 weeks when inoculated with 3AS4. The Shoot/Root ratio was significantly higher when the phosphorus was added along with the actinobacteria. Plants with no phosphorus addition and with 3AS4 showed similar development in the Shoot/Root when compared to inoculated plants with 40 kg.ha-1 of PR and SPT. The evaluation of the enzymatic activity evidenced nine isolates efficient of hydrolyzing chitin and laminarin as the only carbon substrate, three of them; 1AS2a, Caat P5 55 e Caat P8 79 isolates (β-1-4-N-acetylglucosaminidase) was 0.03, 0.22 and 0.16 IU respectively, however, glucanase activity (β-1-3-glucanase) was 0.23, 0.25 and 0.26 IU respectively. Organic extracts of the actinobacteria obtained with dichloromethane (26) and ethyl acetate (15) demonstrated inhibition of fungus growth, only the 1AS2a isolate presented inhibition at the concentration of 165 μg.mL-1. Spectrometric studies have revealed the presence of a compound of the bafilomycins family as the main active compound. The annotation of the genome of the 1AS2a isolate revealed the presence of 33 gene operons involved in biosynthetic pathways for the production of antimicrobial compounds, in which one of them presented 100% similarity to the bafilomycin synthesis cluster. Taxonomic studies have shown that the phylogenetic, morphological and chemical characteristics of the 1AS2c isolate are consistent with the genus Streptomyces. However, some differences in the taxonomic profile, DDH and Multilocus analysis confirmed the isolated 1AS2cT as a type strain for Streptomyces, for which the name Streptomyces rhizosphaericola sp. Nov. With the results obtained we can corroborate: 1. Brazilian biomes possess a great taxonomic and functional diversity 2. Detection of the antagonism of Actinobacteria isolates against S. sclerotiorum fungus 3. Phosphorus solubilization and plant growth stimulation of 3AS4 isolate under conditions of greenhouse 4. The effective antifungal compounds production of the 1AS2a isolate, revealed in the combining use of spectroscopic and bioinformatics tools. Considering all this, there is an enormous potential for biocontrol in isolates of Actinobacteria from different Brazilian biomes that may be a new option for use in agriculture. (AU) | |
| FAPESP's process: | 13/25076-0 - "Actinomycetes isolated from soils of Brazilian ecosystems: potential agents of biological control of Sclerotinia sclerotiorum in cultivation of soybean (Glycine max L. Merrill)" |
| Grantee: | Harold Alexander Vargas Hoyos |
| Support Opportunities: | Scholarships in Brazil - Doctorate |