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Tese
Caracterização de isolados de Bacillus Thuringiensis com potencial inseticida: abordagens genômica e proteômica
Sustainable alternatives that can replace chemical insecticides are necessary for insect pest control. Bacillus thuringiensis (Bt) is a bacterium that produces insecticidal proteins toxic against insect pests of different orders. In this work, we performed bioassays using Bt strains isolated from...
Autor principal: | Alves, Giselly Batista |
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Grau: | Tese |
Idioma: | pt_BR |
Publicado em: |
Universidade Federal do Tocantins
2024
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Assuntos: | |
Acesso em linha: |
http://hdl.handle.net/11612/6329 |
Resumo: |
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Sustainable alternatives that can replace chemical insecticides are necessary for insect pest
control. Bacillus thuringiensis (Bt) is a bacterium that produces insecticidal proteins toxic
against insect pests of different orders. In this work, we performed bioassays using Bt strains
isolated from Tocantins against dipterous and lepidopteran insects. To explore all the genes
related to the pathogenicity of the isolates, the genomes of these bacteria were sequenced.
Furthermore, in chapters two and four, proteomic was combined with genomic analysis to
detect proteins expressed in the spore-crystal mixture. In chapter one, a comparative analysis
of four genomes of Bacillus thuringiensis subsp. israelensis strains with toxicity to Aedes
aegypti and Culex quinquefasciatus revealed high nucleotide sequence identity (>98%), the
same plasmids profile, and equal pesticidal protein content (cry4Ba, cry4Aa, cry11Aa,
cry10Aa, cyt1Aa, cyt2Ba, and cytCa). The genome of the Bt TOD651 strain, presented in
Chapter 2, with toxic activity to A. aegypti and C. quinquefasciatus (CL50 of 0.011 and 0.023
μg/mL, respectively), showed CDS regions highly homologous to cry11Aa3, cry10Aa4,
cry4Aa4, cry4Ba5, cyt1Aa5, cyt1Ca1, cyt2Ba13, mpp60Aa3, and mpp60Ba3 genes. The
expression of Cry11Aa3, Cry10Aa4, Cry4Aa4, Cry4Ba5, Cyt1Aa5, Cyt1Ca1, Cyt2Ba13, and
Mpp60Ba3 proteins was identified in the spore-crystal mixture, of which Cry4Ba5 was more
abundant than Cyt1Aa5. The expression of the enzyme Mppe was the most abundant among
the proteases. In chapter 3, the Bt UFT038, tested for different soybean pests, showed higher
toxicity to Spodoptera cosmioides (CL50=6.8 106/cm2), and its genomic analysis revealed the
presence of cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Finally, in
chapter 4, Bt strain TOL651, phylogenetically close to subspecies kenyae, was more toxic to
Anticarsia gemmatalis (LC50 =1.45 ng.cm-2) compared to Diatraea saccharalis (LC50 = 73.77
ng.cm-2). Its genomic analysis allowed the detection of cry1Aa18, cry1Ia44, cry2Aa9, and
cry1Ac5 genes, while proteomics indicated expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9
proteins, of which Cry1Ac5 was most abundant. In addition, the virulence factor InhA1 was
detected and thus should also contribute to the toxicity of this isolate. In conclusion, the Bt
isolates of this study are alternatives for biological control, and genomic and genomic-
proteomic characterization are important steps that could contribute to the development of
new biocontrol strategies for disease vector mosquitoes and agricultural pests. |