Characterization of the Methyltransferase gene present in the O-antigen gene cluster of the endophytic Burkholderia seminalis TC3.4.2R3, and the role of this gene in the synthesis of antimicrobial compounds
The Burkholderia genus is composed by Gram-negative bacteria, which have high physiological and genetic diversity. Burkholderia is the object of studies that aim at various biotechnological applications as biocontrol agents, plant growth promoters, and bioremediation as producers of major molecules of interest. The Burkholderia seminalis TC3.4.2R3 strain is an endophytic organism isolated from sugarcane roots and shows the ability to inhibit fungal and plant pathogenic bacteria. However, knowledge about the interaction and control mechanisms of the metabolites with antimicrobial properties in Burkholderia spp. is still incipient. Thus, studies to identify the genes of Burkholderia spp. involved in the synthesis of antimicrobial compounds are required. This study aims to evaluate the production of antimicrobial compounds by B. seminalis TC3.4.2R3 through the analysis of the O-antigen cluster of genes responsible for the synthesis of LPS (lipopolysaccharide) as the methyltransferase gene. In a previous study it was observed that a mutation in the methyltransferase gene alters the pattern of interaction between B. seminalis TC3.4.2R3 and phytopathogenic microorganisms, which suggests the possible involvement of this cluster in the production of antibiotics. Therefore, the extraction and characterization of LPS will be held from B. seminalis TC3.4.2R3 by thin-layer chromatography method (CDD) and bioauthography of antimicrobial compounds. Subsequently, O-antigen genes will be studied through the mutant synthesis by the CRISPR/Cas9 system gene knock out and the ability of these mutants to inhibit pathogens of interest will be evaluated. Thus, the main objective of this project is a better understanding of the possible mechanisms involved in the inhibition of plant pathogens by B. seminalis TC3.4.2R3 and the role of the gene in a metiltrasferase present in the O-antigen cluster in this process.
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