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Functional analysis of xylose isomerase in the response to xylose by Xanthomonas citri and X. fuscans- type B and differential proteomic analysis of this response

Grant number: 17/17470-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): February 01, 2018
Effective date (End): August 31, 2020
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Maria Teresa Marques Novo Mansur
Grantee:Evandro Luis Prieto
Home Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Citrus Canker is one of the most devastating diseases for citrus growing, caused by the bacterium Xanthomonas citri subsp. Citri (XAC). Cancerous cancers B and C, milder forms of the disease, are caused by types B and C of Xanthomonas fuscans ssp. Aurantifolii (XauB and XauC, respectively), whose host spectrum is very restricted compared to XAC, which infects all species of Citrus sp. Proteomic analysis between XAC and XauB previously performed by our group detected many differentially expressed proteins, among them, xylose isomerase, whose ORFs present very diverse genomic contexts in both bacteria. The present work aims to investigate if there is a more efficient metabolic response of XAC to xylose that is determinant of its greater virulence relative to XauB, since this carbohydrate is constituent of the cellular wall of the host and can be released during the bacterial invasion. In this work XAC and XauB will be cultured in the absence and presence of xylose, and, based on the resulting growth curves, the conditions under which proteomic analysis (2D-PAGE/MS-MS) will be performed for comparison of periplasmic proteins and Extracellular enzymes induced by this carbohydrate in the two bacteria. Differential expression of xylose isomerase will also be confirmed by means of Western Blot, as well as a deletion mutant for the gene of that enzyme using standardized methodology in our group. Preliminary results of growth of XAC and XauB in xylose are already presented here and show that the bacteria have different responses. The present work aims to contribute to the understanding of the pathogenicity of XAC for purposes of validation and characterization of potential targets for the control of Citrus Canker. (AU)