Análise global da expressão gênica de Xylella fastidiosa submetida a estresses ambientais / Global gene expression analysis of Xylella fastidiosa under environmental stress conditions

AUTOR(ES)
DATA DE PUBLICAÇÃO

2006

RESUMO

Xylella fastidiosa is a phytopathogenic bacterium responsible for diseases in many economically important crops. Although different strains have been studied, little is known about X. fastidiosa stress responses. To investigate X. fastidiosa genes involved in heat, salt and osmotic shock responses, we performed a whole genome microarray analysis in time-course experiments. The expression profiles of the differentially expressed genes were grouped and their expression patterns were validated by quantitative RT-PCR experiments. During heat shock, 261 genes were induced (9.7%) and 222 genes were repressed (8.3%). Among the differentially expressed genes, the ones presenting the highest induction ratios encode heat shock proteins (Hsps), which prevents protein misfolding and aggregation or promote the degradation of the irreversibly denatured polypeptides. We determined the transcription start sites of six heat shock inducible genes and analyzed their promoter regions, which allowed us to propose a putative consensus for sigma32 promoters in X. fastidiosa. We also observed the induction of genes related to the extracytoplasmic stress response, that are regulated by the alternative sigma factor sigmaE. During prolongued exposure to salt and osmotic stress, genes encoding most of the Hsps were repressed, indicating that the response is not mediated by sigma32 or sigmaE. Among the 142 genes induced by both salt and osmotic stress, 57% encode hypothetical or conserved hypothetical proteins, indicating a possible role of these genes in the stress response. In addition, we observed the repression of genes related to protein biosynthesis and intermediary metabolism during the three stresses tested, besides the induction of genes related to virulence such as toxins and adhesins, revealing the complex network of genes that work together in response to environmental stresses. To facilitate the microarray data analysis process, we developed three bioinformatics tools: HTself, which is used to determine the differentially expressed genes; BayGO, which aims at finding over-represented gene categories and SpotWhatR, a system that integrates programs used in different steps of microarray data analysis in a user-friendly interface. These tools were successfully used and are freely available to the research community.

ASSUNTO(S)

genome estresse ambiental genoma expressão gênica phytopathogenic bacterium environmental stresses xylella fastidiosa gene expression xylella fastidiosa bacteria fitopatogênica

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