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Chitin Utilization by the Insect-Transmitted Bacterium Xylella fastidiosa

机译：昆虫传播的细菌Xylella fastidiosa对几丁质的利用

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Xylella fastidiosa is an insect-borne bacterium that colonizes xylem vessels of a large number of host plants, including several crops of economic importance. Chitin is a polysaccharide present in the cuticle of leafhopper vectors of X. fastidiosa and may serve as a carbon source for this bacterium. Biological assays showed that X. fastidiosa reached larger populations in the presence of chitin. Additionally, chitin induced phenotypic changes in this bacterium, notably increasing adhesiveness. Quantitative PCR assays indicated transcriptional changes in the presence of chitin, and an enzymatic assay demonstrated chitinolytic activity by X. fastidiosa . An ortholog of the chitinase A gene ( chiA ) was identified in the X. fastidiosa genome. The in silico analysis revealed that the open reading frame of chiA encodes a protein of 351 amino acids with an estimated molecular mass of 40 kDa. chiA is in a locus that consists of genes implicated in polysaccharide degradation. Moreover, this locus was also found in the genomes of closely related bacteria in the genus Xanthomonas , which are plant but not insect associated. X. fastidiosa degraded chitin when grown on a solid chitin-yeast extract-agar medium and grew in liquid medium with chitin as the sole carbon source; ChiA was also determined to be secreted. The gene encoding ChiA was cloned into Escherichia coli , and endochitinase activity was detected in the transformant, showing that the gene is functional and involved in chitin degradation. The results suggest that X. fastidiosa may use its vectors' foregut surface as a carbon source. In addition, chitin may trigger X. fastidiosa's gene regulation and biofilm formation within vectors. Further work is necessary to characterize the role of chitin and its utilization in X. fastidiosa .
机译：枯草杆菌是一种昆虫传播的细菌，可在大量宿主植物（包括几种具有重要经济价值的作物）的木质部容器中定殖。甲壳质是存在于fast.fastidiosa的叶蝉载体的表皮中的多糖，并且可以用作该细菌的碳源。生物学测定表明，在甲壳质存在下，X。fastidiosa达到了较大的种群。另外，几丁质诱导该细菌的表型变化，特别是增加了粘附性。定量PCR测定法表明在甲壳质存在下转录的变化，而酶法测定法证明X.fastidiosa具有几丁质分解活性。在X. fastidiosa基因组中鉴定了几丁质酶A基因的直系同源物。电脑分析表明，chiA的开放阅读框编码351个氨基酸的蛋白质，估计分子量为40 kDa。 chiA位于一个由与多糖降解有关的基因组成的基因座中。此外，该基因座还存在于Xanthomonas属的密切相关细菌的基因组中，这些细菌与植物相关，但与昆虫无关。当在固态几丁质酵母提取琼脂培养基上生长，并在以几丁质为唯一碳源的液体培养基中生长时，fastidiosa X.降解了几丁质。 ChiA也被确定是被分泌的。将编码ChiA的基因克隆到大肠杆菌中，并在转化子中检测到内切几丁质酶活性，表明该基因具有功能并参与了几丁质的降解。结果表明，X。fastidiosa可能利用其媒介的前肠表面作为碳源。此外，几丁质可能会在载体内触发法氏假单胞菌的基因调控和生物膜形成。表征甲壳素及其在X. fastidiosa中的作用尚需进一步的研究。

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