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鉴定荧光假单胞菌 Pf0-1 中氨基酸的趋化感觉蛋白及其在对番茄根分泌物和根定植的趋化作用中的作用。

Identification of chemotaxis sensory proteins for amino acids in Pseudomonas fluorescens Pf0-1 and their involvement in chemotaxis to tomato root exudate and root colonization.

机构信息

Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1–3–1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739–8530, Japan.

出版信息

Microbes Environ. 2012;27(4):462-9. doi: 10.1264/jsme2.me12005. Epub 2012 Sep 5.

Abstract

Pseudomonas fluorescens Pf0-1 showed positive chemotactic responses toward 20 commonly-occurring l-amino acids. Genomic analysis revealed that P. fluorescens Pf0-1 possesses three genes (Pfl01_0124, Pfl01_0354, and Pfl01_4431) homologous to the Pseudomonas aeruginosa PAO1 pctA gene, which has been identified as a chemotaxis sensory protein for amino acids. When Pf01_4431, Pfl01_0124, and Pfl01_0354 were introduced into the pctA pctB pctC triple mutant of P. aeruginosa PAO1, a mutant defective in chemotaxis to amino acids, its transformants showed chemotactic responses to 18, 16, and one amino acid, respectively. This result suggests that Pf01_4431, Pfl01_0124, and Pfl01_0354 are chemotaxis sensory proteins for amino acids and their genes were designated ctaA, ctaB, and ctaC, respectively. The ctaA ctaB ctaC triple mutant of P. fluorescens Pf0-1 showed only weak responses to Cys and Pro but no responses to the other 18 amino acids, indicating that CtaA, CtaB, and CtaC are major chemotaxis sensory proteins in P. fluorescens Pf0-1. Tomato root colonization by P. fluorescens strains was analyzed by gnotobiotic competitive root colonization assay. It was found that ctaA ctaB ctaC mutant was less competitive than the wild-type strain, suggesting that chemotaxis to amino acids, major components of root exudate, has an important role in root colonization by P. fluorescens Pf0-1. The ctaA ctaB ctaC triple mutant was more competitive than the cheA mutant of P. fluorescens Pf0-1, which is non-chemotactic, but motile. This result suggests that chemoattractants other than amino acids are also involved in root colonization by P. fluorescens Pf0-1.

摘要

荧光假单胞菌 Pf0-1 对 20 种常见的 l-氨基酸表现出正向趋化反应。基因组分析表明,荧光假单胞菌 Pf0-1 拥有三个与铜绿假单胞菌 PAO1 的 pctA 基因同源的基因(Pfl01_0124、Pfl01_0354 和 Pfl01_4431),该基因已被鉴定为氨基酸的趋化感觉蛋白。当 Pf01_4431、Pfl01_0124 和 Pfl01_0354 被引入铜绿假单胞菌 PAO1 的 pctA pctB pctC 三突变体中时,该突变体在氨基酸趋化性方面存在缺陷,其转化体分别对 18、16 和一种氨基酸表现出趋化反应。这一结果表明,Pf01_4431、Pfl01_0124 和 Pfl01_0354 是氨基酸的趋化感觉蛋白,它们的基因分别被命名为 ctaA、ctaB 和 ctaC。荧光假单胞菌 Pf0-1 的 ctaA ctaB ctaC 三突变体仅对 Cys 和 Pro 表现出较弱的反应,而对其他 18 种氨基酸没有反应,表明 CtaA、CtaB 和 CtaC 是荧光假单胞菌 Pf0-1 的主要趋化感觉蛋白。通过无菌竞争根定植试验分析了荧光假单胞菌菌株对番茄根的定殖。发现 ctaA ctaB ctaC 突变体比野生型菌株的竞争力弱,表明对氨基酸的趋化性(根分泌物的主要成分)在荧光假单胞菌 Pf0-1 对根的定殖中起着重要作用。ctaA ctaB ctaC 三突变体比荧光假单胞菌 Pf0-1 的 cheA 突变体更具竞争力,cheA 突变体是非趋化性的,但具有运动性。这一结果表明,除了氨基酸之外,其他趋化剂也参与了荧光假单胞菌 Pf0-1 对根的定殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5278/4103555/e1d7d09ba6e0/27_462f1.jpg

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