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一种LysR型转录调节因子LcrX参与了辣椒青枯菌的毒力、生物膜形成、游动性、铁载体分泌以及在糖源中的生长。

A LysR-Type Transcriptional Regulator LcrX Is Involved in Virulence, Biofilm Formation, Swimming Motility, Siderophore Secretion, and Growth in Sugar Sources in Pv. .

作者信息

Park Hanbi, Do Eunsoo, Kim Minyoung, Park Hye-Jee, Lee Jongchan, Han Sang-Wook

机构信息

Department of Plant Science and Technology, Chung-Ang University, Anseong, South Korea.

Department of Systems Biotechnology, Chung-Ang University, Anseong, South Korea.

出版信息

Front Plant Sci. 2020 Jan 10;10:1657. doi: 10.3389/fpls.2019.01657. eCollection 2019.

Abstract

pv. () is a Gram-negative bacterium that causes bacterial pustule disease in soybean. To acclimate to new environments, the expression of genes in bacteria is controlled directly or indirectly by diverse transcriptional factors. Among them, LysR type transcriptional regulators are well-characterized and abundant in bacteria. In a previous study, comparative proteomic analysis revealed that LysR type carbohydrate-related transcriptional regulator in (LcrX) was more abundant in XVM2, which is a minimal medium, compared with a rich medium. However, the functions of LcrX in have not been characterized. In this study, we generated an LcrX-overexpressing strain, (LcrX), and the knockout mutant strain, (EV), to elucidate the functions of LcrX. Bacterial multiplication of (LcrX) in soybean was significantly impaired, indicating that LcrX is related to virulence. Comparative proteomic analysis revealed that LcrX is mainly involved in carbohydrate metabolism/transport and inorganic ion transport/metabolism. Based on the results of proteomics analysis, diverse phenotypic assays were carried out. A gel electrophoresis mobility shift assay demonstrated that LcrX specifically bound to the putative promoter regions of genes encoding putative fructose 1,6-bisphosphatase and protease. Through a 96-well plate assay under various conditions, we confirmed that the growth of (LcrX) was dramatically affected in the presence of various carbon sources, while the growth of (EV) was only slightly changed. Biofilm formation activity was reduced in (LcrX) but enhanced in (EV). The production of siderophores was also decreased in (LcrX) but not altered in (EV). In contrast, LcrX was not associated with exopolysaccharide production, protease activity, or bacterial motility. These findings provide new insights into the functions of a carbohydrate-related transcriptional regulator in .

摘要

丁香假单胞菌大豆致病变种(Pseudomonas syringae pv. glycinea)是一种革兰氏阴性细菌,可导致大豆细菌性脓疱病。为了适应新环境,细菌中基因的表达直接或间接受多种转录因子的控制。其中,LysR型转录调节因子在细菌中特征明确且数量众多。在之前的一项研究中,比较蛋白质组学分析表明,与丰富培养基相比,在基本培养基XVM2中,丁香假单胞菌大豆致病变种中与碳水化合物相关的LysR型转录调节因子(LcrX)更为丰富。然而,LcrX在丁香假单胞菌大豆致病变种中的功能尚未得到表征。在本研究中,我们构建了LcrX过表达菌株(LcrX)和基因敲除突变菌株(EV),以阐明LcrX 的功能。LcrX过表达菌株在大豆中的细菌增殖显著受损,表明LcrX与毒力有关。比较蛋白质组学分析表明,LcrX主要参与碳水化合物代谢/转运和无机离子转运/代谢。基于蛋白质组学分析结果,进行了多种表型分析。凝胶电泳迁移率变动分析表明,LcrX特异性结合编码推定果糖1,6-二磷酸酶和蛋白酶的基因的推定启动子区域。通过在各种条件下的96孔板分析,我们证实,在各种碳源存在的情况下,LcrX过表达菌株的生长受到显著影响,而基因敲除突变菌株的生长仅略有变化。LcrX过表达菌株的生物膜形成活性降低,而基因敲除突变菌株的生物膜形成活性增强。LcrX过表达菌株的铁载体产量也降低,但基因敲除突变菌株的铁载体产量未改变。相比之下,LcrX与胞外多糖产生、蛋白酶活性或细菌运动性无关。这些发现为丁香假单胞菌大豆致病变种中一种与碳水化合物相关的转录调节因子的功能提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a671/6965072/441a1d795f27/fpls-10-01657-g001.jpg

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