反应调节蛋白GlrR<span>和</span><span>ArcA</span>在<span>嗜线虫致病杆菌</span>CB6菌株中<span>调控</span>xenocoumacins<span>生物合成的研究</span>

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (12): 4703-4714.DOI: 10.1016/j.jia.2024.02.014

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反应调节蛋白GlrR和ArcA在嗜线虫致病杆菌CB6菌株中调控xenocoumacins生物合成的研究

收稿日期:2023-09-09 修回日期:2024-02-29 接受日期:2024-01-02 出版日期:2025-12-20 发布日期:2025-11-13

Studying of the regulatory roles of response regulators GlrR and ArcA in xenocoumacins biosynthesis in Xenorhabdus nematophila CB6

Xiaohui Li, Xiaobing Zheng, Yijie Dong, Youcai Qin, Fenglian Jia, Baoming Yuan, Jiaqi Duan, Beibei Li, Guangyue Li^#

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2023-09-09 Revised:2024-02-29 Accepted:2024-01-02 Online:2025-12-20 Published:2025-11-13
About author:Xiaohui Li, E-mail: xiaohuili0101@163.com; #Correspondence Guangyue Li, Tel: +86-10-82105929, E-mail: liguangyue@caas.cn
Supported by:
We thank the support from the National Natural Science Foundation of China (31972327), the Key Research and Development Program Project of Xinjiang, China (2022B02044), the National Key R&D Program of China (2023YFD1700700), the Agricultural Science and Technology Innovation Program of CAAS and the Chinese Academy of Agriculture Sciences for the fund of Elite Youth Program.

摘要/Abstract

摘要：

Xencoumacins（Xcns）是嗜线虫致病杆菌产生的主要抗菌活性天然产物，在植物病害防治领域具有广阔的应用前景。目前，关于Xcns生物合成的调控研究取得了一些进展，双组分调控系统EnvZ/OmpR和CpxA/CpxR中的反应调节蛋白OmpR和CpxR被报道参与调控Xcns的生物合成，但是对Xcns生物合成的调控机制仍缺乏系统深入的认知。在本研究中，我们首先利用生物信息学方法从嗜线虫致病杆菌CB6菌株的基因组中识别了21个双组分调控系统，通过基因敲除和回补实验表明反应调节蛋白GlrR和ArcA参与调控Xcns的生物合成。RT-qPCR和融合报告基因技术研究表明GlrR和ArcA通过正向调控Xcns生物合成基因簇相关基因的表达来调控Xcns的生物合成，进一步通过生物信息学分析、凝胶阻滞实验（EMSA）和DNase I足迹实验解析了GlrR和ArcA的具体调控机制，实验结果表明GlrR间接调控Xcns的生物合成，ArcA通过直接结合在xcnA和xcnB基因的启动子区域来调控Xcns的生物合成。ArcA在xcnA基因启动子区域的具体结合位点为5′-TCTGATGTTAATGTTTTGTCG-3′，ArcA在xcnB基因启动子区域的具体结合位点为5′-TTTATTAAAAATGAATTGTTACG-3′。此外，研究了基因glrR和arcA的缺失对细胞的形态和生长的影响。扫描电镜结果表明GlrR不影响细胞的形态，但arcA的缺失导致细胞长度明显变长，CB6菌株、ΔglrR和ΔarcA的细胞长度分别为3.23 μm、3.53 μm和7.98 μm。基因glrR和arcA的缺失导致菌株的生长受到了抑制，CB6菌株、ΔglrR和ΔarcA在96 h的OD₆₀₀光密度值分别为17.03、12.23和11.94，表明GlrR和ArcA影响细胞的生长。本研究首次揭示了反应调节蛋白GlrR和ArcA参与调控嗜线虫致病杆菌Xcns的生物合成，并解析了其调控机制，为构建Xcns高产菌株提供理论指导。

Abstract:

Xenocoumacins (Xcns), the major antimicrobial natural products produced by Xenorhabdus nematophila, have gained widespread attention for their potential application in crop protection. However, the regulatory mechanisms involved in the biosynthesis of Xcns remain poorly understood. In this study, we identified 21 potential two-component systems (TCSs) in X. nematophila CB6 by bioinformatic analysis. Among them, the response regulators (RRs), GlrR and ArcA, were proven to positively regulate the production of Xcns based on gene deletion and complementation experiments. In addition, our results showed that GlrR played an important role in cell growth, while ArcA was involved in both cell morphology and growth. Using a variety of molecular biological and biochemical techniques, we found that GlrR controlled the Xcns biosynthesis by indirectly regulating the expression levels of the biosynthetic gene cluster (BGC). ArcA directly binded to the promoter regions of xcnA and xcnB to regulate the transcription of the Xcns BGC, and the binding sites were also identified. This study provides valuable insights into the regulatory network of Xcns biosynthesis, which will contribute to the construction of a high-yielding strain.

Key words: Xenorhabdus nematophila , xenocoumacins , two-component systems , GlrR , ArcA

Xiaohui Li, Xiaobing Zheng, Yijie Dong, Youcai Qin, Fenglian Jia, Baoming Yuan, Jiaqi Duan, Beibei Li, Guangyue Li. 反应调节蛋白GlrR和ArcA在嗜线虫致病杆菌CB6菌株中调控xenocoumacins生物合成的研究[J]. Journal of Integrative Agriculture, 2025, 24(12): 4703-4714.

Xiaohui Li, Xiaobing Zheng, Yijie Dong, Youcai Qin, Fenglian Jia, Baoming Yuan, Jiaqi Duan, Beibei Li, Guangyue Li. Studying of the regulatory roles of response regulators GlrR and ArcA in xenocoumacins biosynthesis in Xenorhabdus nematophila CB6[J]. Journal of Integrative Agriculture, 2025, 24(12): 4703-4714.

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