糖苷水解酶7家族成员VdGH7a通过与GhOLP1相互作用，调控大丽轮枝菌的致病性并诱导寄主防御反应

doi:10.1016/j.jia.2024.03.002

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (11): 4294-4309.DOI: 10.1016/j.jia.2024.03.002

糖苷水解酶7家族成员VdGH7a通过与GhOLP1相互作用，调控大丽轮枝菌的致病性并诱导寄主防御反应

收稿日期:2023-07-21 修回日期:2024-03-21 接受日期:2024-01-08 出版日期:2025-11-20 发布日期:2025-10-13

The glycoside hydrolase 7 member VdGH7a regulates Verticillium dahliae pathogenicity and induces host defenses by interacting with GhOLP1

Junyuan Lü^{1, 2, 4*}, Shichao Liu^{2, 3*}, Jinglong Zhou^{1, 2}, Zili Feng^{1, 2}, Feng Wei^{1, 2}, Lihong Zhao^{1, 2}, Haiqiang Li⁵, Heqin Zhu², Yalin Zhang^{1, 2#}, Hongjie Feng^{1, 2#}

¹ National Nanfan Research Institute (Sanya), Chinese Academy of Agricultural Sciences, Sanya 572024, China
² National Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China
³ Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, China
⁴ College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
⁵ Institute of Plant Protection, Xinjiang Academy of Agricultural Sciences, Urumuqi 830091, China

Received:2023-07-21 Revised:2024-03-21 Accepted:2024-01-08 Online:2025-11-20 Published:2025-10-13
About author:Junyuan Lü, E-mail: ljyuan0426@163.com; Shichao Liu, E-mail: liushichao29@163.com; #Correspondence Hongjie Feng, Tel: +86-371-55912725, E-mail: fenghongjie@caas.cn; Yalin Zhang, E-mail: zhangyalin@caas.cn * These authors contributed equally to this study.
Supported by:
This work was supported by the Project of Sanya Yazhou Bay Science and Technology City, China (SCKJ-JYRC-2022-75), and the Natural Science Foundation of Hainan Province, China (322QN398).

摘要/Abstract

摘要：

病原菌可以分泌多种可以降解细胞壁的酶，从而削弱宿主的细胞壁，便于病原菌侵入植物内部。本研究中，我们鉴定了大丽轮枝菌（Verticillium dahliae）中的一种糖苷酶家族7（GH7）纤维素水解酶VdGH7a，对1,4-β-葡聚糖显示出水解活性。有趣的是，我们发现VdGH7a在存在信号肽的情况下可以诱导烟草（Nicotiana benthamiana）叶片的细胞死亡，在缺失信号肽的情况下丧失这种活性，但是诱导细胞坏死这一现象可以被碳水化合物结合型-1（CBM1）蛋白结构域有效阻止。此外，我们观察到在大丽轮枝菌中敲除VdGH7a基因显著降低了对棉花的致病力，这可以在敲除突变体菌株不能够穿透玻璃纸膜中得到证明。此外，这些敲除突变体菌株利用碳源的能力下降，使其更容易受到渗透胁迫和细胞壁应激的影响。此外，我们通过酵母双杂交技术筛选发现VdGH7a与棉花中的类渗调素蛋白蛋白（GhOLP1）相互作用，并通过双分子荧光互补（BiFC）和荧光素互补成像（LCI）试验进一步确定其存在互作关系。此外，通过利用病毒诱导基因沉默技术在棉花中沉默GhOLP1基因的表达，结果发现沉默GhOLP1的棉花株系中水杨酸（SA）含量和对大丽轮枝菌的抗性都降低了，而在拟南芥中异源过表达GhOLP1则增加了对大丽轮枝菌的抵抗性和水杨酸信号通路相关基因的表达。总的来说，这些发现表明大丽轮枝菌中的分泌蛋白VdGH7a与棉花中GhOLP1相互作用，激活寄主免疫反应，在棉花对大丽轮枝菌的抵抗中发挥重要作用。

Abstract:

Plant pathogens secrete various cell wall-degrading enzymes that compromise host cell wall integrity and facilitate pathogen invasion. This study identified VdGH7a, a glycoside hydrolase family 7 (GH7) cellobiohydrolase from Verticillium dahliae, which demonstrated hydrolytic activity against 1,4-β-glucan. Notably, VdGH7a induced cell death in Nicotiana benthamiana when signal peptides were present, though this effect was inhibited by the carbohydrate-binding type-1 (CBM1) protein domain. The deletion of VdGH7a substantially reduced V. dahliae pathogenicity in cotton plants, as demonstrated by the mutants’ inability to penetrate cellophane membrane. These knockout mutants also exhibited reduced carbon source utilization capacity and increased sensitivity to osmotic and cell wall stresses. Through yeast two-hybrid screening, bi-molecular fluorescence complementation (BiFC), and luciferase complementation imaging (LCI), we identified that VdGH7a interacts with an osmotin-like protein (GhOLP1) in cotton. Virus-induced gene silencing of GhOLP1 resulted in decreased salicylic acid (SA) content and reduced resistance to V. dahliae in cotton, while heterologous overexpression of GhOLP1 in Arabidopsis enhanced both resistance and SA signaling pathway gene expression. These results reveal a virulence mechanism wherein the secreted protein VdGH7a from V. dahliae interacts with GhOLP1 to activate host immunity and contribute significantly to plant resistance against V. dahliae.

Key words: Verticillium dahliae , glycoside hydrolase , pathogenicity , interacting protein , host immunity

Junyuan Lü, Shichao Liu, Jinglong Zhou, Zili Feng, Feng Wei, Lihong Zhao, Haiqiang Li, Heqin Zhu, Yalin Zhang, Hongjie Feng. 糖苷水解酶7家族成员VdGH7a通过与GhOLP1相互作用，调控大丽轮枝菌的致病性并诱导寄主防御反应[J]. Journal of Integrative Agriculture, 2025, 24(11): 4294-4309.

Junyuan Lü, Shichao Liu, Jinglong Zhou, Zili Feng, Feng Wei, Lihong Zhao, Haiqiang Li, Heqin Zhu, Yalin Zhang, Hongjie Feng. The glycoside hydrolase 7 member VdGH7a regulates Verticillium dahliae pathogenicity and induces host defenses by interacting with GhOLP1[J]. Journal of Integrative Agriculture, 2025, 24(11): 4294-4309.

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糖苷水解酶7家族成员VdGH7a通过与GhOLP1相互作用，调控大丽轮枝菌的致病性并诱导寄主防御反应

The glycoside hydrolase 7 member VdGH7a regulates Verticillium dahliae pathogenicity and induces host defenses by interacting with GhOLP1

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