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Journal of Integrative Agriculture
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The glycoside hydrolase 7 member VdGH7a regulates Verticillium dahliae pathogenicity and induces host defenses by interacting with GhOLP1
Junyuan Lv1, 2, 4*, Shichao Liu2, 3*, Jinglong Zhou1, 2, Zili Feng1, 2, Feng Wei1, 2, Lihong Zhao1, 2, Haiqiang Li5, Heqin Zhu2, Yalin Zhang1, 2#, Hongjie Feng1, 2#

1 National Nanfan Research Institute (Sanya), Chinese Academy of Agricultural Sciences, Sanya 572024, Hainan, China

2 National Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang 455000, Henan, China

3 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan, China

4 College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, China

5 Institute of Plant Protection, Xinjiang Academyof Agricultural Sciences, Urumuqi 830091, Xinjiang China

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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7GH7)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-1CBM1)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  Pathogens secrete multiple enzymes that can degrade the cell wall, thereby weakening the host's cell wall and facilitating the penetration of the pathogen into the plant. In this study, we identified VdGH7a, a glycoside hydrolase family 7 (GH7) cellobiohydrolase from Verticillium dahliae, which exhibited hydrolytic activity against 1,4-β-glucan. Interestingly, we found that VdGH7a induced cell death in Nicotiana benthamiana when signal peptides were present. However, this phenomenon was effectively prevented by the carbohydrate-binding type-1 (CBM1) protein domain. Furthermore, we observed that the knockout of VdGH7a significantly reduced the pathogenicity of V. dahliae to cotton plant, as evidenced by the inability of the knockout mutants to penetrate cellophane membrane. Additionally, these knockout mutants displayed diminished ability to exploit carbon sources, rendering them more susceptible to osmotic and cell wall stresses. Moreover, VdGH7a interacted with an osmotin-like protein (GhOLP1) in cotton through yeast two-hybrid screening, and further confirmed using bi-molecular fluorescence complementation (BiFC) and luciferase complementation imaging (LCI). Furthermore, virus-induced gene silencing technology was employed to silence GhOLP1, causing cotton's salicylic acid (SA) content and resistance to V. dahliae were both reduced, whereas heterologous overexpression of GhOLP1 in Arabidopsis increased both resistance and the expression of genes involved in the SA signaling pathway. Collectively, these findings demonstrate a virulence strategy whereby the secreted protein VdGH7a from V. dahliae interacts with GhOLP1 to stimulate host immunity and play a significant role in plant resistance against V. dahliae.
Keywords:  Verticillium dahliae              glycoside hydrolase              pathogenicity              interacting protein              host immunity  
Online: 21 March 2024  
About author:  Junyuan Lv, E-mail: ljyuan0426@163.com; Shichao Liu, E-mail: liushichao29@163.com; #Correspondence Hongjie Feng, Tel: +86-37155912725, E-mail: fenghongjie@caas.cn; Yalin Zhang, E-mail: zhangyalin@caas.cn * Lv Junyuan and Liu Shichao contributed equally to this work.

Cite this article: 

Junyuan Lv, Shichao Liu, Jinglong Zhou, Zili Feng, Feng Wei, Lihong Zhao, Haiqiang Li, Heqin Zhu, Yalin Zhang, Hongjie Feng. 2024. The glycoside hydrolase 7 member VdGH7a regulates Verticillium dahliae pathogenicity and induces host defenses by interacting with GhOLP1. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.03.002

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