效应蛋白XopM靶向丙二烯氧化物合成酶干扰水稻茉莉酸介导的免疫

doi:10.1016/j.jia.2024.08.018

摘要/Abstract

摘要：

稻黄单胞菌（Xanthomonas oryzae pv. oryzae, Xoo）引起水稻白叶枯病（Bacterial blight, BB)是危害水稻安全生产的全球性重要病害。Xoo利用III型分泌系统（type III secretion system, T3SS）分泌的效应蛋白发挥毒力，造成病害的发生发展和产量损失。XopM是黄单胞菌中保守的T3SS效应物之一，然而，XopM的毒性功能在很大程度上是未知的。在本研究中，我们构建了PH-PthXo1菌株用于评估单一non-TALE的毒力贡献。致病性测定结果表明XopM对水稻的Xoo致病力有显著贡献。为了明确XopM在寄主水稻中的靶标进而阐述其致病机理，我们利用酵母双杂交鉴定到一个茉莉酸生物合成的关键酶丙二烯氧化物合酶（allene oxide synthase, OsAOS3）与XopM相互作用。进一步地，通过免疫共沉淀（co-immunoprecipitation, CoIP）、蛋白下拉实验（Pull-down）、分裂荧光素酶互补（split luciferase complementation, SLC）以及荧光双分子（bimolecular fluorescence complementation, BiFC）互补实验证明了XopM与OsAOS3的相互作用。随后，我们发现Xoo侵染水稻后，OsAOS3及其3个OsAOS同源基因（OsAOS1/2/4）均可被诱导上调表达，且在侵染早期2 h时达到顶峰随后下降至正常水平。此外，利用CRISPR/Cas9技术构建了OsAOS3突变体植株，发现OsAOS3基因敲除突变体在侵染和非侵染Xoo情况下JA积累显著减少，对Xoo和Xoc（X. oryzae pv. oryzicola）抗性显著降低；与野生型水稻日本晴相比，OsAOS3突变体中JA相关的防御基因显著下调，这些结果表明OsAOS3参与JA生物合成且正调控JA介导的水稻免疫。JA含量测定结果表明，携带xopM基因菌株能降低水稻中JA含量以及JA相关抗病基因的转录水平。综上所述，本研究结果揭示了一种新的毒力策略，即XopM通过劫持OsAOS3干扰JA介导的防御，从而导致水稻免疫力的抑制。

Abstract:

Bacterial blight (BB) of rice caused by the phytopathogenic bacterium Xanthomonas oryzae pv. oryzae (Xoo) is a disease of global importance. Xoo utilizes the type III secretion system (T3SS) and its effectors for virulence, and XopM is a conserved T3SS effector in Xanthomonas spp. However, the virulence function of XopM is largely unknown. In this study, we show that XopM contributes to Xoo virulence in rice. We demonstrate that XopM interacts with allene oxide synthase OsAOS3, a key enzyme involved in jasmonic acid (JA) biosynthesis. The expression levels of OsAOS3 and three homologues of OsAOS were elevated after Xoo infection. Knockout mutants of OsAOS3 exhibited decreased JA accumulation and reduced resistance to Xoo and X. oryzae pv. oryzicola. Moreover, JA-related defense genes were downregulated in osaos3 mutants during Xoo infection. Based on our results, we propose a model showing how XopM hijacks OsAOS3 to interfere with JA-mediated defenses, leading to a suppression of rice immunity. Our findings reveal a novel virulence strategy where Xanthomonas pathogens interfere with the JA pathway and modulate the host defense response.

Key words: Xanthomonas , XopM , allene oxide synthase , jasmonic acid , plant immunit

Ying Li, Linlin Liu, Qi Wang, Yong Wang, Jiali Yan, Moein Khojasteh, Syed Mashab Ali Shah, Zhengyin Xu, Gongyou Chen. 效应蛋白XopM靶向丙二烯氧化物合成酶干扰水稻茉莉酸介导的免疫[J]. Journal of Integrative Agriculture, 2026, 25(6): 2449-2461.

Ying Li, Linlin Liu, Qi Wang, Yong Wang, Jiali Yan, Moein Khojasteh, Syed Mashab Ali Shah, Zhengyin Xu, Gongyou Chen. The conserved Xanthomonas effector XopM targets allene oxide synthase OsAOS3 and interferes with jasmonate-mediated defense in rice[J]. Journal of Integrative Agriculture, 2026, 25(6): 2449-2461.

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