Chinese Journal OF Rice Science >
OsOPR10 Positively Regulates Rice Blast and Bacterial Blight Resistance
Received date: 2023-12-15
Revised date: 2024-03-10
Online published: 2024-07-11
【Objective】 Jasmonic acid (JA) plays crucial roles in plant growth and development, and responses to both biotic and abiotic stresses. OPR (12-oxo-phytodienoic acid reductase) serves as a pivotal enzyme in the JA biosynthetic pathway. In this study, transgenic rice plants overexpressing OsOPR10 were generated to assess their resistance to rice blast and bacterial blight. The research delved into the molecular mechanisms through which OsOPR10 regulates the defense response to these diseases.【Method】 Methodologically, the study involved the construction of OsOPR10 CRISPR/Cas9 knockout and overexpression vectors. These vectors were then utilized for Agrobacterium-mediated genetic transformation to obtain OsOPR10 knockout (OsOPR10-KO) and overexpressed (OsOPR10-OE) transgenic rice plants, using Nipponbare (NPB) as the wild-type parent. The transgenic plants underwent various assays to evaluate disease resistance, reactive oxygen species (ROS) burst, and the expression of genes related to the salicylic acid (SA) and jasmonic acid (JA) pathways. Additionally, the subcellular localization of OsOPR10 was examined using a laser confocal microscopy, and the interacting protein of OsOPR10 was identified through yeast two-hybrid screening and luciferase complementation experiments.【Result】 The results of the study indicated the successful generation of homozygous plants with OsOPR10 knockout and overexpression. Plants overexpressing OsOPR10 exhibited enhanced resistance to rice blast and bacterial blight. Upon induction with chitin and bacterial flagellin (flg22), ROS accumulation in OsOPR10-OE plants was notably higher than that in the wild type. Furthermore, qRT-PCR analysis revealed up-regulation of JA pathway genes (OsAOS2, OsAOC) and SA pathway genes (OsPR1a, OsPAL1) in OsOPR10-OE plants compared to NPB at 12 hours post-inoculation with Magnaporthe oryzae. Subcellular localization studies demonstrated that the OsOPR10 protein was localized in chloroplasts. The interaction protein OsCYP28 of OsOPR10 was identified through yeast two-hybrid assays and luciferase complementation experiments 【Conclusion】 In conclusion, OsOPR10 plays a significant role in responding to infections by Magnaporthe oryzae and Xanthomonas oryzae pv. oryzea, as well as to the application of exogenous methyl jasmonate (MeJA) and SA. OsOPR10 is involved in the pathogen molecular pattern-triggered immune pathway and positively regulates rice resistance to rice blast and bacterial blight through the JA and SA pathways. Additionally, OsOPR10 protein localizes in chloroplasts and interacts with the OsCYP28 protein.
Key words： rice; jasmonic acid; OsOPR10; CRISPR/Cas9; overexpression; rice blast; bacterial blight
XU Danjie, LIN Qiaoxia, LI Zhengkang, ZHUANG Xiaoqian, LING Yu, LAI Meiling, CHEN Xiaoting, LU Guodong . OsOPR10 Positively Regulates Rice Blast and Bacterial Blight Resistance[J]. Chinese Journal OF Rice Science, 2024 , 38(4) : 364 -374 . DOI: 10.16819/j.1001-7216.2024.231215
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