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CRISPR/Cas9-mediated simultaneous mutation of three salicylic acid 5-hydroxylase (OsS5H) genes confers broad-spectrum disease resistance in rice

文献类型： 外文期刊

作者： Liu, Xiong 1 ; Yu, Yan 1 ; Yao, Wei 1 ; Yin, Zhongliang 1 ; Wang, Yubo 1 ; Huang, Zijian 1 ; Zhou, Jie-Qiang 1 ; Liu, Jinling 1 ; Lu, Xuedan 1 ; Wang, Feng 1 ; Zhang, Guilian 1 ; Chen, Guihua 1 ; Xiao, Yunhua 1 ; Deng, Huabing 1 ; Tang, Wenbang 1 ;

作者机构： 1.Hunan Agr Univ, Coll Agron, Changsha, Peoples R China

2.Hunan Prov Key Lab Rice & Rapeseed Breeding Dis Re, Changsha, Peoples R China

3.Hunan Acad Agr Sci, Hunan Hybrid Rice Res Ctr, Changsha, Peoples R China

4.State Key Lab Hybrid Rice, Changsha, Peoples R China

关键词： rice; salicylic acid 5-hydroxylase; CRISPR; Cas9; disease resistance

期刊名称：PLANT BIOTECHNOLOGY JOURNAL （ 影响因子：13.8； 五年影响因子：13.2 ）

ISSN： 1467-7644

年卷期： 2023 年

页码：

收录情况： SCI

摘要： Salicylic acid (SA) is an essential plant hormone that plays critical roles in basal defence and amplification of local immune responses and establishes resistance against various pathogens. However, the comprehensive knowledge of the salicylic acid 5-hydroxylase (S5H) in rice-pathogen interaction is still elusive. Here, we reported that three OsS5H homologues displayed salicylic acid 5-hydroxylase activity, converting SA into 2,5-dihydroxybenzoic acid (2,5-DHBA). OsS5H1, OsS5H2, and OsS5H3 were preferentially expressed in rice leaves at heading stage and responded quickly to exogenous SA treatment. We found that bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo) strongly induced the expression of OsS5H1, OsS5H2, and OsS5H3. Rice plants overexpressing OsS5H1, OsS5H2, and OsS5H3 showed significantly decreased SA contents and increased 2,5-DHBA levels, and were more susceptible to bacterial blight and rice blast. A simple single guide RNA (sgRNA) was designed to create oss5h1oss5h2oss5h3 triple mutants through CRISPR/Cas9-mediated gene mutagenesis. The oss5h1oss5h2oss5h3 exhibited stronger resistance to Xoo than single oss5h mutants. And oss5h1oss5h2oss5h3 plants displayed enhanced rice blast resistance. The conferred pathogen resistance in oss5h1oss5h2oss5h3 was attributed to the significantly upregulation of OsWRKY45 and pathogenesis-related (PR) genes. Besides, flg22-induced reactive oxygen species (ROS) burst was enhanced in oss5h1oss5h2oss5h3. Collectively, our study provides a fast and effective approach to generate rice varieties with broad-spectrum disease resistance through OsS5H gene editing.

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