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作物学报 ›› 2025, Vol. 51 ›› Issue (2): 334-346.doi: 10.3724/SP.J.1006.2025.32006

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

疣粒野生稻中OgXa13的克隆和功能研究

张正康1,**，苏延红1,**，阮孙美1，张敏1，张攀1，张慧2，曾千春2，罗琼1,*   

  1. 1云南农业大学云南生物资源保护与利用国家重点实验室, 云南昆明650201; 2云南农业大学农学与生物技术学院, 云南昆明650201
  • 收稿日期:2023-02-12 修回日期:2024-10-25 接受日期:2024-10-25 出版日期:2025-02-12 网络出版日期:2024-11-13
  • 基金资助:
    本研究由NSFC-云南联合基金(U2102219, U1302261)资助。

Cloning and functional study of OgXa13 in Oryza meyeriana

ZHANG Zheng-Kang1,**,SU Yan-Hong1,**,RUAN Sun-Mei1,ZHANG Min1,ZHANG Pan1,ZHANG Hui2,ZENG Qian-Chun2,LUO Qiong1,*   

  1. 1 Yunnan State Key Laboratory of Biological Resources Conservation and Utilization, Yunnan Agricultural University, Kunming 650201, Yunnan, China; 2 College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, Yunnan, China
  • Received:2023-02-12 Revised:2024-10-25 Accepted:2024-10-25 Published:2025-02-12 Published online:2024-11-13
  • Supported by:
    This study was supported by NSFC Yunnan United Fund (U2102219, U1302261).

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摘要：

白叶枯病是水稻生产中最严重的细菌性病害。优良抗病基因的挖掘和育种利用是防治该病害最经济有效的方法。疣粒野生稻(Oryza meyeriana)具有对白叶枯病高抗甚至免疫的特性，是白叶枯病抗性基因资源的天然宝库。课题组在疣粒野生稻转录组和基因组测序的基础上，从疣粒野生稻中克隆了水稻白叶枯抗性基因Osxa13的同源基因OgXa13的cDNA和含UTR区域的8908 bp基因组序列。序列分析结果显示，OgXa13基因由5个外显子和4个内含子组成，与水稻中感病基因OsXa13的基因组结构和核心启动子序列均一致。OgXa13与水稻OsXa13的蛋白序列有21个氨基酸的差异，其中4个氨基酸的替换差异位于MtN3.1结构域。过表达OgXa13的感病水稻TP309植株白叶枯病的抗性显著增强，推测氨基酸序列差异导致了OgXa13与OsXa13蛋白功能的不同，OgXa13可作为一个显性白叶枯病抗性基因在育种中利用。利用CRISPR/Cas9敲除日本晴中感病基因OsXa13的T1代纯合株系对白叶枯病的抗性也明显增强，表明通过CRISPR/Cas9编辑感病基因OsXa13是改良水稻对白叶枯病抗性的有效途径。该研究为水稻白叶枯病抗性育种提供了有价值的新基因资源和新信息。

关键词: 水稻, 疣粒野生稻, 白叶枯病抗性, 基因克隆

Abstract:

Bacterial blight is one of the most devastating bacterial diseases in rice production. The identification and utilization of resistance genes in rice breeding is the most economical and effective method for controlling this disease. Oryza meyeriana represents a valuable genetic resource due to its high resistance, and even immunity, to bacterial blight. In this study, we cloned the full-length cDNA and an 8908 bp genomic sequence (3695 bp+2252 bp+2961 bp) of OgXa13, a homolog of OsXa13, from Oryza meyeriana using transcriptome and genome sequencing. Sequence analysis revealed that the gene structure of OgXa13 consists of five exons and four introns, mirroring the structure of rice xa13/OsXa13, and its core promoter sequence is identical to that of the rice susceptibility gene OsXa13. A total of 21 amino acid differences were observed between OgXa13 and xa13/OsXa13, with four key substitutions located in the MtN3.1 domain. Overexpression of OgXa13 and its introduction into TP309 plants via genetic transformation significantly enhanced resistance to bacterial blight. It is hypothesized that the amino acid sequence differences contribute to the distinct functions of the OgXa13 and OsXa13 proteins, suggesting that OgXa13 could be utilized as a dominant resistance gene in rice breeding. Furthermore, knockout of the OsXa13 gene using CRISPR/Cas9 in Nipponbare T1 homozygous lines also significantly enhanced resistance to bacterial blight, demonstrating that editing the susceptibility gene OsXa13 through CRISPR/Cas9 is an effective strategy for improving resistance. This study provides a valuable genetic resource and new insights for breeding rice with enhanced resistance to bacterial blight.

Key words: rice, Oryza meyeriana, bacterial blight resistance, gene cloning, OgXa13

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