水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究

doi:10.3969/j.issn.1001-7216.2010.04.003

中国水稻科学

水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究

刘希玲1,邹华松2,邹丽芳2,陈功友1，2，*

^１南京农业大学植物保护学院／农业部病虫监测与治理重点开放实验室，江苏南京２１００９５； ^２上海交通大学农业与生物学院，上海２００２４０； *通讯联系人， E-mail: gyouchen@njau.edu.cn

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-07-10 发布日期:2010-07-10

Site-Directed Mutagenesis and Functional Analysis of hrcU Gene from Rice Pathogen Xanthomonas oryzae pv. oryzae

LIU Xi-ling¹, ZOU Hua-song², ZOU Li-fang², CHEN Gong-you^1,2,*

¹Key Laboratory for Monitoring and Management of Plant Diseases and Insects, Ministry of Agriculture/College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; ²Department of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China; *Corresponding author, E-mail: gyouchen@njau.edu.cn

Received:1900-01-01 Revised:1900-01-01 Online:2010-07-10 Published:2010-07-10

摘要/Abstract

摘要： 通过基因敲除方式，构建了水稻白叶枯病菌hrcU基因的缺失突变体。突变体丧失了在水稻上的致病性和在非寄主植物上的过敏反应。携带hrcU基因的黏粒完全互补了hrcU突变菌株的表型，互补菌在水稻组织中的生长能力也与野生型菌株相当。免疫杂交结果显示，hrcU基因突变后，水稻白叶枯病菌的Harpin蛋白Hpa1不能从细胞内分泌到胞外。这些结果表明，水稻白叶枯病菌hrcU基因的主要功能是形成Ⅲ型分泌系统，将效应分子分泌到病原细菌胞外，从而决定了在非寄主上的过敏反应和在水稻上的致病性。

关键词: 水稻白叶枯病菌, 定点突变, 致病性, 过敏反应, Ⅲ型分泌系统

Abstract: The hrp gene cluster is composed of 27 members in rice pathogen Xanthomonas oryzae pv. oryzae(Xoo). To analyze the function of hrcU gene, the hrcU gene was knocked out by allelic exchange. The resulting hrcU mutant lost the pathogenicity in rice and the ability to trigger hypersensitive response in nonhost plant tobacco. The sole hrcU gene fully restored the phenotypic alterations in pathogenicity in rice, hypersensitive response in tobacco, and bacterial growth in rice. Western blot displayed that the typeⅢ secretion system was destroyed in hrcU mutant in which the T3S effectors Hpa1 could not be secreted into extracellular. This indicates that the hrcU gene of Xoo was essential for type Ⅲ secretion system to deliver effectors into plants.

Key words: Xanthomonas oryzae pv. oryzae, site-directed mutagenesis, pathogenicity, hypersensitive response, type Ⅲ secretion system

刘希玲,邹华松,邹丽芳,陈功友 . 水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究[J]. 中国水稻科学 2010,24(4): 348-352 . , DOI: 10.3969/j.issn.1001-7216.2010.04.003 .

LIU Xi-ling, ZOU Hua-song,ZOU Li-fang, CHEN Gong-you. Site-Directed Mutagenesis and Functional Analysis of hrcU Gene from Rice Pathogen Xanthomonas oryzae pv. oryzae [J]. Chinese Journal of Rice Science 2010,24(4): 348-352 ., DOI: 10.3969/j.issn.1001-7216.2010.04.003 .

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水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究

Site-Directed Mutagenesis and Functional Analysis of hrcU Gene from Rice Pathogen Xanthomonas oryzae pv. oryzae

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