| 鞭毛素蛋白FliCaaa诱导水稻免疫反应的活性测定 |
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| 引用本文： | 刘莹,陈华民,于清,田芳,鲁国东,何晨阳.鞭毛素蛋白FliCaaa诱导水稻免疫反应的活性测定[J].植物保护学报,2015,42(1):66-71. |
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| 作者姓名： | 刘莹 陈华民 于清 田芳 鲁国东 何晨阳 |
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| 作者单位： | 福建农林大学植物保护学院, 福州 350002;中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193;中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193;中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193;中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193;福建农林大学植物保护学院, 福州 350002;中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 |
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| 基金项目： | 国家"863"计划(2012AA101504),国家"973"计划(2011CB100700) |
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| 摘 要： | 为阐明燕麦嗜酸菌(Acidovorax avenae subsp.avenae,Aaa)鞭毛素蛋白Fli Caaa及其结构域对水稻免疫反应的诱导作用,通过分子生物学和生物信息学方法,对Aaa菌株IPN1的鞭毛素进行基因克隆及其序列分析;在大肠杆菌中对该基因全长、N端和C端片段进行了原核表达,纯化了Fli Caaa蛋白及其截短肽段;采用水稻叶片浸润法测定了表达蛋白对水稻免疫反应的诱导活性。结果表明,通过特异性引物的PCR扩增,获得1 479 bp的鞭毛素基因fli Caaa,其编码产物含有492个氨基酸,分子量为49.4 k D,具有flagellin-N和-C两个保守结构域;原核诱导表达获得了Fli Caaa、Fli Caaa-N和Fli Caaa-C可溶性融合蛋白;3种纯化蛋白均能诱导水稻细胞死亡和H2O2产生等免疫反应,但诱导能力略有差异。
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| 关 键 词： | 燕麦嗜酸菌 鞭毛素 原核表达 纯化 免疫反应 |
| 收稿时间： | 2014/4/27 0:00:00 |
Functional analysis of the flagellin FliCaaa from Acidovorax avenae subsp. avenae in elicitation of immune responses of rice |
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| Liu Ying,Chen Huamin,Yu Qing,Tian Fang,Lu Guodong and He Chenyang.Functional analysis of the flagellin FliCaaa from Acidovorax avenae subsp. avenae in elicitation of immune responses of rice[J].Acta Phytophylacica Sinica,2015,42(1):66-71. |
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| Authors: | Liu Ying Chen Huamin Yu Qing Tian Fang Lu Guodong and He Chenyang |
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| Institution: | College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China |
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| Abstract: | To reveal the functions of flagellin FliCaaa from Acidovorax avenae subsp. avenae (Aaa) in induction of immune responses of rice and its domains, the fliCaaa gene cloned from the strain IPN1 of Aaa was expressed in Escherichia coli strain BL21, and the purified protein FliCaaa and the truncated proteins FliCaaa-N and FliCaaa-C were assayed for induction of immune responses in rice. The results showed that three purified proteins could induce the immune response of the rice Oryza sativa L. cv. Nipponbare, including hypersensitive cell death and hydrogen peroxide (H2O2) production, when the rice leaves were inoculated by infiltration of the tested proteins. Stronger induced defense response was found in FliCaaa-treated rice than in those treated with FliCaaa-N and FliCaaa-C. |
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| Keywords: | Acidovorax avenae subsp avenae flagellin expression purification immune response |
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