| 抑制南方水稻黑条矮缩病毒非结构蛋白P5-1的表达阻碍病毒在昆虫体内的增殖 |
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| 引用本文： | 曹金强,王针针,陈红燕,毛倩卓,贾东升,魏太云.抑制南方水稻黑条矮缩病毒非结构蛋白P5-1的表达阻碍病毒在昆虫体内的增殖[J].植物保护学报,2015,42(3):418-424. |
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| 作者姓名： | 曹金强 王针针 陈红燕 毛倩卓 贾东升 魏太云 |
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| 作者单位： | 福建农林大学植物病毒研究所,福建省植物病毒学重点实验室,福州350002 |
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| 基金项目： | 国家自然科学基金(31130044,31400136,31401712) |
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| 摘 要： | 为明确南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)编码的非结构蛋白P5-1参于SRBSDV在介体白背飞虱体内侵染过程中的作用机制,通过原核表达蛋白制备SRBSDV编码的非结构蛋白P5-1的多克隆抗体,并应用Western blot和免疫荧光标记法检测抗体的特异性,以注射法将来源于P5-1基因的dsRNA(ds P5-1)注入获毒1 d的白背飞虱体内,5 d后通过免疫荧光标记法检测ds P5-1对SRBSDV在白背飞虱体内增殖的影响,同时以注射来源于GFP基因的dsRNA(ds GFP)为对照。结果显示,Western blot和免疫荧光标记分别检测到SRBSDV侵染水稻和白背飞虱表达的P5-1蛋白,表明所制备的P5-1抗体具有特异性。ds GFP处理的对照组白背飞虱带毒率高达81%,而ds P5-1处理的白背飞虱带毒率仅为21%,且P5-1蛋白的表达和SRBSDV在昆虫体内的增殖均受到抑制。表明P5-1蛋白是病毒在昆虫体内增殖的关键因子,可作为阻断病毒在昆虫体内增殖的理想靶标。
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| 关 键 词： | 南方水稻黑条矮缩病毒 P5-1蛋白 抗体制备 RNA干扰 增殖 |
| 收稿时间： | 2014/10/17 0:00:00 |
Knockdown of nonstructural protein P5-1 of Southern rice black-streaked dwarf virus prevents viral multiplication in vector insects |
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| Cao Jinqiang,Wang Zhenzhen,Chen Hongyan,Mao Qianzhuo,Jia Dongsheng and Wei Taiyun.Knockdown of nonstructural protein P5-1 of Southern rice black-streaked dwarf virus prevents viral multiplication in vector insects[J].Acta Phytophylacica Sinica,2015,42(3):418-424. |
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| Authors: | Cao Jinqiang Wang Zhenzhen Chen Hongyan Mao Qianzhuo Jia Dongsheng and Wei Taiyun |
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| Institution: | Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China;Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China |
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| Abstract: | To investigate the function of the nonstructural protein P5-1 of Southern rice black streaked dwarf virus (SRBSDV) in the insect vector white-backed planthopper (WBPH), Sogatella furcifera (Horváth),here the prokaryotic expression of P5-1 was performed, followed by the preparation of polyclonal antibody against P5-1. Then the specificity of polyclonal antibody was tested by western blot and immunofluorescence. WBPH was microinjected with dsRNA targeting P5-1 gene (dsP5-1) of SRBSDV or dsRNA targeting GFP gene (dsGFP) at day one after virus acquisition. Five days later, the effect of dsP5-1 on viral mulitiplication in the insect vector was examined using immunofluorescence assay. The results showed that the protein P5-1 was detected in infected rice plants using western blot, and the specificity of polyclonal antibody against P5-1 was confirmed in viruliferous WBPH by using immunofluorescence. The rate of viruliferous insects in the control was 81%, while that of dsP5-1 microinjection was 21%. dsP5-1 significantly inhibited the expression of P5-1 and multiplication of SRBSDV in the body of WBPH vector. These results suggested that P5-1 was the key factor influencing viral multiplication in insect vector, which may serve as a target for blocking viral multiplication in WBPH. |
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| Keywords: | Southern rice black streaked dwarf virus P5-1 protein antibody preparation RNA interference multiplication |
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