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番木瓜环斑病毒西瓜株系弱毒突变体的筛选与应用

引用本文：黄显德,王玉,闫志勇,耿超,田延平,李向东.番木瓜环斑病毒西瓜株系弱毒突变体的筛选与应用.植物保护学报,2019,46(4):738-744

DOI：10.13802/j.cnki.zwbhxb.2019.2018132

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作者	单位	E-mail
黄显德	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018
王玉	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018
闫志勇	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018
耿超	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018
田延平	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018
李向东	山东农业大学植物保护学院, 植物病毒研究室, 泰安 271018	xdongli@sdau.edu.cn

中文摘要:为防治番木瓜环斑病毒（Papaya ringspot virus，PRSV）在我国葫芦科作物上引起的病毒病害，以PRSV西瓜株系山东分离物（PRSV-SD）侵染性cDNA克隆为基础，采用定点突变方法将辅助成分-蛋白酶保守氨基酸137位和346位的天冬酰胺（N）和417位的缬氨酸（V）突变为丙氨酸（A），应用农杆菌浸润法接种西葫芦叶片并分析突变对PRSV-SD致病力的影响，筛选弱毒突变体，进而评价其交叉保护效果。结果表明，与野生型PRSV-SD相比，获得的3个突变体N137A、N346A和V417A，接种后在西葫芦植株上的症状明显减轻，衣壳蛋白在叶片中的积累水平分别为野生型PRSV-SD的24.0%、13.0%和4.0%，均为弱毒突变体。当保护间隔期为10 d时，弱毒突变体N137A具有完全的交叉保护效果，N346A可延迟发病15 d，而V417A无交叉保护效果。当间隔保护期为15 d时，弱毒突变体N137A和N346A的保护效率分别为100.0%和26.7%，而V417A无交叉保护效果。

中文关键词:番木瓜环斑病毒辅助成分-蛋白酶弱毒突变体葫芦科作物交叉保护

Screening and application of attenuated mutants of Papaya ringspot virus-watermelon strain

Author Name	Affiliation	E-mail
Huang Xiande	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Wang Yu	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Yan Zhiyong	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Geng Chao	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Tian Yanping	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Li Xiangdong	Laboratory of Plant Virology, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China	xdongli@sdau.edu.cn

Abstract:To control the diseases caused by Papaya ringspot virus (PRSV) in cucurbit crops, the asparagine (N) at positions 137 and 346 and valine (V) at position 417 of the helper component-protease (HC-Pro) of a PRSV isolate of watermelon strain from Shandong Province (PRSV-SD) were mutated to alanine (A) in an infectious cDNA clone via site-directed mutagenesis. The virulence of the resultant mutants on Cucurbita pepo plants were analyzed by agroinfiltration, and the cross protection efficiency of attenuated mutants were evaluated. The results indicated that, compared with those of the wild type PRSV-SD, the symptoms of mutants N137A, N346A and V417A on C. pepo plants were significantly attenuated, and their accumulative levels were significantly reduced to 24.0%, 13.0% and 4.0% of the wild type virus, indicating that all of them were attenuated mutants of PRSV-SD. When the protective interval was ten days, the attenuated mutant N137A provided complete cross protection effect, and N346A delayed the appearance of viral symptom by 15 days, while V417A showed no cross-protective effect. When the protective interval was 15 days, the protective efficiencies of attenuated mutants N137A and N346A on C. pepo plants were 100.0% and 26.7%, respectively, while V417A had no cross-protective effect.

keywords:Papaya ringspot virus helper component-protease (HC-Pro) attenuated mutant cucurbitaceous crops cross protection

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