中国小麦花叶病毒弱毒突变体筛选与鉴定  

Screening and identification of attenuated mutants of Chinese wheat mosaic virus

作　　者：许小洁 迟云化 于伟鹏 游朋朋 何梦君 耿超[1,2] 田延平 李向东[1,2] XU Xiao-jie;CHI Yun-hua;YU Wei-peng;YOU Peng-peng;HE Meng-jun;GENG Chao;TIAN Yan-ping;LI Xiang-dong(Department of Plant Pathology,College of Plant Protection,Shandong Agricultural University,Tai'an 271018,China;Shandong Provincial Key Lab of Agricultural Microbiology,Tai'an 271018,China)

机构地区：[1]山东农业大学植物保护学院植物病理学系,泰安271018 [2]山东省农业微生物重点实验室,泰安271018

出　　处：《植物病理学报》2021年第2期217-224,共8页Acta Phytopathologica Sinica

基　　金：农业公益性行业科技项目(201303021);山东省现代农业产业技术体系(SDAIT-02-10);泰山学者建设工程(ts201712023)。

摘　　要：中国小麦花叶病毒(Chinese wheat mosaic virus, CWMV)是我国小麦土传花叶病毒病的重要病原。将CWMV RNA1、RNA2片段分别连接到pCB301-Rz载体,得到重组质粒pCWMV-RNA1和pCWMV-RNA2,这2个质粒可在本氏烟上引起花叶和畸形症状,在小麦上引起黄花叶症状。在pCWMV-RNA2富含半胱氨酸蛋白基因的5′-端融合绿色荧光蛋白基因,可在本氏烟上系统产生绿色荧光。将RNA1 RdRp第1001位Asp(D_(1001))和RNA2 CP第127位Glu(E_(127))的密码子替换为丙氨酸Ala(A)密码子,获得突变体CWMV-RdRp-D1001A、CWMV-CP-E127A、CWMV-RdRp-D1001A/CP-E127A。与野生型CWMV相比,这3个突变体在本氏烟中致病力显著下降,积累量显著降低。Chinese wheat mosaic virus(CWMV) is one of the most economically important virus causing wheat soil-borne mosaic virus disease in China. The RNA 1 and RNA 2 fragments of CWMV were ligated to pCB301-Rz vector, producing recombinant plasmids pCWMV-RNA1 and pCWMV-RNA2, which could induce mosaic and deformation on Nicotiana benthamiana plants and yellow mosaic on wheat leaves. The genes for green fluorescent protein(GFP) was cloned to the 5′-end of the cysteine-rich gene of pCWMV-RNA2 and could produce green fluorescence in N. benthamiana plants. The codons for Asp(D_(1001)) of RdRp in RNA1 and Glu(E_(127)) of CP in RNA2 were mutated to that for Ala. Compared with wild-type CWMV, the resulting mutants, CWMV-RdRp-D1001 A, CWMV-CP-E127 A and CWMV-RdRp-D1001 A/CP-E127 A, showed significantly lower virulence and less CP accumulation in N. benthamiana plants.

关 键 词：中国小麦花叶病毒 侵染性cDNA克隆 致病力 衣壳蛋白 复制酶 

分 类 号：S432.41[农业科学—植物病理学]