灰比诺葡萄病毒侵染性克隆构建及致病性  

Construction and pathogenicity tests of one infectious clone of Grapevine Pinot gris virus

作　　者：刘灿 余尚真 张雪晴 冯晨露 张文镐 王金英 谢印帅 李涵伟 程玉琴 LIU Can;YU Shangzhen;ZHANG Xueqing;FENG Chenlu;ZHANG Wenhao;WANG Jinying;XIE Yinshuai;LI Hanwei;CHENG Yuqin(College of Horticulture,China Agricultural University,Beijing 100193,China;Key Laboratory of Viticulture and Enology of Ministry of Agriculture,Beijing 100083,China)

机构地区：[1]中国农业大学园艺学院,北京100193 [2]农业农村部葡萄酒加工重点实验室,北京100083

出　　处：《中国农业大学学报》2024年第11期74-82,共9页Journal of China Agricultural University

基　　金：国家自然科学基金面上项目(32272646)。

摘　　要：为探索灰比诺葡萄病毒(Grapevine Pinot gris virus,GPGV)在葡萄上的致病性,以感染GPGV山东分离物(GPGV-SD)的‘阳光玫瑰’葡萄植株为试材,利用RT-PCR扩增获得GPGV-SD的基因组全长序列并构建了该病毒的侵染性克隆,随后将其接种本生烟和葡萄植株进行致病性分析。结果表明:GPGV-SD基因组全长为7274个核苷酸(不含poly(A)尾),与国内外已报道的GPGV分离物的核苷酸序列相似度为74.00%~96.90%。GPGV-SD能系统侵染本生烟,分别在接种后5和8 d检测到第1片系统叶和顶部叶中的GPGV,并在接种后约3周观察到本生烟系统叶出现褪绿斑症状。利用农杆菌介导的真空渗透法也可使GPGV-SD系统侵染葡萄植株,‘红地球’和‘马瑟兰’葡萄苗在接种GPGV-SD后未表现出明显的病毒病症状。本研究有助于拓宽对GPGV致病性的认识,所构建的GPGV-SD的侵染性克隆可作为葡萄病毒与寄主互作研究的工具。To explore the pathogenicity of Grapevine Pinot gris virus(GPGV)on grapes,grapevine cultivar Shine Muscat plant infected with GPGV Shandong isolate(GPGV-SD)was taken as the study material.RTPCR was used to amplify the complete genome sequence of GPGV-SD and an infectious cDNA clone of the virus was then constructed.The pathogenicity of GPGV-SD on Nicotiana benthamiana and grapevine plants were also analyzed.The results indicated that:The genome of GPGV-SD was 7274 nucleotides in length excluding the poly(A)tail,which shared 74.00%to 96.90% nucleotide sequence similarity with that of the previous reported isolates of GPGV.GPGV-SD could systemically infect N.benthamiana plants and the virus was detected in the first systemically infected and upper leaves at about 5 to 8 days after inoculation.In addition,the symptoms of chlorotic blotches and spots were observed in systemically infected leaves of N.benthamiana plants at about 3 weeks after inoculation.GPGV-SD was also able to systemically infect grapevine plants via vacuum-agroinfiltration method,and the inoculation of the‘Marselan’and‘Red Globe’grapevine plants with GPGV-SD did not induce obvious symptoms.In conclusion,this study can broaden our knowledge on the pathogenicity of GPGV-SD,and the infectious cDNA clone of GPGV-SD is a power tool for studying the interactions between grapevine viruses and hosts.

关 键 词：葡萄灰比诺病毒 侵染性克隆 致病性 本生烟 葡萄 

分 类 号：S432.41[农业科学—植物病理学]