利用黄瓜花叶病毒m和fny株系的假重组体分析致病-植物病理学报.pdfVIP

植物病理学报 ACTA PHYTOPATHOLOGICA SINICA　 41(1):49-56(2011) 利用黄瓜花叶病毒 M和 Fny株系 的假重组体分析致病关键因子 1* 2 1* 2 * 陈玲娜 , 马云霞 , 高必达 , 朱水芳 1 2 ( 湖南农业大学生物安全科技学院,长沙 410128; 中国检验检疫科学研究院,北京 100029) 摘要: 用RT-PCR扩增黄瓜花叶病毒M株系(CMV-M)全长基因组cDNA,成功构建CMV-M RNA2 和RNA3侵染性克隆 后,与CMV-Fny基因组RNA交换得到3个假重组型病毒 (F1M2F3、F1F2M3、F1M2M3)。 用F1M2F3、F1F2M3、F1M2M3 分别侵染白肋烟,产生坏死环斑、轻微绿斑驳、明脉、黄白化和叶尖线性化等症状。 根据假重组型病毒和野生型病毒的表观 症状,分析引起各种症状的关键因子,初步判定:CP基因是诱导花叶症状的关键因子,CMV-Fny RNA2是诱导叶尖线性化 的关键因子,CMV-MRNA2是诱导叶尖坏死斑关键因子。 实时荧光定量PCR结果显示:野生株CMV-M、CMV-Fny和假重 组体F1M2F3、F1F2M3、F1M2M3侵染烟草后引起的症状差异与病毒基因组RNA累积没有直接关系。 关键词: 黄瓜花叶病毒;侵染性克隆;假重组;实时荧光定量PCR Analysis of keyvirulent factors by pseudo-recombinationbetween Cucumbermosaic virus Mand Fny strains　 1,2 2 1 2 1 CHEN Ling-na , MA Yun-xia , GAO Bi-da , ZHU Shui-fang ( College 2 of Bio-safety Science Technology ,Hunan Agriculture University,Changsha 410128,China; Institute of Animal and Plant Quarantine,Chinese Academy of Inspection and Quarantine,Beijing 100029,China) Abstract: Using reverse transcription-PCR (RT-PCR),full-length cDNA clones of Cucumber mosaic virus M strain (CMV-M) were constructed. Through pseudo-recombination between CMV-M (RNA2 and RNA3) and CMV-FnygenomicRNAs,thepseudo-recombinantsF1M2F3,F1F2M3andF1M2M3 were successfulob- tained. The seedings ofNicotiana tabacum cv. White Burley inoculated with pseudo-recombinants F1M2F3, F1F2M3 and F1M2M3 showed symptoms of necrosis,chlorosis,vein clearing,labefaction,linearization of leaf apex,etc. Basedon comparison of symtomsbetweenpseudo-recombinantsandwildtypevirus,thekeyvir- ulent factorswere speculated. Thekey factor which induced mosaic was