茶毛虫感染核型多角体病毒的快速分子鉴定与应用.pdfVIP

全国茶业科技学术研讨会论文集 茶毛虫感染核型多角体病毒的快速分子鉴定与应用 付建玉，韩宝瑜* (农业部茶叶化学工程重点实验室；中国农业科学院茶叶研究所, 浙江 杭州 310008 ) 摘要：根据茶毛虫核型多角体病毒（EpNPV ）两个核心基因 A13-1 lef-8 和 A13-1 polyhedrin 的核苷酸序列，设计合成了两对特异性引物，应用 PCR 方法分别扩增获得 538 bp 和 281 bp 的两个DNA 片段。克隆至 T 载体测序后与 Genbank 中已知EpNPV 两 基因序列比对，匹配率为 100%。证实所克隆的特异性 DNA 片段可以作为鉴定茶毛虫 感染 EpNPV 的标志性序列。茶园中以茶毛虫性信息素结合 EpNPV 防治茶毛虫，用茶 毛虫性诱剂引来雄蛾，使其阳具感染 EpNPV ，在带毒交尾或产卵过程中，致下代幼虫 感染 EpNPV 。对感病茶毛虫幼虫用此方法鉴定病原，符合率达到 100%。本研究确立 了茶毛虫感染 EpNPV 的分子鉴定方法，并为 EpNPV 防治茶毛虫效果的评价以及 EpNPV 侵染茶毛虫途径等毒理学研究提供依据。 关键词：茶毛虫核型多角体病毒 EpNPV ；PCR ；基因；分子鉴定 Rapidly Molecule Identification of Euproctis pseudoconspersa Infected with EpNPV by PCR Method and Application FU Jian-Yu ，HAN Bao-Yu* (Key Laboratory of Tea Chemical Engineering of Ministry of Agriculture; Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China) Abstract ：Referring to nucleotide sequences of the two key genes, A13-1 lef-8 gene and A13-1 polyhedrin, of Euproctis pseudoconspera nuclear polyhedrosis virus (EpNPV), two special primers were designed. Two typical DNA fragments with the size of 538 bp and 281 bp were amplified by PCR method. The two clone products were sequenced and blasted with the DNA sequence of EpNPV, both of them were matched absolutely. That meant the two DNA fragments can be used as molecule markers to identify the Euproctis pseudoconspersa Strand infected by EpNPV. Sex pheromone was used to attract males of E. pseudoconspera, their phallus were infected with EpNPV, then release them. After the males with EpNPV copulated with f