侵染百合的李属坏死环斑病毒病（PNRSV）的研究.pdfVIP

中国农业科学 2007,40(9):1943-1951 Scientia Agricultura Sinica 侵染百合的李属坏死环斑病毒病（PNRSV）的研究 1 2 韩丽娟 ，刘文洪 1 2 （北京出入境检验检疫局，北京 100025； 浙江中医学院，杭州 310053 ） 摘要：【目的】在分子水平上鉴定百合上发生的李属坏死环斑病毒（Prunus necrotic ringspot virus,PNRSV） 【方法】对待测百合种球随机抽样，在温室内种植并观察，长出叶片后，用RNA Extraction Kit (Sangon, China) 从幼嫩的叶组织中提取核酸，将反转录出的cDNA用 PNRSV的特异性引物进行PCR扩增，电泳PCR产物得到了450bp 的目的片段。低熔点胶回收并克隆至 pGEM-T-easy 载体测序后进行序列比对分析。【结果】 从百合中所扩增的 450bpcDNA 的核苷酸序列与 27 个早期已报道的 PNRSV 分离物的 CP 基因的同源性为 84.5%～99.1%。【结论】百合 是PNRSV的一新寄主。 关键词：百合；李属坏死环斑病毒；RT-PCR；检测 Studies on Prunus Necrotic Ringspot Virus (PNRSV) Occurred on Lily 1 2 HAN Li-juan , LIU Wen-hong 1 2 （Beijing Entry Exit Inspection and Quarantine Bureau, Beijing 100025; Zhejiang College of Traditional Chinese Medicine , Hangzhou 310053 ） Abstract: 【Objective 】Molecular identification of Prunus necrotic ringspot virus (PNRSV) occurred on lily was made. 【Method 】Lily corms were randomly selected and grown in greenhouse. Total RNAs were extracted from young leaves with RNA Extraction Kit (Sangon, China), and partial amplification of CP gene was performed by RT-PCR with primer pairs specific for PNRSV. An expected cDNA fragment of about 450bp was harvested, cloned into pGEM-T-easy vector, sequenced and compared with 27 PNRSV isolates reported previously. 【Result 】The 450 bp cDNAs amplified from lily in this study shared 84.5%-99.1% homology with the 27 PNRSV isolates reported previously at the nucleotide level. 【Conclusion 】Lily is