福建马铃薯A病毒的分子鉴定与检测技术.pdf

农业生物技术学报 Journal of Agricultural Biotechnology 2004,12 1冤院 90~95 ·研究论文· 福建马铃薯A 病毒的分子鉴定及检测技术* 兴泉 陈士华 魏广彪 祖建 谢联辉** （福建农林大学植物病毒研究所，福州，350002 ） 摘要：依据马铃薯A 病毒（ , PVA ）外壳蛋白（coat protein, ） 因核苷酸序列及氨 酸序列对福建省 PVA 进行了鉴定。研究结果表明，利用PVA 序列结合系统进化 分析不但可对PVA 进行准确的鉴定，而且可进行不同分 离物间分子差异性分析。利用病毒特异性引物建立了PVA 的反转录- 聚合酶链式反应（reverse transcription-polymerase chain ，以 （ reaction ，RT-PCR ）技术 DIG 标记的PVA 基因为探针建立了核酸斑点杂交技术 nucleic acid spot hybridization, NASH ），并对它们分别进行了改进。调查结果表明：PVA 在福建省广泛分布，发病率最高可达80%以上。当地农家种可能是田 间PVA 的主要来源。 关键词：马铃薯A 病毒; 分子鉴定; 检测; 发生; 分布 Molecular Identification and Detection Method of WU Xing-Quan CHEN Shi-Hua WEI Guang-Biao WU Zu-Jian XIE Lian-Hui** (Plant Virology Institute, Fuj ian Agriculture and Forestry University ，Fuzhou 350002, China) According to the nucleotide and amino acid sequences of coat protein （CP ）, the (PVA)was identified in Fuji an. Based on the phylogenetic tree and sequence analysis, we could identify PVA and analysze the difference of PVA isolates. Using the PVA specific primers, the RT-PCR detection method was established. Using DIG labeled PVA gene as probe, nucleic acid spot hybridization (NASH) detection method was established 。And these two methods were improved too. The investigation result showed that PVA distributed widely in Fuji an, and the highest disease rate of PVA could reach 80% . The local varity might be the main virus resource in field. ; molecular identification; detection; occurrence; distribution 马铃