马铃薯病毒一步法RT-PCR诊断研究  

A Study on the One-step RT-PCR for Detection of Potato Viruses

作　　者：关翠萍[1] 张鹤龄[1] 门福义[2] 宋艳茹[3] 蒙美莲[2] 陈正华[4] 

机构地区：[1]内蒙古大学生命科学学院 [2]内蒙古农业大学农学院,呼和浩特010018 [3]中国科学院植物研究所,北京100093 [4]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《内蒙古大学学报（自然科学版）》2005年第2期178-185,共8页Journal of Inner Mongolia University：Natural Science Edition

基　　金：国家863中试开发项目-马铃薯微型薯人工种子规模化生产关键技术;内蒙古十五攻关项目:马铃薯新品种选育与种薯产业化研究

摘　　要：运用一步法RT-PCR对马铃薯和烟草中的马铃薯X病毒、马铃薯Y病毒和马铃薯卷叶病毒进行了检测,实验表明:对酚-SDS方法提取的病毒核酸进行扩增,一步法RT-PCR检测到的扩增产物的灵敏度较两步法RT-PCR的高100倍左右.两种提取方法对比,用一步法RT-PCR进行扩增,检测到的PEG法扩增产物的灵敏度较酚-SDS法扩增产物的灵敏度高3倍.建立了快速、简便、灵敏度高、特异性强的马铃薯病毒一步法RT-PCR检测技术.One-step reverse-transcription polymerase chain reaction(RT-PCR) protocol for detection of potato virus X,potato virus Y and potato leafroll virus in infected potatoes and tobaccoes was developed.The results showed that for detection of virus nucleic acid purified by phenol-SDS method, the sensitivity of one-step RT-PCR is about one hundred times higher than that of two-step RT-PCR.Two sample preparation methods:PEG method and phenol-SDS method were compared for one-step RT-PCR detection.The results showed that PEG method is about three times as sensitive as phenol-SDS method.One-step RT-PCR for detecting potato viruses was identified as a rapid,simple,sensitive and specific detecting method.

关 键 词：马铃薯X病毒 马铃薯Y病毒 马铃薯卷叶病毒 反转录-聚合酶链式反应 诊断 

分 类 号：Q939.46[生物学—微生物学] S432.41[农业科学—植物病理学]