苹果茎沟病毒RT-PCR检测技术  

Detection of Apple Stem Grooving Virus by Reverse Transcription-polymerase Chain Reaction(RT-PCR)

作　　者：李文慧[1] 牛建新[1] 

机构地区：[1]石河子大学农学院园艺系,石河子832003

出　　处：《西北农业学报》2006年第4期105-107,共3页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金资助项目(30060053;30360066);国家科技攻关计划引导项目(2003BA546C);兵团科委项目(NKB02SDXNK01SW)

摘　　要：用苹果的叶片作为材料,研究比较了两种提取苹果总RNA方法的效果,确定了较好的提取方法。根据苹果茎沟病毒外壳蛋白基因设计引物,通过RT-PCR扩增在苹果叶片总RNA样品中获得了524 bp的特异片断,通过对该片段的序列分析与苹果茎沟病毒外壳蛋白基因源序列比较,其同源性高达93%。通过多次重复实验验证,该方法能很好地检测苹果茎沟病毒。Using apple leaves as materials ,total RNA was extracted by two methods and these methods were studied, the fittest method for apple were screened out. A pair of primers were designed and synthesized based on the nucleotide sequence of the coat protein gene sequence of apple stem grooving virus. The expected 524 bp fragment was acquired by RT-PCR and the sequence of this fragment by clone and sequence was aligned with the original coat protein gene sequence of apple stem grooving virus. The result showed that the identity of two sequences reach 93%. The method can efficiently detect apple stem grooving virus by more experiences.

关 键 词：苹果茎沟病毒 逆转录-聚合酶链式反应 克隆 序列分析 

分 类 号：S436.611[农业科学—农业昆虫与害虫防治]