地高辛标记cDNA探针检测苹果茎痘病毒  

Detection of Apple stem pitting virus(ASPV) by dot blot hybridization with digoxigenin-labelled viral cDNA probe

作　　者：杨俊玲[1] 董雅凤[1] 李世访[2] 张尊平[1] 何峻[1] 

机构地区：[1]中国农业科学院果树研究所,辽宁兴城125100 [2]中国农业科学院植物保护研究所,北京100094

出　　处：《植物病理学报》2006年第5期470-472,共3页Acta Phytopathologica Sinica

基　　金：植物病虫害生物学国家重点实验室开放基金(Pd1);中国农业科学院科研基金(农科院[2003]168号)

摘　　要：Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP.The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization.The results showed that the probe was sensitive and specific.The probe couldn’t hybridize with total RNA of Apple stem grooving virus,Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control,only hybridized with that extracted from dormant shoot infected with ASPV.The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg.Partial sequence （314 bp） of ASPV was cloned and used as a probe labelled with digoxigenin- 11dUTP. The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization. The results showed that the probe was sensitive and specific. The probe couldn＇ t hybridize with total RNA of Apple stem grooving virus, Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control, only hybridized with that extracted from dormant shoot infected with ASPV. The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg.

关 键 词：苹果茎痘病毒 CDNA探针 地高辛标记 检测 无病毒苗木 病毒病害 virus stem 

分 类 号：S436.611.1[农业科学—农业昆虫与害虫防治]