柑橘类病毒的分子鉴定与分型  

Molecular Detection and Characterization of Citrus Viroids

作　　者：马先锋[1] 韩健[1] 邓子牛[1,2] 舒广平[2] Rizza S Catara A 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]国家柑橘改良中心长沙分中心,长沙410128 [3]Department of Phytosanitary Sciences and Technologies, University of Catania, Italy

出　　处：《中国农业科学》2008年第9期2670-2677,共8页Scientia Agricultura Sinica

基　　金：湖南省重大科技专项(04NK1005)

摘　　要：【目的】检测湖南地区柑橘所携带的柑橘类病毒类型,并鉴定柑橘类病毒的亚型。【方法】以田间柑橘枝条韧皮部为试材,提取总RNA,反转录成cDNA,再用不同类型类病毒的特异引物进行PCR扩增,鉴定出各个样品所携带的柑橘类病毒类型;通过测序分析各类型的序列,并与国内外报道的序列进行同源性分析,确定所测样品的亚型。【结果】建立了CEVd、CVdII、CVdⅢ等柑橘类病毒的RT-PCR检测方法。在5个供试样品中检测出3种类型的柑橘类病毒,即柑橘裂皮病类病毒(citrus exocortis viroid,CEVd)、柑橘类病毒Ⅱ型(citrus viriodⅡ,CVdⅡ,又名hop stunt viroid,HSVd)和柑橘类病毒Ⅲ型(citrus viriodⅢ,CVdⅢ),其中CVdⅡ和CVdⅢ为国内首次发现。亚型分析表明CVdⅡ可能为新的亚型;CVdⅢ为b亚型,可能具有矮化功能。【结论】在中国鉴定出3种类型的柑橘类病毒,CEVd与国内已报道的序列完全一致,其中CVdⅡ和CVdⅢ为首次在中国发现;检测到的CVdⅡ可能为新的亚型。[Objective] Detecting various types of citrus viroids by RT-PCR and characterizing the subtypes by sequencing in Hunan Province, China. [Method] Total RNA was isolated from phloem of young shoots and retro-transcripted to cDNA by RT-PCR with specific primers, and the amplified cDNA fragments were sequenced for identification of subtypes. [Result] An effective RT-PCR technique was established for detecting citrus viroid types, and three types were recognized, i.e. CEVd, CVd Ⅱ and CVdⅢ, and the later two were identified for the first time in China. The sequencing results indicated that the CVd Ⅱ seemed to be a new subtype; CVdⅢb detected in the present study could be further developed as a dwarfing agent. [Conclusion] Three types of citrus viroids were detected, amongst, CVd Ⅱ and CVdⅢ were identified for the first time in China. The sequence of CVd Ⅱ was different from all of the three sub-types reported, implying the identification of a new subtype. The verified CVdⅢb offered a possibility of using it in dwarfing cultivation.

关 键 词：柑橘 类病毒 检测 序列分析 分型 

分 类 号：S436.66[农业科学—农业昆虫与害虫防治]