南瓜蚜传黄化病毒湖北和云南分离物的部分序列分析  

Partial sequence analysis of two isolates of Cucurbit aphid-borne yellows virus from Hubei and Yunnan in China

作　　者：尚巧霞[1] 向海英[2] 韩成贵[2] 李大伟[2] 于嘉林[2] 

机构地区：[1]北京农学院植物科学技术系,北京102206 [2]中国农业大学植物病理学系与农业生物技术国家重点实验室,北京100094

出　　处：《植物病理学报》2008年第1期64-68,共5页Acta Phytopathologica Sinica

基　　金：北京市科技新星计划(2007-B-032);国家“973”计划资助课题(2006CB101903)

摘　　要：本研究从带有黄化症状的南瓜叶片中提取总RNA,用RT-PCR方法扩增得到来自湖北和云南的南瓜蚜传黄化病毒（CABYV）2个分离物的1375nt特异性核苷酸片段。分别将PCR产物插入到克隆载体pMD19-T并转化大肠杆菌DH5α,对筛选到的阳性克隆进行了序列测定和分析（GenBank登录号为EF488996和EF488997）。所获片段含有部分复制酶基因576nt,非编码区199nt和完整的CP基因600nt,编码一个由199个氨基酸组成的分子量约为22kDa的结构蛋白。湖北和云南分离物与法国分离物、意大利分离物、西班牙分离物、北京分离物和上海分离物的CP基因核苷酸序列和推测氨基酸序列的同源性分别为93.1%-98.5%和91.4%-98.5%。Two isolates of Cucurbit aphid-borne yellows virus（CABYV） were obtained from naturally infected cushaws with yellowing symptom from fields in Kunming city of Yunnan and Wuhan city of Hubei. A pair of primers were designed based on a published CABYV sequence, and the expected fragments of about 1375 nt were amplified by RT-PCR and cloned into pMD19-T in DH5α, respectively. The recombinant clones were identified by PCR reaction and subsequently sequenced （GenBank accession number are EF488996 and EF488997）. The obtained sequences started within a single open reading frame which was expected to encode a part of the RdRp of 191 amino acids by 576 nucleotides, an intergenic non-coding region （NCR） of 199 nucleotides and followed by complete capsid protein （CP） of 199 amino acids encoded by 600 nucleotides. The identifies of nucleotide and deduced amino acid sequences of CP gene among CABYV-HB, CABYV-YN and other nine previously reported isolates from France, Italy, Spain, Beijing, Shanghai ranged from 93.1% to 98.5% and 91.4% to 98.5%, respectively.

关 键 词：南瓜蚜传黄化病毒 基因组片段 RT-PCR 克隆和序列分析 

分 类 号：S432.1[农业科学—植物病理学]