西瓜花叶病毒2号南瓜分离物的鉴定及其外壳蛋白基因序列分析  

Identification of Watermelon mosaic virus 2 pumpkin isolate and its coat protein gene sequence

作　　者：杨国慧[1] 张仲凯[1] 崔崇士[2] 

机构地区：[1]云南省农业生物技术重点实验室,昆明650223 [2]东北农业大学园艺学院,哈尔滨150030

出　　处：《植物病理学报》2004年第1期8-13,共6页Acta Phytopathologica Sinica

基　　金：云南省农业生物技术重点实验室开放基金项目资助(2001B-3)

摘　　要：利用电镜和酶联免疫吸附测定法(ELISA)在黑龙江省采集的南瓜病样中检测到西瓜花叶病毒2号(WMV-2)。再利用免疫PCR(IC-PCR)和反转录PCR(RT-PCR)方法,扩增获得其外壳蛋白(CP)基因片段,并克隆到pGEM-T载体中。核苷酸序列测定表明,该分离物CP基因全长为852个核苷酸,编码由284个氨基酸组成的31.8 kDa蛋白。与国外已报道的WMV-2 CP基因相比,其核苷酸序列同源性为92.2％-94.O％,由此推导的氨基酸序列同源性为94.5％-98.1％。与国内2个分离物相比,和山西分离物核苷酸和氨基酸的同源性都达到98.5％,和郑州分离物核苷酸和氨基酸的同源性分别为91.5％和95.0％。Watermelon mosaic virus 2 (WMV-2) was detected in infected pumpkin plant collected from Heilongjiang (HLJ) Province by electron microscopy and enzyme-linked immunosorbent assay (ELISA). A specific fragment about 850 bp in length covering coat protein (CP) gene region was amplified by reverse transcription polymerase chain reaction (RT-PCR) and immuno capture PCR (IC-PCR). The PCR product was cloned into pGEM-T vector. Sequences analysis showed that CP gene of WMV-2 isolate HLJ had 852 nucleotides in length, encoding 284 amino acids with molecular weights of 31. 8 kDa. Comparison showed it shared 92. 2%-94. 0% nucleotide acid identities and 94. 5%-98.1% amino acid identities with the published sequences, respectively. When compared with the two reported Chinese isolates, WMV-2-HLJ CP shared 98. 5% or 91. 5% nucleotide sequence identity and 98. 5% or 95. 0% amino acid sequence identity with that of WMV-2 Shanxi isolate or Zhengzhou isolate.

关 键 词：西瓜花叶病毒2号 南瓜分离物 鉴定技术 外壳蛋白基因 序列分析 

分 类 号：S436.42[农业科学—农业昆虫与害虫防治]