番木瓜环斑病毒南瓜分离物外壳蛋白基因的克隆及序列分析  

Cloning and sequence analysis of CP genes of PRSV from pumpkin

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：《植物保护学报》2007年第3期273-276,共4页Journal of Plant Protection

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法在云南省采集到的5份南瓜病样中检测到番木瓜环斑病毒（Papaya ring spot virus，PRSV）。为了进一步从分子水平确定云南省南瓜病毒病原种类，并为下一步转基因育种提供抗性基因，采用反转录PCR（RT—PCR）方法扩增了5个分离物的外壳蛋白（coat protein，CP）基因片段，并克隆到pGEM—T载体中。核苷酸序列测定表明，番木瓜环斑病毒石屏分离物（PRSV—SP）和番木瓜环斑病毒蒙自分离物（PRSV—MZ）的CP基因长873nt，编码290个氨基酸，番木瓜环斑病毒峨山分离物（PRSV—ES）、番木瓜环斑病毒版纳分离物（PRSV—BN）和番木瓜环斑病毒宾川分离物（PRSV—BC），3个分离物CP基因长867nt，编码288个氨基酸。PRSV5个分离物核苷酸序列的同源性在94％以上，氨基酸序列的同源性在96％以上。与国内外17个分离物相比，核苷酸序列同源性为89．6％～98．7％，氨基酸序列同源性为86．5％～99．6％。其中PRSV—SP和来自于越南分离物PRSV—V47无论是核苷酸序列，还是氨基酸序列同源性都达到了最高，而5个分离物与来自于巴西（PRSV—BR）、美国（PRSV—USA）、墨西哥（PRSV—Y）核苷酸序列同源性均低于90％。Papaya ring spot virus （PRSV） was tested with electron microscopy and ELISA from 5 diseased pumpkin samples collected in Yunnan Province. To further determine pathogen types of pumkin viral disease in Yunnan Province on a molecular level and provide anti-disease genes for trans-gene breeding, CP gene fragment was amplified by RT-PCR from 5 isolates and cloned to pGEM-T vector. Nucleotide sequence showed that CP gene of PRSV Shiping isolate （PRSV-SP） and PRSV Mengzi isolate （PRSV-MZ） contains 873 nt encoding 290 amino acid, while that of PRSV Eshan isolate （PRSV-ES）, PRSV Banna isolate （PRSV-BN） and PRSV Binchuan isolate （PRSV-BC） contains 867 nucleotide encoding 288 amino acid. The nucleotide and amino acid homology of the 5 isolates is higher than 94% and 96% respectively. Compared with sequence of other 17 foreign and domestic isolates, the homology of nucleotide is between 89.6% and 98.7% , the homology of amino acid is between 86.5% and 99.6%. It was also revealed that PRSV-SP was shared highest similarity with sequence of isolate PRSV-V47 from Vietnam, but these five isolates were shared low similarity with sequence of isolate PRSV-BR from Brazil, PRSV-USA from USA and PRSV-Y from Mexico, among which the homology of nucleotide is less than 90%.

关 键 词：南瓜 番木瓜环斑病毒 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]