中国野生柑橘上衰退病毒分离株分子特征分析.pdfVIP

中国农业科学 2007,40 5 :932-939 Scientia Agricultura Sinica 中国野生柑橘上衰退病毒分离株分子特征分析 1 1,2 2 1 2 易 龙 ，周常勇 ，周 彦 ，王志刚 ，唐科志 1 2 （西南大学植物保护学院，重庆 400716 ； 中国农业科学院柑橘研究所，重庆 400712 ） 摘要：【目的】了解中国野生柑橘上携带柑橘衰退病毒（CTV）分离株的CP/HinfⅠRFLP组群构成和分子特征。 【方法】运用 RT-PCR 对采自中国云南、广西、四川、湖南、江西等省 11 个野生柑橘上 CTV 分离株的病毒衣壳蛋 白基因 CPG 进行扩增，利用RFLP和SSCP 对 CPG进行分析，并将测定的CPG序列进行同源性比较和聚类分析。【结 果】发现野生柑橘上11个CTV 分离株以单一组群感染为主；这些CTV分离株CPG的核苷酸序列和相应的氨基酸序 列同源性分别为 92.5％～99.5％和 95.0％～100％；与 GenBank 收录的 9 个全基因组 CTV 分离株的相应基因序列 进行聚类分析，其核苷酸序列和氨基酸序列同源性分别为92.1％～98.8％和94.6％～100％。【结论】序列同源性 比较说明CTV CPG具有高度保守性，同时系统进化分析表明收集到的野生CTV分离株与国外分离株分属5大类群， 具有较复杂的亲缘关系。 关键词：野生柑橘；柑橘衰退病毒；多态性分析；序列比对；系统进化 Molecular Characterization of Citrus Tristeza Virus Isolates in Chinese Wild Type Citrus 2 1,2 1 2 1 YI Long , ZHOU Chang-yong , ZHOU Yan , WANG Zhi-gang , TANG Ke-zhi 1 2 College of Plant Protection, Southwest University, Chongqing 400716; Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712 Abstract: 【Objective 】This study was conducted to investigate the CP/Hinf ⅠRFLP groups and molecular characterization of citrus tristeza virus CTV isolates found in wild Chinese citrus plants. 【Method 】The target sequences of coat protection gene CPG were amplified and sequenced from 11 CTV isolates extracted from the samples of wild citrus plants collected in Yunnan, Guangxi, Sichuan, Hunan and Jiangxi provinces, using RT-PCR and sequencer. CPGs of these isolates were analyzed by RFLP and SSCP, and the CPG sequences of the 11 Chinese isolates and 9 foreign isolates from GenBank were subjected to sequence analysis and compared between each by BioEidt and MEGA software. 【Result 】CPG nucleotide sequences of the 11 Chinese CTV isolates and 9 foreign isolates were found to have identity ranging from 92.5% to 99.5% and 95.0% to 100%, whereas identity of their amino acid sequences ranges from 92.1% to 98.8% and 94.6% to 100%, respectively. RFLP and SSCP analysis