马铃薯纺锤块茎类病毒云南分离物的序列分析  

Sequence Analysis of Potato Spindle Tuber Viroid Isolates in Yunnan

作　　者：孙爱丽 王柱华 高锡春 谭涵月 杨艳丽[1] 蔡红[1] 朱有勇[1] 

机构地区：[1]云南农业大学,农业生物多样性与病虫害控制教育部重点实验室,云南昆明650201 [2]云南省元谋县植保植检站,云南元谋651300 [3]云南省红河州植检植保站,云南蒙自661100

出　　处：《云南农业大学学报（自然科学版）》2017年第5期780-786,共7页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省高校科技创新团队支持计划(云科教[2014]22号);云南省农业产业化专项项目(云财农[2013]364号);云南省科技创新人才计划项目(2011HB026)

摘　　要：【目的】了解云南省马铃薯纺锤块茎类病毒分子变异情况及其致病机制。【方法】以采自云南2个不同地方疑似马铃薯纺锤块茎类病毒病的块茎为材料,对其提取总RNA、反转录为c DNA,利用特异性引物进行RT-PCR检测,扩增产物经纯化后克隆并测序。【结果】扩增产物全长357 bp,分别命名为PSTVd-DasongpingYN(KX159281)和PSTVd-Lianghe YN(KX159282)。将这2个分离物与国内外已报道的PSTVd株系全序列构建系统发育树,并与已报道的PSTVd弱毒、中毒和强毒株系进行序列比对和二级结构预测,表明PSTVd-Dasongping YN和PSTVd-Lianghe YN与PSTVd弱毒株系更接近。【结论】PSTVd-Dasongping YN和PSTVd-Lianghe YN的序列相似性为97.5%,存在9个碱基差异,且主要集中在可变区(V)和右末端区(TR),说明云南不同地方分离物PSTVd也存在差异。PSTVd-Dasongping YN和PSTVd-Lianghe YN属于PSTVd的弱毒株系组。[Purpose]Our aim was to understand the molecular variation and pathogenic mechanism of potato spindle tuber viroid（ PSTVd） of Yunnan Province. [Method] The suspected tubers of PSTVd were collected from two different areas in Yunnan. Total RNA was extracted and reverse transcripted to c DNA. RT-PCR detection was conducted using PSTVd specific primers,and then the amplified products were purified,cloned and sequenced. [Results] The results showed that the full length fragments of PSTVd are 357 bp,which named as PSTVd-Dasongping YN（ KX159281） and PSTVd-Lianghe YN（ KX159282）,respectively. The phylogenetic trees were constructed with two PSTVd isolates from Yunnan and other PSTVd strains reported worldwide. Meanwhile,the sequence comparison and the second structure prediction were analyzed among PSTVd-Dasongping YN,PSTVdLianghe YN and different pathogenic PSTVd strains（ mild,medium and severe）,indicating that thePSTVd-Dasongping YN and PSTVd-Lianghe YN were more close to the mild PSTVd strains. [Conclusion] The similarity of PSTVd-Dasongping YN and PSTVd-Lianghe YN was 97. 5%. There are 9 different bases mainly concentrated in the variable region（ V） and the right terminal region（ TR） which implied that there are obvious differences in two PSTVd Yunnan isolates. PSTVd-Dasongping YN and PSTVd-Lianghe YN belong to the mild strain of PSTVd.

关 键 词：马铃薯纺锤块茎类病毒 反转录-聚合酶链式反应 序列分析 二级结构预测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]