榆林地区马铃薯纺锤块茎类病毒的RT-PCR检测与序列分析  

RT-PCR Detection and Sequence Analysis of Potato Spindle Tuber Viroid in Yulin Area

作　　者：冯光惠[1] 杜虎平[1] 李夏隆[1] 亢福仁[1] 

机构地区：[1]榆林学院生命科学学院,陕西榆林719000

出　　处：《山西农业科学》2014年第10期1047-1050,1059,共5页Journal of Shanxi Agricultural Sciences

基　　金：陕西省教育厅科研计划项目(08JK508;08JK509)

摘　　要：以陕西省榆林地区种植2～3代的马铃薯叶片为材料,首先用Trizol法提取马铃薯叶片总RNA,以已公布的马铃薯纺锤块茎类病毒（PSTVd）基因序列设计合成一对特异引物,反转录合成cDNA,通过RT-PCR扩增目的DNA片段,琼脂糖凝胶电泳检测扩增产物;然后回收纯化扩增产物并进行克隆和测序,采用DNAstar软件分析序列一致性。电泳结果表明,在榆林地区的马铃薯叶片中均扩增到与预期大小一致的目的片段,说明这些马铃薯均感染了PSTVd;序列分析表明,10个不同采样点PSTVd的目的片段大小为250～251 bp,只是单个碱基之间的转换或缺失,PSTVd在榆林地区几乎没有发生变异,与国内外其他15个地区已报道的序列一致性在82.3%～99.5%之间。Taking 2-3 generations of suspected infected potato leaves as material planted in Yulin area of Shaanxi province,total RNA was extracted from potato leaves by Trizol method,and designed specific primers by published potato spindle tuber viriod gene sequence,cDNA was synthesized and the DNA fragment was amplified by RT-PCR,the amplified product was detected by agarose gel electrophoresis,then the purified PCR products was cloned sequenced,and used DNAstar software analysis sequence identity.The electrophoresis results show that the gene sequence amplified from potato leaves are consistent with the expected size,we can know these potatoes are infected with potato spindle tuber viroid;sequence analysis showed that PSTVd fragment size is 250-251 bp in 10 different sampling points,only a single base deletion or conversion,there are no variation of PSTVd in Yulin area,and sequence identity have been reported in the 82.3% to 99.5% between Yulin area and other 15 areas at home and abroad.

关 键 词：马铃薯 纺锤块茎类病毒 RT-PCR检测 序列分析 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]