乌鲁木齐地区马铃薯纺锤块茎类病毒的检测与序列分析

Detection and Sequence Analysis of Potato Spindle Tuber Viroid in Urumqi

根据马铃薯纺锤块茎类病毒（PSTVd）基因序列设计合成1对引物,对新疆乌鲁木齐地区马铃薯田间感病植株进行一步法RT-PCR检测,扩增出251 bp大小的目标片段,而健康植株无此扩增产物。将目的片段克隆到pGM-T载体上并进行序列测定,构建系统进化树分析各分离物间的分子差异性。结果表明马铃薯纺锤块茎类病毒乌鲁木齐分离物与国内外已报道毒株的核酸序列同源性达93.6%~99.2%。

A pair of primers were designed and synthesized based on the nucleotide sequence of potato spindle tuber viroid（PSTVd） gene.The total RNA was directly extracted from the infected potato samples collected from Urumqi,Xinjiang,China,amplified by one-step reverse transcription and polymerase chain reaction（one-step RT-PCR）.A specific RT-PCR amplified fragment was the expected size of PSTVd gene about 251 bp,while no amplified products were obtained from the healthy samples.The unique amplified product was then cloned into the pGM-T vector and sequenced.Based on the phylogenic tree and sequence analysis,it was showed that the homology of nucleotide sequence of the PSTVd isolated from Urumqi was 93.6%-99.2%,comparison with the civil and foreign reported isolates.