河南省马铃薯Y病毒的分子检测与鉴定

The Molecular Detection and Indentification of Potato Virus Y in Henan Province

利用RT-PCR技术对郑州市和信阳市的马铃薯Y病毒(potato virus Y,PVY)进行了鉴定。依据PVY p1基因序列设计合成1对引物,以带毒的马铃薯叶片总RNA为模板,RT-PCR扩增得到0.58kb的目的DNA片段,而健康对照无此片段。对PCR产物进行序列测定,Blast分析表明,该DNA序列与PVY N:O株系p1基因序列相似性可达99%,证明所得DNA片段确为PVY p1基因,从而建立了PVY的RT-PCR检测方法。在此基础上,从节省检测时间、优化反应程序及反应体系等方面进行了改进。

Potato virus Y （PVY） can infect potato and do harm to potato seriously. The PVY in Zhengzhou city and Xinyang city of Henan province was detected and identified by RT-PCR molecular detection technology. The spencific primers PVYP1 and PVYP2 were designed based on PVY pl gene, which amplified a DNA fragment of 0.58kb by RT-PCR using the RNA from the infected plant by PVY as template. However, no DNA fragment was amplified from RNA of healthy potato. The PCR product was sequenced. The results of Blast analysis showed that the similarity of the sequence with the pl gene of PVY strain N ： O reached to 99%, indicating that the PCR product is the partial DNA fragment of PVY pl gene. Then the RT-PCR detection method of PVY was established. The detection method was improved by shortening the reaction time,and optimizing the PCR procedures and PCR reagent system.