马铃薯病毒RT-PCR检测体系的建立

依据马铃薯病毒PVS、PVX、PLRV、PVA的CP保守序列设计特异性引物,从马铃薯病叶组织中提取出病毒总RNA,进行cDNA合成和PCR扩增,得到了与预期片段长度一致的PCR特异扩增产物,建立了能够同步检测PVS、PVX、PLRV、PVA的RT-PCR多重检测体系。该方法对PVS、PVX、PLRV、PVA扩增出的靶带大小分别为435、625、222、300 bp,凝胶电泳易辨别区分。研究结果表明,该方法特异性好、灵敏度高、快速简便,为马铃薯病毒的高效检测提供了有效手段。

Establishment of Potato Virus Detection by RT-PCR

Based on the nucleotide sequence of the coat protein（CP）gene of the four main potato virus（PVS,PVX,PLRV,PVA）to design DNA prlmers.The total RNA was directly extracted from virus infected potato leaves.By the reverse transcription and polymerase chain reaction （RT,-PCR）, and a specific fragment with expected length was obta/ned.The optimized multiplex reverse transcription polymerase chain reaction （m-RT-PCR）can amplify PVS, PVX, PLRV and PVA simultaneously, and the fragments were 435 bp （ PVS ）, 625 bp （ PVX ）, 222 bp （ PLRV ）, 300 bp （ PVA ）.The results showed that it was a specific ,sensitive detecting method and quicker, simpler.It provided an effective method of the detection for potato virus.