马铃薯X病毒荧光定量PCR检测体系的建立及应用

摘要

马铃薯 X 病毒（Potato virus X ，PVX）是危害茄科作物的一种重要病毒，为了建立特异性检测 PVX 的实时荧光定量 PCR 体系，本研究以 PVX-1985分离物中外壳蛋白（coat protein，CP）基因序列为模板，设计引物构建重组质粒并选择扩增效率高、特异性强的引物成功构建出标准曲线。利用建立的体系，成功检测到以含 pCaPVX440侵染性克隆载体的农杆菌 C58C1接种后的本氏烟（Nicotiana benthamiana ）中 PVX 病毒 RNA 的拷贝数。%Potato virus X (PVX)is an important virus infecting many solanaceous crops.In order to develop a re-al-time quantitative PCR system for specific detection of PVX,the standard curve was established successfully based on the recombinant plasmid and high efficient and specific primers derived from coat protein gene sequence of PVX-1985 isolate.We successfully detected the copy quantity of RNA from PVX in the Nicotiana benthamiana which injected by the Agrobacterium C58C1 containing pCaPVX440 plasmid.