辣椒褪绿病毒SYBR Green Ⅰ荧光定量PCR检测方法的建立

Establishment of SYBR Green I real-time PCR assay for detecting capsicum chlorosis virus

辣椒褪绿病毒(Capsicum chlorosis virus, CaCV)可引起辣椒(Capsicum annuum L.)、蝴蝶兰(Phalaenopsis sp.)等重要作物严重病害.根据CaCV核衣壳蛋白基因保守序列设计引物,建立了CaCV SYBR Green Ⅰ荧光定量PCR检测方法.结果表明,以含有CaCV目的基因片段的重组质粒为标准品构建的标准曲线,其循环阈值与模板浓度有良好的线性关系.所建立的SYBR Green Ⅰ荧光定量PCR方法具有特异性、重复性和灵敏性等优点,可用于田间样品中CaCV的定量检测.

Capsicum chlorosis virus(CaCV)can cause serious diseases in several important crops such as peppers and Phalaenopsis orchids.In this study,an SYBR Green Ⅰ real-time PCR assay was established for the detection of CaCV infected samples,and the primers were designed based on the conserved sequence of CaCV nucleocapsid protein gene.The results showed that the standard curve had a good linear relationship between the cycling threshold and the template concentration,which was constructed using the recombinant plasmid containing the CaCV target gene fragment.The established SYBR Green Ⅰ real-time PCR assay has the advantages of specificity,repeatability and sensitivity,which can be used to detect CaCV-infected samples in the field.