摘要
建立了葡萄蚕豆萎蔫病毒(Grapevine fabavirus,GFabV)的实时荧光定量RT-PCR(RT-qPCR)检测技术。该技术标准曲线循环阈值与模板浓度呈良好的线性关系(扩增效率103.8%,相关系数0.998),灵敏度是常规RT-PCR的1 000倍,并具有特异性。采用RT-qPCR和常规RT-PCR方法对葡萄不同发育时期及不同部位的316个样品进行检测,结果表明RT-qPCR方法对GFabV的检出率(96.8%)明显高于RT-PCR(70.8%)。RT-qPCR对休眠枝条中GFabV的检出率达100%,对其他部位样品的检出率均在92%以上,明显高于RT-PCR(42%~97%)。各个发育时期样品GFabV的RT-qPCR检出率(85%~95%)也普遍高于RT-PCR(70%~90%)。该技术对于田间样品的检测适用范围广。
A SYBR GreenⅠRT-qPCR method for GFabV detection was established,an excellent linear correlation(0.998)and amplification efficiency(103.8%)were obtained from standard curve. The sensitivity of the method was 1 000 times higher than conventional RT-PCR,and had a strong specificity. The established RT-qPCR and conventional RT-PCR were used to detect GFabV in 316 grapevine samples from different development stages and different positions,and the results showed that the detection rate of RT-qPCR(96.8%)was apparently higher than conventional RT-PCR(70.8%). The detection rate of dormant branches using RT-qPCR was 100%,and for other positions detection were all above 92%,which were apparently higher than conventional RT-PCR(42%–97%). The detection rates using RT-qPCR(85%–95%)were higher than conventional RT-PCR(70%–90%)in these samples from different development stages. Compared with conventional RT-PCR,the newly established RT-qPCR has obvious superiority in detection of various samples.
作者
张梦妍
张尊平
任芳
胡国君
范旭东
董雅凤
ZHANG Mengyan;ZHANG Zunping;REN Fang;HU Guojun;FAN Xudong;DONG Yafeng(National Center for Eliminating Viruses from Deciduous Fruit Tree,Research Institute of Pomology,Chinese Academy of Agriculture Sciences,Xingcheng,Liaoning 125100,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2020年第1期187-194,共8页
Acta Horticulturae Sinica
基金
国家现代农业产业技术体系建设专项资金项目(CARS-29-bc-1)
国家重点研发计划项目(2018YFD0201301)
中央级公益性科研院所基本科研业务费专项.
作者简介
通信作者:范旭东;通信作者:董雅凤,E-mail:yfdong@163.com
