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Journal of Integrative Agriculture  2020, Vol. 19 Issue (7): 1834-1841    DOI: 10.1016/S2095-3119(19)62784-X
Special Issue: 植物病理合辑Plant Protection—Plant Pathology 植物病毒合辑Plant Virus
Plant Protection Advanced Online Publication | Current Issue | Archive | Adv Search |
A sensitive SYBR Green RT-qPCR method for grapevine virus E and its application for virus detection in different grapevine sample types
REN Fang, ZHANG Zun-ping, FAN Xu-dong, HU Guo-jun, ZHANG Meng-yan, DONG Ya-feng
National Center for Eliminating Viruses from Deciduous Fruit Trees, Research Institute of Pomology, Chinese Academy of Agricultural Sciences, Xingcheng 125100, P.R.China
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To develop a rapid and high-sensitivity method for detection of grapevine virus E (GVE), a SYBR Green based real-time fluorescence quantitative RT-PCR method (RT-qPCR) was established.  This method could be used to detect GVE specifically, and the sensitivity was about 100 times greater than conventional RT-PCR.  An excellent linear correlation (R2=0.997) and a high amplification efficiency (E=97.5%) were obtained from the standard curve of this method.  Reproducibility tests revealed that the coefficients of variation in the intra- and inter-assay results were 0.31–1.03% and 0.82–2.62%, respectively, indicating a good reproducibility.  The RT-qPCR method could be used to detect GVE in a wide range of grapevine sample types.  The detection rates of RT-qPCR for nearly all sample types from different positions and seasons were higher than conventional RT-PCR.  The detection rates in spring, summer, autumn and winter increased gradually.  Samples in autumn and winter were best for detection, and the detection rates of most samples were 80–100%, which were 10 to 40% higher than conventional RT-PCR.  In general, old petioles and branches were the best tissues for GVE detection.  The detection rates of these samples in each season were all 100%, which were 20 to 40% higher than conventional RT-PCR.  The second highest rates were in the old leaf, with detection rates for RT-qPCR of 80–100% in all seasons, which were 20 to 40% higher than conventional RT-PCR.  GVE could be difficultly detected in young leaves by conventional RT-PCR, and the detection rates were only 0–50%, while by RT-qPCR the rates could increase to 0–80%.  A total of 33 out of 363 samples (belonging to 68 cultivars) from 20 regions in China were detected to be positive by RT-qPCR (9.1%), which was more than twice the rate of the conventional RT-PCR (3.9%). 
Keywords:  grapevine       grapevine virus E        detection        RT-qPCR        conventional RT-PCR  
Received: 29 April 2019   Accepted:
Fund: This research was supported by the earmarked fund for China Agriculture Research System (CARS-29-bc-1).
Corresponding Authors:  Correspondence DONG Ya-feng, Tel: +86-429-3598278, E-mail:    
About author:  REN Fang, Tel: +86-429-3598137, E-mail:;

Cite this article: 

REN Fang, ZHANG Zun-ping, FAN Xu-dong, HU Guo-jun, ZHANG Meng-yan, DONG Ya-feng. 2020. A sensitive SYBR Green RT-qPCR method for grapevine virus E and its application for virus detection in different grapevine sample types. Journal of Integrative Agriculture, 19(7): 1834-1841.

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