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A simplified RT-PCR assay for the simultaneous detection of tomato chlorosis virus and tomato yellow leaf curl virus in tomato

文献类型： 外文期刊

作者： Tu, Liqin 1 ; Wu, Shuhua 1 ; Gan, Shexiang 1 ; Zhao, Wenhao 1 ; Li, Shuo 1 ; Cheng, Zhaobang 1 ; Zhou, Yijun 1 ; Zhu, Yuelin 2 ; Ji, Yinghua 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Plant Protect, Key Lab Food Qual & Safety Jiangsu Prov, State Key Lab Breeding Base, Nanjing 210014, Peoples R China

2.Nanjing Agr Univ, Coll Hort, Nanjing 210095, Peoples R China

关键词： Tomato chlorosis virus; Tomato yellow leaf curl virus; Simultaneous detection via RT-PCR

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.623； 五年影响因子：2.239 ）

ISSN： 0166-0934

年卷期： 2022 年 299 卷

页码：

收录情况： SCI

摘要： Tomato chlorosis virus (ToCV), a species of single-stranded RNA virus belonging to the Crinivirus genus, and Tomato yellow leaf curl virus (TYLCV), a species of single-stranded circular DNA virus belonging to the Begomovirus genus, are two major emerging viruses transmitted by whiteflies and are causing huge losses to tomato production worldwide. To facilitate the simultaneous detection of both viruses in co-infected plants for disease control, a duplex reverse-transcription PCR assay was developed. The assay used three primers, a degenerate reverse primer targeting a conserved region of TYLCV and the RNA2 of ToCV, and two virus-specific forward primers targeting the minor coat protein gene of ToCV and the C3 gene of TYLCV, respectively, to amplify a 762bp and a 338-bp fragment from ToCV and TYLCV, respectively, in a single reaction. The concentration of the primers, annealing temperature and amplification cycles used in the assay were optimized, and the sensitivity of the assay was assessed. Using this assay, 150 tomato leaf samples collected from the field during 2018 were tested. The results showed that both viruses could be detected simultaneously in co-infected field samples. The assay should benefit the rapid detection of these two viruses in tomato crops and would facilitate early warning of infections for the control of the two virus diseases.

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