doi: 10.5423/PPJ.OA.12.2019.0298 BibTeX RIS Mendeley


Rapid and Sensitive Detection of Lettuce Necrotic Yellows Virus and Cucumber Mosaic Virus Infecting Lettuce (Lactuca sativa L.) by Reverse Transcription Loop-Mediated Isothermal Amplification

Rapid and Sensitive Detection of Lettuce Necrotic Yellows Virus and Cucumber Mosaic Virus Infecting Lettuce (Lactuca sativa L.) by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Zhang,Yubao(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,
  • Xie,Zhongkui(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,
  • Fletcher,John D(The New Zealand Institute for Plant and Food Research) ,
  • Wang,Yajun(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,
  • Wang,Ruoyu(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,
  • Guo,Zhihong(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,
  • He,Yuhui(Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ,

초록

Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM $Mg^{2+}$ at $65^{\circ}C$ for LNYV and $60^{\circ}C$ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.


키워드

cucumber mosaic virus;lettuce;lettuce necrotic yellows virus;RT-LAMP amplification;RT-PCR;


발행 정보

서지정보
The Plant Pathology JournalThe Plant Pathology Journal / v.36, no.1, 2020년, pp.76-86
발행기관
한국식물병리학회
저널 DOI
doi: 10.5423
pISSN
1598-2254
eISSN
2093-9280
발행일
2020.02.01
언어
영어

이력 정보

2020.02.01 발행
2020.02.10 DOI발급

위치 정보


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