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Ultra-rapid Real-time PCR for the Detection of Tomato yellow leaf curl virus

초고속 Real-time PCR을 이용한 Tomato yellow leaf curl virus의 신속진단

  • Kim, Tack-Soo (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Choi, Seung-Kook (Horticultural and Herbal Crop Environment Division, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Ko, Min-Jung (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Lee, Minho (Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Choi, Hyung Seok (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Lee, Se-Weon (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Kyungseok (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Jin-Woo (Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration)
  • 김택수 (국립농업과학원 농업미생물과) ;
  • 최승국 (국립원예특작과학원 원예특작환경과) ;
  • 고민정 (국립농업과학원 농업미생물과) ;
  • 이민호 (국립농업과학원 유기농업과) ;
  • 최형석 (국립농업과학원 농업미생물과) ;
  • 이세원 (국립농업과학원 농업미생물과) ;
  • 박경석 (국립농업과학원 농업미생물과) ;
  • 박진우 (국립농업과학원 농업미생물과)
  • Received : 2012.11.08
  • Accepted : 2012.11.30
  • Published : 2012.12.31

Abstract

Tomato yellow leaf curl virus (TYLCV), transmitted exclusively by the whitefly (Bemisia tabaci) in a circulative manner is one of the most important virus in tomato. Since the first report of TYLCV incidence in Korea in 2008, the virus has rapidly spread nationwide. TYLCV currently causes serious economic losses in tomato production in Korea. Early detection of TYLCV is one of the most important methods to allow rouging of infected tomato plants to minimize the spread of TYLCV disease. We have developed an ultra-rapid and sensitive real-time polymerase chain reaction (PCR) using a new designed real-time PCR system, GenSpectorTM TMC-1000 that is a small and portable real-time PCR machine requiring only a $5{\mu}l$ reaction volume on microchips. The new system provides ultra-high speed reaction (30 cycles in less than 15 minutes) and melting curve analysis for amplified TYLCV products. These results suggest that the short reaction time and ultra sensitivity of the GenSpector$^{TM}$-based real-time PCR technique is suitable for monitoring epidemics and pre-pandemic TYLCV disease. This is the first report for plant virus detection using an ultra-rapid real-time PCR system.

Acknowledgement

Supported by : National Academy of Agricultural Science, Rural Development Administration

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