Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification

Autor: Bishwo P. Mainali, Yeong-Hoon Lee, Hang-Won Kang, Dae-Hyeon Bae, Soon-Do Bae, Bong-Sub Kim, Young-Nam Yoon, In-Hee Park, Su-Heon Lee, Hyun-Joo Kim
Rok vydání: 2015
Předmět:
Zdroj: Research in Plant Disease, Vol 21, Iss 4, Pp 315-320 (2015)
ISSN: 1598-2262
DOI: 10.5423/rpd.2015.21.4.315
Popis: Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at 58°C for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.
Databáze: OpenAIRE