Poster: Diseases of Plants: Disease Detection & Diagnosis
Detection of Apple scar skin viroid in apple fruits by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
S. KWON (1), I. Cho (2), S. Choi (1), J. Yoon (1), G. Choi (1) (1) National Institute of Horticultural and Herbal Science, South Korea; (2) National Institute of Horticultural and Herbal Scicence, South Korea
Apple scar skin viroid (ASSVd) is one of the most important pathogen causing scar skin, dappling or cracking on the surface of apple fruits. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of ASSVd. A set of two primers were designed inner primers (FIP and BIP) and outer primers (F3 and B3) from sequences of the Korean strain of ASSVd. The LAMP reaction was optimized using Bst DNA polymerase for 30 min at 65°C. The RT-LAMP products were evaluated as ladder-like bands by electrophoresis and visualized in the reaction tube by staining with SYBR Green I dye. RT-LAMP was more sensitive than that of conventional RT-PCR for the detection of ASSVd. The RT-LAMP assay developed in this study is a simple, rapid and sensitive and can be applied as a practical molecular diagnostic tool.