Establishment and Application of Real-time Fluorescent LAMP Rapid Detection of Fusarium solani of Panax notoginseng Root Rot
CHE Xiao-Li1,2, LIANG Ting-Ting1,2, QU Yuan1,2, LIU Di-Qiu1,2,*
1 Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China; 2 Yunnan Key Laboratory of Panax notoginseng, Kunming 650500, China
Abstract：Root rot is a serious disease of Panax notoginseng in China, and Fusarium solani is the main fungal pathogen causing root rot of P. notoginseng. In this study, a loop mediated isothermal amplification (LAMP) system was established to rapidly detect F. solani from P. notoginseng. Firstly, the D-amino acid oxidase gene (DAO) was screened by PCR as a specific gene locus for detection of F. solani, and the specific primer pairs of LAMP reaction were designed and the reaction conditions were optimized. The optimum reaction temperature was 65 ℃; the optimum concentration of MgSO4 was 6 mmol/L; the optimum concentration of dNTPs was 0.6 mmol/L; the amplification reaction was more efficient in the presence of cyclic primers. Sensitivity of LAMP to F. solani was tested, and the template concentration could be as low as 0.009 pg/μL. At last, the on-site rapid detection technology of F. solani was established based on the combination of DNA extraction-free and calcein coloration. This study provides technical support for the rapid diagnosis of F. solani of root rot in P. notoginseng cultivation and seedling trade.
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