农业生物技术学报2024，Vol.32Issue(4)：939-948,10.DOI:10.3969/j.issn.1674-7968.2024.04.018
三七根腐病菌茄腐镰刀菌的实时荧光LAMP快速检测方法的建立与应用
Establishment and Application of Real-time Fluorescent LAMP Rapid Detection of Fusarium solani of Panax notoginseng Root Rot
摘要
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.关键词
三七/茄腐镰刀菌/实时荧光LAMP/快速分子检测Key words
Panax notoginseng/Fusarium solani/Real-time fluorescent LAMP/Rapid molecular detection分类
农业科技引用本文复制引用
车晓莉,梁婷婷,曲媛,刘迪秋..三七根腐病菌茄腐镰刀菌的实时荧光LAMP快速检测方法的建立与应用[J].农业生物技术学报,2024,32(4):939-948,10.基金项目
国家自然科学基金(82060693) （82060693）
云南省重大科技专项计划(202102AA100022) （202102AA100022）
