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

doi:10.3969/j.issn.1674-7968.2024.04.018

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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.

Key words： Panax notoginseng Fusarium solani Real-time fluorescent LAMP Rapid molecular detection

Received: 29 May 2023

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Corresponding Authors: *diqiuliu@126.com

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	CHE Xiao-Li
	LIANG Ting-Ting
	QU Yuan
	LIU Di-Qiu

Cite this article:

CHE Xiao-Li,LIANG Ting-Ting,QU Yuan, et al. Establishment and Application of Real-time Fluorescent LAMP Rapid Detection of Fusarium solani of Panax notoginseng Root Rot[J]. 农业生物技术学报, 2024, 32(4): 939-948.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2024.04.018 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2024/V32/I4/939

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