Nested Multiplex PCR to Detect Two Major Fungal Pathogens of Fritillaria thunbergii

doi:10.16409/j.cnki.2095-039x.2023.02.009

Abstract

Abstract: Anthracnose and dry rot diseases are common diseases in Fritillaria thunbergii, which seriously affect the quality and output of F. thunbergii. It is urgent to establish a rapid and accurate detection method of the two diseases, which is of great significance for early diagnosis, effective prevention and control of the disease. In this study, diseased F. thunbergii were collected from Zhangshui Town, Haishu District, Ningbo City. On the basis of previous results, two pairs of specific primers FO1-F/FO1-R and BMTJ4-F/BMTJ4-R were designed according to the ITS sequence of Colletotrichum lineola and Fusarium oxysporum on NCBI, respectively. The nested multiplex PCR method for detection of anthracnose and dry rot of F. thunbergii was established. The results showed that the nested multiplex PCR system had a high specificity and could detect C. lineola and F. oxysporum in infected plant tissues rapidly with a sensitivity of DNA 100 pg/μL. To verify the feasibility and accuracy of the detection system, 10 diseased samples were randomly collected from the field. The results showed that C. lineola was detected in four samples, F. oxysporum was detected in two samples, and two pathogens were simultaneously detected in four samples, the detection rate was as high as 100%, indicating that this method can be used to identify anthracnose and dry rot in F. thunbergii quickly, accurately and sensitively. Our research results provided technical support for field detection of anthracnose and dry rot diseases of F. thunbergii.

Key words: Fritillaria thunbergia, Colletotrichum lineola, Fusarium oxysporum, nested multiplex PCR

CLC Number:

S476

WANG Xinlong, ZHANG Nuo, CAO Zenyan, SONG Yang, YU Xiaopin, SHENTU Xupin. Nested Multiplex PCR to Detect Two Major Fungal Pathogens of Fritillaria thunbergii[J]. Chinese Journal of Biological Control, 2023, 39(2): 493-498.

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