CRISPR‐based platforms for the specific and dual detection of defoliating/nondefoliating strains of Verticillium dahliae
CRISPR‐based platforms for the specific and dual detection of defoliating/nondefoliating strains of Verticillium dahliae
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DOI：
10.1002/ps.7940
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Yonglin Wang
中科院分区：
文献类型：
--
作者：
Qi Chen;Jin Wu;Chen Tang;Yonglin Wang
AbstractBackgroundVerticillium dahliae is a soil‐borne pathogenic fungus that causes Verticillium wilt disease on more than 400 plant species worldwide. Due to its broad host range and its ability to survive long term in the soil, there are few effective control measures for V. dahliae once it has become established. Accurate, sensitive, and rapid detection of V. dahliae is crucial for limiting pathogen entry into new regional environments and early management of Verticillium wilt.ResultsIn this study, we developed a method to detect V. dahliae based on recombinase polymerase amplification (RPA) and CRISPR/Cas technology and used fluorescence and lateral flow test strips to monitor the outcomes. Through the establishment and optimization of RPA‐CRISPR/Cas13a detection, the sensitivity of the fluorescence method was 1 aM for genomic DNA (gDNA) within 20 min, while the sensitivity of the lateral flow strip method was 100 aM for gDNA in 30 min. The field applicability of RPA‐CRISPR/Cas13a was also validated by the detection of V. dahliae on smoke trees (Cotinus coggygria) in Xiangshan Park, Beijing, China. Finally, diplex detection for defoliating and nondefoliating pathotypes of V. dahliae was established by combining CRISPR‐Cas12a/Cas13a with specific target genes.ConclusionTaken together, this study achieved rapid, sensitive, and accurate detection of V. dahliae and the differentiation of defoliating and nondefoliating pathotypes and provides potential for field‐deployable diagnostic tools for rapid and ultrasensitive detection.This article is protected by copyright. All rights reserved.
