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Development of a Recombinase Polymerase Amplification and CRISPR-Cas12a-Based Assay for Rapid Detection of Rice Bakanae Disease Caused by Fusarium fujikuroi

文献类型： 外文期刊

作者： Li, Hongyu 1 ; Qiu, Yue 3 ; Zhang, Anpeng 3 ; Hu, Yingxiong 3 ; Cheng, Can 2 ; Zhou, Jihua 2 ; Niu, Fuan 2 ; Sun, Bin 2 ; Dai, Yuting 3 ; Xie, Kaizhen 3 ; Feng, Zhizun 3 ; Ding, Xiaorui 3 ; Hu, Bilian 1 ; Zhang, Xueqing 3 ; Cao, Liming 2 ; Chu, Huangwei 2 ;

作者机构： 1.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China

2.Shanghai Acad Agr Sci, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

3.Shanghai Acad Agr Sci, Minist Agr & Rural Affairs, Crop Breeding & Cultivat Res Inst, Key Lab Germplasm Innovat & Genet Improvement Grai, Shanghai 201403, Peoples R China

4.Shanghai Normal Univ, Coll Life Sci, Dev Ctr Plant Germplasm Resources, Shanghai 200234, Peoples R China

关键词： CRISPR-Cas12a; RPA; Fusarium fujikuroi; rice bakanae disease; detection

期刊名称：AGRONOMY-BASEL （ 影响因子：3.4； 五年影响因子：3.8 ）

ISSN：

年卷期： 2025 年 15 卷 3 期

页码：

收录情况： SCI

摘要： Fusarium fujikuroi is the primary causal agent of rice bakanae disease, which can lead to substantial yield losses. Developing a rapid, highly specific, and accurate method for detecting F. fujikuroi is crucial for effective surveillance, prevention, and control of rice bakanae disease. In this study, a novel detection assay, RPA-Cas12a-F, was developed by integrating recombinase polymerase amplification (RPA) and Cas12a for the detection of F. fujikuroi. This assay demonstrated a limit of detection (LOD) of 1 copy/mu L of reference plasmid or 0.1 fg/mu L of F. fujikuroi genomic DNA (gDNA). Furthermore, to enable on-site detection, the RPA-Cas12a technique was combined with a lateral flow strip (LFS) for visual readout, thereby developing the RPA-Cas12a-LFS assay. The LOD of the RPA-Cas12a-LFS assay was 1000 copies/mu L of plasmid or 10 fg/mu L of F. fujikuroi gDNA. The RPA-Cas12a-based assays developed in this study enable rapid, highly accurate, sensitive, and specific detection of F. fujikuroi, making them a promising tool for on-site detection without the need for expensive equipment and time-consuming methodologies.

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