Rapid and Sensitive On-Site Nucleic Acid Detection of Three Main Fusarium Pathogens of Maize Stalk Rot Based on RPA-CRISPR/Cas12a
文献类型： 外文期刊
作者： Jiang, Fan 1 ; Ding, Xinhua 2 ; Wang, Xiaowu 2 ; Fu, Kaiyun 2 ; Jia, Zunzun 2 ; Liang, Liang 4 ; Guo, Wenchao 2 ;
作者机构： 1.Chinese Acad Inspect & Quarantine, Beijing 100176, Peoples R China
2.Xinjiang Acad Agr Sci, Inst Plant Protect, Key Lab Integrated Pest Management Crops Northwest, Xinjiang Key Lab Agr Biosafety, Urumqi 830091, Peoples R China
3.CAIQ Ctr Biosafety, Sanya 572025, Hainan, Peoples R China
4.Acad Agr Planning & Engn, MARA, Beijing 100125, Peoples R China
关键词：
CRISPR/Cas12a;
期刊名称：PLANT DISEASE （ 影响因子：4.4； 五年影响因子：4.8 ）
ISSN： 0191-2917
年卷期： 2025 年 109 卷 2 期
页码：
收录情况： SCI
摘要： Maize stalk rot is a soilborne disease that poses a serious threat to maize production worldwide, with the most significant cause being fungal stalk rot. The development of a visual and rapid detection method for the maize stalk rot pathogen is significant for its prompt and accurate identification, enhancing agricultural production efficiency, and implementing timely preventive measures. These measures will help safeguard the maize yield and quality, ultimately reducing agricultural losses. In this study, we aimed to develop an efficient method to detect maize stalk rot pathogens. We focused on three pathogenic fungi commonly found in maize-producing regions worldwide: Fusarium verticillioides, F. proliferatum, and F. graminearum. Based on translation elongation factor 1-alpha, we developed a rapid detection technique using recombinase polymerase amplification-CRISPR/Cas12a, combined with test strips to develop an on-site rapid visual detection test for these pathogens. The method showed detection sensitivity for F. verticillioides, F. proliferatum, and F. graminearum within 20 min at concentrations of 7.8 pg/mu l, 0.11 ng/mu l, and 0.13 ng/mu l, respectively. The sensitivity increased with increasing reaction time. Testing of field disease samples indicated that the method is effective in detecting nucleic acids obtained through crude extraction methods. In conclusion, we developed a visually rapid detection technology that does not rely on complex instruments and equipment for the on-site early detection of F. verticillioides, F. proliferatum, and F. graminearum in the field to implement effective control measures, ensuring stable and high maize yields.
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