Rapid and quantitative detection of Aspergillus niger Van Tieghem using loop-mediated isothermal amplification assay
文献类型： 外文期刊
作者： Dai, Xiaodong 1 ; Cao, Yanyong 2 ; Yu, Minghui 4 ; Hou, Meiwei 2 ; Li, Huimin 2 ; Li, Jie 4 ; Li, Hangyu 4 ; Li, Peipei 4 ; Wang, Zhenyu 5 ; Zhang, Xinyou 1 ;
作者机构： 1.Henan Acad Crops Mol Breeding, Henan Prov Key Lab Oil Crops Improvement, Natl & Prov Joint Engn Lab Peanut Genet Improvemen, Key Lab Oil Crops Huang Huai Hai Plains,Minist Agr, Zhengzhou 450002, Peoples R China
2.Henan Acad Agr Sci, Inst Cereal Crops, Zhengzhou 450002, Peoples R China
3.Shennong Lab, Zhengzhou 450002, Peoples R China
4.Xinyang Acad Agr Sci, Xinyang 464000, Peoples R China
5.Henan Acad Agr Sci, Inst Plant Protect, Zhengzhou 450002, Peoples R China
关键词： Aspergillus niger; GOD; Loop-mediated isothermal amplification; Rapid and quantitative detection
期刊名称：JOURNAL OF PLANT PATHOLOGY （ 影响因子：2.2； 五年影响因子：2.5 ）
ISSN： 1125-4653
年卷期： 2024 年
页码：
收录情况： SCI
摘要： Peanut (Arachis hypogaea L.) crown rot and root rot are common diseases caused by Aspergillus niger Van Tieghem. Early and accurate detection of A. niger is key to disease management. In this study, the design of two to five sets of loop-mediated isothermal amplification (LAMP) primers was based on the EglA, GOD, Tub, NRPS, Tan, CbhA, and CbhB genes of A. niger. Of these, primer set GOD-91 was selected for optimization of the three-factor LAMP system: the Bst DNA polymerase concentration, the concentration ratio of the inner and outer primers, and the concentration of Mg2+. In addition, the optimized LAMP reaction system for A. niger detection was validated for specificity, sensitivity, and on-site feasibility. The specificity test showed that A. niger could be specifically detected with the proposed method without cross-amplification of other pathogenic fungi DNA. Moreover, based on the sensitivity test, the lowest detection limit of this reaction system was 5.1 x 10(-7) ng/mu L pAN01 plasmid DNA, after which a standard curve was generated for the quantitative detection of A. niger. The LAMP method was further applied for field sample assessment before and after A. niger infection, successfully detecting A. niger presence in the samples collected in the field. This study yielded a sensitive, specific, and reproducible LAMP system that can be used to assess on-site samples within 45 min. It is an effective approach for the rapid and quantitative detection of A. niger.
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