Abstract
Root rot, caused by several pathogens, is one of the most devastating diseases on Astragalus membranaceus, and can lead to serious yield loss. Fusarium acuminatum and F. solani are known to be major pathogens causing root rot of A. membranaceus in Shanxi Province, China. It is well-known that accurate identification of the pathogen and early diagnosis of the symptoms play a significantly important role to control this disease in the early or mid-term stage. Here, two new loop-mediated isothermal amplification (LAMP) assays were developed to detect F. acuminatum and F. solani based on the partial translation elongation factor-1α (TEF-1α) gene region. The LAMP products were successfully assessed by visual assessment using SYBR Green I, as well as electrophoresis on 2.0% agarose gel. The LAMP reactions for F. acuminatum and F. solani were performed under the optimized conditions of 63 °C and 62 °C for 60 min, respectively. The detection limit of LAMP assay was 100 fg/μL for F. acuminatum and 1 pg/μL for F. solani. Additionally, the two LAMP assays are highly specific to target Fusarium species, and could differentiate F. acuminatum and F. solani from other Fusarium species. In the field, the suspected diseased samples were used to verify the feasibility of the developed assays. As a result, the two rapid and specific LAMP assays could be applied for direct detection of F. acuminatum and F. solani on A. membranaceus, and precise diagnosis of A. membranaceus root rot caused by F. acuminatum and F. solani.
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This work was financially supported by Research Program Sponsored by State Key Laboratory of Sustainable Dryland Agriculture (in preparation), Shanxi Agricultural University (No. 202105D121008–3-3), the Sub-project of National Key R&D Program “Study on Modernization of Traditional Chinese Medicine” (2019YFC1710802–05), the Earmarked Fund for Modern Agro-industry Technology Research System (2021-11), Special Project of Academic Restoration of Shanxi Agricultural University (2020xshf05), and Science and Technology Innovation Funds of Shanxi Agricultural University (2016YJ11 and 2016YJ12).
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Yan Wang conceived the ideas and wrote the manuscript. Chunwei Wang designed the methodology, and made the corresponding revisions. Yurong Ma and Xiqian Zhang performed the assays. Hongbin Yang, Guangxin Li, and Xinfeng Li collected the samples. Meiqin Wang, Xiaojun Zhao, Jianming Wang and Xianhong Zhang analyzed the data.
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Wang, Y., Wang, C., Ma, Y. et al. Rapid and specific detection of Fusarium acuminatum and Fusarium solani associated with root rot on Astragalus membranaceus using loop-mediated isothermal amplification (LAMP). Eur J Plant Pathol 163, 305–320 (2022). https://doi.org/10.1007/s10658-022-02478-x
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DOI: https://doi.org/10.1007/s10658-022-02478-x