A Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Didymella segeticola Causing Tea Leaf Spot

文献类型： 外文期刊

第一作者： Tu, Yiyi

作者： Tu, Yiyi;Wang, Yuchun;Jiang, Hong;Ren, Hengze;Wang, Xinchao;Lv, Wuyun;Wang, Yuchun;Wang, Xinchao;Lv, Wuyun

作者机构：

关键词： tea plant; Didymella segeticola; diagnosis; specificity; efficiency

期刊名称：JOURNAL OF FUNGI （ 影响因子：4.2； 五年影响因子：4.5 ）

ISSN：

年卷期： 2024 年 10 卷 7 期

页码：

收录情况： SCI

摘要： Tea leaf spot caused by Didymella segeticola is an important disease that threatens the healthy growth of tea plants (Camellia sinensis) and results in reductions in the productivity and quality of tea leaves. Early diagnosis of the disease is particularly important for managing the infection. Loop-mediated isothermal amplification (LAMP) assay is an efficient diagnostic technique with the advantages of simplicity, specificity, and sensitivity. In this study, we developed a rapid, visual, and high-sensitivity LAMP assay for D. segeticola detection based on sequence-characterized amplified regions. Two pairs of amplification primers (external primers F3 and B3 and internal primers FIP and BIP) were designed based on a specific sequence in D. segeticola (NCBI accession number: OR987684). Compared to common pathogens of other genera in tea plants and other species in the Didymella genus (Didymella coffeae-arabicae, Didymella pomorum, and Didymella sinensis), the LAMP method is specific for detecting the species D. segeticola. The assay was able to detect D. segeticola at a minimal concentration of 1 fg/mu L genomic DNA at an optimal reaction temperature of 65 degrees C for 60 min. When healthy leaves were inoculated with D. segeticola in the laboratory, the LAMP method successfully detected D. segeticola in diseased tea leaves at 72 h post inoculation. The LAMP assays were negative when the DNA samples were extracted from healthy leaves. Leaf tissues with necrotic lesions from 18 germplasms of tea plants tested positive for the pathogen by the LAMP assay. In summary, this study established a specific, sensitive, and simple LAMP method to detect D. segeticola, which provides reliable technical support for estimating disease prevalence and facilitates sustainable management of tea leaf spot.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序