Development of the Droplet Digital PCR to Detect the Teliospores of Tilletia controversa Kuhn in the Soil With Greatly Enhanced Sensitivity

文献类型： 外文期刊

第一作者： Liu, Jianjian

作者： Liu, Jianjian;Li, Chao;Muhae-Ud-Din, Ghulam;Liu, Taiguo;Chen, Wanquan;Gao, Li;Liu, Jianjian;Zhang, Jianmin

作者机构：

关键词： Tilletia controversa Kuhn; dwarf bunt; droplet digital PCR; soil born disease; detection

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.64； 五年影响因子：6.32 ）

ISSN： 1664-302X

年卷期： 2020 年 11 卷

页码：

收录情况： SCI

摘要： Background and Aims The dwarf bunt disease of wheat is caused by Tilletia controversa Kuhn. This pathogen is primarily involved in the stunted growth of wheat and affects seed quality. Many countries in the world have therefore imposed quarantine bans to prevent the spread of T. controversa. Morphological observations are the main method of detecting teliospores in soil. However, this is a lengthy and laborious process; this method is thus unable to quickly meet the demand for detection of teliospores in the soil. Methods We compared PCR, real-time PCR and droplet digital PCR (ddPCR) for the qualitative and quantitative measurement of the teliospores of T. controversa in soil. Results We suggest the use of ddPCR for detection of the soil samples, which was demonstrated to have the most sensitive detection at 2.1 copies/mu L. In contract, SYBR Green I real-time PCR could detect 7.97 copies/mu L of T. controversa in soil, and this sensitivity was 100 times more sensitive than that of simple PCR. Conclusion This study was the first report using ddPCR techniques to detect T. controversa teliospores in soil with greatly enhanced sensitivity.

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