AUTHOR=Liu Jianjian , Li Chao , Muhae-Ud-Din Ghulam , Liu Taiguo , Chen Wanquan , Zhang Jianmin , Gao Li 

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

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.00004

DOI=10.3389/fmicb.2020.00004

ISSN=1664-302X

ABSTRACT=<sec><title>Background and Aims</title><p>The dwarf bunt disease of wheat is caused by <italic>Tilletia controversa</italic> Kühn. 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 <italic>T. controversa</italic>. 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.</p></sec><sec><title>Methods</title><p>We compared PCR, real-time PCR and droplet digital PCR (ddPCR) for the qualitative and quantitative measurement of the teliospores of <italic>T. controversa</italic> in soil.</p></sec><sec><title>Results</title><p>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/μL. In contract, SYBR Green I real-time PCR could detect 7.97 copies/μL of <italic>T. controversa</italic> in soil, and this sensitivity was 100 times more sensitive than that of simple PCR.</p></sec><sec><title>Conclusion</title><p>This study was the first report using ddPCR techniques to detect <italic>T. controversa</italic> teliospores in soil with greatly enhanced sensitivity.</p></sec>