AUTHOR=Ren Zhaoyu , Chen Rongzhen , Muhae-Ud-Din Ghulam , Fang Mingke , Li Tianya , Yang Yazheng , Chen Wanquan , Gao Li 

TITLE=Development of real-time PCR and droplet digital PCR based marker for the detection of Tilletia caries inciting common bunt of wheat

JOURNAL=Frontiers in Plant Science

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2022.1031611

DOI=10.3389/fpls.2022.1031611

ISSN=1664-462X

ABSTRACT=<p>This is the first study reporting droplet digital PCR and quantitative real time PCR for detection of <italic>Tilletia caries</italic> (syn. <italic>T. tritici</italic>), which causes common bunt of wheat and leads to yield losses of 80% in many wheat growing areas worldwide. To establish an accurate, rapid and quantifiable detection method, we tested 100 inter simple sequence repeats (ISSR) primers and obtained a species-specific fragment (515 bp) generated by ISSR 827. Then, a specific 266 bp band for the sequence characterized amplified region (SCAR) marker was produced from <italic>T. caries</italic>. The detection limit reached 50 pg/μL. Based on the SCAR marker, we further developed a higher sensitivity of quantitative real time-polymerase chain reaction (qRT-PCR) with a detection limit of 2.4 fg/μL, and droplet digital PCR (ddPCR) with a detection limit of 0.24 fg/μL. Both methods greatly improved the detection sensitivity of <italic>T. caries</italic>, which will be contribute a lot for quickly and accurately detection of <italic>T. caries</italic>, which causes wheat common bunt.</p>