Detection of Tilletia controversa Kühn by real time quantitative PCR.
Objective: To detection of Tilletia controversa Kühn (TCK) sensitively and accurately, real-time PCR systems were developed. Method: The species-specific primer pair CQUTCK4/CQUTCK5 and probe CQUP1 were designed based on a selected specific fragment (1 322 bp) specific for TCK, and the SYBR Green I and TaqMan quantitative PCR detection systems were established with optimized reaction conditions. Result: The detection limit of the two systems were 0.1 fg, equal to 2.31×104 copies, which was 102-103 fold higher than conventional PCR. By the constructed detection systems, the TCK and Tilletia caries (DC) Tul (TCT) could be distinguished. The teliospore and mycelium of TCK in the infected wheat plant tissue also could be identified accurately and rapidly. Conclusion: The earlier diagnosis approaches of wheat durwf bunt pathogen were set up using the two real-time PCR systems.