Establishment of recombinase polymerase amplification detection method for Puccinia striiformis f. sp. Tritici

Wheat stripe rust caused by Puccinia striiformis f. sp. tritici ( Pst )is an airborne disease that endangers the entire growth period of wheat. In this study, the effect of gene PS4593 of Pst was used as the target gene, that PS4593 effector genes have significant virulence functions in wheat, this effector gene could suppress wheat ETI against an avirulent Pst isolate, and the primers PS-RPA-F and PS-RPA-R, as well as the probe PS-LF-Probe, were designed for recombinase polymerase amplification (RPA) technology. Flow chromatography was combined with the process to establish an RPA detection method for Pst . This method could establish visual detection within 10 min under a constant temperature of 39°C, and the detection results were consistent with those of ordinary PCR analysis. This method only had high specificity for Pst , and the detection limit was 10 fg/µL. In addition, this rapid method could detect Pst from wheat leaves during the field incubation period, which has strong benefits in application. In sum, the RPA detection method established in this study has the favorable characteristics of high efficiency, simple functionality, and rapid and universal practicability, which provides a theoretical basis for the early detection and prevention of Pst .