Qinghai Provincial Key Laboratory of Agricultural Integrated Pest ManagementScientific Observing and Experimental Station of Crop Pest in Xining, Ministry of Agriculture, Academy of Agriculture and Forestry Science, Qinghai University, Xining, 810016, Qinghai, People's Republic of China.
Sci Rep. 2023 Sep 26;13(1):16133. doi: 10.1038/s41598-023-42663-4.
Wheat stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is an airborne disease that endangers wheat during its entire growth period. In this study, the Pst134EA_003354 uncharacterized protein (GenBank: XM_047941824.1) of Pst was used as the target sequence, 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 successfully established visual detection within 10 min under a constant temperature of 39 °C, and the detection results were consistent with those of ordinary PCR analysis. However, it only had high specificity for Pst, and the detection limit was 10 fg/μL. In addition, this rapid method successfully detected Pst from wheat leaves during the field incubation period, indicating substantial benefits for applied use. In summary, the RPA detection method established in this study has the favourable characteristics of high efficiency, simple functionality, and rapid and universal practicability, providing a theoretical basis for the early detection and prevention of Pst.
小麦条锈病由条形柄锈菌小麦专化型（Pst）引起，是一种在小麦整个生长期间危害小麦的气传病害。本研究以 Pst 的一个未知功能蛋白 Pst134EA_003354（GenBank: XM_047941824.1）为靶标序列，设计了引物 PS-RPA-F 和 PS-RPA-R 以及探针 PS-LF-probe，用于重组酶聚合酶扩增（RPA）技术。通过结合流色谱，建立了 Pst 的 RPA 检测方法。该方法在恒温 39°C 下成功实现了 10 min 内的可视化检测，且检测结果与普通 PCR 分析一致。然而，该方法仅对 Pst 具有高度特异性，检测限为 10 fg/μL。此外，该快速方法成功检测到田间孵育期小麦叶片中的 Pst，表明其在实际应用中具有很大的优势。综上所述，本研究建立的 RPA 检测方法具有高效、简便、快速、通用的特点，为 Pst 的早期检测和预防提供了理论依据。
