Qinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai University, Xining, 810016, Qinghai, People's Republic of China.
Sci Rep. 2024 Oct 23;14(1):25079. doi: 10.1038/s41598-024-76921-w.
Hulless barley sheath rot is a spike disease caused by Dactylobotrys graminicola. In recent years, it has generally occurred in hulless barley growing areas in China, resulting in reduced hulless barley yields. In this study, primers and probes were designed based on conserved genome sequences, and a method was established using recombinant enzyme polymerase amplification-lateral flow burette (RPA-LFD) technology for the rapid diagnosis of sheath rot in hulless barley. The method can be successfully established in five minutes at a constant temperature of 39℃, and the results are consistent with those of normal PCR analysis. The method demonstrated high sensitivity, with a detection limit of 10 fg/µL. Furthermore, the rapid method was able to successfully detect D. graminicola in hulless barley during field incubation, which highlighted the significant advantage of the method in practical applications. In conclusion, the RPA method established in this study exhibited several advantageous characteristics, including high efficiency, simplicity, rapidity and practicality, which provide a theoretical basis for the early detection and prevention of D. graminicola.
青稞无壳秕粒病是一种由 Dactylobotrys graminicola 引起的穗部病害。近年来，在中国青稞种植区普遍发生，导致青稞产量下降。本研究基于保守的基因组序列设计引物和探针，建立了一种利用重组酶聚合酶扩增-侧流层析（RPA-LFD）技术快速诊断青稞无壳秕粒病的方法。该方法在 39℃恒温下可在五分钟内成功建立，且结果与常规 PCR 分析一致。该方法具有较高的灵敏度，检测限为 10 fg/μL。此外，该快速方法能够成功检测到田间培养过程中的青稞无壳秕粒病，突出了该方法在实际应用中的显著优势。综上所述，本研究建立的 RPA 方法具有高效、简便、快速和实用等优点，为 D. graminicola 的早期检测和防治提供了理论依据。
