Real-time Fluorescent RPA Detection Method for Rhizoctonia Solani Based on ITS Sequences

Rhizoctonia solani, a pathogen with a broad host range, propagates through its hyphae in the soil, invading plant roots or seeds, and subsequently affecting surrounding plants. Given that the pathogenic fungus continues to develop within the crop population after infection, early detection of the pathogen and timely prevention measures are crucial for reducing losses and preventing the spread of the disease. In this study RPA (Recombinase polymerase amplification) specific primers and fluorescent probe were designed based on the ITS sequence of Rhizoctonia solani and an RPA-real time detection method for Rhizoctonia solani was established. By optimizing the reaction conditions of RPA, the optimal conditions for RPA detection method were 39℃ for 30 min. This assay can distinguish Rhizoctonia solani from other related species. The sensitivity of RPA-real time detection method for Rhizoctonia solani can be achieved at 6×10-2 ng/μL. The method for Rhizoctonia solani diagnosis using RPA-real time, which is rapid convenient and sensitive can provide a new rapid assay for the detection of Rhizoctonia solani.