Establishment of LFD-RPA Rapid Detection Technique for Tobacco Target Spot

Target spot caused by Rhizoctonia solani Kühn is a tobacco leaf disease. In recent years, it has occurred seriously in some provinces of China, which endangers tobacco production. In this study, the recombinant enzyme polymerase technology related primers RsRPA-F3/R3 were designed according to the internal transcribed spacer (ITS) of the pathogen, and a method of recombinase polymerase amplification combined with a lateral flow dipstick (LFD-RPA) was developed to establish early and rapid diagnosis technique for tobacco target spot disease. The LFD-RPA detection technology could detect plasmid DNA of R. solani (1.40×10²copies/μL) within 15 min at 37 ℃. The sensitivity was consistent with that of conventional PCR. The detection technology can be applied to the real-time detection of tobacco target spot disease in the field with a 95.83% accuracy rate. The rapid detection technology established in this study has the characteristics of fast, simple and visual, and provides a new method for the rapid field diagnosis of tobacco target spot.