Rapid Detection of Rhizoctonia solani in Turfgrass and Soil Using a Loop-Mediated Isothermal Amplification-Based Point-of-Care Testing System

Abstract

Climate change is increasing the frequency and scale of turf disease outbreaks, yet systems for rapid and accurate disease diagnosis in the field remain limited. Existing microscopic examination methods require skilled professionals. And by the time visible symptoms appear, the optimal window for prevention is often missed. Rhizoctonia solani is a destructive pathogen causing severe turfgrass diseases. To address this, we report a rapid, field-deployable loop-mediated isothermal amplification (LAMP) assay targeting a fungal-specific ITS-5.8S rDNA sequence. The amplification reaction was optimized at 65 °C for 20 minutes, achieving an analytical sensitivity with a limit of detection (LoD) of 1 fg per reaction. The assay exhibited 100% analytical specificity with no cross-reactivity, validated across a panel of 31 fungal isolates comprising target Rhizoctonia species and non-target taxa, as well as successfully tested on artificially contaminated turf and soil matrices. Overall, this highly sensitive and specific LAMP-based system provides a reliable and practical tool for the rapid, on-site detection of R. solani in turf management.