Loop mediated isothermal amplification (LAMP) as a rapid and portable diagnostic tool for the detection of pea root rot pathogens

Abstract

Abstract Pea root rot, caused by soilborne pathogens such as Aphanomyces euteiches, various Fusarium species and Pythium ultimum, represents a significant challenge to global pea production. Current diagnostic methods are costly, time-consuming and require specialised laboratory setups, limiting their application for rapid decision-making in agricultural settings. To address this, we developed a Loop-Mediated Isothermal Amplification (LAMP) assay targeting the Internal Transcribed Spacer 1 (ITS1) region of these pathogens for rapid and in-field detection. The assay demonstrated high specificity and sensitivity, with detection limits as low as 0.02 ng of gDNA and an initial inoculum of 10 spores per sample in both soil and infected root tissue. Results were generated within 60 min, providing a practical alternative to conventional diagnostic methods. Validation with qPCR showed LAMP to be comparable or superior in accuracy, and field deployment with a portable LAMP device confirmed its effectiveness for on-farm diagnostics. This LAMP assay offers a robust, cost-effective and user-friendly tool for the early detection of pea root rot pathogens, enhancing disease management and potentially reducing crop losses.