A Loop-Mediated Isothermal DNA Amplification (LAMP) Assay for Detection of the Grape White Rot Pathogen Coniella...

Grape white rot, caused by Coniella vitis, is a devastating disease that affects grape production in China and worldwide, resulting in substantial yield and quality losses. Early and accurate detection of C. vitis is critical for effective disease management. In this study, we developed a rapid and highly sensitive loop-mediated isothermal amplification (LAMP) assay targeting the translation elongation factor 1-alpha (TEF-1α) gene of C. vitis. The reaction temperature and time were optimized, and the ideal conditions for the LAMP assay were 63°C for 30 min. Specificity tests against 22 fungal pathogens, including eight C. vitis strains and two C. granati isolates closely related to C. vitis, confirmed the assay's exclusivity to C. vitis. The LAMP assay demonstrated a detection limit of 100 fg genomic DNA per reaction, which was 10-fold more sensitive than conventional PCR. Field validation using infected grape tissues successfully identified C. vitis in symptomatic samples. This LAMP assay offers a rapid and cost-effective method for the early diagnosis and surveillance of grape white rot in vineyards.