LAMP Assay for Rapid Fusarium verticillioides Detection on Corn Plant

Objective A rapid, sensitive, and visual loop-mediated isothermal amplification (LAMP)-based method to detect Fusarium verticillioides of corn stalk rot was developed.

Method A set of primers based on the Elongation factor-1α sequence of F. verticillioides with the template DNA from infected corn leaves and optimized temperature and time were applied for the LAMP assay. Specificity and sensitivity of the prototype method were tested to verify the assay applicability for accurate pathogen detection on clinic specimens.

Result Allowing a reaction time of 60m at 65℃, the assay effectively and specifically detected F. verticillioides on the diseased corn plants. It had a detection limit of 10 fg·μL⁻¹ and provided results consistent with the morphological diagnosis. On 35 field-collected corn plants that were suspected of stalk rot infection, the LAMP assay as well as the physiological identification found 17 to be F. verticillioides-positive and 18 negative.

Conclusion The newly established LAMP assay could visually, rapidly, specifically, and accurately detect F. verticillioides on the infected corn plants. It was adequate for monitoring and early diagnosis to facilitate effective control of the disease.