Development of loop-mediated isothermal amplification (LAMP) assay for rapid detection of Fusarium proliferatum causing ear and kernel rot on maize

Abstract

Fusarium proliferatum, one of the predominant causal agents associated with ear and kernel rot in China, reduces maize yield and quality, and produces various mycotoxins that are harmful to animals and humans. A key challenge for the integrated management strategy of this disease is rapid and accurate detection of F. proliferatum on maize. Therefore, a new loop-mediated isothermal amplification (LAMP) assay was developed for the detection of F. proliferatum from the diseased maize. The primers were selected and designed using the translation elongation factor-1α (TEF-1α) gene region as target DNA sequence. The amplicons were successfully inspected by visual assessment using SYBR Green I as well as electrophoresis on agarose gel. The target gene was efficiently amplified at 62 °C for 30 min. The minimum detection concentration was 10 pg μL−1. The LAMP assay was found to be highly specific for F. proliferatum from the other fungal species. In field trial, F. proliferatum could be detected from suspected diseased maize samples. The results showed that the new and readily available LAMP assay has potential to apply successfully for the detection of F. proliferatum causing ear and kernel rot on maize in the field.