Rapid diagnosis of wheat head blight caused by Fusarium asiaticum using a loop-mediated isothermal amplification assay

摘要

Fusarium asiaticum is an important fungal pathogen that causes wheat head blight or scab in China. In this study, we describe a loop-mediated isothermal amplification assay that directly detects F. asiaticum in diseased wheat head tissues, thus allowing rapid diagnosis of wheat head blight caused by this pathogen. We selected the CYP51C gene sequence to design loop-mediated isothermal amplification (LAMP) primers specific for F. asiaticum. The target gene (CYP51C) was efficiently amplified in 60 min at 63 A degrees C using the LAMP assay. The sensitivity of the assay was 100 pg center dot mu L-1. Specificity was compared with those for other Fusarium spp. and other fungal species. The result was directly visualized by adding hydroxynaphthol blue prior to amplification; a color change (sky blue) was observed only in the presence of F. asiaticum, whereas all other isolates showed no color change. We successfully applied the LAMP assay to diagnose the suspected disease in wheat head samples collected in the field. This study provides a new tool for the diagnosis of Fusarium head blight caused by F. asiaticum.