Rapid diagnosis of rice bakanae caused by Fusarium fujikuroi and F-proliferatum using loop-mediated isothermal amplification assays

摘要

Rice bakanae is an important disease that causes serious rice production loss worldwide. We describe a new method for rapid diagnosis of rice bakanae caused by Fusarium fujikuroi and F.proliferatum, based on loop-mediated isothermal amplification (LAMP) assays. After screening, primers were selected to target FusariumDNA sequences, that is, the intergenic spacer (IGS) region of the nuclear ribosomal operon and reductase-coding region (RED1) in F.fujikuroi and F.proliferatum, respectively. Both LAMP assays efficiently amplified target genes in 70min at 62 degrees C. A colour change from purple to sky blue (visible to the unaided eye) was observed in the presence of the DNA of the targeted pathogens only, by adding hydroxynaphthol blue to the reaction system prior to amplification. The minimum of genomic DNA needed in the assays was 67 and 346pg/l for F.fujikuroi and F.proliferatum, respectively. Using the two assays described here, we successfully and rapidly diagnosed suspected diseased rice plant and seed samples collected from Jiangsu Province.