Development and evaluation of a novel and rapid detection assay for Blumeria graminis f. sp. tritici based on Loop-Mediated Isothermal Amplification
Shuangjun Gong: Institute of Plant Protection and Soil Science, Hubei Academy of Agricultural Sciences
<div>Powdery mildew, caused by <em>Blumeria </em><em>graminis</em> f. sp. <em>tritici</em>, is the most common foliar disease of wheat in China. In this study, we developed a rapid method, loop-mediated isothermal amplification (LAMP), to detect <em>B</em>. <em>graminis</em> f. sp. <em>Tritici</em>(Bgt) in field. Eight pairs of primers were designed to amplify Bgt specific sequences, two pairs of primers showed both high efficiency and high specificity with 35 Bgt strains from wheat and 7 powdery mildew strains from other plant host. Less than 300fg genomic DNA is needed for detection by this method, showed comparable sensitivity with normal PCR method. The LAMP reaction could be quickly done within 60 minutes at 65℃, and color change in magnesium pyrophosphate precipitation after adding calcein could be observed with naked eye. Since its technique simplicity, time efficiency and does not require expensive equipment, this LAMP-based diagnostic method has potential to be used under field conditions to make disease forecasting more accurate and efficient.</div>
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