D3229: Loop-mediated isothermal amplification of PBAN gene for molecular diagnosis of Bemisia tabaci (Hemiptera: Aleyrodidae)

Tobacco whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a serious pest that transmits Tomato yellow leaf curl virus (TYLCV) in tomato plantations. Greenhouse whitefly Trialeurodes vaporariorum (Hemiptera: Aleyrodidae), a closely allied species of tobacco whiteflies, shows morphological differences, but it is difficult to distinguish them visually in agricultural fields. Therefore, when whiteflies appear in the field, it is very important to quickly and accurately diagnose the species of whiteflies in the pest management strategy. In this study, we used a loop-mediated isothermal amplification (LAMP) assay to quickly and effectively detect B. tabaci. Primer sets were investigated for the specificity of B. tabaci by visible detection of target DNA templates. Two primer sets for pheromone biosynthesis activating neuropeptide (PBAN) gene were obtained from transcriptomic analysis of B. tabaci biotype Q. The PBAN-based LAMP primer set showed specific amplification of the target region. The optimal conditions for B. tabaci detection were 60°C for 60 min with four LAMP primers of PBAN. The minimum amount of genomic DNA required for visible detection was 100 ng. These results suggest that the PBAN-based LAMP assay can be applicable for field monitoring of B. tabaci.