D3319: Development of LAMP-based molecular species diagnosis method for three major planthopper pests in rice

Molecular-based species identification tools are helpful not only to tiny insect pests but also to planthopper pests because of their morphological similarity. Rapid and accurate species diagnosis is essential in quarantine and agricultural research fields. In this study, we focused on three major planthopper pests, Nilaparvata lugens, Laodelphax striatellus, and Sogatella furcifera. Based on partial mt genome sequence alignment, a species-specific sequence region was identified in the tRNAs coding region between NADH dehydrogenase, including N. lugens (GenBank: JX880069.1), L. striatellus (GenBank: MK292897.1), S. furcifera (GenBank: KC512914.1). Species diagnostic primers were designed using these unique regions and applied in LAMP and multiplex PCR　to identify the field collected. The most optimal incubation condition of the LAMP assay was 65℃ for 60 minutes with 4 LAMP primers. Additional loop primers increased the amplification efficiency in LAMP assay in Nilaparvata lugens and Laodelphax striatellus, but increased nonspecific amplification in Sogatella furcifera. This new molecular diagnostic method is easy to use, accurate, and possibly applied in monitoring and ecological studies.