DNA-barcoding based identification of potential plant hosts of phytoplasmas from leafhoppers

Description:DNA barcoding is a process for rapid and precise species identification. Currently available DNA barcoding markers makes it possible to simultaneously identify the insect, the plants the insect ingests, and microbial species within a single insect specimen. In this project, the potential plant hosts of phytoplasmas harbored by seven leafhopper species (Leofa dispar, Pravistylus exquadratus, Neoaliturus opacipennis, Macrosteles sordidipennis, Mayawa capitata, Mayawa affinifacialis and Acharis ussuriensis) were identified. The findings will provide a basis for further study on phytoplasma, plant, and insect associations in their natural habitat.Method:The plant barcoding marker, rbcL gene was amplified by semi nested PCR from the leafhoppers that tested positive for phytoplasmas. PCR amplicons were cloned into TOPO TA cloning vector and five clones were randomly picked up and followed by Sanger sequencing. The obtained sequence from the individual clone was blasted against NCBI database to identify the plant species.