LAUSR

Identifying dead insect larvae by morphology is mostly challenging but often required in ecological studies, which entails a molecular method. In this study, we developed a protocol of Cytochrome Oxidase c Subunit I (COI) DNA barcoding that could quickly identify larval tephritid flies in a Tibetan alpine meadow. The protocol includes two major operations. The first is to build up a comprehensive reference library after identifying adults of tephritid flies to species by morphology and by COI DNA barcoding sequences. The second is to calculate species-specific delimitation criteria using the sequences in the library, such that COI DNA sequences of insect larvae could be assigned to species using local blast. This protocol was proved successful in the species identification of larval flies among 27 tephritid species belonging to nine genera of subfamily Tephritinae. We suggest that such a protocol can be widely applied to fast identification of insect larvae.