Application of DNA barcoding to species identification of an unknown insect larva
Objective: To establish the method of applying DNA Barcoding techniques to the identification of unknown insect larva, and discuss the practicality of DNA barcoding in the species identification. Methods: PCR was performed with the metazoan invertebrates degenerate Mitochondrial Cytochrome Oxidase I (COI) primers. 709 bp fragments of mithochondrial cytochrome oxidase I (COI) were amplified from both adult and larva of the same species. The PCR products were cloned into pGM-T Vector for sequencing, and then blasted the sequences in Genbank. Results: There was only one base pair difference out of these 709 bp between the adult and larva, identity was as high as 99.9％. It was 99.6％ identical to the COI sequences of Sitophilus oryzae, and 86.6％ to Sitophilus zeamais published in Genbank. Phylogeny trees were built with ClustalW - 2 online as well. Tested with the classification morphology characteristics of the adult, the unknown species was Sitophilus oryzae. Conclusion: All these results show that application of DNA Barcoding to species identification of unknown insect larva is desirable and with high confidence.
NETL
NSTL
万方数据
维普
