The genus Aphidius is the most commercially available species in greenhouse control, especially for aphids. Although twelve species were recorded in Korea, we collected 7 species including 2 unrecorded and 1 unidentified species. We aligned 16 sequences, except for duplicates, from 31 individuals of 14 species including GenBank sequences, and made phylogenetic tree based on Neighbor-Joining method. Surprisingly, 3 species were clustered into a clade and some species was not monophyletic. In this study, we reconstructed phylogenetic tree of Korean Aphidius species using COI marker, and compared them with GenBank sequences. In addition, we try to experiment a new marker and check its ability of discrimination.

Lysiphlebus orientalis is previously recorded in 2010, which is a more recently recorded than Lysiphlebia japonica. Host range of Lysiphlebus orientalis range is narrow, while that of Lysiphlebia japonica is very broad. Although two species, Lysiphlebus orientalis and Lysiphlebia japonica, are belonged to different genus, respectively, they are morphologically similar each other, which make us confused. Therefore, we have to identify these two cryptic species using COI DNA barcode. We used the ‘NCBI-BLAST (National center for biotechnology information-Basic Local Alignment Search Tool)’ to perform COI DNA barcode identification, and introduce preliminary results in this presentation.

DNA barcoding uses a 650 bp segment of the mitochondrial cytochrome c oxidase I (COI) gene as the basis for an identification system for members of the animal kingdom and some other groups of eukaryotes. PCR amplification of the barcode region is a key step in the analytical chain, but it sometimes fails because of a lack of homology between the standard primer sets and target DNA. Two forward PCR primers were developed following analysis of all known arthropod mitochondrial genome arrangements and sequence alignment of the tRNA-W gene which was usually located within 200 bp upstream of the COI gene. These two primers were combined with a standard reverse primer (LepR1) to produce a cocktail which generated a barcode amplicon from 125 of 141 species that included representatives of 121 different families of Hexapoda. High quality sequences were recovered from 79% of the species including groups, such as scale insects, that invariably fail to amplify with standard primers. The current results show that primers designed to bind to highly conserved gene regions upstream of COI will aid the amplification of this gene region in species where standard primers fail and provide valuable information to design a primer for problem groups.