Differentiation of Some Indigenous and Exotic Trichogrammatids (hymenoptera: Trichogrammatidae) from India Based on Internal Transcribed Spacer-2 and Cytochrome Oxidase-I Markers and Their Phylogenetic Relationship

Trichogrammatids (Hymenoptera: Trichogrammatidae) are the most important egg parasitoids widely used in different biocontrol programs worldwide. Since there are several species, strains, and ecotypes available, correct identification is the first step for successful biological control programs. Morphological identification remains complex due to subtle difference in male genetalia, therefore alternative molecular techniques were employed for rapid and reliable identification of this group of parasitoids. Furthermore, many of the field collected specimens are females, which are not identifiable using morphological keys. This study was carried out to unravel the discrimination success in the two molecular marker loci cytochrome oxidase I (COI) and internal transcribed spacer-2 (ITS-2) region of trichogrammatids. In achieving these objectives, the single specimen of 22 trichogrammatids belonging to 19 species were subjected to DNA extraction, PCR amplification, and sequencing. Additionally, we obtained homologous sequences from GenBank and sequences in BOLD database to understand internal relationships between the trichogrammatids. Bayesian inference phylogenetic analysis conducted with 84 and 76 sequences of COI and ITS-2 loci respectively and studied discrimination among the different species. Based on trees in comparison, we observed that there was a total of 10 and 6 out of 19 species correctly discriminate with COI and ITS-2 respectively. Our result revealed that the ITS-2 gene was less divergent than the COI gene in the majority of species and failed to differentiate all terminal clades. Therefore, we recommend that COI is suitable as the primary DNA barcode locus in trichogrammatids. Overall, we suggest that COI gene has higher discrimination power and can be considered as an appropriate molecular marker for species identification in trichogrammatids. (C) 2016 Elsevier Inc. All rights reserved.