DNA Based Identification and Genetic Diversity Studies of Antlion Species of Pakistan

Abstract

The present study was conducted during 2008-2014. This study initiate molecular taxonomy and reference library build up for future studies of antlion. The results revealed that Pakistan has a rich antlion fauna. Still lot of it needs to be explored. The current study have increased the species count from 28 species to 44 species belonging to 2 subfamilies 8 tribes and 20 genera. Current study has added 17 species to the local fauna out of which 2 are expected new species. Current study have not only revised the number of species but also indicated that there are so many genera which inhibit this country which have been neglected or missed somehow. Current study also synonymized some species like Palpers cepholate as Palpres trichogaster. Also two new species Stenares clarus and Centroclisis thari. Current study also report either Tomatares genus does not exists in this region or may have been raised due to mistaken identification of any other species. This study has established a reference library with specific Barcode Index Number (BIN) for each of 39 species consisting 703 COI DNA barcode sequences. While for one case Palpares astarte sequence length was less about 447bp so no Barcode index number was assigned to it. The integrity of sequences was tested using BLAST. Sequence having similarity to close relatives in Neuroptra were considered right and rest were discarded. The data regarding specimen’s collection, morphology (pictures) and DNA sequences were added to BOLD. Species discrimination was tested using online tool from BOLD and using MEGA6 (free tool). Specimens indicating more than 2% difference were considered different species. The species differentiation results from molecular data were similar to morphological data results. The analysis revealed that antlion species area not completely monophyletic. Still lot of data needs to be added for resolving antlions phylogeny. The sequence from DNA barcode region was very useful for species discrimination at 2% difference. But there was confusion in phylogenetic tree construction. When large number of taxa were used. Results showed that DNA barcode region can lead to some transgenic taxon arrangement as nearby neighbor. Hence it can be inferred that antlions morphological taxonomic hierarchy needs improvement. Also DNA barcode region alone may not be a good option to be used for phylogenetic analysis.