Phylogeny and molecular studies of twelve Butterfliesspecies genus using RAPD-PCR technique from the Tiruchirappalli district, Tamil Nadu region in India
The present study reveals that RAPD-PCR is extremely useful for rapid identification of genetic polymorphisms in lepidopteron because of reproducibility of the result for each of the species.Mitochondrial genome is one of the most frequently used loci in phylogenetic and phylogeographic analysis, and it is becoming increasingly possible to sequence and analyse this genome in its entirety from diverse taxa. The mitochondrial gene coding for Cytochrome oxidase subunit I (COI) was extensively used for studying the molecular diversity of eukaryotic organisms, and hence the same has been adopted in this study as well. From the 12 selected butterfly’s species, the genomic DNA was isolated by CTAB-NaCl method, and their quality and quantity were tested through agarose gel electrophoresis. The COI gene was amplified using the custom-designed degenerate primer set which yielded around ~650 bp amplicons. The genus and species names of the closest match from the database were identified based on the BLAST parameters like identity, query coverage and E value. The identity of all the sequences were close to 100 % (99-98 %) and it is obvious as the COI gene is highly conserved in nature for the same purpose it has been used as a marker to study evolutionary relationships. In this study, a phylogenetic tree was constructed for the 12 COI gene sequences of different butterflies by Neighbor-Joining method employing Bootstrap type. Dendogram constructed using the UPGMA of NTSYS spc2.2 software divided the butterflies species into two clades. There is a difference in the branching pattern between the morphological and molecular data, which signifies the need for using molecular tools for taxonomic classification as well as in understanding the evolutionary relationship.