Molecular Phylogeny of Myllocerus viridanus (Fabricius) by mitochondrial Cytochrome B gene Sequencing

Abstract

In order to create accurate control techniques against insect pests, correct taxonomic identification is critical. This paper describes the molecular phylogeny of Grey weevil, Myllocerus viridanus (Fabricius) by the sequencing of their mitochondrial cytochrome B gene. The development of a simple and most elegant tools of DNA barcoding has made taxonomy and phylogeny studies a much facile process. M. viridanus is among the important polyphagous pests of plants of agricultural and horticultural importance. The sequencing of Cyt B gene yielded a 273 bp nucleotide product. Phylogentic analysis was carried out in Mega 11 software. Gen Bank deposition indicated the novel and first time records of cytochromeb gene barcode of M. viridanus and first time deposit of a species from the genus Myllocerus. The deposit was provided with an accession number OM223090. Further analysis showed that the partial sequence of mitochondrial cytochrome B of M.viridanus has 88.89% similarity with that of Romualdius bifoveolatus (KX087356) .The identity of M.viridanus(OM223090)ranged from 88.17 to 83.68% when nucleotide BLAST was carried out with nearest 21 species of curculionids (NCBI). The result indicated that cytochrome b gene barcoding can be successfully used to resolve species level identification of the members of the family Curculionidae and the genus Myllocerus of the order Coleoptera. Thus mitochondrial Cytochrome b gene like mitochondrial COX 1 gene, can be used as an efficient tool for the species level identification of the members of Coleopterans. Further interpretation on phylogeny was done by constructing neighbor joining tree using Kimura -2 Parameter. This approach was very informative and it helped in the better understanding of genetic variation among closely related species of Curculionids.