Identification of pumpkin beetle (Aulacophora spp.) through mtDNA-COI barcode approach in cucurbitaceous vegetable crops

Abstract

Cucurbitaceous vegetable crops are infested by various Aulacophora spp. and management of this pest requires a proper identification approach. But the identification of Aulacophora spp. through morphological study has few limitations. These issues are overcome by molecular barcoding approach because it offers great potential for identification of Aulacophora spp. Therefore, genomic DNA was extracted from legs and wings rich in mitochondria of 10 samples of Aulacophora spp. which were collected from different zones of Bihar, during 2022 and 2023. And then amplification of mitochondrial cytochrome c oxidase subunit I gene was done using universal primer pair at the toxicology laboratory of the Department of Entomology, Bihar Agricultural University, Sabour, Bihar, India. PCR amplified Mt-COI barcode region from Aulacophora spp. were sequenced and submitted to NCBI-GeneBank and received NCBI-GeneBank accession number of 10 entries. BLAST analysis confirmed two Aulacophora spp. namely Aulacophora lewissi and Aulacophora indica by comparing sequence identity between the obtained sequences of 10 samples of Aulacophora spp. and existing sequences of different countries from NCBI GenBbank. At 0.020 bar scale, phylogenetic tree was clustered into three major groups. Additionally, haplotype analysis revealed Aulacophora lewisii and Aulacophora indica were discreted from their common ancestors by distinct mutational steps. Further, some strong codons were found in each Aulacophora spp. which can be further utilized in translation process.