Development of species-specific marker and molecular diversity of Conogethes punctiferalis and Conogethes sahyadriensis (Lepidoptera: Crambidae)

Aim: The aim of this study was to develop species specific marker for quick identification of two Conogethes species and to study their molecular diversity. Methodology: Species-specific markers were developed using mitochondrial cytochrome oxidase 1 (COI) partial gene sequences of Conogethes punctiferalis and C. sahyadriensis and they were validated. Phylogenetic tree was constructed using MEGA X program, tree robustness was evaluated by bootstrapping with 1000 replicates with all sequences of C. punctiferalis and C. sahyadriensis. Results: Bioinformatic analysis of C. punctiferalis and C. sahyadriensis COI sequences revealed that they have sufficient genetic variability to develop species specific marker. The COI based species-specific markers amplified an expected fragment size of 333 bp for C. punctiferalis and 522 bp for C. sahyadriensis, which clearly differentiate these two species. Interpretation: The species specific marker developed in this study will be useful in quick identification of C. punctiferalis and C. sahyadriensis. The use of COI gene as a marker can provide a better estimate of genetic diversity, as it is maternally inherited and can be helpful to understand evolutionary processes. Key words: Conogethes punctiferalis, C. sahyadriensis, MtCOI, Molecular diversity, Species-specific marker