The present study on DNA barcoding of spider mite (Tetranychus urticae) associated with okra was conducted during 2025-2026 using morphological and mitochondrial COI gene analyses. Morphological observations showed only minor variations among populations from seven agro-climatic regions of Gujarat, with stable taxonomic characters across all populations. Molecular characterization using COI I and CGI II genes confirmed the identity of Tetranychus species and revealed low genetic divergence, and high genetic homogeneity among populations. Phylogenetic and SNP analyses indicated slight regional mitochondrial variation in the Narmada and Navsari populations while maintaining overall genetic similarity. Fourteen COI sequences were submitted to the Barcode of Life Data System (BOLD), establishing reliable DNA barcodes and demonstrating the effectiveness of COl-based DNA barcoding for accurate species identification, genetic characterization, and future pest management studies.
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