The present investigation entitled "Molecular characterization and DNA barcoding of stingless bee worker (Tetragonula spp.) populations from Gujarat, India using mitochondrial COI gene" was carried out at the Department of Plant Molecular Biology and Biotechnology, N. M. College of Agriculture, Navsari Agricultural University (NAU), Navsari, Gujarat during the period from 2025-2026. Genomic DNA extracted from adult worker bees using a modified CTAB method showed high quality, with A 260 /A 280 ratios ranging from 1.78 to 1.92. Mitochondrial COI and COII genes were successfully amplified and sequenced, producing fragments of 700 bp and 750 bp, respectively. BLASTn analysis confirmed the sequences as belonging to genus Tetragonula with most populations showing 99-100% similarity to Tetragonula iridipennis, while the Anand COI sequence showed 99.41% similarity with Tetragonula shubhami. Overall, this study revealed limited genetic variation among populations and suggesting the possible occurrence of T. shubhami in the state, pending confirmation through male morphometry and genitalia-based studies.
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