Accurate taxonomic identification of stingless bees is critical for their conservation and sustainable management, yet morphological methods are often limited by species complexity and lack of expertise. This study employed DNA barcoding targeting a 650 bp fragment of the mitochondrial cytochrome oxidase I (COI) gene to identify stingless bee specimens collected from 19 locations across mainland Tanzania. A total of 28 specimens from wild colonies were analyzed, yielding reliable species-level identification (>97 % similarity to reference sequences in the BOLD database) for 53.6 % of the samples. Identified species included Plebeina armata, Hypotrigona gribodoi, Axestotrigona ferruginea, and Dactylurina schmidti. Specimens with barcode similarity between 95 and 97 % were assigned to the genus Axestotrigona, with their taxonomic status remaining unresolved. Despite reliable identification, we observed significant morphological diversity among P. armata and H. gribodoi specimens, with average within-group genetic distances of 3.5 and 4.1 %, respectively. Phylogenetic analysis corroborated these identifications and revealed potential cryptic speciation and genetic structuring consistent with geographical locations. The findings underscore the utility of DNA barcoding to complement traditional taxonomic approaches, and highlight the underexplored diversity of Afrotropical stingless bees. Further molecular and morphological studies are recommended to clarify species boundaries within the genus Axestotrigona, resolve taxonomic discrepancies in Afrotropical stingless bees, and improve regional biodiversity assessments.
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