DNA Barcoding applied to the identification of Meliponini in the Lençóis Maranhenses microregion, Brazil

Stingless bees (Meliponini) are essential pollinators widely distributed in tropical regions, playing a crucial role in ecosystem maintenance. In Brazil, they are found throughout the national territory. DNA barcoding has emerged as an effective tool for species identification and genetic diversity assessment, especially in areas affected by anthropogenic activities. This study aimed to identify and characterize local stingless bee species at the molecular level. A total of 32 specimens were collected from three communities in Tutóia, Maranhão (MA). Samples were processed at the LabWick/UEMA, with DNA extracted using the phenol-chloroform method and amplification of the COI region via PCR. Sequence analyses were performed using dedicated software. The mitochondrial COI marker enabled the identification of eight stingless bee species distributed across seven distinct genera. The DNA fragments analyzed averaged 450 bp in length, with a predominance of adenine (A) and thymine (T). Sequence similarity with the NCBI database ranged from 96.33% to 98.93%, with Melipona fuliginosa showing the highest match. A total of 25 haplotypes were identified, 10 belonging to the Melipona genus and 15 to other genera. Phylogenetic analysis revealed four groupings within Melipona and six among the remaining genera. The geographic isolation of the sampled communities may have limited species distribution across the study area. Despite the effectiveness of COI, the absence of a clear barcode gap and the scarcity of reference data in databases such as GenBank limit identification accuracy for certain genera. This study underscores the importance of DNA barcoding in contributing genetic data to repositories like NCBI and enhancing the known species diversity in the Lençóis Maranhenses microregion.