Disclosing the hidden nucleotide sequences: a journey into DNA barcoding of raptor species in public repositories

Background In nucleotide public repositories, studies discovered data errors which resulted in incorrect species identification of several accipitrid raptors considered for conservation. Mislabeling, particularly in cases of cryptic species complexes and closely related species, which were identified based on morphological characteristics, was discovered. Prioritizing accurate species labeling, morphological taxonomy, and voucher documentation is crucial to rectify spurious data. Objective Our study aimed to identify an effective DNA barcoding tool that accurately reflects the efficiency status of barcodes in raptor species (Accipitridae). Methods Barcode sequences, including 889 sequences from the mitochondrial cytochrome c oxidase I ( COI ) gene and 1052 sequences from cytochrome b ( Cytb ), from 150 raptor species within the Accipitridae family were analyzed. Results The highest percentage of intraspecific nearest neighbors from the nearest neighbor test was 88.05% for COI and 95.00% for Cytb , suggesting that the Cytb gene is a more suitable marker for accurately identifying raptor species and can serve as a standard region for DNA barcoding. In both datasets, a positive barcoding gap representing the difference between inter-and intra-specific sequence divergences was observed. For COI and Cytb , the cut-off score sequence divergences for species identification were 4.00% and 3.00%, respectively. Conclusion Greater accuracy was demonstrated for the Cytb gene, making it the preferred primary DNA barcoding marker for raptors.