Phylogenetic Relationships Classification of Bufotes pewzowi (Bedriaga, 1898) Inhabiting near Aquatic Basins of Central and South Uzbekistan

This study analyzed partial nucleotide sequences of the mitochondrial DNA (mtDNA) 16S rRNA gene region in Bufotes pewzowi (Bedriaga, 1898). Over 580 nucleotides were amplified using PCR and subsequently sequenced. The resulting sequence data were compared with entries in the NCBI database and showed a 99.83% match with B. pewzowi. During the study, the nucleotide sequence of the mtDNA 16S rRNA gene for the B. pewzowi_Uzb specimen was submitted to NCBI and assigned the accession number PV124861. Molecular analysis confirmed that this specimen is a distinct species within the family Bufonidae, identified as B. pewzowi. Future research will enhance understanding of this species’ morphological traits and contribute to amphibian taxonomy. Phylogenetic analysis revealed that species within the Bufonidae family form four monophyletic groups, with bootstrap support values ranging from 50% to 100%. The monophyletic group containing the genus Bufotes, including B. pewzowi_Uzb, showed bootstrap support values between 75% and 100%, confirming its placement within the phylogenetic tree.