We aim to characterize the diversity of amphibians and reptiles associated with plantations of avocado cv. Hass (Persea americana) in the department of Risaralda, Colombia from an ecological, molecular, and evolutionary perspective. Two sampling periods were implemented between 2019 and 2020 using visual encounter surveys; rarefaction curves and species richness estimators were used to evaluate the completeness of the sampling. Identification of the material was based on both morphological characters as well as DNA barcoding methods sequencing the mitochondrial cytochrome C oxidase subunit I (COI) marker. We also used the COI sequences to construct phylogenies and evaluate the phylogenetic diversity (PD) of the community. Seven amphibian and 18 reptile species were recorded, for which 49 COI sequences were obtained, 15 of which represent the first sequence barcoding records for Colombia. We also found 12 instances of incongruence between the morphological and DNA barcoding identification methods, and we provide a discussion on the correct identification. Finally, although we commonly observed a positive relationship between species richness and PD, we also found that PD can provide valuable information when species richness values are uninformative at comparing two communities. Our results support the need for a greater sampling effort in the area to recognize its true diversity, as this will allow a better understanding of the complex dynamics present in modified landscapes. Also, we conclude with the great contribution of DNA barcoding studies at a small geographical scale and support the use of the COI marker as a suitable source of information for species identification and evaluating the diversity of communities from a historical perspective.