Biodiversity loss is occurring on a large scale and the need to monitor it is becoming more and more necessary. The use of morphological techniques can be enhanced with the use of molecular tools to help resolve diversity and evolutionary history knowledge gaps. The objective of this research was to evaluate the use of the mitochondrial cytochrome c oxidase 1 (COI) gene as a DNA barcode marker to characterize possible pollinators of Colombian cacao crops. The study was implemented in the departments of Meta and Northern Santander, Colombia. Taxa sampled directly from flowers were analyzed to assess their diversity and phylogenetic relationships using Maximum-Likelihood (ML) and Bayesian Inference (BI). We generated sequences of approximately 656 bp for 25 Culicomorpha individuals, 13 from Meta and 12 from Northern Santander and downloaded 388 sequences of the Ceratopogonidae and Chironomidae families from GenBank. Analysis of the COI sequences reveals that our sequences were placed in three Ceratopogonidae lineages (Forcipomyia, Dasyhelea and Stilobezzia) and five unresolved lineages of Chironomidae. We also found that species that visited cacao flowers in Meta and Northern Santander plantations represented two separate guilds, which could have been influenced by the orogenic processes of the Andes Mountains. Phylogenetic reconstruction indicated that most (n=17) of our sequences were resolved in the Forcipomyia group. Additionally, none of our sequences were identical to any GenBank sequences, reflecting an investigative bias towards northern temperate regions and the need for more molecular studies on tropical species. Our data offer new DNA sequences of Colombian Ceratopogonidae for the development of a global inventory of pollinating species of plants of economic interest, such as cacao. The need to continue sampling tropical taxa is also suggested in order to clarify the evolutionary history of these families of flies.
