Yellow pitahaya is an exotic fruit that is rich in essential nutrients and antioxidants. In Colombia, it grows naturally in warm areas, but it is not clear which species exist because this genus presents a lot of intra and interspecific hybridization. More studies are needed in this field along with characterizations of the genotypes. This study aimed to undertake an in situ morphoagronomic evaluation of yellow pitahaya genotypes from five municipalities in Boyacá, Colombia. Measurements were taken in a completely random design. Qualitative and quantitative descriptors for cladodes, fruits and covered production systems were evaluated with a descriptive analysis, Spearman correlation variance, and multivariate and cluster analysis. The results showed that cladode characteristics such as cladode width, distance between areoles, number of spines, length of areoles, margin ribs of cladode and waxiness could be used to identify Megalanthus spp. Characteristics such as weight (270–274 g), size (100 mm), color of the fruit and pulp as well as acidity (0.18) and soluble solids (15.7) were highly variable between the genotypes. Genotypes with good morphological and fruit characteristics were identified (Gen2 and Gen9), which can provide the bases for the selection of pitahaya that satisfy the needs of farmers and consumers.