We report here the karyotype characterization of five Physalis peruviana L. ecotypes. Three wild types were collected from the areas around Villa de Leyva (Boyaca, Colombia), La Calera and Choachi municipalities (Cundinamarca, Colombia) and two were from cultivated cape gooseberry crops. One of the latter was the native Colombia ecotype found near the Subachoque municipality (Cundinamarca) and the other was a foreign ecotype from Kenya, which had been cultivated near Paipa municipality (Cundinamarca). Karyotypes were obtained after standardizing a protocol involving obtation of radical apexes from seeds and leaves, using in vitro auxinenriched culture medium. The chromosomes protocol was standardized using different pretreatment, fixation, hydrolysis and samplemounting techniques. Duration of the cellular cycle was also evaluated for determining the mitotic hour. A wide genetic variability among the ecotypes was observed. Wild ecotypes presented 2n=24 chromosomes while Colombia and Kenya ecotypes 2n=32 and 2n=48, respectively. Morphological and anatomical characterization also showed variation among the ecotypes reflecting ploidy level, according to the literature, in terms of plant height, foliate area, number of stoma/mm2, chloroplast number in stoma guard cells, fruit and seed diameter and dry mass content.