The Passifloraceae family is one of the most representative in tropical America, with food, pharmaceutical, and ornamental importance. This study evaluated seed morphometry and germination of eight accessions of four Passiflora edible species, P. edulis; P. ligularis; P. quadrangularis; and P. tripartita var. mollissima, by studying accessions conserved several years in the gene bank (−15 °C) and recently collected accessions. Four experimental phases were carried out as follows: (1) morphometric characterization of seeds with qualitative and quantitative variables; (2) evaluation of germination under two thermal regimes (20 °C/30 °C and 25 °C); (3) application of six pre-germination treatments to overcome dormancy; and (4) tetrazolium tests. In phase 1, P. quadrangularis stood out for its unique morphological characteristics according to multivariate analysis. In phase 2, the alternating thermal regime (20 °C/30 °C) promoted the highest germination. In phase 3, the germination response was specific to each species: mechanical scarification in P. edulis (85.7%), KNO3 (0.5%) in P. ligularis (35.7%), control in P. quadrangularis (71.1%), and gibberellic acid (GA3 400 ppm) in P. tripartita (71.4%). The tetrazolium phase 4 identified the viability status of the seeds. It is concluded that the differences in morphometry and germination reflect the intrinsic characteristics of each species, highlighting the importance of specific protocols for their germination. This study provides tools to optimize the conservation and regeneration of Passiflora spp. germplasm under ex situ conditions, as a genetic base to be utilized in the future.