In the present study, the PSY1 and PSY2 homologous genes were identified in five accessions of three pepper species Capsicum annuum, Capsicum chinense, and Capsicum frutescens, differing in fruit pigmentation pattern. Within the group of studied accessions, the variability of PSY1 and PSY2 genomic and cDNA sequences was determined. The PSY1 and PSY2 sequences were 82% similar and differed in the N- and C-terminal NAGLRYSD and KLTSSSL indels, as well as in the conserved motifs characteristic of the PSY2 and PSY1 homologs. The expression of PSY1 and PSY2 was analyzed in leaves, sepals, petals, and ovaries, as well as in the peel and pulp of the fruits at the three stages of ripening in all five analyzed accessions. The maximum level of PSY1 expression was shown in the petals and in the pericarp of mature fruits of C. annuum and C. frutescens accessions. In the C. chinense cv. Pimenta da Neyde, PSY1 was expressed only in leaves. PSY2 transcripts were found in all analyzed organs of all pepper accessions; the maximum level was in the leaves, and the minimum level was in the fruit pericarp. The obtained data suggest that the PSY1 and PSY2 homologs of pepper species retained conserved key functions in the carotenoid synthesis in fruits (PSY1) and photosynthetic tissues (PSY2).