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Loss of biodiversity and ecosystem degradation become major concerns worldwide, making the conservation process an important strategy for maintaining biodiversity. Capsicum chinense Jacq. is the most Brazilian species of the genus, with representatives in different biomes. Anthropic pressure, such as burns, real estate speculation, and changing cultivation habit has led to risks of genetic erosion. Conservation and characterization of conserved accessions are paramount to ensure genetic diversity, useful for the bioeconomy and for genetic improvement. We report the characterization of 55 C. chinense accessions from four different regions of Brazil and one accession from Peru. The accessions were characterized based on 37 morpho-agronomic variables, Inter Simple Sequence Repeats—ISSR and Simple Sequence Repeat—SSR. Qualitative descriptors were analyzed using a descriptive statistical, while the quantitative descriptors were analyzed via F test and significant differences in mean values were separated using Scott-Knott test. The relative contribution of each quantitative trait was determined. A correlation between morphological and molecular distances was calculated. Color of ripe fruit and fruit shape had the largest number of observed classes. Six distinct groups and the joint analysis presented an entanglement rate of 0.58, evidencing the divergence of accessions between the groups of both dendrograms. Bayesian analysis allowed the distinction of two clusters for SSR. A significant variability was observed among accession with potential to integrate several breeding programs. [ABSTRACT FROM AUTHOR]