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Guatemala’s wild avocado germplasm holds vital genetic value, but lacking conservation strategies imperils it. Studying its diversity is pivotal for conservation and breeding. The study aimed to comprehensively assess the wild avocado germplasm in Guatemala by combining phenotypic and genotypic data and to create a core collection for conservation and future breeding programs. A total of 189 mature avocado trees were sampled across Guatemala’s northern, southern, and western regions. Morphological characteristics were documented, and genetic diversity was assessed using 12 SSR loci. The investigated germplasm revealed three distinct genetic clusters, exhibiting an average gene diversity of 0.796 and a 7.74% molecular variation among them. The samples showed various morphological characteristics that indicate the presence of three avocado races in Guatemala. The weak correlation between phenotypic and genotypic distances highlighted their independence and complementary nature. The joint matrix effectively integrated and captured genotypic and phenotypic data for comprehensive genetic diversity analysis. A core collection comprising 20% of total accessions that captured maximum genetic diversity was formed. This study exposed wild Guatemalan avocados’ genetic diversity, morphological traits, and conservation significance. Integrated data capture via clustering validates holistic genetic insight for conservation and breeding strategies.