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Background/Objectives: Lebanon, one of the oldest centers of grapevine (Vitis vinifera L.) cultivation, is home to a rich diversity of local grape varieties. This biodiversity is linked to the country's unique topography and millennia of cultural history. However, the wine industry primarily utilizes international varieties, putting many local varieties at risk of extinction. Methods: In this study, we analyzed 202 samples from old vineyards, home gardens, and private collections using 21 microsatellite markers to assess their identity and genetic diversity. Results: A total of 67 different genotypes were identified, with 34 not matching any existing profiles in the consulted databases, based on comparisons with the European Vitis Database, the Vitis International Variety Catalogue (VIVC), and the databases established in two previous studies conducted in Armenia and Lebanon. Cluster analyses revealed Lebanon's rich diversity of local grape varieties, highlighting cases of synonymy, homonymy, and misnaming. All loci were polymorphic, with 228 alleles and an average of 11.4 alleles being detected. The highest number of alleles was observed at the VVIV67 locus (19 alleles), while the lowest was found at the VVIQ52 and VVIN73 loci (5 alleles). The observed heterozygosity was 0.732, slightly below the expected value of 0.757, with gene diversity varying among the markers. Conclusions: Of the 67 genetic profiles identified, 34 are absent from national and international databases, underscoring Lebanon as a hotspot for grapevine genetic diversity. This unique genetic variation, which includes several synonyms due to geographic isolation, could provide valuable opportunities for producing distinctive wines and emphasizes the need for further research and documentation. [ABSTRACT FROM AUTHOR]