The objective of this study was to characterize the arbuscular mycorrhizal fungi (AMF) community associated with local varieties of Pinot noir, Sauvignon blanc, and Chardonnay grown in the Malleco and Caut & iacute;n valleys, in La Araucan & iacute;a region, Chile. The research attempted to answer the following question: How do soil and climatic conditions, as well as cultivar characteristics could influence root colonization, spore abundance, and AMF composition in vineyards in this emerging region? Rhizosphere samples were collected from two locations and six grapevine cultivars. Root colonization rates and AMF spore abundance were measured, and the spores present were morphologically identified. In addition, differences in the AMF community were evaluated regarding cultivar and plant age. The results showed root colonization rates higher than 50%, with no significant differences between sites. However, variations in spore abundance and AMF community composition were observed among cultivars. Twelve AMF genera were identified, including Glomus, Sclerocystis, Dominikia, Rhizoglomus, Oehlia, and Paraglomus. Overall, Glomus rubiforme, Sclerocystis sp. CL1, Rhizoglomus irregulare, and Diversispora versiformis were the most abundant morphotypes. Additionally, R. irregulare, G. rubiforme, and Paraglomus occultum were consistently detected across nearly all analyzed rhizospheres. The presence of AMF genera varies according to cultivar, but not according to clones or plant age. It is hypothesized that differences in root architecture and root exudates of different grapevine cultivars are responsible for the observed variations in the composition of native AMF. These factors should be further investigated in future studies.