Tuber itzcuinzapotl is described as a new species to science. This species is characterized by its pale brown ascomata, finely granular peridium, pale brown to gray gleba, and 22–52 × 15–40 µm alveolate ascospores. The new species is putatively associated with Pinus patula in conifer mixed forests in the state of Veracruz, located in eastern Mexico. Phylogenetic analysis based on the nrITS region places the new species in the Maculatum clade, closely related, but morphologically distinctive, to T. miquihuanense and T. mexiusanum. The new species consumed by the Nahua people, traditionally named “Itzcuinzapotl”, constitutes the first edible truffle with biocultural importance in Mexico. Macro- and micromorphological characterization, results of phylogenetic analysis, and photographs are presented. Ethnomycological aspects related to the species are also briefly discussed.