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Charophytes (stoneworts and bassweeds) are a typical macrophytic component of inland water ecosystems. Well-developed submerged meadows of charophytes are expression of clear water and rather low phytoplankton concentrations. Consequently, among aquatic macroscopic primary producers, charophytes are one of the most threatened groups being very sensitive to phosphorous availability, turbidity and water level perturbations. Accordingly, charophytes have been suffering a massive diversity loss worldwide over the last century, mainly because of human-induced pressures. During summer 2011, detailed field surveys were carried out with the main purpose of filling knowledge gaps concerning aquatic flora and vegetation of the lake Garda – the largest lake in Italy and one of the deepest in Europe. Along randomly selected transects, floristic data were collected following standard procedures, as imposed by the Water Framework Directive. Overall, 12 different species of charophytes were recorded, which accounts for 36% of total Italian charophytes and 19% of European species. The most diffuse species were Chara globularis and C. intermedia; whereas, the most interesting taxa were Chara polyacantha and Nitella hyalina, two species with narrow distribution in Italy. Overall in the lake Garda, dense stands of charophytes covered almost homogeneously the littoral sectors at a water depth between 3 and 12 m. The deepest species was C. globularis, which reached a maximum depth of colonisation of about 17.5 m. Charophytes represent a major element among the primary producers in the lake Garda. The high local charophyte diversity and the rather wide most colonised areas (~1000-1200 ha) confirm that the lake Garda is an important reserve for many rare and threatened charophytes. For the first time, these results highlight the key role of the lake Garda for charophyte diversity at a national and European level.