Phylogenetic analyses of SSU-ITS-LSU nrDNA sequences and morphological studies of spores and mycorrhizae confirmed our supposition of finding two new species of arbuscular mycorrhizal fungi of the genus Septoglomus in the phylum Glomeromycota. Morphologically, the first species, named S. jasnowskae, is distinguished by its pale yellow to brownish yellow, small spores with a 2-layered spore wall, of which the colourless outer layer 1 stains dark in Melzer's reagent and layer 2 is laminate. The spores usually arise in loose clusters. The structures most distinguishing S. turnauae are its two coloured laminate layers in the 4-layered spore wall. In the field S. jasnowskae was associated with roots of Ammophila arenaria and an unrecognized plant species colonizing maritime dunes of the Mediterranean Sea near Thessalonica (Greece) and Calella (Spain), respectively, and S. turnauae formed mycorrhiza with a Cistus sp. (Cistaceae) growing in the soil of a mine located in Sulcis-Iglesiente, SW-Sardinia, Italy. In single-species cultures with Plantago lanceolata as host plant, the mycorrhiza of S. jasnowskae consisted of arbuscules, hyphae and vesicles, and that of S. turnauae comprised arbuscules and hyphae only.