The genus Nephroselmis (Nephroselmidophyceae), which had been placed in the Prasinophyceae, is one of the primitive green flagellates that are important to our understanding of the early evolution of green plants. We studied a new species of Nephroselmis isolated from Japan, Fiji and South Africa. This species has been known for a long time as undescribed species 'N. viridis.' N. viridis possesses some ultrastructural characters shared with only the freshwater type species N. olivacea, including a disc-like structure beneath the pyrenoid and bipolar spiny body scales with 1-5-8-5-1 spines. Molecular phylogenetic analysis based on 18S rDNA also supports a sister relationship between N. viridis and N. olivacea. However, N. viridis is distinguishable from N. olivacea by the shape of its starch sheath, its scales, its pigment composition and its habitat. In this paper, we designate the formal description of N. viridis sp. nov. We also describe variability in the 18S rDNA introns of various N. viridis strains. This detailed study of N. viridis provides some insights into the evolution of Nephroselmis.