A new Oxalis (Oxalidaceae) species from the Hantamsberg (South Africa) is described and typified as Oxalis noctiflora using morphological, palynological, and molecular characters. Superficially it closely resembles Oxalis aridicola, Oxalis callosa, Oxalis odorata, and Oxalis primuloides, and it shares a unique supra-areolate pollen type with most of these species. No single vegetative morphological character defines this species, but a unique combination of bulb and leaflet characters distinguishes it from all other known species. The flowers of Oxalis noctiflora are unique for the genus in displaying typical moth-pollination characters. The white flowers open at night, have a long, unusually narrow, corolla tube, and produce a strong sweet scent. All previously described Oxalis species have flowers that open during the day. Phylogenetic analyses based on nuclear Internal Transcribed Spacer data placed this species sister to O. aridicola within a clade of morphologically and palynologically similar taxa.