A new species of Amblysomus, related to A. hottentotus (A. Smith, 1829) and A. septentrionalis Roberts, 1913, and a new subspecies of A. hottentotus with affinities to A.h. pondoliae Thomas & Schwann, 1905 and A.h. iris Thomas & Schwann, 1905, are described from the eastern Mpumalanga highveld, South Africa. The new species, found only in high altitude grasslands near Dullstroom and Belfast, is unique in having a diploid chromosome number (2n) of 36, and a fundamental number of 68. It has a more robust build than most other Amblysomus species, and in colour and size closely resembles A. septentrionalis (2n = 34) and A.h. longiceps (Broom, 1907) from montane grasslands further south in Mpumalanga and KwaZulu-Natal (2n = 30). While no single measurement unequivocally distinguishes between these taxa, they can be diagnosed with at least 90% efficiency using a multiple discriminant functions technique based on 16 cranial measurements. The new subspecies, recorded thus far from only the Graskop and White River districts of the northern Drakensberg escarpment, has a mid-dorsal band of reddish-black fur which distinguishes it from all other forms on the highveld, as well as from A.h. pondoliae and A.h. iris, coastal subspecies which it resembles both karyotypically and craniometrically.