A comparative leaf anatomy study was undertaken on Johannesteijsmannia H.E. Moore in Peninsular Malaysia in order to assess leaf anatomical variation which may be useful in species identification. All four species representatives of Malaysian Johannesteijsmannia were studied. Idioblast cells, sclereids, sclerenchymatous inner bundle sheaths, parenchymatous outer bundle sheaths, division of phloem in the vascular bundles, sub-epidermal fibrous strands below abaxial and adaxial epidermis are commonly present in all the species studied. The outline of the leaf margins, types of stomata, shaped of epidermis cells on the adaxial side, the position of vascular bundles in the leaf lamina and the presence of simple unicellular trichome, fibres at the tip of leaf margins, sub-epidermal fibrous strands in the mesophylls cells, hypodermis layer below adaxial epidermis and stegmata are useful in distinguishing individual species. In conclusion, anatomical characters have systematic significance in Johannesteijsmannia, especially at the species level.