Diospyros L. (Ebenaceae) is an important source of ebony, a precious wood used for several economically important timber products. Species are overexploited in many regions, including Madagascar, for both the national and international trade, but little is known about their wood anatomy, despite its importance for forensic identification. Wood anatomy has a major role to play in ensuring the sustainable and equitable utilization of Diospyros species that are not threatened by extinction, and in law enforcement to protect threatened species from illegal logging. This study aims to identify, describe, and test the usefulness of anatomical features to support a taxonomic revision of the genus in Madagascar and to enrich databases for wood identification. Ninety-nine wood specimens were collected from the various bio-geographical regions of Madagascar, representing 15 endemic species (twelve previously described and three new) of large trees (reaching DBH ⩾ 20 cm and/or height ⩾ 20 m) were investigated. Standard methods for wood anatomical studies were used. Statistical analysis of the data using Factorial Analysis on Mixed Data was performed for 14 wood anatomical characters. Detailed descriptions and comparisons of the wood anatomy of the 15 species are provided, along with a wood identification key. Analyses showed that all the characters are highly significant () in the separation of the species studies.