Because the embryological features of Jeffersonia dubia are poorly understood, we conducted the first embryological study comparing it to other related genera of Berberidaceae. Important embryological features of J. dubia are as follows: the anther is tetrasporangiate, anther wall formation confirms basic type, glandular tapetum cells are two nucleate, the epidermis persistent, and the endothecium develops fibrous thickenings, anther dehiscence by two valves, meiosis in a microspore mother cell is accompanied by simultaneous cytokinesis, microspore tetrads are usually tetrahedral, pollen grains two cells at the time of anthesis. The ovule is bitegmic, anatropous and crassinucellate, archesporium single celled, development of the embryo sac Polygonum type, a mature embryo sac is ellipsoidal in shape. Endosperm formation is of Nuclear type and embryogeny Onagrad type. Seeds are arillate and seed coat exotestal type. Embryological comparisons showed that Jeffersonia resemble to Epimedium and Vancouveria rather than Berberis and Mahonia in some features, like as number of tapetal cells, cytokinesis in meiosis, and thickness of exotesta. It also resembles to Gymnospermium in mode of anther wall formation, number of tapetal cells, formation of nucellar cap, and nature of antipodal cells. Nevertheless, Jeffersonia and Gymnospermium differ from several other embryological features and molecular data too. Therefore, embryological evidences support that Jeffersonia is closely related with Epimedium and Vancouveria.