A callus induction and plant regeneration protocol was developed from leaf and petiole explants of the endemic Astragalus nezaketae
. Explants were cultured on Murashige and Skoog medium (MS) supplemented with different plant growth regulators (PGRs) [a-naphthaleneacetic acid (NAA), benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kin), thidiazuron (TDZ)]. The combinations and concentrations of PGRs were shown significant variations for the frequency of callus formation, appearence of callus and the potential of callus differentiation. NAA x BA have been found highly affective in callusing and plant regeneration. Other PGRs have not resulted in callus differentiation for shoot formation. The highest number of shoots (6/explants) was obtained from leaf explants cultured on MS with 0.5 mg/l NAA and 4 mg/l BA. The regenerated shoots transferred to rooting medium (MS with 0.5 mg/l indole-3-butyric acid) were successfully rooted (100%) and showed rapid elongation. Rooted plantlets were acclimatized in pots containing 1:1 mixture of peat and perlite.