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Hildegardia species are an important resource for fiber industry. This investigation was conducted to develop a plant regeneration protocol for Hildegardia populifolia (Roxb.) Schott & Endl. via indirect organogenesis Callus was obtained from leaf, internode and petiole explants, among these explants internode explant gave best result on MS medium supplemented with different concentrations of 2,4-Dichlorophenoxy acetic acid (2,4-D). The highest percentage (100%) of regeneration was obtained with benzyladenine (BA) (2.0 mg/l) + indole-3-acetic acid (IAA) (0.1 mg/l) + glutamine (25 mg/l) + thidiazuron (TDZ) (0.5 mg/l) from internode explants. Shootlets were highly rooted on MS medium supplemented with 3.0 mg/l indole-3-butyric acid (IBA). In vitro rooted seedlings were successfully acclimatized. This in vitro regeneration system will facilitate further development of reliable procedures for this genus.