Abstract
Hypocotyl and internodal explants were collected from in vitro grown seedlings and in vitro proliferated shoots, respectively. Callus induction and subsequent plant regeneration were established from those explants. Friable callus with somatic embryo-like structure (ELS) developed from hypocotyl and internodal explants on Murashige and Skoog (MS) medium supplemented with 0.89-4.44μM 6-benzylaminopurine (BAP) and 2.26-9.05μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 2.69-10.74μM α-naphthaleneacetic acid (NAA). The maximal frequency of callus and ELS formation were obtained when the MS medium was amended with 0.89μM BAP and 4.52μM 2,4-D. After callus induction, they were cultured on the MS medium supplemented with 0.98-4.44μM BAP and 0.54-2.69μM NAA or 0.49-2.46μM IBA for the regeneration of the shoots. Among different hormonal conditions, a combination of 2.22μM BAP and 1.07μM NAA produced the highest percentage of shoot differentiation from calli. In vitro grown shoots were rooted on the MS medium with either of 0.5-4.0μM indole-3-butyric acid (IBA), NAA, or indole-3-acetic acid (IAA). Regenerants were transferred to Kanuma soil and successfully established under ex vitro environment.
