Micropropagation of plantlets through callus in kihada (Phellodendron amurense Rupr.)

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 0.89-4.44 micro M 6-benzylaminopurine (BAP) and 2.26-9.05 micro M 2, 4-dichloro-phenoxyacetic acid (2,4-D) or 2.69-10.74 micro M a-naphthaleneacetic acid (NAA). The maximal frequency of callus and ELS formation were obtained when the MS medium was amended with 0. 89 micro M BAP and 4.52 micro M 2, 4-D. After callus induction, they were cultured on the MS medium supplemented with 0.98-4.44 micro M BAP and 0.54-2.69 micro M NAA or 0.49-2.46 micro M IBA for the regeneration of the shoots. Among different hormonal conditions, a combination of 2.22 micro M BAP and 1.07 micro M NAA produced the highest percentage of shoot differentiation from calli. In vitro grown shoots were rooted on the MS medium with either of 0.5-4.0 micro M 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.