Somatic embryogenesis in four tree legumes

Somatic embryogenesis was achieved in four leguminous tree species, that is, Acacia catechu, Acacia arabica, Hardwickia binata, and Dalbergia sissoo using immature zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 0.25-1.0 mg/l Kn (kinetin) and 2.0-3.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) or NAA (1-napthaleneacetic acid) and 3% sucrose. MS medium containing 2.0 mg/l 2,4-D and 1.0-1.5 mg/l Kn was noted to be most effective in inducing friable embryogenic callus (FEC); the number of somatic embryos per culture varied in MS medium supplemented with 1.0-2.0 mg/l 2,4-D or NAA and 0.25-1.5 mg/l kinetin. The maximum number of somatic embryos was obtained in MS medium containing 1.5-2.0 mg/l 2,4-D or NAA and 1.0-1.5 mg/l kinetin; proliferation of embryogenic calli was enhanced in cultures having 1.0-2.0 mg/l 2,4-D, 1.0-1.5 mg/l kinetin, and 400-600 mg/l L-Proline. The somatic embryos in various shapes and sizes after the first subculture on MS medium supplemented with 0.1 mg/l IAA and 0.25 mg/l BA; developed shoots and rooted in 1/2 strength MS medium supplemented with 0.1 mg/l IBA or IAA. The somatic embryo-derived plantlets were transferred to the field after being hardened in the climate-controlled hardening chamber.