Kim T.D. et al., In Vitro Cell. Dev. Biol.—Plant 41:253–257 (2005)
Summary
Regeneration of plants via somatic embryogenesis was achieved from zygotic embryo explants isolated from mature seeds of Schisandra chinensis. Merkle and Sommer’s medium, fortified with 2,4-dichlorophenoxyacetic acid (2,4-D; 9.04mM) and zeatin (0.09 mM), was effective for induction of embryogenic callus. The development of a proembryogenic mass and somatic embryos occurred on Murashige and Skoog medium (MS) free of plant growth regulators. The embryogenic callus induced on Merkle and Sommer’s medium supplemented with 2,4-D (9.04 mM) and zeatin (0.09 mM) showed development of the maximum number of somatic embryos when transferred to MS medium free of plant growth regulators. The maximum maturation and germination of cotyledonary somatic embryos (46.3%) occurred on MS medium supplemented with 2,4-D (0.45mM) and N6-benzyladenine (1.11 mM). The somatic embryo-derived plants were successfully hardened, with a survival rate of approximately 67%, and established in the field.
Objectives
Development of a repeatable protocol for mass propagation of plants through somatic embryogenesis from zygotic embryos of S. chinensis.
Materials
Mature seeds of S. chinensis (bulked seed lot from several different source trees), obtained from Heung Nong seed company, Jangi-ri, Anseong-myun, Muju-gun, Chollabuk-do, Korea,
Explant
The seeds were rinsed five times more with sterile distilled water and soaked in distilled water for 48 h at 48C. The seeds were then surface-sterilized with 5% sodium hypochlorite solution for 5 min, followed by 1 min treatment in 70% ethyl alcohol, and a further five washes in sterile distilled water. The seeds were then further surface-disinfected with 0.1% mercuric chloride solution for 10 min and rinsed five times with sterile distilled water.
Initial culture
Merkle and Sommer’s medium, fortified with 2,4-dichlorophenoxyacetic acid (2,4-D; 9.04 µM) and zeatin (0.09 µM), was effective for induction of embryogenic callus.
Shoot multiplication
The maximum maturation and germination of cotyledonary somatic embryos (46.3%) occurred on MS medium supplemented with 2,4-D (0.45 µM) and N6-benzyladenine (1.11 µM).