참나물(Pimpinella brachycaroa)의 경단절편체로부터 다량의 이차체세포배 발생과 식물체 재분화

Multi-Secondary Somatic Embryogenesis and Plant Regeneration from Shoot-Tip Cultures of Pimpinella brachycarpa

For the induction of somatic embryogenic callus, shoot-tip explants of Pimpinella brachycarpa were cultured on MS medium supplemented with 0.5mg/L 2.4-D and 0.1mg /L BA for four weeks. Embryogenic calli were cluster of small and dense cells termed proembryogenic masses(PEMs), whereas non-embryogenic calli were friable. These PEMs developed into multi-somatic embryo when transferred to MS medium with 0.1mg /L NAA. After three weeks of culture, globular, heart and torpedo-stage embryos appeared and these embryos were excised and transferred to MS liquid medium with OJ mg/L NAA at a density of 20 embryos/mL. After two weeks of culture in the liquid medium, these somatic embryos developed into plantlets with cotyledons, hypocotyls and radicles. Upon transfer to MS medium with 2.0 mg/L GA₃ plantlets were proliperated and these regenerants were acclimated to soil under greenhouse conditions. Secondary somatic embryogenesis took placed in shoot-tip explants of in vitro PEM-derived regenerants on MS medium with 0.1 mg/L NAA at a frequency of 72 %