摘要
党参下胚轴愈伤组织原生质体在附加１．２ｍｇ／Ｌ２，４－Ｄ，０．２ｍｇ／Ｌ ＮＡＡ，０．２ｍｇ／Ｌ ＢＡＰ和０．１ｍｇ／Ｌ ＺＴ的ＭＳ，Ｃ８１Ｖ，ＤＰＤ及ＫＭ８ｐ培养基中进行液体体层培养。在ＫＭ８ｐ中获得了最高的分裂频率。葡萄糖作渗透剂优于甘露醇，两者结合使用效果更好。在合适的条件下，原生质体培养３天出现第１次分裂，４周内形成大细胞团，培养６周后形成０．５－１．０ｍｍ大小的小愈伤组织。
Embryogenic cell line was established from hypocotyl segments of Codonopsis pilosula (Franch. )Nannf. 4—8 day old embryogenic callus was used to isolate protoplasts in an enzyme solution containing 1.5% cellulase Onozuka R-10 and 3% pectinase. Protoplasts were cultured in MS,C81V,DPD and KM8p basal medium supplemented with 1.2 mg/L 2,4-D, 0. 2 mg/L NAA,0.2 mg/L BAP,0.1 mg/L ZT,and different combinations of glucose and mannitol . Protoplast-derived cells underwent sustained divisions in KM8p medium. As an osmoticum,glucose was more beneficial to protoplast division. A combination of 0. 30 mol/L glucose with 0. 10 mol/L mannitol gave the best result,Under proper conditions,protoplasts underwent the first division on the 3rd day of culture,formed colonies within 30 days ,and developed into microcalli in 6 weeks . Plantlets were regenerated from protoplast-derived calli through somatic embryogenesis. 0. 2% activated charcoal promoted embryoid formation and root development.
基金
国家自然科学基金资助课题
关键词
党参
原生质体培养
植株再生
Codonopsis pilosula
Protoplast culture
Somatic embryogenesis
Plant regeneration
