短毛独活悬浮细胞的原生质体培养

PLANT REGENERATION EROM CELL SUSPENSION DERIVED PROTOPLASTS OF HERACLEUM MOELLENDORFFII

短毛独活(Heracleum moellendorf fii Hance)是伞形科植物,其根可作药用。我们曾以多种伞形科植物进行原生质体培养获得体细胞胚及再生植株。

Heracleum moellendor f fiz Hance is a herb belonging to Umbelliferae used in traditional medicine in China.The young stem-nodes were induced for callus formation on MS medium containging 1 mg/L 2,4-D.After subcultured for about five months,the embryogenic calli were used for cell suspension culture.The protoplasts were prepared from this suspension by digestion with enzyme mixture containing 1.5% cellulase Onozuka R-10 + 0.3% macerozyme R-10 + 0.5% snailase + 5 mmol CaCl_2+ 0.6 mol/L mannitol,at pH 5.8, and cultured in modified MS and modified N_6 media with 0.3% agarose.They divided after 3 days and developed into small cell colonies after about 2 weeks.From this time on,the glucose concentration in the culture media was decreased to 0.2mol/L,which led to futher growth of the colonies to small cali .After a period of proliferation on solid medium with 0.5mg/L 2,4-D,the calli were transferred to a medium with 0.1 mg/L zeatin on which somatic embryos differentiated and developed to plantlets