走马胎的离体培养与快速繁殖

Culture in vitro and Rapid Propagation of Ardisia gigantifolia

【目的】走马胎是我国传统药用植物,长期依赖自然采挖,以致野生资源枯竭,急需发展驯化种植。采用走马胎的茎作为外植体,建立组织培养快速繁殖方法。【方法】采用70%酒精和0.1%的升汞溶液为消毒液,设置不同处理时间对外植体进行消毒,得到无菌外植体后接种在含有6-BA、KT和IBA的改良MS培养基进行丛芽增殖;采用1/2改良MS培养基作为基础培养基,添加NAA和IBA不定芽进行生根诱导,再生植株经过炼苗后,移栽。【结果】走马胎外植体消毒采用70%酒精处理30 s结合0.1%的升汞溶液消毒处理7~9 min最合适,诱导不定芽的增殖培养基配方为改良MS培养基+6-BA 1.0 mg/L+KT 1 mg/L+IBA 0.05 mg/L,生根培养基配方为1/2改良MS培养基+NAA 0.2 mg/L+IBA 0.3 mg/L。【结论】通过对快速繁殖体系中外植体的消毒方法,优化增殖和生根培养基,建立走马胎的离体培养与快速繁殖方法,为人工种植提供种苗基础。

【Objective】Ardisia gigantifolia is an important traditional Chinese herbal medicine,which has relied on harvesting from nature for a long time,resulting that resources are almost exhausted.Therefore,it is urgent to domesticate and plant such medicine.In this study,stems of A.gigantifolia were took as the explants to establish a rapid propagation method by tissue culture in vitro.【Method】The explants were sterilized with 70% alcohol and 0.1% mercury chloride solution for different treatment time.The obtained aseptic explant were inoculated in the modified MS medium containing 6-BA,KT and IBA for cluster buds proliferation.The 1/2 modified MS medium(as basal medium)with NAA and IBA were used to induce rooting for the adventitious buds.After seedling refining,the regenerated plants were transplanted.【Result】The result showed that disinfecting the stems via 70% alcohol treatment for 30 seconds with 0.1%mercuric chloride solution treatment for 7-9 mins was the best method.The modified MS medium with 1.0 mg/L 6-BA KT and 0.05 mg/L IBA was the optimal combination for inducing and reproduction.The 1/2 MS medium with 0.2 mg/L NAA and 0.3 mg/L IBA was suitable for rooting.【Conclusion】By optimizing the sterilization method,proliferation and rooting medium of explants in rapid propagation system,the method for in vitro culture and rapid propagation of A.gigantifolia was established,which provided seedling foundation for artificial planting.