【作者】 林龙云；

【导师】 潘大仁；

【作者基本信息】 福建农林大学 ， 作物栽培学与耕作学， 2006， 硕士

【摘要】 本研究以太子参的幼叶为外植体，诱导培养获得太子参愈伤组织，从而建立太子参细胞悬浮系并从中提取、分离出太子参皂苷；研究不同水平或组合的激素、氮源、磷源、碳源、肌醇、细胞接种量等因素对太子参细胞生长及其皂苷合成的影响，从而建立太子参细胞生长及其皂苷合成的动态曲线，通过太子参细胞悬浮获得最高太子参皂苷产量。研究结果如下： 1．用MS／NB+BA0.2mg·L^-1+2，4-D1.0mg·L^-1+KT1.0mg·L^-1液体培养基可获得大量繁殖速度快、生长均匀一致的悬浮细胞。 2．由细胞悬浮培养获得的太子参皂苷的HPLC色谱峰值与常规种植及组培苗的相同，但纯度较好。细胞悬浮培养约30d时，培养1g干重细胞的培养液可提取总皂苷量为2.13-2.92mg，略低于大田常规种植所收获的每克干重太子参块根内的总皂苷含量（3.6-4.3mg），与组培苗收获的太子参块根内的总皂苷含量相近。 3．太子参细胞悬浮培养培养条件优化结果表明：以5.0mmol·L^-1KH₂PO₄+0.5mmol·L^-1（NH₄）₂SO₄+2.06mmol·L^-1CaCl₂+200mg·L^-1肌醇+40g·L^-1蔗糖改良的NB为基本培养基，激素配方为2，4-D1.0mg·L^-1+NAA3.0mg·L^-1+KT0.5mg·L^-1，配合每20mL培养液接种0.03g（DW）左右的细胞液，为最适培养条件。 4．以最适培养条件进行培养，建立太子参细胞生长与皂苷合成的动态曲线结果表明：在第24d太子参细胞生长到达曲线顶峰，即可获最大的太子参细胞生物量，0.3578g（DW）·20mL^-1。此后，太子参细胞生长量开始下降，而太子参皂苷含量开始迅速增加，在第45d达到皂苷产量的最大值329.7147mg·20mL^-1。更多还原

【Abstract】 In this study, the saponin was obtained from the suspension-cells cultured in the established cell suspension culture system which was induced from the leaflets of Pseudostellaria heterophyll （Miq.）. On the study of influence of different combinations of hormone,nitrogen source,phosphorus source,carbon cource,inostol and the cell inculated amount on the Pseudostellaria Heterophyll’cell growth and synthesization of the saponin, the dynamic curves were established in order to the highest saponin yield though cell suspension of Pseudostellaria heterophyll （Miq.）.1. Plenty of more uniformly and quickly growing suspension cells were harvested from the MS/NB liquid medium with 0.2mg·L^-1 BA, 1.0mg·L^-1 2,4-D and 1.0mg·L^-1 KT.2. The HPLC chromatograms of saponin extracted from suspension cell were consistent with those from conventional cultivation or tissue culture seedlings. At about 30 days, the total content of saponin was 2.13-2.92mg·g^-1（DW） cells, slightly lower than those from the roots of conventional cultivation seedlings, and from the roots of tissue culture plantlets.3. After culture condition optimized for cell suspension culture, the results showed that the best condition was the modified NB medium with 5.0 mmol·L^-1 KH₂PO₄+0.5 mmol·L^-1 （NH₄）₂SO₄+2.06mmol·L^-1 CaCl₂+200 mg·L^-1 Inositol +40 g·L^-1 sugar and 1.0 mg·L^-12,4-D +3.0 mg·L^-1NAA+ 0.5 mg·L^-1 KT ,and added 20 mL liquid medium with 0.03 g（DW）cell·L-14. Under the optimal culture condition, the dynamic curves between the Pseudostellaria Heterophyll’s cell growth and synthesization of the saponin indicated that the Pseudostellaria Heterophyll’s cell growth reached to the top at the 24th day and the biomass was 0.3578 g（DW）·20mL^-1, and from then the cell’s growth quantum was reducing, but the saponin content increased rapidly which reached to the top of 329.7147 mg·20mL^-1 in the 45th day.更多还原