【作者】 高桂珍；

【导师】 周吉源；

【作者基本信息】 华中师范大学 ， 植物学， 2003， 硕士

【摘要】 喜树碱是一种色氨酸-萜类生物碱，具有抗癌、清热、杀虫功能，是继紫杉醇之后的第二个从植物提取的衍生抗癌药物，具有积极的开发前景。但目前的研究情况和国内外的研究趋势来看，喜树碱及类似物主要是从喜树中分离获得，其产量必受到喜树生长发育和环境因素的影响，因此通过高等植物细胞培养生产次生代谢物质喜树碱将是一个很好的选择。本文以喜树(Camptotheca acuminata Decsne.CPA)愈伤组织为材料，比较系统的研究了悬浮培养条件下光照、激素以及多种理化因子喜树细胞悬浮培养生长、生理生化特性的影响，使喜树细胞培养生长速度有明显提高，为进一步研究积累了资料。 1．植物生长调节物质的影响 不同种类、不同浓度的激素及其组合对细胞生长效应不相同。NAA浓度为0.5mg／1时最有利于细胞生长：在NAA与KT组合中，以NAA0.5mg／1+KT0.1mg／1的效果最佳，细胞生长速度最快。 2．生理生化特性的研究 在光照条件下，细胞生长周期为30天；而在黑暗条件下，生长周期为27天，细胞增殖曲线均呈“S”型，可大致分为延迟期、指数生长期和静止期：在两种光照条件下，其培养液的pH先下降后都呈上升趋势：培养细胞中可溶性蛋白质含量变化曲线相似，在12天和21天出现两个高峰，黑暗条件下的含量低于光照条件下的含量；培养细胞的过氧化物酶活性变化曲线相似，均出现两个高峰，第一个高峰均出现在第3天，光照条件下的第二个高峰出现在15天，而黑暗条件下出现在18天；培养细胞超氧化物歧化酶活性变化曲线相似，均出现两个峰值，光照条件下的第一个高峰出现在18天，而黑暗条件下出现在21天，两种情况下的第二个高峰均出现在第27天。 d＼硕士学位论文 啥啼动妒M从几rsul0JS 3．不同理化因子的影响 在喜树细胞悬浮培养中，BS培养基和仆h：培养基较有利于细胞的生长：适宜的接种量为3－蛇鲜重八00ml培养基：随着蔗糖和葡萄糖浓度的升高，细胞生长速度呈上升趋势，且在浓度为3％～4池八时最有利于细胞生长；添加适宜的琼脂糖能促进细胞生长：NO．；和N比“的比例不同，细胞生长效应也不同。 最后，对作为药用资源的植物细胞悬浮培养进行了总结，对细胞悬浮培养的开发，出现的问题以及对这些问题的探索进行了讨论。同时对今后的研究方向也作了展望。更多还原

【Abstract】 Camptothecin is a kind of anticancer medicine having good prospect of exploitation, CPT and analogue are get from CPA. at the moment, output of CPT was effected by the growth and environment of CPA. so production of CPT by cell culture may be a promising choice .In the paper ,based on callus of Camptotheca acuminata Decsne., the influences of light, hormones and several factors on the growth, physiological, biochemical characters of suspension culture of Camptotheca acuminata Decsne. Were systematically studied, cell growth speed was obviously improved, it would give references for future studies.1.Effects of hormonesDifferent kinds, different concentration and combinations of hormones had different effects on cell growth. 0.5mg/l NAA was optimal for the cell growth ; Among the combination of NAA and KT ,0.5mg/lNAA+0.1mg/lKT was optimal for the cell growth.2.Study of physiological and biochemical charactersWhen CPA was culture in the light and in the dark ,both of their cell growth curves were like "S", but their growth circles were about 30 and 27 days respectively ,It could be divided into lag phase, exponential growth phase and stationary phase .The cell grew more rapidly in the white light than in the dark ;The PH values of culture mediums lowered and then showed up gradually ;Two peaks of soluble protein contents and peroxides and superoxide dismutase activities of culture cell were showed up ,but they were on the different days ;the soluble protein content was higher in the white light.3.Effects of different physical and chemical factorsIn the suspension culture ,B5medium and White medium were optimal for cell growth ;The inoculums quantity of 3-4g @ FW/1001 was better for the cellgrowth ;The growth rate increased with the increasing concentrations of Sucrose and Glucose ;The medium with certain concentration of Agarose could increase the growth rate ; Effect of cultural cell growth was different with different proportion of NO3-and NH4+.Finally, the conditions in cell suspension culture of some medicine plants were analyzed ;research and development -, problems and resolutions were discussed; prospects of future researches were also given.更多还原