利用红花培养细胞生产维生素E的研究
甘烦远
学位类型硕士
导师郑光植
1989
学位授予单位中国科学院昆明植物研究所
学位授予地点中国科学院昆明植物研究所
学位专业植物学
摘要通过TLc和HpLC的初步分析和鉴定,发现从红花无菌苗的不同部位(根、茎和子叶)及其花蕾中诱导出的愈伤组织均具有产生维生素E族中具有最强活性的α-生育酚的能力,其中以茎愈伤组织的生长及维生素E含量为最高。不同培养基、有机附加物和C源等对红花愈伤组织的生长及维生系E的含量都有很大的影响。为了防止维生素E被氧化掉,在培养基中加入不同的抗氧化剂。发现100ppm的角鲨烯显著提高了维生素E的含量而且不影响细胞的生长。培养于高浓度(5%)的CO_2培养箱中也有效地提高了维生素E的含量。培养基中加入较高浓度的前体(50 ~ 125ppm植醇)可以显著增加维生素E的含量,而且不抑制甚至增加细胞的生长。加入诱导因子(植物细胞壁寡糖)也同时促进了生长及产生的含量,其中以寡糖F效果最好。利用细胞平板培养技术结合紫外线诱发,对红花的细胞进行了初步的筛选。在分析的218个克隆系中,发现细胞在维生素E合成能力上存在着极大的变异。第一代的初步分析获得了含量高达13.60和13.89mg/100gDW的两个细胞系,但所有的高产系(高于5mg/100gDW)在已进行的4代稳定性观察中,其平均产物含量均比第一代的低,并表现出不稳定的状态。利用紫外线处理未获得满意结果。通过改变培养条件或者进行细胞克隆筛选都使细胞的生长和含量提高了。
其他摘要Preliminary identification by TLC and HPLC methods demonstrated that the calli induced from different parts of Carthamus tinctorius shoot (i.e., stem, root and leaf) and flower bud had the capability to synthesize - toccpherol which has the strongest vitamin E activity among tocopherol analogues. Among them, the stem-callus was better than others both in growth and in vitamin E content. Callus growth rate and vitamin E content of C. tinctorius was influenced by different culture conditions such as media, organic additives and carbon sources. For preventing VE from oxidation, different anti-oxidizers were administered to liquid medium, Results found that the VE content was effectively increased by 100 ppm squalelene. VE content was also increased by incubating in high CO_2 conc. (5%) condition. For selection more advanced cell lines both in growth and product content. Plating culture technique combination with irradiation of UV-light was use. Through preliminary selection, the cell lines which produce high content VE among 218 cell lines had been selected. Among them, the cell lines CT-033 and CT-094 were found to synthesize high content of VE and reached 13.60 and 13.89mg/100gDW. respectively. (in the first generation). But the VE content was variable in period successive subcultures. (testing has been proceeded for 4 generations). Experiment demonstrated that the better lines in term of both growth rate and product content could be obtained by selection using various culture conditions and cell clone technique.
页数31
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/850
专题昆明植物所硕博研究生毕业学位论文
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甘烦远. 利用红花培养细胞生产维生素E的研究[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所,1989.
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