稳定高产黄酮的水母雪莲细胞系的建立  

Establishment of stable and high-yield Saussurea cell line for producing flavonoids

作　　者：胡显文[1] 高丽华[1] 胥照平[1] 陈惠鹏[1] 李佐虎[2] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]中国科学院过程工程研究所,北京100080

出　　处：《高技术通讯》2006年第6期610-614,共5页Chinese High Technology Letters

基　　金：军事医学科学院生物工程研究所开发课题（200001）资助项目.

摘　　要：利用^60Co-γ射线照射、高温诱变和紫外光线照射等手段诱变雪莲愈伤组织，结果表明：剂量为40Gy的60Co-γ射线照射原始雪莲细胞系RP-1得到的IRP-5细胞系，黄酮含量可从细胞干重的4．34％提高至6％。分别采用温度和紫外光线等手段诱变IRP-5细胞系得到的细胞系TIP-33和UIP-17，相应的黄酮含量可提高至10．1％和8．8％。用上述三种诱变方法处理原始雪莲细胞系，获得了一株稳定高产雪莲细胞系TUIP-8，黄酮含量大于12％，比天然植株高20-30倍，连续传代培养100代，细胞的培养密度和黄酮含量一直维持在20g（干重）／L和12％-14％左右。考察了温度对TUIP-8细胞悬浮培养的影响，该细胞系可以在20-30℃环境中生长，最适培养温度范围为20-25℃。The methods of establishing stable and high-yield Saussurea medusa （ S. medusa） cell lines and cell culture techniques were investigated in order to increase production of flavonoids. Three transforming means including γ-ray irradiation, induction of mutant with high-temperature and ultraviolet irradiation were used to establish stable and high-yield S. medusa cell lines. The results showed that the content of flavonoids in the dry cells increased from 4.34 % to 6 % when the initial S. medusa cell lines, RP-1, was irradiated by ^60Co-γ ray with the dose of 40Gy and the acquired cell line was named IRP-5. When IRP-5 cell line was transformed by means of temperature and ultraviolet irradiation, the flavonoid contents in the acquired cell lines, TIP-33 and UIP-17, were raised to 10.1% and 8.8% respectively. Especially, when the initial cell line was treated by three transformed methods herein, a stable and high-yield cell hne, TUIP-8, was obtained, in which the flavonoid content was above 12% and 20 - 30 times higher than that in natural S. medusa. Because of its genetical stability with respect to the cell growth and the flavonoid biosynthesis, the cell culture density and flavonoid content always kept about 20g DW/L and 12% - 14% during the period of TUIP-8 cell subculture for 100 generations. Effects of culture temperature on TUIP-8 cell growth and flavonoid biosynthesis were studied. The results showed that TUIP-8 cells could grow at 20 - 30℃, and the temperature ranged from 20℃ to 25℃ was more suitable for TUIP-8 cell growth and flavonoid formation.

关 键 词：水母雪莲 植物细胞培养 黄酮 筛选 高产细胞系 

分 类 号：R28[医药卫生—中药学]