Preliminary Study on Two-phase Culture of Tripterygium wilfordii Adventitious Root
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摘要: 以雷公藤(Tripterygium wilfordii)不定根为材料,通过两相培养技术,研究有机溶剂邻苯二甲酸二丁酯(DBP)不同浓度及培养时间对雷公藤不定根生长及次生代谢产物含量的影响。结果显示,DBP浓度为6%、培养至第6d时,不定根增长量达1.14g/瓶,为对照(1.08g/瓶)的1.06倍;DBP浓度为2%、培养至第8d时,内酯醇含量达74.96μg/g,为对照(53.67μg/g)的1.40倍;DBP浓度为2%、培养至第2d时,所收获的内酯醇总产量最高(0.40mg/瓶),且为对照(0.27mg/瓶)的1.48倍。本研究结果表明,在培养基中添加一定浓度DBP,虽然适合雷公藤内酯醇的形成,但不适合雷公藤生物碱的形成;不论添加DBP浓度大小、培养时间长短,不定根中吉碱和次碱含量及每瓶总产量均低于对照。Abstract: Through two-phase culture, we studied the effects of culture time extension and volume percentages of dibutylphthalate (DBP) on growth and secondary metabolite content in the adventitious roots of Tripterygium wilfordii Hook. f.. Results showed that adventitious root growth increased 1.06-fold when the volume percentage of DBP was 6% and addition of organic solvent was on day 6. Triptolide content in the root was 1.40-fold higher when the volume percentage of DBP was 2% and addition of organic solvent was on day 8. Triptolide yield in the two-phase culture was 1.48-fold higher than that of the control when the volume percentage of DBP was 2% and addition of organic solvent was on day 2. Thus, DBP in the medium was suitable for triptolide accumulation, but alkaloid accumulation was inhibited significantly. The contents of wilforgine and wilforine in the adventitious roots and yields of wilforgine and wilforine per flask were all lower than that of the control, regardless of the concentration of DBP added to the medium or the extension of culture time.
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[1] 黄文华,张蕊,郭宝林, 斯金平. 雷公藤药材总生物碱含量及影响因素的研究[J]. 中国中药杂志,2008, 33(1): 15-18. Huang WH, Zhang R, Guo BL, Si JP. Study on total alkaloids content and its influential factors in medicinal materials of Tripterygium[J]. China Journal of Chinese Materia Medica, 2008, 33(1): 15-18.
[2] Brinker AM, Raskin I. Determination of triptolide in root extracts of Tripterygium wilfordii by solid-phase extraction and reverse-phase high-performance liquid chromatography[J]. J of Chromatogr A, 2005, 1070 (1-2): 65-70.
[3] Ni L, Ma J, Li CJ, Li L, Guo JM, Yuan SP, Hou Q, Guo Y, Zhang DM. Novel rearranged and highly oxygenated abietane diterpenoids from the leaves of Tripterygium wilfordii[J]. Tetrahedron Letters, 2015, 56(10): 1239-1243.
[4] Gao C, Huang XX, Bai M, Wu J, Li JY, Liu QB, Li LZ, Song SJ. Anti-inflammatory sesquiterpene pyridine alkaloids from Tripterygium wilfordii[J]. Fitoterapia, 2015, 105: 49-54.
[5] Lu Y, Zhang Y, Li L, Feng XJ, Ding S, Zheng W, Li JX, Shen PP. TAB1: A target of triptolide in macrophages[J]. Chem Biol, 2014, 21: 246-256.
[6] 姜泉,曹炜,唐晓颇,焦娟. 雷公藤提取物治疗类风湿关节炎的系统评价[J].中国中药杂志,2009,34(20):2637-2643. Jiang Q, Cao W, Tang XP, Jiao J. Tripterygium wilfordii extract for treating rheumatoid arthritis:Systematic review[J]. China Journal of Chinese Materia Medica, 2009, 34(20): 2637-2643.
[7] Li Y, Wang JH, Xiao YC, Wang YH, Chen SS, Yang YF, Lu AP, Zhang SW. A systems pharmacology approach to investigate the mechanisms of action of Semen Strychni and Tripterygium wilfordii Hook. f. for treatment of rheumatoid arthritis[J]. J Ethnopharmacol, 2015, 175(4): 301-314.
[8] Lin N, Liu C, Xiao C, Jia H, Imada K, Wu H, Ito A. Triptolide, a diterpenoid triepoxide, suppresses inflammation and cartilage destruction in collagen induced arthritis mice [J]. Biochem Pharmacol, 2007, 73 (1): 136-146.
[9] 李琰,崔蕾,杨钰琪,赵磊,雷嘉敏,张兴.雷公藤不定根培养体系的建立及中试放大研究[J].中国中药杂志,2015,40(1): 53-58. Li Y, Cui L, Yang YQ, Zhao L, Lei JM, Zhang X. Establishment of adventitious root culture system and scale-up fermentation of Tripterygium wilfordii[J]. China Journal of Chinese Materia Medica,2015,40(1): 53-58.
[10] 李琰,崔蕾,雷嘉敏,李群,张兴.不同浓度有机物对雷公藤不定根生长和次生代谢产物含量的影响[J].植物科学学报,2014,32(2):174-180. Li Y, Cui L, Lei JM, Li Q, Zhang X. Effects of different concentrations of organic affixture on the growth and se-condary metabolites contents in adventitious roots of Tripterygium wilfordii[J]. Plant Science Journal,2014,32(2):174-180.
[11] 张兴,李琰,冯俊涛,王智辉,王永宏. 一种由雷公藤悬浮细胞诱导不定根的方法[P].中国,2008,ZL200810017697.8. Zhang X, Li Y, Feng JT, Wang ZH, Wang YH. A method of suspension cells induced adventitious root of Triptery-gium wilfordii[P]: China, ZL200810017697.8. [12] 李琰,冯俊涛,史晓燕,吴勇延,张兴.雷公藤组培产物中雷公藤甲素和总生物碱含量的测定[J].农药学学报,2009,11(3):367-372. Li Y, Feng JT, Shi XY, Wu YY, Zhang X. Study on the determination of triptolide and total alkaloids in Tripterygium wilfordii Hook. f. cultures[J]. Chinese Journal of Pesticide Science, 2009, 11(3): 367-372.
[13] Yabannavar VM, Wang DIC. Extractive fermentation for lactic acid production[J]. Biotechnol Bioeng, 1991, 37(11): 1095-1100.
[14] 李祝,任秀秀,周礼红,刘吴娟.两相培养用于提高植物次生代谢物产量的研究进展[J].食品工程,2011,(1):10-13. Li Z, Ren XX,Zhou LH,Liu WJ, Study on two-phase culture techniques for improving the secondary metabolites production of the plants[J]. Food Engineering, 2011(1): 10-13.
[15] 吴兆亮,元英进,胡萍,胡宗定.东北红豆杉细胞两液相培养生产紫杉醇的研究[J].中草药, 1998, 29(9): 589-592. Wu ZL, Yuan YJ, Hu P, Hu ZD. Studies on the two-liquid-phase culture of Japanese yew (Taxus cuspidata) for the production of paclitaxel[J]. Chinese Traditional and Herbal Dregs, 1998, 29(9): 589-592.
[16] Xu QM, Cheng JS, Ge ZQ, Yuan YJ. Eects of organic solvents on membrane of Taxus cuspidata cells in two-liquid-phase cultures[J]. Plant Cell Tiss Organ Cult, 2004, 79(1): 63-69.
[17] Wu ZL, Yuan YJ, Ma ZH, Hu ZD. Kinetics of two-liquid-phase Taxus cuspidata cell culture for production of Taxol[J]. Biochem Eng J, 2000, 5(2): 137-142.
[18] Miao GP, Zhu CS, Feng JT, Han J, Song XW, Zang X. Aggregate cell suspension cultures of Tripterygium wilfordii Hook. f. for triptolide, wilforgine, and wilforine production[J]. Plant Cell Tiss Organ Cult, 2013, 112(1): 109-116.
[19] Buitelaar RM, Langenhoff AAM, Heidstra R, Tramper J. Growth and thiophene production by hairy root cultures of Tagetes patula in various two-liquid-phase bioreactors[J]. Enzyme Microb Technol, 1991, 13(6): 487-494.
[20] Kim DJ, Chang HN. Enhanced shikonin production from Lithospermum erythrorhizon by in situ extraction and cal-cium alginate immobilization[J]. Biotechnol Bioeng, 1990, 36(5): 460-466.
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