柴胡生物培养技术与柴胡皂苷合成生物学的研究进展
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摘要
合成生物学是解析中药活性成分生物合成途径、发掘参与生物合成功能基因的一门新兴学科。柴胡是我国常用大宗中药材,药用价值显著。柴胡皂苷是其指标性成分,具有抗炎、抗病毒、抗肿瘤等显著活性。但目前野生柴胡资源遭到破坏,人工栽培过程中存在一定问题。生物培养技术及合成生物学的应用可扩大柴胡皂苷的来源,保护柴胡资源。不同植物的培养条件没有固定模式可遵循,不同药用活性成分的生物合成途径也是不一致的。目前,尚无柴胡组织培养技术及柴胡皂苷生物合成途径研究的综述报道。该文拟介绍合成生物学过程中用到的愈伤组织培养、不定根培养、毛状根培养与悬浮细胞培养等生物培养技术与柴胡皂苷生物合成途径及其关键酶功能基因的研究进展,建议通过借鉴其他传统根类药材的组织培养技术,对柴胡的生物培养技术进行优化,为深入研究柴胡皂苷的生物合成途径与代谢调控提供参考。
Synthetic biology is an emerging discipline that analyzes the biosynthesis pathways of active constituents in traditional Chinese medicine and explores genes involved in biosynthesis. Bupleuri Radix is one of the most commonly used Chinese medicinal materials with remarkable medicinal value,its index component is saikosaponins,which has significant anti-inflammatory,anti-viral and anti-tumor activities. However,the current wild resources of Bupleuri Radix have been destroyed,and there were some problems in the process of artificial cultivation. The application of biological culture technology and synthetic biology can expand the sources of saikosaponins and protect resources of Bupleuri Radix. The culture conditions of different plants can be followed without a fixed pattern,and the biosynthetic pathways of different medicinal active ingredients are also inconsistent.At present,there is no review report on the culture technology of Bupleuri Radix and the research on the biosynthesis pathway of saikosaponins. This paper introduces the research progress of biological culture techniques,such as callus culture,adventitious root culture,hairy root culture and suspension cell culture used in synthetic biology,and the biosynthesis pathway of saikosaponins and its key enzyme functional genes. It is suggested to optimize the biological culture technology of Bupleuri Radix by referring to the tissue culture technology of other traditional root medicinal materials,so as to provide a reference for the in-depth study on the biosynthesis pathway and metabolic regulation of saikosaponins.
Synthetic biology is an emerging discipline that analyzes the biosynthesis pathways of active constituents in traditional Chinese medicine and explores genes involved in biosynthesis. Bupleuri Radix is one of the most commonly used Chinese medicinal materials with remarkable medicinal value,its index component is saikosaponins,which has significant anti-inflammatory,anti-viral and anti-tumor activities. However,the current wild resources of Bupleuri Radix have been destroyed,and there were some problems in the process of artificial cultivation. The application of biological culture technology and synthetic biology can expand the sources of saikosaponins and protect resources of Bupleuri Radix. The culture conditions of different plants can be followed without a fixed pattern,and the biosynthetic pathways of different medicinal active ingredients are also inconsistent.At present,there is no review report on the culture technology of Bupleuri Radix and the research on the biosynthesis pathway of saikosaponins. This paper introduces the research progress of biological culture techniques,such as callus culture,adventitious root culture,hairy root culture and suspension cell culture used in synthetic biology,and the biosynthesis pathway of saikosaponins and its key enzyme functional genes. It is suggested to optimize the biological culture technology of Bupleuri Radix by referring to the tissue culture technology of other traditional root medicinal materials,so as to provide a reference for the in-depth study on the biosynthesis pathway and metabolic regulation of saikosaponins.
引文
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[4] ZHOU Y J,GAO W,RONG Q,et al. Modular pathway engineering of diterpenoid synthases and the mevalonic acid pathway for miltiradiene production[J]. J Am Chem Soc,2012,134(6):3234-3241.
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[17]余马,左文松,刘茹,等.北柴胡新品种川北柴1号组织培养快繁初步研究[J].种子,2017,36(9):27-34.
[18]王梅,郝建平,赵暘,等.乙酸钠、水杨酸和Cu2+对北柴胡不定根培养合成皂苷的影响[J].天然产物研究与开发,2012,24(S1):87-91.
[19]战晴晴,金钺,魏建和,等.北柴胡不定根培养及茉莉酸甲酯处理对柴胡皂苷含量的影响[J].生物技术通讯,2011,22(1):57-60.
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[21] Qavami N,Azizi M,Yazdian F,et al. Effective induction of hairy roots in persian poppy(Papaver bracteatum Lindl.)using sonication method[J]. J Med Plants,2018,17(65):73-82.
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[24]孙晶,高珂,王玲,等.不同培养基、外源激素和真菌诱导子对北柴胡毛状根生长及柴胡皂苷含量的影响[J].生物技术通讯,2015,26(4):546-550.
[25] YANG D F,FANG Y M,XIA P G,et al. Diverse responses of tanshinone biosynthesis to biotic and abiotic elicitors in hairy root cultures of Salvia miltiorrhiza and Salvia castanea Diels f. tomentosa[J]. Gene,2018,643:61-67.
[26]程玉鹏,林进华,高宁,等.不同因素对狭叶柴胡悬浮细胞生长的影响[J].山西农业大学学报:自然科学版,2016,36(5):305-309.
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[29] Ramos-Valdivia A C,van der Heijden R,Verpoorte R.Isopentenyl diphosphate isomerase:a core enzyme in isoprenoid biosynthesis:a review of its biochemistry and function[J]. Nat Prod Rep,1997,14(6):591-603.
[30] Ebizuka Y,Katsube Y,Tsutsumi T,et al. Functional genomics approach to the study of triterpene biosynthesis[J]. Pure Appl Chem,2003,75(2):369-374.
[31] Nakamura A,Shimada H,Masuda T,et al. Two distinct isopentenyl diphosphate isomerases in cytosol and plastid are differentially induced by environmental stresses in tobacco[J]. FEBS Lett,2001,506(1):61-64.
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[34]隋春,魏建和,战晴晴.北柴胡鲨烯合酶基因及其编码区c DNA克隆与序列分析[J].园艺学报,2010,37(2):283-290.
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[36]徐洁森.北柴胡皂苷合成相关P450、UGT基因的筛选、克隆及遗传转化初步研究[D].北京:北京协和医学院,2013.
[37] Bowles D, Isayenkova J, Lim E K, et al.Glycosyltransferases:managers of small molecules[J].Curr Opin Plant Biol,2005,8(3):254-263.
[38] WANG X Q. Structure,mechanism and engineering of plant natural product glycosyltransferases[J]. FEBS Lett,2009,583(20):3303-3309.
[39] YANG N,ZHOU W P,SU J,et al. Overexpression of Sm MYC2 increases the production of phenolic acids in Salvia miltiorrhiza[J]. Front Plant Sci,2017,doi:10. 3389/fpls. 2017. 01804.
[40]于一凡,郭娟,苏平,等.灯盏细辛转基因毛状根体系建立[J].中国中药杂志,2018,43(10):2048-2052.
[41] Murthy H N,Dandin V S,Paek K Y. Tools for biotechnological production of useful phytochemicals from adventitious root cultures[J]. Phytochem Rev,2016,15(1):129-145.
[42]左北梅,高文远,王娟,等.鼓泡式生物反应器培养人参不定根的研究[J].中国中药杂志,2012,37(24):3706-3711.
[43]毛亚平,马莹,卜俊玲,等.丹参植株原位侵染RNAi毛状根体系的建立及其干扰效果[J].世界中医药,2018,13(2):256-258.
[44]黄志成,任加惠,沈丽红,等.丹参毛状根和丹参药材HPLC指纹图谱的比较研究[J].浙江理工大学学报:自然科学版,2016,35(3):444-449.
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[2] Ro D K,Paradise E M,Ouellet M,et al. Production of the antimalarial drug precursor artemisinic acid in engineered yeast[J]. Nature,2006,440(7086):940-943.
[3] Ajikumar P K,XIAO W H,Tyo K E J,et al. Isoprenoid pathway optimization for taxol precursor overproduction in Escherichia coli[J]. Science,2010,330(6000):70-74.
[4] ZHOU Y J,GAO W,RONG Q,et al. Modular pathway engineering of diterpenoid synthases and the mevalonic acid pathway for miltiradiene production[J]. J Am Chem Soc,2012,134(6):3234-3241.
[5]董乐萌,刘玉军,魏建和.柴胡皂苷合成途径中三个关键酶基因片段的克隆与序列分析[J].世界科学技术—中医药现代化,2008,10(5):56-60.
[6]李晓伟,王玉庆,杜国军,等.药用柴胡资源调查及市场现状分析[C]//中国自然资源学会.海峡两岸暨CSNR全国中药及天然药物资源学术研讨会论文集:2012年卷.北京:中国自然资源学会,2012:159-165.
[7]孟杰,姚入宇,陈兴福,等.柴胡属植物分类研究进展[J].中国中药杂志,2012,37(11):1523-1526.
[8]程玉鹏,李天聪,林进华,等.狭叶柴胡愈伤组织诱导的研究[J].中医药学报,2015,43(6):46-48.
[9] Castro A H F,Braga K D Q,Sousa F M D,et al. Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia(L.)DC.(Malpighiaceae)[J]. Rev Cienc Agron,2016,47(1):143-151.
[10]姚智,高文远,李克峰,等.柴胡愈伤组织生长和不定根诱导研究[J].中草药,2007,38(2):275-278.
[11]丛登立,鞠会艳,张婷婷.北柴胡高频愈伤诱导优化技术研究[J].人参研究,2018,30(2):14-17.
[12]黄贤荣,向凤宁,夏光敏,等.胡萝卜与柴胡的体细胞杂交及其愈伤组织的同工酶鉴别[J].中国中药杂志,2002,27(12):887-890.
[13]向凤宁,黄贤荣,王颖,等.柴胡与高寒藏药———川西獐芽菜科间体细胞杂交[J].山东大学学报:理学版,2003,38(2):93-96.
[14]程玉鹏,李天聪,林进华,等.狭叶柴胡愈伤组织原生质体的制备及纯化[J].山西农业大学学报:自然科学版,2017(4):254-257.
[15]关倩.北柴胡不定根培养中生物转化与皂苷合成研究[D].太原:山西大学,2011.
[16]杨东方,郝建平.北柴胡愈伤组织及不定根诱导研究[J].山西农业大学学报:自然科学版,2012,35(1):140-144.
[17]余马,左文松,刘茹,等.北柴胡新品种川北柴1号组织培养快繁初步研究[J].种子,2017,36(9):27-34.
[18]王梅,郝建平,赵暘,等.乙酸钠、水杨酸和Cu2+对北柴胡不定根培养合成皂苷的影响[J].天然产物研究与开发,2012,24(S1):87-91.
[19]战晴晴,金钺,魏建和,等.北柴胡不定根培养及茉莉酸甲酯处理对柴胡皂苷含量的影响[J].生物技术通讯,2011,22(1):57-60.
[20] Hilo A,Shahinnia F,Druege U,et al. A specific role of iron in promoting meristematic cell division during adventitious root formation[J]. J Exp Bot,2017,68(15):4233-4247.
[21] Qavami N,Azizi M,Yazdian F,et al. Effective induction of hairy roots in persian poppy(Papaver bracteatum Lindl.)using sonication method[J]. J Med Plants,2018,17(65):73-82.
[22]孙晶,杨洪一,隋春.不同因子对药用植物毛状根产量和次生代谢产物积累影响的研究进展[J].中国现代中药,2014,16(11):945-952.
[23] Renouard S,Corbin C,Drouet S,et al. Investigation of Linum flavum(L.)hairy root cultures for the production of anticancer aryltetralin lignans[J]. Int J Mol Sci,2018,19(4):990-1008.
[24]孙晶,高珂,王玲,等.不同培养基、外源激素和真菌诱导子对北柴胡毛状根生长及柴胡皂苷含量的影响[J].生物技术通讯,2015,26(4):546-550.
[25] YANG D F,FANG Y M,XIA P G,et al. Diverse responses of tanshinone biosynthesis to biotic and abiotic elicitors in hairy root cultures of Salvia miltiorrhiza and Salvia castanea Diels f. tomentosa[J]. Gene,2018,643:61-67.
[26]程玉鹏,林进华,高宁,等.不同因素对狭叶柴胡悬浮细胞生长的影响[J].山西农业大学学报:自然科学版,2016,36(5):305-309.
[27]周永强.狭叶柴胡悬浮细胞及内生真菌活性成分抗肿瘤研究[D].哈尔滨:黑龙江中医药大学,2014.
[28] Choi D,Ward B L,Bostock R M. Differential induction and suppression of potato 3-nydroxy-3-methylglutaryl coenzyme A reductase genes in response to Phytophthora infestans and to its elicitor arachidonic acid[J]. Plant Cell,1992,4(10):1333-1344.
[29] Ramos-Valdivia A C,van der Heijden R,Verpoorte R.Isopentenyl diphosphate isomerase:a core enzyme in isoprenoid biosynthesis:a review of its biochemistry and function[J]. Nat Prod Rep,1997,14(6):591-603.
[30] Ebizuka Y,Katsube Y,Tsutsumi T,et al. Functional genomics approach to the study of triterpene biosynthesis[J]. Pure Appl Chem,2003,75(2):369-374.
[31] Nakamura A,Shimada H,Masuda T,et al. Two distinct isopentenyl diphosphate isomerases in cytosol and plastid are differentially induced by environmental stresses in tobacco[J]. FEBS Lett,2001,506(1):61-64.
[32] Okada K,Kasahara H,Yamaguchi S,et al. Genetic evidence for the role of isopentenyl diphosphate isomerases in the mevalonate pathway and plant development in Arabidopsis[J]. Plant Cell Physiol,2008,49(4):604-616.
[33]隋春,战晴晴,魏建和,等.北柴胡皂苷生物合成途径关键酶IPPI的全长c DNA克隆及其序列分析[J].中草药,2010,41(7):1178-1184.
[34]隋春,魏建和,战晴晴.北柴胡鲨烯合酶基因及其编码区c DNA克隆与序列分析[J].园艺学报,2010,37(2):283-290.
[35]董乐萌.北柴胡柴胡皂苷合成相关基因的分子克隆与组织表达分析[D].北京:北京林业大学,2008.
[36]徐洁森.北柴胡皂苷合成相关P450、UGT基因的筛选、克隆及遗传转化初步研究[D].北京:北京协和医学院,2013.
[37] Bowles D, Isayenkova J, Lim E K, et al.Glycosyltransferases:managers of small molecules[J].Curr Opin Plant Biol,2005,8(3):254-263.
[38] WANG X Q. Structure,mechanism and engineering of plant natural product glycosyltransferases[J]. FEBS Lett,2009,583(20):3303-3309.
[39] YANG N,ZHOU W P,SU J,et al. Overexpression of Sm MYC2 increases the production of phenolic acids in Salvia miltiorrhiza[J]. Front Plant Sci,2017,doi:10. 3389/fpls. 2017. 01804.
[40]于一凡,郭娟,苏平,等.灯盏细辛转基因毛状根体系建立[J].中国中药杂志,2018,43(10):2048-2052.
[41] Murthy H N,Dandin V S,Paek K Y. Tools for biotechnological production of useful phytochemicals from adventitious root cultures[J]. Phytochem Rev,2016,15(1):129-145.
[42]左北梅,高文远,王娟,等.鼓泡式生物反应器培养人参不定根的研究[J].中国中药杂志,2012,37(24):3706-3711.
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