诱导子对颠茄毛状根生长及托品烷类生物碱质量分数的影响
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摘要
目的:研究不同浓度茉莉酸甲酯(MJ)、硝酸银(AgNO_3)、水杨酸(SA)、酵母提取物(YE)4种诱导子对颠茄毛状根生长及托品烷类生物碱合成的影响.方法:将0.5g新鲜颠茄毛状根接种于B5液体培养基中,黑暗培养12d后,将原有培养基换成加有不同浓度诱导子的4种新培养基,同样条件下再培养2d,收集颠茄毛状根材料,测定其鲜质量与干质量及托品烷类生物碱的含量.结果:MJ抑制了毛状根生长与托品烷类生物碱的积累,而AgNO_3虽然抑制了颠茄毛状根的生长,但却显著提高了托品烷类生物碱的积累,托品烷类生物碱的产量在100μmol/L AgNO_3处理下达到26.191mg,是对照的219.9%.在SA处理下,较小程度地抑制了毛状根的生长,对东莨菪碱的积累有促进作用.在YE处理下,随试验浓度升高,对毛状根生长及托品烷类生物碱的积累的促进作用也逐渐增强.结论:AgNO_3是提高毛状根中托品烷类生物碱的最佳诱导子,而YE是对颠茄毛状根生长的最佳诱导子.
Objective:To study the influences of elicitors(MJ,AgNO_3,SA and YE)on the growth and tropane alkaloids in the hairy roots of Atropa belladonna L.Methods:Fresh hairy roots of A.belladonna L.0.5 g in weight were inoculated into B5 liquid medium and cultured for 12 days in dark.Then the original media were poured out and replaced by four kinds of new media containing different concentrations of MJ,AgNO_3,SA or YE and cultured under the same conditions for 2 days.Finally,the hairy roots were gathered and their fresh weight,dry weight and contents of tropic alkane alkaloids were determined.Results:MJ restrained the growth and alkaloid accumulation of the hairy roots.Though AgNO_3 inhibited the growth of the hairy roots,it significantly improved the accumulation of tropane alkaloids in them.With a concentration of 100μmol/L,AgNO_3 treatment increased the contents of tropane alkaloids by 119.9% as compared with CK.SA had little influence on the growth of the hairy roots and stimulated the accumulation of scopolamine.In YE treatments,hairy root growth and contents of tropane alkaloids increased with the increasing of YE concentration;Conclusions:Elicitors have selective influence on the growth of the hairy roots of A.belladonna L.,and AgNO_3 is the best elicitor for trapane alkaloid accumulation in the hairy roots,and YE is the best elicitor for improving hairy root growth of A.belladonna L..
Objective:To study the influences of elicitors(MJ,AgNO_3,SA and YE)on the growth and tropane alkaloids in the hairy roots of Atropa belladonna L.Methods:Fresh hairy roots of A.belladonna L.0.5 g in weight were inoculated into B5 liquid medium and cultured for 12 days in dark.Then the original media were poured out and replaced by four kinds of new media containing different concentrations of MJ,AgNO_3,SA or YE and cultured under the same conditions for 2 days.Finally,the hairy roots were gathered and their fresh weight,dry weight and contents of tropic alkane alkaloids were determined.Results:MJ restrained the growth and alkaloid accumulation of the hairy roots.Though AgNO_3 inhibited the growth of the hairy roots,it significantly improved the accumulation of tropane alkaloids in them.With a concentration of 100μmol/L,AgNO_3 treatment increased the contents of tropane alkaloids by 119.9% as compared with CK.SA had little influence on the growth of the hairy roots and stimulated the accumulation of scopolamine.In YE treatments,hairy root growth and contents of tropane alkaloids increased with the increasing of YE concentration;Conclusions:Elicitors have selective influence on the growth of the hairy roots of A.belladonna L.,and AgNO_3 is the best elicitor for trapane alkaloid accumulation in the hairy roots,and YE is the best elicitor for improving hairy root growth of A.belladonna L..
引文
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[19]NOVAIS J M,MAVITUNA F,PAIS M S S.Plant Cell Biotechnology[M].Berlin:Springer-Verlag,1994.
[2]张敏敏,陈玉梁,赵瑛,等.药用植物组织培养生产有效成分的影响因素研究进展[J].甘肃农业科技,2013(7):43-46.
[3]任志华,李玲.植物细胞培养技术提高次生代谢物产量的方法(综述)[J].亚热带植物科学,2003,32(3):64-67.
[4]曲伟红,赵建国.毛状根培养在药用植物中的应用概况[J].九江学院学报(自然科学版),2006,21(4):85-87.
[5]李琳琳.茉莉酸甲酯对颠茄毛状根的生长及次生代谢产物产生的影响[D].重庆:西南大学,2015.
[6]ZARATE R,HERMOSIN B,CANTOS M,et al.Tropane Alkaloid Distribution in Atropa baetica Plants[J].Journal of Chemical Ecology,1997,23(8):2059-2066.
[7]肖春桥,高洪,池汝安.诱导子促进植物次生代谢产物生产的研究进展[J].天然产物研究与开发,2004,16(5):473-476.
[8]任振兴.黄芩愈伤组织培养及其次级代谢物合成调控的研究[D].太原:山西大学,2006.
[9]孙际薇.茉莉酸甲酯对曼陀罗毛状根的生长及次生代谢产物产生的影响[D].重庆:西南大学,2014.
[10]肖莹.丹参酚酸类成分生源合成的调控研究[D].上海:第二军医大学,2009.
[11]VASCONSUELO A,BOLAND R.Molecular Aspects of the Early Stages of Elicitation of Secondary Metabolites in Plants[J].Plant Science,2007,172(5):861-875.
[12]CLEMENS S.Toxic Metal Accumulation,Responses to Exposure and Mechanisms of Tolerance in Plants[J].Biochimie,2006,88(11):1707-1718.
[13]YING X,GAO S H,PENG D,et al.Lithospermic Acid B is More Responsive to Silver Ions(Ag+)than Rosmarinic Acid in Salvia miltiorrhiza Hairy Root Cultures[J].Bioscience Reports,2009,30(1):33-40.
[14]虞珍珍.诱导子对水母薛莲次生代谢的影响及雪莲查尔酮合成酶基因克隆[D].北京:中国植物科学院,2006.
[15]梅兴国,张舟宁,苏湘鄂,等.水杨酸对红豆杉细胞的诱导作用[J].生物技术,2000,10(6):18-20.
[16]CHEN H,CHEN F.Effect of Yeast Elicitor on the Secondary Metabolism of Ti-Transformed Salvia Mihiorrhiza Cell Suspension Cultures[J].Plant Cell Reports,2000,19(7):710-717.
[17]徐春明,赵兵,欧元.酵母提取物对水母雪莲悬浮培养合成紫丁香苷的影响[J].食品科学技术学报,2009,27(3):1-4.
[18]FAHRENDORF T,NI W,SHORROSH B S,et al.Stress Responses in Alfalfa(Medicago sativa L.)XIX.Transcriptional Activation of Oxidative Pentose Phosphate Pathway Genes at the Onset of the Isoflavanoid Phytoalexin Response[J].Plant Molecular Biology,1995,28(5):885-900.
[19]NOVAIS J M,MAVITUNA F,PAIS M S S.Plant Cell Biotechnology[M].Berlin:Springer-Verlag,1994.