生物反应器中金线莲丛生芽的增殖及酚和黄酮积累的研究
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摘要
利用气升式生物反应器研究了反应器培养方式和培养天数对金线莲(Anoectochilus roxburghii(Wall.)Lindl.)丛生芽增殖及总酚和总黄酮含量的影响,同时对诱导子促进酚和黄酮积累的影响也进行了调查。结果表明:在反应器内球体底部搭建"载桥"后进行培养的方式有利于金线莲丛生芽的增殖及酚和黄酮的积累,总酚和总黄酮的生产量最多。金线莲丛生芽在生物反应器培养30d时,酚和黄酮积累最大。因此,30d为最佳的培养周期。水杨酸、茉莉酸甲酯及酵母提取物对金线莲总酚和总黄酮含量均有提高作用,采用525μM的茉莉酸甲酯处理16d的效果最佳。
The effects of bioreactor culture methods and culture days on the proliferation and the accumulation of phenolics and flavonoids of multiple buds of Anoectochilus roxburghii(Wall.)Lindl.as well as the elicitor on enhancement of phenolic and flavonoid accumulation were were studied.The results showed that making a supporter(bridge)on the sphere bottom of the bioreactor was propitious to the proliferation of multiple buds and the accumulation of phenolics and flavonoids.The maximum accumulation of both total phenolics and flavonoids was obtained at 30 days after culture,indicating that 30 dwas the optimum culture period for multiple bud culture in bioreactors.Salicylic acid,methyl jasmonate(MeJA),and the yeast extract were favorable to increase the accumulation of phenolics and flavonoids;among these,the optimum treatment was that 30-day-old bioreactor cultured multiple buds were treated with 525 μM for16 days.
The effects of bioreactor culture methods and culture days on the proliferation and the accumulation of phenolics and flavonoids of multiple buds of Anoectochilus roxburghii(Wall.)Lindl.as well as the elicitor on enhancement of phenolic and flavonoid accumulation were were studied.The results showed that making a supporter(bridge)on the sphere bottom of the bioreactor was propitious to the proliferation of multiple buds and the accumulation of phenolics and flavonoids.The maximum accumulation of both total phenolics and flavonoids was obtained at 30 days after culture,indicating that 30 dwas the optimum culture period for multiple bud culture in bioreactors.Salicylic acid,methyl jasmonate(MeJA),and the yeast extract were favorable to increase the accumulation of phenolics and flavonoids;among these,the optimum treatment was that 30-day-old bioreactor cultured multiple buds were treated with 525 μM for16 days.
引文
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[18]Folin O,Ciocalteu V.On tyrosine and tryptophane determination in proteins[J].J Biol Chem,1927,73(2):627-650.
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[20]MuranakaT,Kazuki S.Production of pharmaceuticals by plant tissue cultures[J].Compr Nat Prod II:Chem Biol,2010(3):615-628.
[21]韩璐,廉美兰,金美玉,等.几种因素对白鹤芋组培苗增殖生长的影响[J].延边大学农学学报,2016,38(4):313-316.
[22]张万博,朴炫春,杨帆,等.植物生长调节剂和蔗糖浓度对狼爪瓦松愈伤组织生物量及有效物质积累的影响[J].北方园艺,2016(19):171-175.
[23]焦蒙丽,曹蓉蓉,陈红艳,等.水杨酸对丹参培养细胞中迷迭香酸生物合成及其相关酶的影响[J].生物工程学报,2012,28(3):320-328.
[24]Bulgakov V P,Tchernoded G K,Mischenko N P,et al.Effect of salicylic acid,methyl jasmonate,ethephon and cantharidin on anthraquinone production by Rubia cordifolia callus cultures transformed with the rolBand rolC genes[J].J Biotechnol,2002,97(3):213-221.
[25]Afitlhile M M,Fukushige H,Nishimura M,et al.A defect in glyoxysomal fatty acid beta-oxidation reduces jasmonic acid accumulation in Arabidopsis[J],Plant Physiol Bioch,2005,43:603-609.
[26]杜晓映.葡萄细胞的悬浮培养及诱导子对悬浮细胞多酚产量影响的研究[D].咸阳:西北农林科技大学,2008.
[27]郭双,杨怡,刘兴,等.酵母提取物对颠茄毛状根氮代谢及次生代谢影响的机制探究[J].中国中药杂志,2018,43(08):1610-1617.
[2]沈廷明,吴仲玉,黄春情,等.金线莲化学成分、药理、组培及栽培研究进展[J].海峡药学,2016,28(12):26-30.
[3]刘辉辉,沈岚,毛碧增.金线莲化学成分、药理及组织培养研究进展[J].药物生物技术,2015,22(06):553-556.
[4]李执坤,陈虹竹,冯艳钰,等.5种芽苗菜黄酮提取物的体外抗氧化活性研究[J].黑龙江八一农垦大学学报,2017,29(02):48-51.
[5]朱红霞,李鹏,朱英慧,等.欧亚旋覆花总黄酮质量分数及抗菌活性初步研究[J].河南科技学院学报(自然科学版),2017,45(02):15-18,22.
[6]陈宇,董映雪,郑璐,等.竹叶黄酮类物质的抗氧化性能研究[J].赤峰学院学报(自然科学版),2015,31(12):4-8.
[7]于帅,杜彬,杨越冬,等.植物中多酚的研究进展[J].河北科技师范学院学报,2011,25(03):68-74.
[8]高洁,盛婴,金爱萍,等.丹参多酚酸盐对冠状动脉内支架置入术后患者血清脂联素与抵抗素的影响[J].内蒙古民族大学学报(自然科学版),2014,29(05):593-595.
[9]陈会良,鲍广刚,凡敏,等.茶多酚对营养性肥胖大鼠肝脏组织结构的影响[J].安徽科技学院学报,2009,23(06):6-9.
[10]王博,伏劲松,蔡光华,等.香柏总酚酸的含量测定[J].西南民族大学学报(自然科学版),2010,36(04):600-602.
[11]孙紫薇.藤茶扦插育苗技术和组培苗快速繁殖技术研究[D].恩施:湖北民族学院,2017.
[12]刘春朝,王玉春,欧阳藩,等.生物反应器技术应用于植物组织培养的研究进展[J].化工冶金,1999(03):329-336.
[13]杨帆,廉美兰,李美兰,等.金线莲丛生芽培养及总酚与总黄酮生产研究[J].中草药,2016,47(18):3284-3288.
[14]吴荣哲.生物反应器在金线莲扩繁中的应用研究[J].安徽农业科学,2010(36):20571-20573,20591.
[15]廉美兰,朴炫春,王颖,等.间歇浸没式生物反应器在辰星草培养苗扩繁中的应用[J].园艺学报,2004(04):532.
[16]李慧娟,朴炫春,代月,等.人参不定根生物反应器培养及其动态学研究[J].延边大学农学学报,2011,33(02):94-98,107.
[17]吴晓玲,邓光存,丁向真,等.植物细胞次级代谢产物生产条件的调控[J].生物学通报,2002(12):17-18.
[18]Folin O,Ciocalteu V.On tyrosine and tryptophane determination in proteins[J].J Biol Chem,1927,73(2):627-650.
[19]吴荣志,徐莲英,陶建生.银杏叶总黄酮提取工艺探讨[J].中成药研究,1987(10):13-14.
[20]MuranakaT,Kazuki S.Production of pharmaceuticals by plant tissue cultures[J].Compr Nat Prod II:Chem Biol,2010(3):615-628.
[21]韩璐,廉美兰,金美玉,等.几种因素对白鹤芋组培苗增殖生长的影响[J].延边大学农学学报,2016,38(4):313-316.
[22]张万博,朴炫春,杨帆,等.植物生长调节剂和蔗糖浓度对狼爪瓦松愈伤组织生物量及有效物质积累的影响[J].北方园艺,2016(19):171-175.
[23]焦蒙丽,曹蓉蓉,陈红艳,等.水杨酸对丹参培养细胞中迷迭香酸生物合成及其相关酶的影响[J].生物工程学报,2012,28(3):320-328.
[24]Bulgakov V P,Tchernoded G K,Mischenko N P,et al.Effect of salicylic acid,methyl jasmonate,ethephon and cantharidin on anthraquinone production by Rubia cordifolia callus cultures transformed with the rolBand rolC genes[J].J Biotechnol,2002,97(3):213-221.
[25]Afitlhile M M,Fukushige H,Nishimura M,et al.A defect in glyoxysomal fatty acid beta-oxidation reduces jasmonic acid accumulation in Arabidopsis[J],Plant Physiol Bioch,2005,43:603-609.
[26]杜晓映.葡萄细胞的悬浮培养及诱导子对悬浮细胞多酚产量影响的研究[D].咸阳:西北农林科技大学,2008.
[27]郭双,杨怡,刘兴,等.酵母提取物对颠茄毛状根氮代谢及次生代谢影响的机制探究[J].中国中药杂志,2018,43(08):1610-1617.