吲哚美辛调控小球藻生长及代谢的初步研究
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摘要
本研究通过在小球藻培养液中添加不同浓度的吲哚美辛(0mg/L,50mg/L,75mg/L,100mg/L,112.5mg/L,125 mg/L,150 mg/L)来研究其对小球藻生长及代谢的影响,并探索其影响机制。研究表明:吲哚美辛对小球藻生长和代谢的影响与其浓度有关,低浓度吲哚美辛(≤75 mg/L)可促进小球藻的生长,而高浓度(≥100 mg/L)则会抑制其生长,且在75 mg/L处理组中,吲哚美辛对小球藻生长的促进作用最明显,培养液p H值的变化最小,小球藻可溶性糖含量最高(3.12μg/106 cells)。而在高浓度(150 mg/L)处理组中,吲哚美辛可显著促进小球藻细胞内叶绿素a和可溶性蛋白的积累,可分别达到0.38μg/10~6 cells和7.09μg/106 cells,油脂含量也相对于对照组提高了60%。研究将为探索吲哚美辛调控小球藻生长及代谢的作用机制提供理论基础和技术资料,也将为小球藻的产业化培养及其代谢产物的调控提供思路。
Effects of indomethacin(IM) with different concentrations(0 mg/L, 50 mg/L, 75 mg/L, 100 mg/L,112.5 mg/L, 125 mg/L, 150 mg/L) on the growth and metabolism of Chlorella sorokiniana were investigated in this study, while the effect mechanism was explored. Research showed that the effects of IM on the growth and metabolism of C. sorokiniana were related to its concentrations and presented a dose-dependent manner. The growth of C. sorokiniana was enhanced in low IM concentrations(≤75 mg/L), but inhibited in high IM concentrations(≥100 mg/L). In 75 mg/L treatment, stimulatory effects of IM on the growth of C. sorokiniana were most remarkable. And the highest content of polysaccharide(3.12 μg/106 cells) in C. sorokiniana and the minimum change of p H value in the culture of C. sorokiniana were also observed in 75 mg/L IM treatment. However,indomethacin could obviously promote the accumulation of chlorophyll a and soluble protein in C. sorokiniana with 150 mg/L IM treatment, which were 0.38 μg/106 cells and 7.09 μg/10~6 cells, respectively. The lipid content also increased by 60% compared with that of the controls in 150 mg/L IM treatment. This study might provide theoretical basis and technical data to explore mechanism of action on the growth and metabolism of C.sorokiniana regulated by Indomethacin. It might also provide insight for the industrialization cultivation and metabolites regulation of C. sorokiniana.
Effects of indomethacin(IM) with different concentrations(0 mg/L, 50 mg/L, 75 mg/L, 100 mg/L,112.5 mg/L, 125 mg/L, 150 mg/L) on the growth and metabolism of Chlorella sorokiniana were investigated in this study, while the effect mechanism was explored. Research showed that the effects of IM on the growth and metabolism of C. sorokiniana were related to its concentrations and presented a dose-dependent manner. The growth of C. sorokiniana was enhanced in low IM concentrations(≤75 mg/L), but inhibited in high IM concentrations(≥100 mg/L). In 75 mg/L treatment, stimulatory effects of IM on the growth of C. sorokiniana were most remarkable. And the highest content of polysaccharide(3.12 μg/106 cells) in C. sorokiniana and the minimum change of p H value in the culture of C. sorokiniana were also observed in 75 mg/L IM treatment. However,indomethacin could obviously promote the accumulation of chlorophyll a and soluble protein in C. sorokiniana with 150 mg/L IM treatment, which were 0.38 μg/106 cells and 7.09 μg/10~6 cells, respectively. The lipid content also increased by 60% compared with that of the controls in 150 mg/L IM treatment. This study might provide theoretical basis and technical data to explore mechanism of action on the growth and metabolism of C.sorokiniana regulated by Indomethacin. It might also provide insight for the industrialization cultivation and metabolites regulation of C. sorokiniana.
引文
Bertozzini E.,Galluzzi L.,Penna A.,and Magnani M.,2011,Application of the standard addition method for the absolute quantification of neutral lipids in microalgae using Nile red,J.Microbiol.Methods.,87(1):17-23
Cao X.M.,and Du L.M.,2007,Spectroscopic study on binding of indomethacin to bovine serum albumin,Guangpuxue Yu Guangpu Fenxi(Spectroscopy and Spectral Analysis),27(5):973-977(曹玺珉,杜黎明,2007,吲哚美辛与牛血清白蛋白结合作用的研究,光谱学与光谱分析,27(5):973-977)
Chen Y.,Li W.B.,and Sun Y.R.,1998,Status and prospects of researches and applications of Chlorellasp.Biotechnology,Shengwu Gongcheng Jinzhan(Progress in Biotechnology),18(6):12-16(陈颖,李文彬,孙勇如,1998,小球藻生物技术研究应用现状及展望,生物工程进展,18(6):12-16)
Czerpak R.,and Bajguz A.,1997,Stimulatory effect of auxins and cytokinins on carotenes,with differential effects on xanthophylls in the green alga Chlorella pyrenoidosa Chick,Acta Societatis Botanicorum Poloniae,66(1):41-46
Deng X.Y.,Cheng J.,Gao K.,and Wang C.H.,2015,Effects of1-octyl-3-methylimidazolium bromide on the growth,photosynthetic activity and antioxidant enzymes of Chlorella pyrenoidosa,International Journal of Ecological Science and Environmental Engineering,1(1):1-10
Deng X.Y.,Gao K.,and Sun J.L.,2012,Physiological and biochemical responses of Synechococcus sp.PCC7942 to Irgarol 1051 and diuron,Aquat.Toxicol.,122-123:113-119
Deng X.Y.,Hu X.L.,Cheng J.,Ma Z.X.,and Gao K.,2016,Effects of nanoparticle Ce O2 on the physiology of Chlorella pyrenoidosa,Shengtai Duli Xuebao(Asian Journal of Ecotoxicology),11(5):111-116(邓祥元,胡小丽,成婕,马志欣,高坤,2016,纳米二氧化铈对蛋白核小球藻的生物学效应研究,生态毒理学报,11(5):111-116)
Ehsan H.,Reichheld J.P.,Roef L.,Witters E.,Lardon F.,Van Bockstaele D.,Van Montagu M.,InzéD.,and Van Onckelen H.,1998,Effect of indomethacin on cell cycle dependent cyclic AMP fluxes in tobacco BY-2 cells,FEBS Letters,422(2):165-169
Ehsan H.,Roef L.,Witters E.,Reichheld J.P.,Van Bockstaele D.,InzéD.,and Van Onckelen H.,1999,Indomethacin-induced G1/S phase arrest of the plant cell cycle,FEBS Letters,458(3):349-353
Lai J.P.,Lu C.Y.,and He X.W.,2003,Preparation of molecularly imprinted microspheres against acidic indometacin and its chromatographic characterization by aqueous microsuspension polymerization,Gaodeng Xuexiao Huaxue Xuebao(Chemical Research In Chinese Universities),24(7):1175-1179(赖家平,卢春阳,何锡文,2003,水溶液微悬浮聚合法制备酸性药物吲哚美辛分子印迹微球及其色谱表征,高等学校化学学报,24(7):1175-1179)
Li L.X.,and Zhou C.J.,2007,Biogas slurry comprehensive utilization technology,Nongye Ziyuan Yu Huanjing Xuebao(Agro-Environment and Development),24(1):45-46(黎良新,周丛钜,2007,沼液分层综合利用技术,农业资源与环境学报,24(1):45-46)
Li S.W.,Li H.Q.,and Zhang J.J.,1998,Progress in studies on large-scale culture of Chlorella,Zhiwu Xuebao(Chinese Bulletin of Botany),15(4):45-50(李师翁,李虎乾,张建军,1998,小球藻大规模培养研究的进展,植物学报,15(4):45-50)
Liang Y.,and Wang S.,2009,Current status and prospect of studies on microalgae stress by heavy metals,Haiyang Huzhao Tongbao(Transactions of Oceanology and Limnology),4:72-82(梁英,王帅,2009,重金属对微藻胁迫的研究现状及前景,海洋湖沼通报,4:72-82)
Luo L.Y.,Li J.Y.,Chen Z.,Liu H.,Ming C.B.,Liu R.,and Wang Q.,2016,Research progress in the application of aquatic animal feed of Chlorella,Liangshi Yu Siliao Gongye(Cereal and Feed Industry),12(6):55-57(罗柳茵,李家泳,陈卓,刘晖,明彩兵,刘锐,王琴,2016,小球藻在水产动物饲料中的应用研究进展,粮食与饲料工业,12(6):55-57)
Mazur H.,Konop A.,and Synak R.,2001,Indole-3-acetic acid in the culture medium of two axenic green microalgae,Journal of Applied Phycology,13(1):35-42
Piotrowska A.,Czerpak R.,Pietryczuk A.,Olesiewicz A.,and W誰do覥owska M.,2008,The effect of indomethacin on the growth and metabolism of green alga Chlorella vulgaris Beijerinck,Plant Growth Regulation,55(2):125-136
Song L.N.,Zheng X.Y.,Gu Y.J.,and Li L.,2010,Effect of Pconcentrations on chlorophyll fluorescence and growth of Chlorella sp.,Huanjing Wuran Yu Fangzhi(Environmental Pollution and Control),32(8):20-24(宋丽娜,郑晓宇,顾詠洁,李伶,2010,磷浓度对海洋小球藻叶绿素荧光及生长的影响,环境污染与防治,32(8):20-24)
Stanier R.Y.,Kunisawa R.,Mandel M.,and Cohen-Bazire G.,1971,Purification and properties of unicellular blue-green algae(order Chroococcales),Bacteriol.Rev.,35(2):171-205
Sun Q.G.,and Gao Q.Y.,2002,Effect of indomethacin on the stem rooting after removal of hypocotyl and root of mung bean(Phaseolus radiates),Zhiwu Shengli Xuebao(Plant Physiology Communications),38(2):133-134(孙全根,高庆义,2002,吲哚美辛对去除下胚轴和根后的绿豆茎段生根的影响,植物生理学报,38(2):133-134)
Tang M.H.,Sun L.,Tang L.H.,Yang T.Y.,and Zhou H.Y.,2013,The effect of compound sodium nitrophenolate on the growth and photosynthetic pigment of Chlorella vulgaris,Shuichan Yangzhi(Journal of Aquaculture),34(10):5-7(唐明虎,孙莉,汤连华,杨庭玉,周海洋,2013,复硝酚钠对小球藻生长及光合色素含量的影响,水产养殖,34(10):5-7)
Wilkie A.C.,Edmundson S.J.,and Duncan J.G.,2011,Indigenous algae for local bioresource production:phycoprospecting,Energy for Sustainable Development,15(4):365-371
Young E.B.,and Beardall J.,2003,Rapid ammonium-and nitrate-induced perturbations to Chl a fluorescence in nitrogen-stressed Dunaliella tertiolecta(Chlorophyta),Journal of Phycology,39(2):332-342
Zha R.B.,Zhang H.,and Sun J.M.,2015,Active components and bioactivities of microalgae,Jilin Zhongyiyao(Jilin Journal of Traditional Chinese Medicine),35(1):64-67(查荣博,张辉,孙佳明,2015,微藻的有效成分及生物活性,吉林中医药,35(1):64-67)
Zhang G.Y.,Wen X.B.,Liang F.Ouyang Z.R.,Geng Y.H.,Mei H.,and Li Y.G.,2011,The effects of physical and chemical factors on the growth and lipid production of Chlorella,Shengtai Xuebao(Acta Ecologica Sinica),31(8):2076-2085(张桂艳,温小斌,梁芳,欧阳峥嵘,耿亚红,梅洪,李夜光,2011,重要理化因子对小球藻生长和油脂产量的影响,生态学报,31(8):2076-2085)
Zhang S.R.,1999,A discussion on chlorophyll fluorescence kinetics parameters and their significance,Zhiwuxue Tongbao(Chinese Bulletin of Botany),16(4):444-448(张守仁,1999,叶绿素荧光动力学参数的意义及讨论,植物学通报,16(4):444-448)
Zhou W.L.,Qiao X.T.,Xiao H.,Wang Y.,Qu L.,Tang X.X.,and Zhang P.Y.,2009,Effects of antibiotics on chlorophyll a level in marine microalga,Haiyang Huanjing Kexue(Marine Environmental Science),28(3):268-271(周文礼,乔秀亭,肖慧,王悠,曲良,唐学玺,张培玉,2009,三种抗生素对几种海洋微藻叶绿素a含量影响的初步研究,海洋环境科学,28(3):268-271)
Cao X.M.,and Du L.M.,2007,Spectroscopic study on binding of indomethacin to bovine serum albumin,Guangpuxue Yu Guangpu Fenxi(Spectroscopy and Spectral Analysis),27(5):973-977(曹玺珉,杜黎明,2007,吲哚美辛与牛血清白蛋白结合作用的研究,光谱学与光谱分析,27(5):973-977)
Chen Y.,Li W.B.,and Sun Y.R.,1998,Status and prospects of researches and applications of Chlorellasp.Biotechnology,Shengwu Gongcheng Jinzhan(Progress in Biotechnology),18(6):12-16(陈颖,李文彬,孙勇如,1998,小球藻生物技术研究应用现状及展望,生物工程进展,18(6):12-16)
Czerpak R.,and Bajguz A.,1997,Stimulatory effect of auxins and cytokinins on carotenes,with differential effects on xanthophylls in the green alga Chlorella pyrenoidosa Chick,Acta Societatis Botanicorum Poloniae,66(1):41-46
Deng X.Y.,Cheng J.,Gao K.,and Wang C.H.,2015,Effects of1-octyl-3-methylimidazolium bromide on the growth,photosynthetic activity and antioxidant enzymes of Chlorella pyrenoidosa,International Journal of Ecological Science and Environmental Engineering,1(1):1-10
Deng X.Y.,Gao K.,and Sun J.L.,2012,Physiological and biochemical responses of Synechococcus sp.PCC7942 to Irgarol 1051 and diuron,Aquat.Toxicol.,122-123:113-119
Deng X.Y.,Hu X.L.,Cheng J.,Ma Z.X.,and Gao K.,2016,Effects of nanoparticle Ce O2 on the physiology of Chlorella pyrenoidosa,Shengtai Duli Xuebao(Asian Journal of Ecotoxicology),11(5):111-116(邓祥元,胡小丽,成婕,马志欣,高坤,2016,纳米二氧化铈对蛋白核小球藻的生物学效应研究,生态毒理学报,11(5):111-116)
Ehsan H.,Reichheld J.P.,Roef L.,Witters E.,Lardon F.,Van Bockstaele D.,Van Montagu M.,InzéD.,and Van Onckelen H.,1998,Effect of indomethacin on cell cycle dependent cyclic AMP fluxes in tobacco BY-2 cells,FEBS Letters,422(2):165-169
Ehsan H.,Roef L.,Witters E.,Reichheld J.P.,Van Bockstaele D.,InzéD.,and Van Onckelen H.,1999,Indomethacin-induced G1/S phase arrest of the plant cell cycle,FEBS Letters,458(3):349-353
Lai J.P.,Lu C.Y.,and He X.W.,2003,Preparation of molecularly imprinted microspheres against acidic indometacin and its chromatographic characterization by aqueous microsuspension polymerization,Gaodeng Xuexiao Huaxue Xuebao(Chemical Research In Chinese Universities),24(7):1175-1179(赖家平,卢春阳,何锡文,2003,水溶液微悬浮聚合法制备酸性药物吲哚美辛分子印迹微球及其色谱表征,高等学校化学学报,24(7):1175-1179)
Li L.X.,and Zhou C.J.,2007,Biogas slurry comprehensive utilization technology,Nongye Ziyuan Yu Huanjing Xuebao(Agro-Environment and Development),24(1):45-46(黎良新,周丛钜,2007,沼液分层综合利用技术,农业资源与环境学报,24(1):45-46)
Li S.W.,Li H.Q.,and Zhang J.J.,1998,Progress in studies on large-scale culture of Chlorella,Zhiwu Xuebao(Chinese Bulletin of Botany),15(4):45-50(李师翁,李虎乾,张建军,1998,小球藻大规模培养研究的进展,植物学报,15(4):45-50)
Liang Y.,and Wang S.,2009,Current status and prospect of studies on microalgae stress by heavy metals,Haiyang Huzhao Tongbao(Transactions of Oceanology and Limnology),4:72-82(梁英,王帅,2009,重金属对微藻胁迫的研究现状及前景,海洋湖沼通报,4:72-82)
Luo L.Y.,Li J.Y.,Chen Z.,Liu H.,Ming C.B.,Liu R.,and Wang Q.,2016,Research progress in the application of aquatic animal feed of Chlorella,Liangshi Yu Siliao Gongye(Cereal and Feed Industry),12(6):55-57(罗柳茵,李家泳,陈卓,刘晖,明彩兵,刘锐,王琴,2016,小球藻在水产动物饲料中的应用研究进展,粮食与饲料工业,12(6):55-57)
Mazur H.,Konop A.,and Synak R.,2001,Indole-3-acetic acid in the culture medium of two axenic green microalgae,Journal of Applied Phycology,13(1):35-42
Piotrowska A.,Czerpak R.,Pietryczuk A.,Olesiewicz A.,and W誰do覥owska M.,2008,The effect of indomethacin on the growth and metabolism of green alga Chlorella vulgaris Beijerinck,Plant Growth Regulation,55(2):125-136
Song L.N.,Zheng X.Y.,Gu Y.J.,and Li L.,2010,Effect of Pconcentrations on chlorophyll fluorescence and growth of Chlorella sp.,Huanjing Wuran Yu Fangzhi(Environmental Pollution and Control),32(8):20-24(宋丽娜,郑晓宇,顾詠洁,李伶,2010,磷浓度对海洋小球藻叶绿素荧光及生长的影响,环境污染与防治,32(8):20-24)
Stanier R.Y.,Kunisawa R.,Mandel M.,and Cohen-Bazire G.,1971,Purification and properties of unicellular blue-green algae(order Chroococcales),Bacteriol.Rev.,35(2):171-205
Sun Q.G.,and Gao Q.Y.,2002,Effect of indomethacin on the stem rooting after removal of hypocotyl and root of mung bean(Phaseolus radiates),Zhiwu Shengli Xuebao(Plant Physiology Communications),38(2):133-134(孙全根,高庆义,2002,吲哚美辛对去除下胚轴和根后的绿豆茎段生根的影响,植物生理学报,38(2):133-134)
Tang M.H.,Sun L.,Tang L.H.,Yang T.Y.,and Zhou H.Y.,2013,The effect of compound sodium nitrophenolate on the growth and photosynthetic pigment of Chlorella vulgaris,Shuichan Yangzhi(Journal of Aquaculture),34(10):5-7(唐明虎,孙莉,汤连华,杨庭玉,周海洋,2013,复硝酚钠对小球藻生长及光合色素含量的影响,水产养殖,34(10):5-7)
Wilkie A.C.,Edmundson S.J.,and Duncan J.G.,2011,Indigenous algae for local bioresource production:phycoprospecting,Energy for Sustainable Development,15(4):365-371
Young E.B.,and Beardall J.,2003,Rapid ammonium-and nitrate-induced perturbations to Chl a fluorescence in nitrogen-stressed Dunaliella tertiolecta(Chlorophyta),Journal of Phycology,39(2):332-342
Zha R.B.,Zhang H.,and Sun J.M.,2015,Active components and bioactivities of microalgae,Jilin Zhongyiyao(Jilin Journal of Traditional Chinese Medicine),35(1):64-67(查荣博,张辉,孙佳明,2015,微藻的有效成分及生物活性,吉林中医药,35(1):64-67)
Zhang G.Y.,Wen X.B.,Liang F.Ouyang Z.R.,Geng Y.H.,Mei H.,and Li Y.G.,2011,The effects of physical and chemical factors on the growth and lipid production of Chlorella,Shengtai Xuebao(Acta Ecologica Sinica),31(8):2076-2085(张桂艳,温小斌,梁芳,欧阳峥嵘,耿亚红,梅洪,李夜光,2011,重要理化因子对小球藻生长和油脂产量的影响,生态学报,31(8):2076-2085)
Zhang S.R.,1999,A discussion on chlorophyll fluorescence kinetics parameters and their significance,Zhiwuxue Tongbao(Chinese Bulletin of Botany),16(4):444-448(张守仁,1999,叶绿素荧光动力学参数的意义及讨论,植物学通报,16(4):444-448)
Zhou W.L.,Qiao X.T.,Xiao H.,Wang Y.,Qu L.,Tang X.X.,and Zhang P.Y.,2009,Effects of antibiotics on chlorophyll a level in marine microalga,Haiyang Huanjing Kexue(Marine Environmental Science),28(3):268-271(周文礼,乔秀亭,肖慧,王悠,曲良,唐学玺,张培玉,2009,三种抗生素对几种海洋微藻叶绿素a含量影响的初步研究,海洋环境科学,28(3):268-271)