褪黑素对雨生红球藻(Haematococcus pluvialis)生理和虾青素积累的影响
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摘要
本实验初步研究了不同浓度褪黑素(melatonin,MLT)对胁迫条件下雨生红球藻Haematococcus pluvialis LUGU生长、虾青素、相关生理指标及基因表达的影响。结果表明,10μmol/LMLT可显著提高藻细胞中虾青素产量(31.32mg/g),是对照组(13.27mg/g)的2.36倍;碳水化合物和蛋白含量同比提高了1.36、1.37倍。此外,虾青素合成相关基因pds和bkt的表达水平分别提高了5.79和12.16倍。研究表明,MLT可显著促进雨生红球藻中虾青素的积累,为利用微藻生产虾青素提供了一种有效的策略。
In this study,the effects of different concentrations of melatonin(MLT) on the growth,astaxanthin content,physiological indexes and gene expression of Haematococcus pluvialis LUGU were investigated.Results showed that the astaxanthin content significantly increased in algal cells at 10μmol/L MLT,the maximum astaxanthin content was 31.32 mg/g,and enhanced by 2.36 times compared with that of the control.The carbohydrate and protein contents increased by 1.36 and 1.37 fold,respectively.Furthermore,the expression levels of the astaxanthin synthesis genes pds and bkt improved by 5.79 and 12.16 times,respectively.These results indicated that MLT could significantly enhance astaxanthin accumulation in H.pluvialis.This study proposed a strategy to efficiently produce astaxanthin by using microalgae.
In this study,the effects of different concentrations of melatonin(MLT) on the growth,astaxanthin content,physiological indexes and gene expression of Haematococcus pluvialis LUGU were investigated.Results showed that the astaxanthin content significantly increased in algal cells at 10μmol/L MLT,the maximum astaxanthin content was 31.32 mg/g,and enhanced by 2.36 times compared with that of the control.The carbohydrate and protein contents increased by 1.36 and 1.37 fold,respectively.Furthermore,the expression levels of the astaxanthin synthesis genes pds and bkt improved by 5.79 and 12.16 times,respectively.These results indicated that MLT could significantly enhance astaxanthin accumulation in H.pluvialis.This study proposed a strategy to efficiently produce astaxanthin by using microalgae.
引文
丁巍,尚敏敏,余旭亚等,2017.茴香醚对雨生红球藻(Haematococcus pluvialis LUGU)虾青素积累和脂肪酸合成的影响.食品与发酵工业,43(2):26-32
巩彪,史庆华,2017.园艺作物褪黑素的研究进展.中国农业科学,50(12):2326-2337
梁竹,魏玮,陶利,2014.褪黑素在治疗睡眠障碍中的研究进展.解放军药学学报,30(1):82-85
梁英,冯力霞,尹翠玲等,2007.叶绿素荧光技术在微藻环境胁迫研究中的应用现状及前景.海洋科学,31(1):71-76
梁英,陈书秀,2009.温度对雨生红球藻叶绿素荧光特性及虾青素含量的影响.海洋湖沼通报,(3):112-120
韩春梅,刘建国,张勇,2014.不同激素配伍对雨生红球藻(Haematococcus pluvialis)细胞生长和虾青素累积的调节作用、藻株差异及应用.海洋与湖沼,40(4):430-436
Arnon D I,1949.Copper enzymes in isolated chloroplasts.Polyphenoloxidase in Beta vulgaris.Plant Physiology,24(1):1-15
Berges J A,Fisher A E,Harrison P J,1993.A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana.Marine Biology,115(2):187-193
Chen G Q,Wang B B,Han D X et al,2015.Molecular mechanisms of the coordination between astaxanthin and fatty acid biosynthesis in Haematococcus pluvialis(Chlorophyceae).The Plant Journal,81(1):95-107
Ebrahimian A,Kariminia H R,Vosoughi M,2014.Lipid production in mixotrophic cultivation of Chlorella vulgaris in a mixture of primary and secondary municipal wastewater.Renewable Energy,71:502-508
Fasstt R G,Coombes J S,2012.Astaxanthin in cardiovascular health and disease.Molecules,17(2):2030-2048
Gao Z Q,Meng C X,Zhang X W et al,2012a.Differential expression of carotenogenic genes,associated changes on astaxanthin production and photosynthesis features induced by JA in H.pluvialis.PLoS One,7(8):e42243
Gao Z Q,Meng C X,Zhang X W et al,2012b.Induction of salicylic acid(SA)on transcriptional expression of eight carotenoid genes and astaxanthin accumulation in Haematococcus pluvialis.Enzyme and Microbial Technology,51(4):225-230
Guerin M,Huntley M E,Olaizola M,2003.Haematococcus astaxanthin:applications for human health and nutrition.Trends in Biotechnology,21(5):210-216
Gwak Y,Hwang Y S,Wang B et al,2014.Comparative analyses of lipidomes and transcriptomes reveal a concerted action of multiple defensive systems against photooxidative stress in Haematococcus pluvialis.Journal of Experimental Botany,65(15):4317-4334
Han D X,Li Y T,Hu Q,2013.Astaxanthin in microalgae:pathways,functions and biotechnological implications.Algae,28(2):131-147
Huang J C,Chen F,Sandmann G,2006.Stress-related differential expression of multipleβ-carotene ketolase genes in the unicellular green alga Haematococcus pluvialis.Journal of Biotechnology,122(2):176-185
Huang W P,Ye J R,Zhang J J et al,2016.Transcriptome analysis of Chlorella zofingiensis to identify genes and their expressions involved in astaxanthin and triacylglycerol biosynthesis.Algal Research,17:236-243
Kobayashi M,Todoroki Y,Hirai N,et al,1998.Biological activities of abscisic acid analogs in the morphological change of the green alga Haematococcus pluvialis.Journal of Fermentation and Bioengineering,85(5):529-531
Li D F,Zhao Y T,Ding W et al,2017.A strategy for promoting lipid production in green microalgae Monoraphidium sp.QLY-1 by combined melatonin and photoinduction.Bioresource Technology,235:104-112
Li J,Zhu D L,Niu J F et al,2011.An economic assessment of astaxanthin production by large scale cultivation of Haematococcus pluvialis.Biotechnology Advances,29(6):568-574
Liang C,Zhai Y,Xu D et al,2015.Correlation between lipid and carotenoid synthesis and photosynthetic capacity in Haematococcus pluvialis grown under high light and nitrogen deprivation stress.Grasas Aceites,66(2):e077
Lu Y D,Jiang P,Liu S F et al,2010.Methyl jasmonate-or gibberellins A3-induced astaxanthin accumulation is associated with up-regulation of transcription ofβ-carotene ketolase genes(bkts)in microalga Haematococcus pluvialis.Bioresource Technology,101(16):6468-6474
Ma X N,Liu J,Liu B et al,2016.Physiological and biochemical changes reveal stress-associated photosynthetic carbon partitioning into triacylglycerol in the oleaginous marine alga Nannochloropsis oculata.Algal Research,16:28-35
Poonkum W,Powtongsook S,Pavasant P,2015.Astaxanthin induction in microalga H.pluvialis with flat panel airlift photobioreactors under indoor and outdoor conditions.Preparative Biochemistry and Biotechnology,45(1):1-17
Raman V,Ravi S,2011.Effect of salicylic acid and methyl jasmonate on antioxidant systems of Haematococcus pluvialis.Acta Physiologiae Plantarum,33(3):1043-1049
Reiter R J,Tan D X,Osuna C et al,2000.Actions of melatonin in the reduction of oxidative stress.Journal of Biomedical Science,7(6):444-458
Shang M M,Ding W,Zhao Y T et al,2016.Enhanced astaxanthin production from Haematococcus pluvialis using butylated hydroxyanisole.Journal of Biotechnology,236:199-207
Shi H,Jiang C,Ye T et al,2015.Comparative physiological,metabolomic and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass(Cynodon dactylon(L).Pers.)by exogenous melatonin.Journal of Experimental Botany,66(3):681-694
Su Y X,Wang J X,Shi M L et al,2014.Metabolomic and network analysis of astaxanthin-producing Haematococcus pluvialis under various stress conditions.Bioresource Technology,170:522-529
Tan D X,Manchester L C,Hardeland R et al,2003.Melatonin:a hormone,a tissue factor,an autocoid,a paracoid,and an antioxidant vitamin.Journal of Pineal Research,34(1):75-78
Tan D X,Hardeland R,Manchester L C et al,2012.Functional roles of melatonin in plants,and perspectives in nutritional and agricultural science.Journal of Experimental Botany,63(2):577-597
Wei W,Li Q T,Chu Y N et al,2015.Melatonin enhances plant growth and abiotic stress tolerance in soybean plants.Journal of Experimental Botany,66(3):695-707
Wen Z W,Liu Z Y,Hou Y Y et al,2015.Ethanol induced astaxanthin accumulation and transcriptional expression of carotenogenic genes in Haematococcus pluvialis.Enzyme and Microbial Technology,78:10-17
Zhao Y T,Shang M M,Xu J W et al,2015.Enhanced astaxanthin production from a novel strain of Haematococcus pluvialis using fulvic acid.Process Biochemistry,50(12):2072-2077
巩彪,史庆华,2017.园艺作物褪黑素的研究进展.中国农业科学,50(12):2326-2337
梁竹,魏玮,陶利,2014.褪黑素在治疗睡眠障碍中的研究进展.解放军药学学报,30(1):82-85
梁英,冯力霞,尹翠玲等,2007.叶绿素荧光技术在微藻环境胁迫研究中的应用现状及前景.海洋科学,31(1):71-76
梁英,陈书秀,2009.温度对雨生红球藻叶绿素荧光特性及虾青素含量的影响.海洋湖沼通报,(3):112-120
韩春梅,刘建国,张勇,2014.不同激素配伍对雨生红球藻(Haematococcus pluvialis)细胞生长和虾青素累积的调节作用、藻株差异及应用.海洋与湖沼,40(4):430-436
Arnon D I,1949.Copper enzymes in isolated chloroplasts.Polyphenoloxidase in Beta vulgaris.Plant Physiology,24(1):1-15
Berges J A,Fisher A E,Harrison P J,1993.A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana.Marine Biology,115(2):187-193
Chen G Q,Wang B B,Han D X et al,2015.Molecular mechanisms of the coordination between astaxanthin and fatty acid biosynthesis in Haematococcus pluvialis(Chlorophyceae).The Plant Journal,81(1):95-107
Ebrahimian A,Kariminia H R,Vosoughi M,2014.Lipid production in mixotrophic cultivation of Chlorella vulgaris in a mixture of primary and secondary municipal wastewater.Renewable Energy,71:502-508
Fasstt R G,Coombes J S,2012.Astaxanthin in cardiovascular health and disease.Molecules,17(2):2030-2048
Gao Z Q,Meng C X,Zhang X W et al,2012a.Differential expression of carotenogenic genes,associated changes on astaxanthin production and photosynthesis features induced by JA in H.pluvialis.PLoS One,7(8):e42243
Gao Z Q,Meng C X,Zhang X W et al,2012b.Induction of salicylic acid(SA)on transcriptional expression of eight carotenoid genes and astaxanthin accumulation in Haematococcus pluvialis.Enzyme and Microbial Technology,51(4):225-230
Guerin M,Huntley M E,Olaizola M,2003.Haematococcus astaxanthin:applications for human health and nutrition.Trends in Biotechnology,21(5):210-216
Gwak Y,Hwang Y S,Wang B et al,2014.Comparative analyses of lipidomes and transcriptomes reveal a concerted action of multiple defensive systems against photooxidative stress in Haematococcus pluvialis.Journal of Experimental Botany,65(15):4317-4334
Han D X,Li Y T,Hu Q,2013.Astaxanthin in microalgae:pathways,functions and biotechnological implications.Algae,28(2):131-147
Huang J C,Chen F,Sandmann G,2006.Stress-related differential expression of multipleβ-carotene ketolase genes in the unicellular green alga Haematococcus pluvialis.Journal of Biotechnology,122(2):176-185
Huang W P,Ye J R,Zhang J J et al,2016.Transcriptome analysis of Chlorella zofingiensis to identify genes and their expressions involved in astaxanthin and triacylglycerol biosynthesis.Algal Research,17:236-243
Kobayashi M,Todoroki Y,Hirai N,et al,1998.Biological activities of abscisic acid analogs in the morphological change of the green alga Haematococcus pluvialis.Journal of Fermentation and Bioengineering,85(5):529-531
Li D F,Zhao Y T,Ding W et al,2017.A strategy for promoting lipid production in green microalgae Monoraphidium sp.QLY-1 by combined melatonin and photoinduction.Bioresource Technology,235:104-112
Li J,Zhu D L,Niu J F et al,2011.An economic assessment of astaxanthin production by large scale cultivation of Haematococcus pluvialis.Biotechnology Advances,29(6):568-574
Liang C,Zhai Y,Xu D et al,2015.Correlation between lipid and carotenoid synthesis and photosynthetic capacity in Haematococcus pluvialis grown under high light and nitrogen deprivation stress.Grasas Aceites,66(2):e077
Lu Y D,Jiang P,Liu S F et al,2010.Methyl jasmonate-or gibberellins A3-induced astaxanthin accumulation is associated with up-regulation of transcription ofβ-carotene ketolase genes(bkts)in microalga Haematococcus pluvialis.Bioresource Technology,101(16):6468-6474
Ma X N,Liu J,Liu B et al,2016.Physiological and biochemical changes reveal stress-associated photosynthetic carbon partitioning into triacylglycerol in the oleaginous marine alga Nannochloropsis oculata.Algal Research,16:28-35
Poonkum W,Powtongsook S,Pavasant P,2015.Astaxanthin induction in microalga H.pluvialis with flat panel airlift photobioreactors under indoor and outdoor conditions.Preparative Biochemistry and Biotechnology,45(1):1-17
Raman V,Ravi S,2011.Effect of salicylic acid and methyl jasmonate on antioxidant systems of Haematococcus pluvialis.Acta Physiologiae Plantarum,33(3):1043-1049
Reiter R J,Tan D X,Osuna C et al,2000.Actions of melatonin in the reduction of oxidative stress.Journal of Biomedical Science,7(6):444-458
Shang M M,Ding W,Zhao Y T et al,2016.Enhanced astaxanthin production from Haematococcus pluvialis using butylated hydroxyanisole.Journal of Biotechnology,236:199-207
Shi H,Jiang C,Ye T et al,2015.Comparative physiological,metabolomic and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass(Cynodon dactylon(L).Pers.)by exogenous melatonin.Journal of Experimental Botany,66(3):681-694
Su Y X,Wang J X,Shi M L et al,2014.Metabolomic and network analysis of astaxanthin-producing Haematococcus pluvialis under various stress conditions.Bioresource Technology,170:522-529
Tan D X,Manchester L C,Hardeland R et al,2003.Melatonin:a hormone,a tissue factor,an autocoid,a paracoid,and an antioxidant vitamin.Journal of Pineal Research,34(1):75-78
Tan D X,Hardeland R,Manchester L C et al,2012.Functional roles of melatonin in plants,and perspectives in nutritional and agricultural science.Journal of Experimental Botany,63(2):577-597
Wei W,Li Q T,Chu Y N et al,2015.Melatonin enhances plant growth and abiotic stress tolerance in soybean plants.Journal of Experimental Botany,66(3):695-707
Wen Z W,Liu Z Y,Hou Y Y et al,2015.Ethanol induced astaxanthin accumulation and transcriptional expression of carotenogenic genes in Haematococcus pluvialis.Enzyme and Microbial Technology,78:10-17
Zhao Y T,Shang M M,Xu J W et al,2015.Enhanced astaxanthin production from a novel strain of Haematococcus pluvialis using fulvic acid.Process Biochemistry,50(12):2072-2077