温、光、盐对硅藻STR01生长、总脂、脂肪酸的影响
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摘要
为了优化新分离STR01的生态培养条件,采用单因子试验和正交试验研究了不同温度、光照强度、盐度和温、光、盐三因素三水平对该藻的生长、总脂和脂肪酸组成影响。结果表明:温、光、盐对STR01的生长、总脂和脂肪酸组成影响显著(P<0.05)。生长的适宜温度为15—35℃,最适25—30℃(K值达0.679—0.682),总脂含量积累的最适温度是25℃(总脂可达17.23%),温度20℃时有利于该藻PUFA的积累,可达34.23%。STR01生长的适宜光照强度为40—120μmol/(m~2·s),最适光强为60μmol/(m~2·s),光照强度40μmol/(m~2·s)有利于该藻的PUFA积累,可达34.29%。STR01生长的适宜盐度为10—35,最适盐度25,盐度25时PUFA含量较高(43.42%)。正交试验结果表明温度对STR01的平均相对生长速率和总脂含量影响显著,生长的最优组合:温度30℃、光照强度60μmol/(m~2·s)、盐度25,该组合下的生长速率达0.756;总脂含量积累的最优组合:温度30℃、光照强度60μmol/(m~2·s)、盐度20,该组合下的总脂含量为20.00%。PUFA的最优组合:温度25℃、光照强度60μmol/(m~2·s)、盐度20,该组合下PUFA的含量为35.37%。综上所述:该藻生长迅速,总脂含量较高,PUFA丰富,是一种可开发利用的耐高温浮游硅藻。
To optimize the cultivation condition of microalgae, single-factor design and orthogonal experiments were performed to assess the growth rate, total lipid content and fatty acid composition of STR01 by temperature, light intensity and salinity. The results showed that all these ecological factors exerted significant effects on the growth, total lipid content and fatty acid composition of STR01(P< 0.05) with the appropriate temperature for growth at 15—35℃,the optimum temperature at 25—30℃(K-value 0.679—0.682) and the optimum temperature for total lipid content accumulation at 25℃(17.23%). The highest PUFA content at 34.23% appeared at 20℃. The appropriate light intensity for growth was 40—120 μmol/(m~2·s) and the optimum one was 60 μmol/(m~2·s). The light intensity of 40 μmol/(m~2·s)had the highest PUFA accumulation at 34.29%. The appropriate salinity for growth was 10—35 and the optimum salinity was 25. The orthogonal experiments showed that the optimum condition for growth was temperature 30℃, light intensity 60 μmol/(m~2·s), and salinity 25, and that the optimum total lipid accumulation was temperature 30℃, light intensity 60 μmol/(m~2·s) and salinity 20, and that the optimum PUFA content was temperature 25℃, light intensity60 μmol/(m~2·s), and salinity 20 at 37%. In conclusion, STR01 was promising prospect in future aquaculture with fast growth rate and high total lipid and PUFA contents.
To optimize the cultivation condition of microalgae, single-factor design and orthogonal experiments were performed to assess the growth rate, total lipid content and fatty acid composition of STR01 by temperature, light intensity and salinity. The results showed that all these ecological factors exerted significant effects on the growth, total lipid content and fatty acid composition of STR01(P< 0.05) with the appropriate temperature for growth at 15—35℃,the optimum temperature at 25—30℃(K-value 0.679—0.682) and the optimum temperature for total lipid content accumulation at 25℃(17.23%). The highest PUFA content at 34.23% appeared at 20℃. The appropriate light intensity for growth was 40—120 μmol/(m~2·s) and the optimum one was 60 μmol/(m~2·s). The light intensity of 40 μmol/(m~2·s)had the highest PUFA accumulation at 34.29%. The appropriate salinity for growth was 10—35 and the optimum salinity was 25. The orthogonal experiments showed that the optimum condition for growth was temperature 30℃, light intensity 60 μmol/(m~2·s), and salinity 25, and that the optimum total lipid accumulation was temperature 30℃, light intensity 60 μmol/(m~2·s) and salinity 20, and that the optimum PUFA content was temperature 25℃, light intensity60 μmol/(m~2·s), and salinity 20 at 37%. In conclusion, STR01 was promising prospect in future aquaculture with fast growth rate and high total lipid and PUFA contents.
引文
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[15]Aaronson S.Effect of incubation temperature on the macromolecular and lipid content of the phytoflagellate Ochromonas danica[J].Journal of Phycology,1973,9(1):111-113
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[17]Xu N,LüS H,Chen J F,et al.The influence of water temperature and salinity on the growth of Scrippsiella trochoidea[J].Marine Environmenal Science,2004,3(23):36-38[徐宁,吕颂辉,陈菊芳,等.温度和盐度对锥状斯氏藻生长的影响.海洋环境科学,2004,3(23):36-38]
[18]Ye L,Jiang X M,Mao X X,et al.Effects of temperature,light intensity and salinity on the growth total lipid and fatty acid of Phaeodactylum tricornutum mutant[J].Chinese Journal of Ecology,2015,34(2):454-462[叶丽,蒋霞敏,毛欣欣,等.温、光、盐对三角褐指藻紫外诱变株生长、总脂及脂肪酸的影响.生态学杂志,2015,34(2):454-462]
[19]Cao C H,Sun S C,Mai K S,et al.Effect of light intensity on the total lipid contents and fatty acid composition in 4 strains of marine green algae[J].Acta Ecologica Sinica,2010,30(9):2347-2353[曹春晖,孙世春,麦康森,等.光照强度对四株海洋绿藻总脂含量和脂肪酸组成的影响.生态学报,2010,30(9):2347-2353]
[20]Xu N J,Zhang X C.Effects of temperature,light intensity and pH on the Growth and fatty acid compositions of Ellipsoidion sp.[J].Journal of Ocean University of Qingdao,2001,31(4):541-547[徐年军,张学成.温度、光照、pH值对后棘藻生长及脂肪酸含量的影响.青岛海洋大学学报,2001,31(4):541-547]
[21]Anning T,Macintyre H,PrattS M,et al.Photoacclimation in the marine diatom Skeletonema costatum[J].Limnology and Oceanography,2000,45(18):1807-1817
[22]Gao X Z,Jing X M,Ye L.Effects of temperature,light intensity and salinity on the growth and fatty acid composition of Skeletlnema munzelii SM-1 and SM-2[J].Journal of Biology,2014,31(6):64-69[高秀芝,蒋霞敏,叶丽.温度、光照和盐度对2株曼氏骨条藻生长及脂肪酸组成的影响.生态学杂志,2014,31(6):64-69]
[23]Miao J L,Wang B,Kan G F,et al.The influence of environment factors on lipid content and fatty acid composition in two species of Antarctic green microalga[J].Marine Sciences,2005,29(1):4-11[缪锦来,王波,阚光锋,等.环境因子对2种南极绿藻脂肪含量和脂肪酸组成的影响.海洋科学,2005,29(1):4-11]
[24]Bao J,Ting X L,Dong S L,et al.Effect of temperature salinity and light intensity on nitrogen and phosphorus uptake by Sargassum thunbergii[J].Journal of Fishery Sciences of China,2008,2(15):293-300[包杰,田相利,董双林,等.温度、盐度和光照强度对鼠尾藻氮、磷吸收的影响.中国水产科学,2008,2(15):293-300]
[25]Guo J,Yang W D,Liu J S,et al.Effects of salinity,temperature and light intensity on the growth and toxin production of Phaeocystis globose[J].Acta Scientiae Circumstantiae,2007,8(27):1341-1346[郭瑾,杨维东,刘洁生.温度、盐度和光照对球形棕囊藻生长和产毒的影响研究.环境科学学报,2007,8(27):1341-1346]
[26]Kuwada Y,Ohta Y.Effect of salinity on hydrogen production and growth of Lyngbya sp[J].Journal of the Faculty of Applied Hiroshima University,1991,30(1):13-17
[27]Zhang Q T,Hu G K.Study on effect of salinity on the growth of micro algae Isochrysis[J].Journal of Salt and Chemical Industry,2009,6(38):8-10[张青田,胡桂坤.盐度对等鞭金藻生长的影响研究.盐业与化工,2009,6(38):8-10]
[28]Du X F,Zou N,Sun D H,et al.Optimization of oil-rich cultural conditions of Nannochloropsis sp[J].China Oils and Fats,2012,37(12):66-69[杜晓凤,邹宁,孙东红,等.微绿球藻富集油脂的培养条件优化.微生物油脂,2012,37(12):66-69]
[2]Tang H.Removal of Nitrogen and phosphorus in sewage by immobilized microalgae[D].Nanjing Agricultural University.2016[唐皓.固定化微藻去除污水中氮磷的研究.南京农业大学.2016]
[3]Shi J,Pan K H.Effects of different culture conditions and growth phases on lipid of microalgae[J].Marine Fisheries Research,2004,25(6):79-85[石娟,潘克厚.不同培养条件对微藻总脂含量和脂肪酸组成的影响.海洋水产研究,2004,25(6):79-85]
[4]Jiang X M.Effects of temperatures,light intensity and nitrogen concentrations on the growth and fatty acid composition of Nannochloropsis oculata[J].Marine Sciences,2002,26(8):9-13[蒋霞敏.温度、光照、氮含量对微绿球藻生长及脂肪酸组成的影响.海洋科学,2002,26(8):9-13]
[5]Yang X X,Yu H,Zeng X Q.Summarization of the factors affecting the fatty acid composition of microalgae[J].Transactions of Oceanology and Limnology,2001,1:76-80[杨秀霞,于浩,曾晓起.影响微藻脂肪酸组成因素概述.海洋湖沼通报,2001,1:76-80]
[6]Alonso D L,Grima E M,Perez J A S,et al.Isolation of clones of Isochrysis galbanarich in eicosapentaenoic acid[J].Aquaculture,1992,102(4):363-371
[7]Bligh E G,Dyer W J.A rapid method of total lipid extraction and purification[J].Canadian Journal of Biochemistry and Physiology,1959,37(8):911-917
[8]Xu J L,Yan X J,Zhou C X,et al.The effects of air bubbling condition on fatty acid composition in 19 strains of marine microalgae[J].Journal of Ningbo University(NSEE),2006,19(2):180-185[徐继林,严小军,周成旭,等.19种(株)海洋微藻脂肪酸组成及充气产生的影响.宁波大学学报:理工版,2006,19(2):180-185]
[9]Reiser S,Somerville C.Isolation of mutants of Acinetobacter calcoaceticus deficient in wax ester synthesis and complementation of one mutation with a gene encoding afatty acyl-coenzyme A reductase[J].Journal of Bacteriology,1997,179:2969-2975
[10]Jiang X M,Liu M H and Xing C G.Effect of different ecological conditions on the growth and fatty acid composition of Pavloav viridis[J].Acta Hydrobiologica Sinica,2007,31(1):88-93[蒋霞敏,柳敏海,邢晨光.不同生态条件对绿色巴夫藻生长及脂肪酸组成的影响.水生生物学报,2007,31(1):88-93]
[11]Pan J,Yu J Z,Ma X L,et al.Effect of temperature on the growth and fatty acid composition of Phaeodactylum tricornutum[J].Progress in Fishery Sciences,2010,31(6):91-94[潘瑾,俞建中,马晓磊,等.温度影响三角褐指藻生长和脂肪酸组成的初步探讨.渔业科学进展,2010,31(6):91-94]
[12]Woldman J C,Mann R.Temperature influenced variations in speciation and the chemical composition of marine phytoplankton in outdoormass cultures[J].Journal of Experimental Marine Biology and Ecology,1980,46:29-40
[13]Jin D X,Chen D X,Liu S C,et al.Influence of temperature and salinity on the growth of planktonic Diatoms[J].Oceanologia et Limnologia Sinnica,1964,7(4):373-383[金德祥,陈真奋,刘师成,等.温度和盐度对三种海洋浮游硅藻生长繁殖的影响.海洋与湖沼,1964,7(4):373-383]
[14]Zhu M,Zhang X C,Mao Y X,et al.Effects of temperature,salinity and illumination on the growth of Thalassiosira sp.[J].Marine Sciences,2003,27(12):58-60[朱明,张学成,茂云翔,等.温度、盐度及光照强度对海链藻(Thalassiosira sp.)生长的影响.海洋科学,2003,27(12):58-60]
[15]Aaronson S.Effect of incubation temperature on the macromolecular and lipid content of the phytoflagellate Ochromonas danica[J].Journal of Phycology,1973,9(1):111-113
[16]Opute,F.I.Studies on fat accumulation in Nitzschia palea[J].Annals of Botany,1974,38(4):889-902
[17]Xu N,LüS H,Chen J F,et al.The influence of water temperature and salinity on the growth of Scrippsiella trochoidea[J].Marine Environmenal Science,2004,3(23):36-38[徐宁,吕颂辉,陈菊芳,等.温度和盐度对锥状斯氏藻生长的影响.海洋环境科学,2004,3(23):36-38]
[18]Ye L,Jiang X M,Mao X X,et al.Effects of temperature,light intensity and salinity on the growth total lipid and fatty acid of Phaeodactylum tricornutum mutant[J].Chinese Journal of Ecology,2015,34(2):454-462[叶丽,蒋霞敏,毛欣欣,等.温、光、盐对三角褐指藻紫外诱变株生长、总脂及脂肪酸的影响.生态学杂志,2015,34(2):454-462]
[19]Cao C H,Sun S C,Mai K S,et al.Effect of light intensity on the total lipid contents and fatty acid composition in 4 strains of marine green algae[J].Acta Ecologica Sinica,2010,30(9):2347-2353[曹春晖,孙世春,麦康森,等.光照强度对四株海洋绿藻总脂含量和脂肪酸组成的影响.生态学报,2010,30(9):2347-2353]
[20]Xu N J,Zhang X C.Effects of temperature,light intensity and pH on the Growth and fatty acid compositions of Ellipsoidion sp.[J].Journal of Ocean University of Qingdao,2001,31(4):541-547[徐年军,张学成.温度、光照、pH值对后棘藻生长及脂肪酸含量的影响.青岛海洋大学学报,2001,31(4):541-547]
[21]Anning T,Macintyre H,PrattS M,et al.Photoacclimation in the marine diatom Skeletonema costatum[J].Limnology and Oceanography,2000,45(18):1807-1817
[22]Gao X Z,Jing X M,Ye L.Effects of temperature,light intensity and salinity on the growth and fatty acid composition of Skeletlnema munzelii SM-1 and SM-2[J].Journal of Biology,2014,31(6):64-69[高秀芝,蒋霞敏,叶丽.温度、光照和盐度对2株曼氏骨条藻生长及脂肪酸组成的影响.生态学杂志,2014,31(6):64-69]
[23]Miao J L,Wang B,Kan G F,et al.The influence of environment factors on lipid content and fatty acid composition in two species of Antarctic green microalga[J].Marine Sciences,2005,29(1):4-11[缪锦来,王波,阚光锋,等.环境因子对2种南极绿藻脂肪含量和脂肪酸组成的影响.海洋科学,2005,29(1):4-11]
[24]Bao J,Ting X L,Dong S L,et al.Effect of temperature salinity and light intensity on nitrogen and phosphorus uptake by Sargassum thunbergii[J].Journal of Fishery Sciences of China,2008,2(15):293-300[包杰,田相利,董双林,等.温度、盐度和光照强度对鼠尾藻氮、磷吸收的影响.中国水产科学,2008,2(15):293-300]
[25]Guo J,Yang W D,Liu J S,et al.Effects of salinity,temperature and light intensity on the growth and toxin production of Phaeocystis globose[J].Acta Scientiae Circumstantiae,2007,8(27):1341-1346[郭瑾,杨维东,刘洁生.温度、盐度和光照对球形棕囊藻生长和产毒的影响研究.环境科学学报,2007,8(27):1341-1346]
[26]Kuwada Y,Ohta Y.Effect of salinity on hydrogen production and growth of Lyngbya sp[J].Journal of the Faculty of Applied Hiroshima University,1991,30(1):13-17
[27]Zhang Q T,Hu G K.Study on effect of salinity on the growth of micro algae Isochrysis[J].Journal of Salt and Chemical Industry,2009,6(38):8-10[张青田,胡桂坤.盐度对等鞭金藻生长的影响研究.盐业与化工,2009,6(38):8-10]
[28]Du X F,Zou N,Sun D H,et al.Optimization of oil-rich cultural conditions of Nannochloropsis sp[J].China Oils and Fats,2012,37(12):66-69[杜晓凤,邹宁,孙东红,等.微绿球藻富集油脂的培养条件优化.微生物油脂,2012,37(12):66-69]