不同海洋微藻对石油污染的响应差异
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摘要
微藻对油污染物的不同响应,势必造成其群落种类组成和结构的变化,进一步影响海洋生态系统的变迁。由于油污染对于浮游植物的影响具有污染源及生物响应的多样性、多维度性和复杂性,产生影响的时效长但可显示度低等特点,造成该研究领域的极大限制。作者以石油标准品20-3和3种石油化学品(双酚、对苯二甲酸、对二甲苯)作为油污染源,利用多孔板高通量检测方法及大数据处理分析软件Simca-P,对中国沿海常见单种微型藻类的种群消长受油污染影响进行了研究。以石油标准品20-3为油污染源,分析了31株微藻增殖对不同油标浓度的不同响应;利用Simca-P软件对3种不同浓度的石油化学品对4种典型微藻(柔弱角刺藻(Chaetoceros debilis)、球等鞭金藻(Isochrysis galbana)、东海原甲藻(Prorocentrum donghaiense)、剧毒卡罗藻(Karlodinum veneficum))增殖的影响进行了比较分析。研究结果显示:不同种类微藻因油污种类及其浓度不同,其响应存在显著差异。利用多孔板以及微藻的吸光特征,可以高通量快速筛查不同微藻对不同油污及其不同浓度的响应差异,并利用计算机软件对大数据的分析能力,挖掘和比较不同微藻与不同油污之间的相互关系,从而就油污染对微藻影响这种多维度复杂关系进行研究。
Microalgae are a fundamental component of aquatic food web. Oil spills or the release of other sources of hydrocarbons into the marine environments account for most of the organic pollution that occurs in coastal areas. Different species of microalgae will respond differently to oil pollution, producing different downstream effects on phytoplankton communities and possibly reshaping marine ecosystems. Research into the response of different marine microalgae to oil pollution has largely been held back by the high degree of complexity that exists in the relationship between various phytoplankton species and the many different oil pollution scenarios that are possible. The present study uses a high-throughput screening method to examine the responses of different microalgae to oil pollution. This is accomplished using oil standard 20-3 and 3 oil chemical products: bisphenol, terephthalic acid, and paraxylene, as sources of oil pollution together with 30 strains of microalgae isolated from the coastal area of China. For the first time, principle component analysis(PCA) based on Simca-P software was used to analyze a large data set measuring population growth of microalgae over 72 h in the presence of various oil pollutants. The resultsshow that the growth of different strains of microalgae was affected differently by the oil pollutants and the effect of oil pollutants was concentration-dependent. Multiwell culture plates, e.g., 24-well plate, were efficient growth vehicles for algae growth when used in the high-throughput screening procedure. Analysis of this large data set showing microalgal population growth in response to various oil pollutants of different concentration can be performedeffectively by applying the method of PCA based on the computer software Simca-P.
Microalgae are a fundamental component of aquatic food web. Oil spills or the release of other sources of hydrocarbons into the marine environments account for most of the organic pollution that occurs in coastal areas. Different species of microalgae will respond differently to oil pollution, producing different downstream effects on phytoplankton communities and possibly reshaping marine ecosystems. Research into the response of different marine microalgae to oil pollution has largely been held back by the high degree of complexity that exists in the relationship between various phytoplankton species and the many different oil pollution scenarios that are possible. The present study uses a high-throughput screening method to examine the responses of different microalgae to oil pollution. This is accomplished using oil standard 20-3 and 3 oil chemical products: bisphenol, terephthalic acid, and paraxylene, as sources of oil pollution together with 30 strains of microalgae isolated from the coastal area of China. For the first time, principle component analysis(PCA) based on Simca-P software was used to analyze a large data set measuring population growth of microalgae over 72 h in the presence of various oil pollutants. The resultsshow that the growth of different strains of microalgae was affected differently by the oil pollutants and the effect of oil pollutants was concentration-dependent. Multiwell culture plates, e.g., 24-well plate, were efficient growth vehicles for algae growth when used in the high-throughput screening procedure. Analysis of this large data set showing microalgal population growth in response to various oil pollutants of different concentration can be performedeffectively by applying the method of PCA based on the computer software Simca-P.
引文
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[3]王修林,李克强.渤海主要化学污染物海洋环境容量[M].北京:科学出版社,2006:12-113.Wang Xiulin,Li Keqiang.The marine environmental capacity of mainly chemical pollutants in the Bohai Sea[M].Beijing:Science Press,2006:12-113.
[4]沈南南,李纯厚,王晓伟.石油污染对海洋浮游生物的影响[J].生物技术通报,2006(增刊):95-99.Shen Nannan,Li Chunhou,Wang Xiaowei.Effects of oil pollution on marine planktons[J].Biotechnology Bulletin,2006(s):95-99.
[5]黄逸君,陈全震,曾江宁,等.石油污染对海洋浮游植物群落生长的影响[J].生态学报,2011,31(2):513-521.Huang Yijun,Chen Quanzhen,Zeng Jiangning,et al.The impact of oil pollution on marine phytoplankton community growth chang[J].Acta Ecologica Sinica,2011,31(2):513-521.
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[7]高尚德.集油对几种海藻光合作用、呼吸作用影响的初步研究[J].环境科学学报,1986,6(8):11.Gao Shangde.Oil to the influence of several kinds of algal photosynthesis,respiration,a preliminary study[J].Acta Scientiae Circumstantiae,1986,6(8):11.
[8]陈中伟,徐韧,刘材材,等.不同季节轻质油浓度对浮游植物的影响[J].上海海洋大学学报,2015,24(2):272-279.Chen Zhongwei,Xu Ren,Liu Caicai,et al.The effect of light oil concentration on microalgae in different season[J].Journal of Shanghai Ocean University,2015,24(2):272-279.
[9]Ozhan K,Bargu S.Distinct responses of Gulf of Mexico phytoplankton communities to crude oil and the dispersant corexit?Ec9500A under different nutrient regimes[J].Ecotoxicology,2014,23:370-385.
[10]Selala C,Botha A M,Klerk L P de,et al.Using phytoplankton diversity to determine wetland resilience,one year after a vegetable oil spill[J].Water Air Soil Pollut,2014,225:2051.
[11]程振波.渤海饵料生物培养[M].北京:农业出版社,1992:69-75.Cheng Zhenbo.The Bohai Sea bait biological cultivation[M].Beijing:Agriculture Press,1992:69-75.
[12]王伟杰,吴长江.我国海洋石油污染对渔业的危害及其防治[J].海洋信息,1993,3:2-27.Wang Weijie,Wu Changjiang.China's marine oil pollution and its prevention countermeasures to the harm of fisheries[J].Marine Information,1993,3:2-27.
[13]王晓伟,李纯厚,沈南南.石油污染对海洋生物的影响[J].南方水产,2006,2(2):76-80.Wang Xiaowei,Li Chunhou,Shen Nannan.Effect of oil pollution on marine organism[J].South China Fisheries Science,2006,2(2):76-80.
[14]倪朝辉,翟良安.石油对鱼类等水生生物的毒性[J].淡水渔业,1997,27(6):38-40.Ni Zhaohui,Zhai Liangan.Oil toxicity to fish and other aquatic organisms[J].Fresh Water Fisheries,1997,27(6):38-40.
[15]宋光彪,王伟洪,张杰,等.石油烃污染对海洋浮游植物的影响[J].环境科学与管理,2013,38(3):84-87.Song Guangbiao,Wang Weihong,Zhang Jie,et al.Influence of petroleum hydrocarbon pollution on marine phytoplankton[J].Environmental Scienceand Management,2013,38(3):84-87.
[16]王君丽,刘春光,冯剑丰,等.石油烃对海洋浮游植物生长的影响研究进展[J].环境污染与防治,2011,4(4):81-86.Wang Junli,Liu Chunguang,Feng Jianfeng,et al.Literature review of the effects of petroleum hydrocarbons on marine phytoplankton[J].Environmental Pollution&Control,2011,4(4):81-86.
[17]张雷,王修林,韩秀荣,等.石油烃污染对海洋浮游植物生长的影响——实验与模型[J].青岛海洋大学学报,2002,32(5):804-810.Zhang Lei,Wang Xiulin,Han Xiurong,et al.Effects of petroleum hydrocarbon on the growth of marine algae experiment versus model[J].Journal of Ocean University of Qingdao,2002,32(5):804-810.
[18]Hing L S,Ford T,Finch P,et al.Laboratory stimulation of oil-spill effects on marine phytoplankton[J].Aquatic Toxicology,2011,103:32-37.
[19]Hjorth M.Plankton stress responses from PAH exposure and nutrient enrichment[J].Marine Ecology Progress Series,2008,363:121-130.
[20]Carrera-martinez D,Mateos-sanz A,López-rodas V,et al.Microalgae response to petroleum spill:An experimental model analyzing physiological and genetic response of Dunaliella tertiolecta(Chlorophyceae)to oil samples from the tanker Prestige[J].Aquatic Toxicology,2010,97:151-159.
[21]Siron R,Pelletier E R,Roy S.Effects of dispersed and adsorbed crude oil on microalgal and bacterial communities of cold seawater[J].Ecotoxicology,1996,5(4):229-251.
[22]Fabregas J,Herreroe C,Veiga M.Effect of oil and dispersant on growth and chlorophyll a content of the marine microalga Tetraselmis suecica[J].Applied and Environmental Microbiology,1984,47(2):445-447.
[23]Megharej M,Singleton I,Mcclure N C,et al.Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil[J].Archives of Environmental Contamination Toxicology,2000,38:439-445.
[24]黄逸君,江志兵,曾江宁,等.石油烃污染对海洋浮游植物群落的短期毒性效应[J].植物生态学报,2010,34(9):1095-1106.Huang Yijun,Jiang Zhibing,Zeng Jiangning,et al.Short-term toxic effects of crude oil pollution on marine phytoplankton community[J].Chinese Journal of Plant Ecology,2010,34(9):1095-1106.
[25]刘瑀,姚敬元,李颖,等.溢油胁迫下海洋微藻脂肪酸合成过程中碳稳定同位素分馏效应[J].海洋环境科学,2015,34(1):54-65.Liu Yu,Yao Jingyuan,Li Ying.Carbon stable isotopic fractionation associated with biosynthesis of fatty acids by marine microalgae under the stress of oil spills[J].Marine Environmental Science,2015,34(1):54-65.
[26]Radix P,Leonard M,Papantoniou C,et al.Comparison of four chronic toxicity tests using algae,bacteria,and invertebrates assessed with sixteen chemicals[J].Ecotoxicology and Environmental Safety,2000,47:186-194.
[27]O’brien P Y,Dixon P S.The effects of oils and oilcomponents on algae:A review[J].British Phycological Journal,1976,11:115-142.
[28]?zhan K,Miles S M,Gao H,et al.Relative phytoplankton growth responses to physically and chemically dispersed South Louisiana sweet crude oil[J].Environ Monit Assess,2014,186:3941-3956.
[29]王摆,高士博,董颖,等.6种苯系物对球等鞭金藻和新月菱形藻的生长抑制[J].生态毒理学报,2014,9(2):233-238.Wang Bai,Gao Shibo,Dong Ying,et al.Inhibitory effect of six kinds of BTEXs on growth of the Chrysophyceae isochrysisgalbana and the diatom Nitzschia closterium[J].Asian Journal of Ecotoxicology,2014,9(2):233-238.
[30]Pareons M L,Turner R E,Overton E B.Sediment-preserved diatom assemblages can distinguish a petroleumactivity signal separately from the nutrient signal of the Mississippi River in coastal Louisiana[J].Marine Pollution Bulletin,2014,85:164-171.
[2]李贵珍,赖其亮,闫培生,等.海洋石油污染及其微生物修复研究进展[J].生物技术进展,2015,5(3):164-169.Li Guizhen,Lai Qiliang,Yan Peisheng,et al.Advance on marine petroleum pollution and microbial remediation[J].Current Biotechnology,2015,5(3):164-169.
[3]王修林,李克强.渤海主要化学污染物海洋环境容量[M].北京:科学出版社,2006:12-113.Wang Xiulin,Li Keqiang.The marine environmental capacity of mainly chemical pollutants in the Bohai Sea[M].Beijing:Science Press,2006:12-113.
[4]沈南南,李纯厚,王晓伟.石油污染对海洋浮游生物的影响[J].生物技术通报,2006(增刊):95-99.Shen Nannan,Li Chunhou,Wang Xiaowei.Effects of oil pollution on marine planktons[J].Biotechnology Bulletin,2006(s):95-99.
[5]黄逸君,陈全震,曾江宁,等.石油污染对海洋浮游植物群落生长的影响[J].生态学报,2011,31(2):513-521.Huang Yijun,Chen Quanzhen,Zeng Jiangning,et al.The impact of oil pollution on marine phytoplankton community growth chang[J].Acta Ecologica Sinica,2011,31(2):513-521.
[6]Smith J E.Torrey canyon pollution and marine life[M].London:Cambridge University Press,168-196.
[7]高尚德.集油对几种海藻光合作用、呼吸作用影响的初步研究[J].环境科学学报,1986,6(8):11.Gao Shangde.Oil to the influence of several kinds of algal photosynthesis,respiration,a preliminary study[J].Acta Scientiae Circumstantiae,1986,6(8):11.
[8]陈中伟,徐韧,刘材材,等.不同季节轻质油浓度对浮游植物的影响[J].上海海洋大学学报,2015,24(2):272-279.Chen Zhongwei,Xu Ren,Liu Caicai,et al.The effect of light oil concentration on microalgae in different season[J].Journal of Shanghai Ocean University,2015,24(2):272-279.
[9]Ozhan K,Bargu S.Distinct responses of Gulf of Mexico phytoplankton communities to crude oil and the dispersant corexit?Ec9500A under different nutrient regimes[J].Ecotoxicology,2014,23:370-385.
[10]Selala C,Botha A M,Klerk L P de,et al.Using phytoplankton diversity to determine wetland resilience,one year after a vegetable oil spill[J].Water Air Soil Pollut,2014,225:2051.
[11]程振波.渤海饵料生物培养[M].北京:农业出版社,1992:69-75.Cheng Zhenbo.The Bohai Sea bait biological cultivation[M].Beijing:Agriculture Press,1992:69-75.
[12]王伟杰,吴长江.我国海洋石油污染对渔业的危害及其防治[J].海洋信息,1993,3:2-27.Wang Weijie,Wu Changjiang.China's marine oil pollution and its prevention countermeasures to the harm of fisheries[J].Marine Information,1993,3:2-27.
[13]王晓伟,李纯厚,沈南南.石油污染对海洋生物的影响[J].南方水产,2006,2(2):76-80.Wang Xiaowei,Li Chunhou,Shen Nannan.Effect of oil pollution on marine organism[J].South China Fisheries Science,2006,2(2):76-80.
[14]倪朝辉,翟良安.石油对鱼类等水生生物的毒性[J].淡水渔业,1997,27(6):38-40.Ni Zhaohui,Zhai Liangan.Oil toxicity to fish and other aquatic organisms[J].Fresh Water Fisheries,1997,27(6):38-40.
[15]宋光彪,王伟洪,张杰,等.石油烃污染对海洋浮游植物的影响[J].环境科学与管理,2013,38(3):84-87.Song Guangbiao,Wang Weihong,Zhang Jie,et al.Influence of petroleum hydrocarbon pollution on marine phytoplankton[J].Environmental Scienceand Management,2013,38(3):84-87.
[16]王君丽,刘春光,冯剑丰,等.石油烃对海洋浮游植物生长的影响研究进展[J].环境污染与防治,2011,4(4):81-86.Wang Junli,Liu Chunguang,Feng Jianfeng,et al.Literature review of the effects of petroleum hydrocarbons on marine phytoplankton[J].Environmental Pollution&Control,2011,4(4):81-86.
[17]张雷,王修林,韩秀荣,等.石油烃污染对海洋浮游植物生长的影响——实验与模型[J].青岛海洋大学学报,2002,32(5):804-810.Zhang Lei,Wang Xiulin,Han Xiurong,et al.Effects of petroleum hydrocarbon on the growth of marine algae experiment versus model[J].Journal of Ocean University of Qingdao,2002,32(5):804-810.
[18]Hing L S,Ford T,Finch P,et al.Laboratory stimulation of oil-spill effects on marine phytoplankton[J].Aquatic Toxicology,2011,103:32-37.
[19]Hjorth M.Plankton stress responses from PAH exposure and nutrient enrichment[J].Marine Ecology Progress Series,2008,363:121-130.
[20]Carrera-martinez D,Mateos-sanz A,López-rodas V,et al.Microalgae response to petroleum spill:An experimental model analyzing physiological and genetic response of Dunaliella tertiolecta(Chlorophyceae)to oil samples from the tanker Prestige[J].Aquatic Toxicology,2010,97:151-159.
[21]Siron R,Pelletier E R,Roy S.Effects of dispersed and adsorbed crude oil on microalgal and bacterial communities of cold seawater[J].Ecotoxicology,1996,5(4):229-251.
[22]Fabregas J,Herreroe C,Veiga M.Effect of oil and dispersant on growth and chlorophyll a content of the marine microalga Tetraselmis suecica[J].Applied and Environmental Microbiology,1984,47(2):445-447.
[23]Megharej M,Singleton I,Mcclure N C,et al.Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil[J].Archives of Environmental Contamination Toxicology,2000,38:439-445.
[24]黄逸君,江志兵,曾江宁,等.石油烃污染对海洋浮游植物群落的短期毒性效应[J].植物生态学报,2010,34(9):1095-1106.Huang Yijun,Jiang Zhibing,Zeng Jiangning,et al.Short-term toxic effects of crude oil pollution on marine phytoplankton community[J].Chinese Journal of Plant Ecology,2010,34(9):1095-1106.
[25]刘瑀,姚敬元,李颖,等.溢油胁迫下海洋微藻脂肪酸合成过程中碳稳定同位素分馏效应[J].海洋环境科学,2015,34(1):54-65.Liu Yu,Yao Jingyuan,Li Ying.Carbon stable isotopic fractionation associated with biosynthesis of fatty acids by marine microalgae under the stress of oil spills[J].Marine Environmental Science,2015,34(1):54-65.
[26]Radix P,Leonard M,Papantoniou C,et al.Comparison of four chronic toxicity tests using algae,bacteria,and invertebrates assessed with sixteen chemicals[J].Ecotoxicology and Environmental Safety,2000,47:186-194.
[27]O’brien P Y,Dixon P S.The effects of oils and oilcomponents on algae:A review[J].British Phycological Journal,1976,11:115-142.
[28]?zhan K,Miles S M,Gao H,et al.Relative phytoplankton growth responses to physically and chemically dispersed South Louisiana sweet crude oil[J].Environ Monit Assess,2014,186:3941-3956.
[29]王摆,高士博,董颖,等.6种苯系物对球等鞭金藻和新月菱形藻的生长抑制[J].生态毒理学报,2014,9(2):233-238.Wang Bai,Gao Shibo,Dong Ying,et al.Inhibitory effect of six kinds of BTEXs on growth of the Chrysophyceae isochrysisgalbana and the diatom Nitzschia closterium[J].Asian Journal of Ecotoxicology,2014,9(2):233-238.
[30]Pareons M L,Turner R E,Overton E B.Sediment-preserved diatom assemblages can distinguish a petroleumactivity signal separately from the nutrient signal of the Mississippi River in coastal Louisiana[J].Marine Pollution Bulletin,2014,85:164-171.