洋河水库微囊藻毒素及产毒株种群丰度的时空分布特征
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摘要
在洋河水库设置6个采样点,于2015年5—10月采集水样,利用高效液相色谱测定了微囊藻毒素(MC-LR、MC-RR、MC-YR)的浓度,并采用基于mcy A和mcy B基因的荧光定量PCR技术检测产毒蓝藻与产毒微囊藻种群.同时,分析了水体中微囊藻毒素浓度和组成的时空分布特征,以及产毒蓝藻与产毒微囊藻丰度的动态变化.结果表明,洋河水库微囊藻毒素(Microcystins,MCs)浓度范围为0.24~10.99μg·L~(-1);水体中产毒蓝藻占总蓝藻种群丰度的7%~55%,产毒微囊藻占总微囊藻种群丰度的8%~59%,是主要的产毒蓝藻;微囊藻毒素浓度与产毒株种群丰度时空分布特征一致,季节变化规律为夏季>秋季>春季,空间分布表现为西洋河口附近区域高于东洋河口附近区域;而微囊藻毒素组成无显著时空差异,均以MC-RR和MC-LR 2种亚型为主.Pearson相关分析显示,产毒微囊藻种群丰度及其比例是影响微囊藻毒素浓度变化的主要因素,且其与总磷和叶绿素a浓度呈显著正相关(p<0.01).
Water samples were collected from six sites located in the Yanghe Reservoir of Qinhuangdao city to characterize the spatio-temporal distribution and dynamics of microcystins and microcystin-producing cells from May to October in 2015. The concentrations of MC-RR,MC-LR and MC-YR were analyzed using high-pressure liquid chromatography( HPLC). And the abundance of microcystin-producing cyanobacteria and Microcystis spp. were subsequently detected by real-time PCR assays,targeting the microcystin synthetase gene mcy A and mcy B respectively. The results show that the concentration of microcystins ranged from 0.24 to 10.99 μg·L~(-1)during the study period. The ratio of toxic cyanobacteria to total cyanobacteria population,and toxic Microcystis to total Microcystis population ranged in 7% ~ 55%,and 8% ~ 59% respectively. Microcystis was the dominant microcystins producer in the Yanghe Reservoir. Microcystins concentration in the reservoir corresponded well with the toxic strains abundance in the spatial and temporal scale,presenting a decreased trend from summer towards spring with higher concentration and abundance occurred in west area than that in east area of the reservoir. However,the microcystins composition show no significant difference in spatio-temporal patterns,which was made of MC-LR and MC-RR.Correlation analysis by Pearson indicates that the abundance and proportion of toxic Microcystis were the determine factors affecting the variation of microcystins concentration,whose growth were correlated with the concentrations of total phosphorus and chlorophyll a in the water column( p<0.01).
Water samples were collected from six sites located in the Yanghe Reservoir of Qinhuangdao city to characterize the spatio-temporal distribution and dynamics of microcystins and microcystin-producing cells from May to October in 2015. The concentrations of MC-RR,MC-LR and MC-YR were analyzed using high-pressure liquid chromatography( HPLC). And the abundance of microcystin-producing cyanobacteria and Microcystis spp. were subsequently detected by real-time PCR assays,targeting the microcystin synthetase gene mcy A and mcy B respectively. The results show that the concentration of microcystins ranged from 0.24 to 10.99 μg·L~(-1)during the study period. The ratio of toxic cyanobacteria to total cyanobacteria population,and toxic Microcystis to total Microcystis population ranged in 7% ~ 55%,and 8% ~ 59% respectively. Microcystis was the dominant microcystins producer in the Yanghe Reservoir. Microcystins concentration in the reservoir corresponded well with the toxic strains abundance in the spatial and temporal scale,presenting a decreased trend from summer towards spring with higher concentration and abundance occurred in west area than that in east area of the reservoir. However,the microcystins composition show no significant difference in spatio-temporal patterns,which was made of MC-LR and MC-RR.Correlation analysis by Pearson indicates that the abundance and proportion of toxic Microcystis were the determine factors affecting the variation of microcystins concentration,whose growth were correlated with the concentrations of total phosphorus and chlorophyll a in the water column( p<0.01).
引文
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Horst G P,Sarnelle O,White J D,et al.2014.Nitrogen availabilit increases the toxin quota of a harmful cyanobacterium,Microcysti aeruginosa[J].Water Research,54:188-198
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Nonneman D,Zimba P V.2002.A PCR-based test to assess the potentia for microcystin occurrence in channel catfish production ponds[J]Journal of Phycology,38:230-233
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Sevilla E,Martin Luna B,Vela L,et al.2008.Iron availability affects mcy Dexpression and microcystin-LR synthesis in Microcystis aeruginosa PCC7806[J].Environmental Microbiology,10(10):2476-2483
Singh S,Rai P K,Chau R,et al.2015.Temporal variations in microcystinproducing cells and microcystin concentrations in two fresh water ponds[J].Water Research,69:131-142
Song L R,Chen W,Peng L,et al.2007.Distribution and bioaccumulation of microcystins in water columns:A systematic investigation into the environmental fate and the risk associated with microcystins in Meiliang Bay,Lake Taihu[J].Water Research,41:2853-2864
Tillett D,Dittmann E,Erhard M,et al.2000.Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806:an integrated peptide-polyketide synthetase system[J].Chemistry and Biology,7(10):753-764
Wang X Y,Sun M J,Xie M J,et al.2013.Differences in microcystin production and genotype composition among Microcystis colonies of different sizes in Lake Taihu[J].Water Research,47(15):5659-5669
王菲凤,仝川,杨芳,等.2011.福州山仔水库水华微囊藻毒素时空分布特征[J].环境科学学报,31(3):533-546
王丽平,郑丙辉.2013.洋河水库富营养化发展趋势及其关键影响因素[J].生态学报,33(3):1011-1017
Yang Z,Kong F X,Shi X L,et al.2015.Effects of UV-B radiation on microcystin production of a toxic strain of Microcystis aeruginosa and its competitiveness agianist a non-toxic strain[J].Journal of Hazardous Materials,283:447-453
Yoshida M,Yoshida T,Takashima Y,et al.2007,Dynamics of microcystin-producing and non-microcystin-producing Microcystis populations is correlated with nitrate concentration in a Japanese lakes[J].FEMS Microbiology Letters,266(1):48-53
Yoshida M,Yoshida T,Kashima A,et al.2008,Ecological dynamics of the toxic bloom-forming cyanobacterium Microcystis aeruginosa and its cyanophages in freshwater[J].Applied and Environmental Microbiology,74(10):3269-3273
张丽媛,王圣瑞,储昭升,等.2010.洋河水库流域土壤与库区沉积物中磷形态特征研究[J].中国环境科学,30(11):1529-1536
赵伟,李涵,许秋瑾,等.2011.洋河水库水华优势种演替的原因分析[J].环境工程学报,5(9):1989-1993
Davis T W,Berry D L,Boyer G L,et al.2009.The effects of temperatur and nutrients on the growth and dynamics of toxic and non-toxi strains of Microcystis during cyanobacterial bloom[J].Harmfu Algae,8(5):715-725
高天珩,钱新,储昭升,等.2010.气候、水文和生态过程对洋河水库富营养化影响研究[J].水生态学杂志,3(3):28-31
国家环境保护总局.2002.水和废水监测分析方法(第四版)[M].北京:中国环境科学出版社
Hisbergues M,Christiansen G,Rouhiainen L,et al.2003.PCR-based identification of microcystin producing genotypes of differen cyanobacterial genera[J].Archives of Microbiology,180(6)402-410
Horst G P,Sarnelle O,White J D,et al.2014.Nitrogen availabilit increases the toxin quota of a harmful cyanobacterium,Microcysti aeruginosa[J].Water Research,54:188-198
Kaebernick M,Neilan B A,Borner T,et al.2000.Light and th transcriptional response of the microcystin biosynthesis gene cluste[J].Applied and Environmental Microbiology,66(8):3387-3392
Kim S G,Joung S H,Ahn C Y,et al.2010.Annual variation of Microcysti genotypes and their potential toxicity in water and sediment from eutrophic reservoir[J].FEMS Microbiology Ecology,74(1):93-102
Kuniyoshi T M,Sevilla E,Bes M T,et al.2013.Phosphate deficiency(N/P40∶1)induces mcy D transcription and microcystin synthesis in Microcystis aeruginosa PCC 7806[J].Plant Physiology and Biochemistry,65:120-124
Kurmayer R,Kutzenberger T.2003.Application of real-time PCR fo quantification of microcystin genotypes in a population of the toxi cyanobacterium Microcystis sp.[J].Applied and Environmenta Microbiology,69(11):6723-6730
Li R N,Chen Q W,Zhang X Q,et al.2015.Effects of temperature and macronutrients on phytoplankton communities across three largel different lakes identified by a time-space trade-off approach[J]Ecological Informatics,29:174-181
李大命,孔繁翔,阳振,等.2011.巢湖夏季和冬季有毒微囊藻和无毒微囊藻种群丰度研究[J].环境科学学报,31(12):2672-2680
李凤彬.2003.洋河水库富营养化的现状和对策探讨[J].给水排水,29(8):16-19
Pineda Mendoza R M,Zuniga G,Martinez Jeronimo F,et al.2014Infochemicals released by Daphnia magna fed on Microcysti aeruginosa affect mcy A gene expression[J].Toxin,80:78-86
Messineo V,Bogialli S,Melchiorre S,et al.2009.Cyanobacterial toxins in Italian freshwater[J].Limnologica,39:95-106
Neilan B A,Pearson L A,Muenchhoff J,et al.2013.Environmenta conditions that influence toxin biosynthesis in cyanobacteria[J]Environmental Microbiology,15(5):1239-1253
Nonneman D,Zimba P V.2002.A PCR-based test to assess the potentia for microcystin occurrence in channel catfish production ponds[J]Journal of Phycology,38:230-233
Oh H M,Lee S J,Jang M H,et al.2000.Microcystin production b Microcystis aeruginosa in a phosphorous-limited chemostat[J]Applied and Environmental Microbiology,66:176-179
O'Neil J M,Davis T W,Burford M A,et al.2012.The rise of harmfu cyanobacteria blooms:The potential roles of eutrophication and climate change[J].Harmful Algae,14:313-334
Otten T G,Crosswell J R,Mackey S,et al.2015.Application of molecular tools for microbial source tracking and public health risk assessment of a Microcystis bloom traversing 300km of the Klamath River[J].Harmful Algae,46:71-81
Pearson L,Mihali T,Moffitt M,et al.2010.On the chemistry,toxicology and genetics of the cyanobacterial toxins,microcystin,nodularin,saxitoxin and cylindrospermopsin[J].Marine Drugs,8(5):1650-1680
Rinta Kanto J M,Ouellette A J A,Boyer G L,et al.2005.Quantification of toxic Microcystis spp.during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR[J].Environmental Science&Technology,39(11):4198-4205
Sevilla E,Martin Luna B,Vela L,et al.2008.Iron availability affects mcy Dexpression and microcystin-LR synthesis in Microcystis aeruginosa PCC7806[J].Environmental Microbiology,10(10):2476-2483
Singh S,Rai P K,Chau R,et al.2015.Temporal variations in microcystinproducing cells and microcystin concentrations in two fresh water ponds[J].Water Research,69:131-142
Song L R,Chen W,Peng L,et al.2007.Distribution and bioaccumulation of microcystins in water columns:A systematic investigation into the environmental fate and the risk associated with microcystins in Meiliang Bay,Lake Taihu[J].Water Research,41:2853-2864
Tillett D,Dittmann E,Erhard M,et al.2000.Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806:an integrated peptide-polyketide synthetase system[J].Chemistry and Biology,7(10):753-764
Wang X Y,Sun M J,Xie M J,et al.2013.Differences in microcystin production and genotype composition among Microcystis colonies of different sizes in Lake Taihu[J].Water Research,47(15):5659-5669
王菲凤,仝川,杨芳,等.2011.福州山仔水库水华微囊藻毒素时空分布特征[J].环境科学学报,31(3):533-546
王丽平,郑丙辉.2013.洋河水库富营养化发展趋势及其关键影响因素[J].生态学报,33(3):1011-1017
Yang Z,Kong F X,Shi X L,et al.2015.Effects of UV-B radiation on microcystin production of a toxic strain of Microcystis aeruginosa and its competitiveness agianist a non-toxic strain[J].Journal of Hazardous Materials,283:447-453
Yoshida M,Yoshida T,Takashima Y,et al.2007,Dynamics of microcystin-producing and non-microcystin-producing Microcystis populations is correlated with nitrate concentration in a Japanese lakes[J].FEMS Microbiology Letters,266(1):48-53
Yoshida M,Yoshida T,Kashima A,et al.2008,Ecological dynamics of the toxic bloom-forming cyanobacterium Microcystis aeruginosa and its cyanophages in freshwater[J].Applied and Environmental Microbiology,74(10):3269-3273
张丽媛,王圣瑞,储昭升,等.2010.洋河水库流域土壤与库区沉积物中磷形态特征研究[J].中国环境科学,30(11):1529-1536
赵伟,李涵,许秋瑾,等.2011.洋河水库水华优势种演替的原因分析[J].环境工程学报,5(9):1989-1993