草鱼养殖池塘蓝藻暴发时水体细菌群落特征分析
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摘要
为了解蓝藻暴发池塘中细菌群落特征,采集3个地区(广东、云南、贵州)4个淡水养殖场的蓝藻暴发池塘和非蓝藻暴发池塘(对照池塘)水样,并检测其理化因子及生物指标,采用PCR-DGGE技术分析其细菌群落结构差异。依据PCR-DGGE指纹谱带的丰度对养殖水体细菌群落多样性进行了分析,并对水体细菌群落结构进行了UPGMA聚类分析。结果发现:蓝藻暴发池塘水体的PO_4-P含量均显著高于对照池塘(P<0.01);线性回归分析表明,PO_4-P与代表蓝藻暴发程度的叶绿素a存在正相关关系(R~2=0.869,P<0.01);而且理化因子与细菌群落的RDA分析表明,PO_4-P与蓝藻暴发池塘细菌群落关系密切。蓝藻暴发池塘与对照池塘水体的细菌群落结构间存在显著差异;进一步测序分析显示,蓝藻暴发池塘特定的细菌为Flexibacter,其可能对蓝藻有裂解作用;而Synechococcsus在蓝藻暴发池塘的含量明显低于对照池塘,可能是Microcystis的大量暴发抑制了Synechococcsus的生长。
In order to reveal the characteristics of bacterial community in the cyanobacterial blooming pond,the water samples in the cyanobacterial blooming ponds and the control ponds were collected from four aquaculture farms located in Guangdong, Yunnan and Guizhou Provinces, respectively. Both the physicochemical factors and biological indicators were analyzed. By applying polymerase chain reactiondenaturing gradient gel electrophoresis( PCR-DGGE) technique,the differences in bacterial community structure in these water bodies were found. The bacterial community diversity in aquaculture water were analyzed according to the abundance of PCR-DGGE fingerprint bands,and conducted an unweighted pair group method with arithmetic mean( UPGMA) cluster analysis on the bacterial community structure in the water body. The results indicated that the PO_4-P content was significantly higher in cyanobacterial blooming pond than that in the control pond( P < 0. 01). Linear regression analysis revealed a positive correlation between PO_4-P and the content of chlorophyll a( Chl. a),which represents the degree of cyanobacterial blooming( R~2= 0. 869,P < 0. 01). The analysis of physicochemical factors in combination with the redundancy analysis( RDA) in bacterial community indicated a close relationship between PO_4-P and bacterial community in aquaculture water. While no significant difference in bacterial community diversity was seen between cyanobacterial blooming pond and control pond( P > 0. 05),significant differences in bacterial community structure between the two ponds existed. Further sequencing analysis revealed that the characteristic bacteria in the cyanobacterial blooming pond was Flexibacter,which might play a role in algal pyrolysis. The Synechococcsus content was significantly lower in the cyanobacterial blooming pond than that in the control pond. This reduced Synechococcsus content might be related to the inhibition of Synechococcsus growth by the substantial blooming of Microcystis.
In order to reveal the characteristics of bacterial community in the cyanobacterial blooming pond,the water samples in the cyanobacterial blooming ponds and the control ponds were collected from four aquaculture farms located in Guangdong, Yunnan and Guizhou Provinces, respectively. Both the physicochemical factors and biological indicators were analyzed. By applying polymerase chain reactiondenaturing gradient gel electrophoresis( PCR-DGGE) technique,the differences in bacterial community structure in these water bodies were found. The bacterial community diversity in aquaculture water were analyzed according to the abundance of PCR-DGGE fingerprint bands,and conducted an unweighted pair group method with arithmetic mean( UPGMA) cluster analysis on the bacterial community structure in the water body. The results indicated that the PO_4-P content was significantly higher in cyanobacterial blooming pond than that in the control pond( P < 0. 01). Linear regression analysis revealed a positive correlation between PO_4-P and the content of chlorophyll a( Chl. a),which represents the degree of cyanobacterial blooming( R~2= 0. 869,P < 0. 01). The analysis of physicochemical factors in combination with the redundancy analysis( RDA) in bacterial community indicated a close relationship between PO_4-P and bacterial community in aquaculture water. While no significant difference in bacterial community diversity was seen between cyanobacterial blooming pond and control pond( P > 0. 05),significant differences in bacterial community structure between the two ponds existed. Further sequencing analysis revealed that the characteristic bacteria in the cyanobacterial blooming pond was Flexibacter,which might play a role in algal pyrolysis. The Synechococcsus content was significantly lower in the cyanobacterial blooming pond than that in the control pond. This reduced Synechococcsus content might be related to the inhibition of Synechococcsus growth by the substantial blooming of Microcystis.
引文
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[2]ZHONG F,GAO Y N,YU T,et al.The management of undesirable cyanobacteria blooms in channel catfish ponds using a constructed wetland:contribution to the control of offflavor occurrences[J].Water Research,2011,45(19):6479-6488.
[3]SEVRIN-REYSSAC J,PLETIKOSIC M.Cyanobacteria in fish ponds[J].Aquaculture,1990,88(1):1-20.
[4]SCHRADER K K,DENNIS M E.Cyanobacteria and earthy/musty compounds found in commercial catfish(Ictalurus punctatus)ponds in the Mississippi Delta and MississippiAlabama Blackland Prairie[J].Water Research,2005,39(13):2807-2814.
[5]张永生,孔繁翔,于洋,等.蓝藻伪空胞的特性及浮力调节机制[J].生态学报,2010,30(18):5077-5090.ZHANG Y S,KONG F X,YU Y,et al.The characteristics and buoyancy regulations of cyanobacterial gas vesicles[J].Acta Ecologica Sinica,2010,30(18):5077-5090.
[6]秦伯强,杨柳燕,陈非洲,等.湖泊富营养化发生机制与控制技术及其应用[J].科学通报,2006,51(16):1857-1866.QIN B Q,YANG L Y,CHEN F Z,et al.Mechanism and control of lake eutrophication[J].Chinese Science Bulletin,2006,51(19):2401-2412.
[7]赵孟绪,韩博平.汤溪水库蓝藻水华发生的影响因子分析[J].生态学报,2005,25(7):1554-1561.ZHAO M X,HAN B P.Analysis of factors affecting cyanobacteria bloom in a tropical reservoir(Tangxi Reservoir,China)[J].Acta Ecologica Sinica,2005,25(7):1554-1561.
[8]NIU Y,SHEN H,CHEN J,et al.Phytoplankton community succession shaping bacterioplankton community composition in Lake Taihu,China[J].Water Research,2011,45(14):4169-4182.
[9]郑小红,肖琳,任晶,等.玄武湖微囊藻水华暴发及衰退期细菌群落变化分析[J].环境科学,2008,29(10):2956-2962.ZHENG X H,XIAO L,REN J,et al.Variation of bacterial community composition in the outbreak and decline of Microcystis spp.Bloom in Lake Xuanwu[J].Environmental Science,2008,29(10):2956-2962.
[10]陈威.富营养化水体不同水华阶段细菌群落结构的变化[D].新乡:河南师范大学,2010.CHEN W.Variation of bacteria community stuctures in eutrophic water body during water bloom[D].Xinxiang:He'nan Normal University,2010.
[11]叶文瑾.太湖富营养化水体和底泥中微生物群落的分子生态学研究[D].上海:上海交通大学,2009.YE W J.Molecular ecology research on microbial community in the water and sediment of eutrophic Lake Taihu[D].Shanghai:Shanghai Jiao Tong University,2009.
[12]奚万艳,吴鑫,叶文瑾,等.太湖梅梁湾水域蓝藻水华前与水华末期细菌群落结构的变化[J].应用与环境生物学报,2007,13(1):97-103.XI W Y,WU X,YE W J,et al.Changes in bacterial community structure during preceding and degraded period of cyanobacterial bloom in a bay of the Taihu Lake[J].Chinese Journal of Applied and Environmental Biology,2007,13(1):97-103.
[13]田川.太湖浮游细菌分子生态学及溶藻细菌的研究[D].上海:上海交通大学,2012.TIAN C.Research on bacterioplankton diversity and the Algicidal bacteria in Lake Taihu[D].Shanghai:Shanghai Jiao Tong University,2012.
[14]CHEN M J,CHEN F Z,XING P,et al.Microbial eukaryotic community in response to Microcystis spp.bloom,as assessed by an enclosure experiment in Lake Taihu,China[J].FEMS Microbiology Ecology,2010,74(1):19-31.
[15]翁笑艳,林美爱,严颖.地表水浮游植物叶绿素a测定方法比较研究[J].中国环境监测,2009,25(3):36-38,76.WENG X Y,LIN M A,YAN Y.Comparison of determination of phytoplankton chlorophyll a by spectroscopic methods in freshwater[J].Environmental Monitoring in China,2009,25(3):36-38,76.
[16]桂佳,辛艳萍,韩博平,等.Beacon公司微囊藻毒素检测试剂盒的性能评价[J].生态科学,2009,28(5):428-432.GUI J,XIN Y P,HAN B P,et al.Performance evaluation of Beacon Microcystin Plate Kit[J].Ecological Science,2009,28(5):428-432.
[17]TORSBIK V,DAAE F L,SANDAA R A,et al.Review article:novel techniques for analysing microbial diversity in natural and perturbed environments[J].Journal of Biotechnology,1998,64(1):53-62.
[18]KENNEDY A C.Bacterial diversity in agroecosystems[J].Agriculture,Ecosystems&Environment,1999,74(1/3):65-76.
[19]胡韧,顾继光,聂祥,等.3座南亚热带串联调水水库浮游植物群落的CCA分析[J].生态科学,2008,27(5):315-319.HU R,GU J G,NIE X,et al.A comparison of phytoplankton assemblages in three subtropical pumped storage reservoirs based on CCA analysis[J].Ecological Science,2008,27(5):315-319.
[20]XU Y,WANG G X,YANG W B,et al.Dynamics of the water bloom-forming Microcystis and its relationship with physicochemical factors in Lake Xuanwu(China)[J].Environmental Science and Pollution Research,2010,17(9):1581-1590.
[21]刘庆,郭亮,邢鹏,等.富营养化浅水湖泊蓝藻的种群结构和多样性研究[J].农业环境科学学报,2012,31(3):570-579.LIU Q,GUO L,XING P,et al.Cyanobacterial community composition and diversity in shallow lakes of different trophic status[J].Journal of Agro-Environment Science,2012,31(3):570-579.
[22]LI N,ZHANG L,LI F C,et al.Metagenome of microorganisms associated with the toxic Cyanobacteria Microcystis aeruginosa analyzed using the 454 sequencing platform[J].Chinese Journal of Oceanology and Limnology,2011,29(3):505-513.
[23]李勤生,黎尚豪,王大耜.中华屈挠杆菌的分离和鉴定[J].微生物学报,1984,24(1):7-13.LI Q S,LI S H,WANG D S.Isolation and characterization of Flexibacter chinenses sp.nov.[J].Acta Microbiology Sinica,1984,24(1):7-13.
[24]李勤生,利群,蔡庆华.东湖屈挠杆菌的分离及其生物学特性研究[J].水生生物学报,1990,14(3):216-223.LI Q S,LI Q,CAI Q H.Study on isolation and biological characters of flexibacter strains from Donghu lake[J].Acta Hydrobiologica Sinica,1990,14(3):216-223.
[25]RASHIDAN K K,BIRD D F.Role of predatory bacteria in the termination of a cyanobacterial bloom[J].Microbial Ecology,2001,41(2):97-105.
[26]胡智泉,刘永定.微囊藻毒素在细长聚球藻中的积累及其毒性效应[J].中国环境科学,2005,25(1):23-27.HU Z Q,LIU Y D.Accumulation of microcystin-RR in Synechococcus elongatus and its toxic effects[J].China Environmental Science,2005,25(1):23-27.
[27]RAVEN J A.The twelfth tansley lecture.Small is beautiful:the picophytoplankton[J].Functional Ecology,1998,12(4):503-513.
[28]KONDO R,YOSHIDA T,YUKI Y,et al.DNA-DNA reassociation among a bloom-forming cyanobacterial genus,Microcystis[J].International Journal of Systematic and Evolutionary Microbiology,2000,50(2):767-770.
[29]胡智泉,刘永定,肖波.微囊藻毒素对几种淡水微藻的生长和光合活性的影响[J].生态环境,2008,17(3):885-890.HU Z Q,LIU Y D,XIAO B.Effects of microcystin on the growth and photosynthetic activity of algae in freshwater[J].Ecology and Environment,2008,17(3):885-890.
[30]ZENG J,YANG L Y,DU H W,et al.Bacterioplankton community structure in a eutrophic lake in relation to water chemistry[J].World Journal of Microbiology and Biotechnology,2009,25(5):763-772.
[31]孙鑫鑫,刘惠荣,冯福应,等.乌梁素海富营养化湖区浮游细菌多样性及系统发育分析[J].生物多样性,2009,17(5):490-498.SUN X X,LIU H R,FENG F Y,et al.Diversity and phylogenetic analysis of planktonic bacteria in eutrophic zone of Lake Wuliangsuhai[J].Biodiversity Science,2009,17(5):490-498.
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