三十六脚湖水库沉积物聚磷菌多样性及群落组成
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摘要
采用末端限制性片段长度多态性(Terminal restriction fragment length polymorphism,T-RFLP)技术结合克隆文库构建,分析福建省平潭岛三十六角湖水库夏冬季特定样点沉积物聚磷菌(PAO)多样性及群落组成.结果显示,三十六脚湖沉积物中夏季聚磷菌的多样性明显高于冬季,夏季优势聚磷菌末端限制性片段(Terminal restriction fragment,T-RF)集中在200-300 bp,而冬季聚磷菌T-RF片段集中在150-200 bp.克隆测序和系统发育分析表明,三十六脚湖水库沉积物样品中检测到优势的聚磷菌类群为变形菌门(Proteobacteria)、放线菌门(Actinobacteria)和酸杆菌门(Acidobacteria).夏季丰度大于10%优势的菌属为厌氧粘细菌属(Anaeromyxobacter)、固氮螺菌属(Azospirillum)、聚磷小月菌(Microlunatus phosphovorus)和伯克氏菌属(Burkholderia),而冬季丰度大于14%优势的菌属主要有厌氧粘细菌属、甲基杆菌属(Methylobacterium)、Solibacter属、固氮螺菌属和聚磷小月菌.上述结果表明三十六脚湖沉积物中聚磷菌多样性及群落组成均呈现出一定的季节变化特征,沉积物各形态磷对聚磷菌多样性有一定影响,且与铁铝结合态磷相关性最显著,这可为今后揭示水库沉积物磷代谢循环的微生物学机制提供科学基础.
The diversity and community composition of phosphorus accumulating organisms(PAOs) in the sediments of the Sanshiliujiao Lake reservoir in Pingtan island, Fujian Province, were investigated by terminal restriction fragment length polymorphism(T-RFLP) methods combined with the construction of clone libraries. The diversity of PAOs in summer samples was significantly higher than that in winter. The dominant terminal restriction fragments(T-RFs) of PAOs were in the range of 200 to 300 bp in summer, while they were in the range of 150 to 200 bp in winter. Cloning sequencing and phylogenetic analysis showed that the main phyla of PAOs in the sediment of the Sanshiliujiao Lake reservoir were Proteobacteria, Actinobacteria, and Acidobacteria. Anaeromyxobacter, Azospirillum, Microlunatus phosphovorus, and Burkholderia were the dominant genera in the summer samples. Anaeromyxobacter, Methylobacterium, Solibacter, Azospirillum, and Microlunatus phosphovorus were the dominant genera in sediment samples in winter. The results of this study demonstrated that the diversity and community composition of PAOs in the sediments of the Sanshiliujiao Lake reservoir had specific seasonal characteristics. Various forms of phosphorus also showed their influence on the diversity of PAOs, especially Fe/Al-P. Our findings lays a scientific foundation for the future disclosure of microbial mechanisms involved in the phosphorus metabolic cycle in the sediments of reservoirs.
The diversity and community composition of phosphorus accumulating organisms(PAOs) in the sediments of the Sanshiliujiao Lake reservoir in Pingtan island, Fujian Province, were investigated by terminal restriction fragment length polymorphism(T-RFLP) methods combined with the construction of clone libraries. The diversity of PAOs in summer samples was significantly higher than that in winter. The dominant terminal restriction fragments(T-RFs) of PAOs were in the range of 200 to 300 bp in summer, while they were in the range of 150 to 200 bp in winter. Cloning sequencing and phylogenetic analysis showed that the main phyla of PAOs in the sediment of the Sanshiliujiao Lake reservoir were Proteobacteria, Actinobacteria, and Acidobacteria. Anaeromyxobacter, Azospirillum, Microlunatus phosphovorus, and Burkholderia were the dominant genera in the summer samples. Anaeromyxobacter, Methylobacterium, Solibacter, Azospirillum, and Microlunatus phosphovorus were the dominant genera in sediment samples in winter. The results of this study demonstrated that the diversity and community composition of PAOs in the sediments of the Sanshiliujiao Lake reservoir had specific seasonal characteristics. Various forms of phosphorus also showed their influence on the diversity of PAOs, especially Fe/Al-P. Our findings lays a scientific foundation for the future disclosure of microbial mechanisms involved in the phosphorus metabolic cycle in the sediments of reservoirs.
引文
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2 Stokholmbjerregaard M,Mcilroy SJ,Nierychlo M,Karst SM,Albertsen M,Nielsen PH.A critical assessment of the microorganisms proposed to be important to enhanced biological phosphorus removal in full-scale wastewater treatment systems[J].Front Microbiol,2017,8:doi 10.3389/fmicb.2017.00718
3 Wong PY,Cheng KY,Kaksonen AH,Sutton DC,Ginige MP.A novel post denitrif ication conf iguration for phosphor us recover y using polyphosphate accumulating organisms[J].Water Res,2013,47(17):6488-6495
4 Weissbrodt DG,Holliger C,Morgenroth E.Modeling hydraulic transport and anaerobic uptake by PAOs and GAOs during wastewater feeding in EBPR granular sludge reactors[J].Biotechnol Bioeng,2017,114(8):1688-1702
5 Chan C,Guisasola A,Baeza JA.Enhanced biological phosphor us removal at low sludge retention time in view of its integration in a-stage systems[J].Water Res,2017,118:217-226
6 Valverdepérez B,Wágner DS,Lóránt B,Gülay A,Smets BF,Plósz BG.Short-sludge age ebpr process-microbial and biochemical process characterisation during reactor start-up and operation[J].Water Res,2016,104:320-329
7陈楠.太湖沉积物微生物群落组成与物质循环及藻华爆发的相关性[D].北京:中国农业大学,2015[Chen N.Relationship between microbial community structure and nutrient cycles and algal bloom in Taihu Lake[D].Beijing:China Agricultural University,2015]
8胡颖,江和龙,何超群.太湖草藻型沉积物的物理化学特征及其对沉积物中聚磷菌的影响[J].生态环境学报,2012,21(11):1859-1864[Hu Y,Jiang HL,He CQ.Characteristic of physics and chemistry of grass/algae type zone sediments in Lake Taihu and effect on polyphosphate accumulation[J].Acta Ecol Environ Sin,2012,21(11):1859-1864]
9 Hupfer M,Gloess S,G rossar t H P.Poly phosphate-accu mulating microorganisms in aquatic sediments[J].Aquat Microb Ecol,2007,47(3):299-311
10 García Martín H,Ivanova N,Kunin V,Warnecke F,Barry KW,McHardy AC,Yeates C,He S,Salamov AA,Szeto E,Dalin E,Putnam NH,Shapiro HJ,Pangilinan JL,Rigoutsos I,Kyrpides NC,Blackall LL,McMahon KD,Hugenholtz P.Metagenomic of two enhanced biological phosphorus removal(EBPR)sludge communities[J].Nat Biotechnol,2006,24(10):1269-1263
11 Camejo PY,Owen BR,Martirano J,Ma J,Kapoor V,Domingo JS,McMahon KD,Noguera DR.Candidatus Accumulibacter phosphatis clades enriched under cyclic anaerobic and microaerobic conditions simultaneously use different electron acceptors[J].Water Res,2016,102:125-137
12张丽敏,曾薇,王安其,张洁,彭永臻.城市污水处理厂Candidatus Accumulibacter的菌群结构及定量分析[J].环境科学学报,2016,36(4):1226-1235[Zhang LM,Zeng W,Wang AQ,Zhang J,Peng YZ.Community structures and quantitative analyses of Candidatus Accumulibacter in municipal wastewater treatment plants[J].Acta Sci Circumstant,2016,36(4):1226-123]
13东野脉兴,樊竹青,张灼,夏学惠,田升平,周建民.滇池微生物解磷与聚磷作用的实验研究及磷的现代沉积与微生物成矿作用[J].吉林大学学报,2003,33(3):282-289[Dongye MX,Fan ZQ,Zhang Z,Xia XH,Tian SP,Zhou JM.Experimental reseaches on phosphorus decomposing and concentrating of microorganism in Dianchi lake:an example of modern phosphorus deposition and microorganism mineralization[J].J Jilin Univ(Earth Sci Ed),2003,33(3):282-289]
14夏学惠,东野脉兴,周建民,田升平,张灼,彭彦华.滇池现代沉积物中磷的地球化学及其对环境影响[J].沉积学报,2002,20(3):416-420[Xia XH,Dongye MX,Zhou JM,Tian SP,Zhang Z,Peng YH.Geochemistry and influence to environment of phosphorus in modern sediment in Dianchi Lake[J].Acta Sedimentol Sin,2002,20(3):416-420]
15 He S,Gall DL,McMahon KD.Candidatus Accumulibacter population structure in enhanced biological phosphorus removal sludges as revealed by polyphosphate kinase genes[J].Appl Environ Microbiol,2007,73(18):5865-5874
16 Kunin V,He S,Warnecke F,Peterson SB,Martin HG,Haynes M,Ivanova N,Blackall LL,Breitbar t M,Rohwer F,McMahon K D,Hugenholtz P.A bacterial metapopulation adapts locall to phase predation despite global dispersal[J].Genome Res,2008,18(2):293-297
17朱艳霞.太湖入水口底泥微生物宏基因组及聚磷菌多样性研究[D].苏州:苏州科技学院,2012[Zhu YX.Research an metagenomie DNA and biodiversity of phosphor us-accumulating organisms in the sediments from water inlet of Taihu Lake[D].Suzhou:Suzhou University of Science and Technology,2012]
18翁彩云.平潭县三十六脚湖水环境问题及周边工程治理对策浅析[J].能源与环境,2009(3):11-13[Weng CY.Analyses on water environment problems and engineering treating counter measure of surrounding in Sanshiliujiao Lake in Pingtan County[J].Energy Environ,2009(3):11-13]
19 Ruban V,López-Sánchez JF,Pardo P,Rauret G,Muntau H,Quevauviller P.Har monized protocol and cer tif ied reference material for the determination of extractable contents of phosphorus in freshwater sediments-a synthesis of recent works[J].Fresenius J Anal Chem,2001,370(2-3):224-228
20苏玉萍,林佳,何灵,林婉珍,王家乐.福建省山仔水库沉积物磷对水体磷浓度贡献的估算[J].湖泊科学,2008,20(6):748-754[Su YP,Lin J,He L,Lin WZ,Wang JL.Calculation of the contribution of the sediment phosphorus to the phosphorus concentration of the water body of Shanzi deep reservoir in Fujian Province[J].J Lake Sci,2008,20(6):748-754]
21 Ludwig F,Mehlig L,Eschenhagen M,R?ske I.Development of new primer systems for the detection of the polyphosphate kinase gene in activated sludge[J].Water Sci and Technol,2009,30(2):213-218
22 Fierer N,Schimel JP,Holden PA.Inf luence of dr ying-rewetting frequency on soil bacterial community structure[J].Microb Ecolol,2003,45(1):63-71
23 Manzano M,Moran AC,Tesser B,Gonzalez B.Role of eukaryotic microbiota in soil survival and catabolic Performance of the 2,4-Dherbicide degrading bacteria Cupriavidus necator JMP134[J].Antonie van Leeuwenhoek,2007,91(2):115-126
24尹承苗,王功帅,李园园,陈学森,吴树敬,毛志泉.连作苹果园土壤真菌的T-RFLP分析[J].生态学报,2014,34(4):837-846[Yin CM,Wang GS,Li YY,Chen XS,Wu SJ,Mao ZQ.Assessment of fungal diversity in apple replanted orchard soils by T-RFLP analysis[J].Acta Ecol Sin,2014,34(4):837-846]
25佘晨兴,仝川.闽江口芦苇沼泽湿地土壤产甲烷菌群落结构的垂直分布[J].生态学报,2012,32(17):5299-5308[She CX,Tong C.Vertical distribution of methanogen community str uctures in Phragmites australis marsh soil in the Min River estuary[J].Acta Ecol Sin,2012,32(17):5299-5308]
26佘晨兴,仝川,王维奇.互花米草沼泽湿地产甲烷古菌的多样性及垂向分布[J].环境科学学报,2014,34(1):186-193[She CX,Tong C,Wang WQ.Vertical distribution and diversity of methanogenic archaea in estuarine marsh dominated by Spartina alternif lora[J].Acta Sci Circumstant,2014,34(1):186-193]
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