Fe (Ⅱ)对反硝化过程及其功能微生物群落的影响

摘要
图/表
参考文献
相关文章 (4)

全文: PDF (729 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要以华南稻田土壤为研究对象，通过室内模拟控制实验，研究了Fe （Ⅱ）对厌氧稻田土壤中反硝化过程及其功能微生物群落结构组成与相对丰度的影响.结果表明，加入Fe （Ⅱ）减缓了土壤中NO₃^-的还原，但促进了NO₂^-的还原和N₂O的生成；同时Fe （Ⅱ）只在Soil+Fe （Ⅱ）+NO₃^-处理中发生氧化.通过定量PCR结果发现，Fe （Ⅱ）的加入提高了亚硝酸盐还原基因nirS和N₂O还原基因nosZ的拷贝数；但降低了细胞膜硝酸盐还原基因narG的拷贝数.通过高通量和克隆文库分析发现，Fe （Ⅱ）的加入主要对napA-周质硝酸盐还原微生物群落结构有明显影响，Soil+NO₃^-处理中优势菌是Dechloromonas，而Soil+Fe （Ⅱ）+NO₃^-处理中为Azonexus、Dechloromonas和Azospira.Fe （Ⅱ）对厌氧稻田中的反硝化过程及其功能微生物群落具有显著影响，这对了解华南红壤地区稻田体系中的氮元素循环与铁元素转化的关系具有重要意义.

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	李爽
	李晓敏
	李芳柏

关键词 ： Fe (Ⅱ), 反硝化过程, 功能基因, 功能微生物, 稻田土壤

Abstract：Paddy soil was collected from Southern China and used as an inoculum in an anoxic experiment to investigate the effect of Fe(Ⅱ) oxidation on denitrification and associated microbial communities. The results showed that the presence of Fe(Ⅱ) slowed down the NO₃^- reduction, but facilitated the NO₂^- reduction and N₂O formation; meanwhile Fe(Ⅱ) oxidation was observed only in the Soil+Fe(Ⅱ)+NO₃^- treatment. The real-time quantitative PCR analysis demonstrated that the presence of Fe(Ⅱ) increased the copy numbers of nitrite reduction gene (nirS) and nitrous oxide (N₂O) reduction gene (nosZ), but inhibited the activity of membrane nitrate reduction gene (narG). The results of high-throughput sequencing and clone library exhibited that the presence of Fe(Ⅱ) significantly influences the microbial communities affiliated with periplasmic nitrate reduction gene (napA). The most abundant napA-based nitrate reducer was Dechloromonas in the Soil+NO₃^- treatment, while those were Azonexus, Dechloromonas and Azospira in the Soil+Fe(Ⅱ)+NO₃^- treatment. In summary, Fe(Ⅱ) significantly influences the denitrification process and the associated microbial communities in anoxic paddy soil. This can provide better understanding of the relationship between the nitrogen cycling and the iron transformation in the red paddy soil from South China.

Key words： Fe (Ⅱ) denitrification functional gene functional microbial communities paddy soil

收稿日期: 2017-06-05

PACS:

X172

基金资助:国家自然科学基金资助项目（41571130052，41330857）；省院创新平台建设专项（2016GDASPT-0105）

通讯作者: 李芳柏,研究员,cefbli@soil.gd.cn E-mail: cefbli@soil.gd.cn

作者简介: 李爽(1989-),女,河南驻马店,中国科学院广州地球化学研究所博士研究生,主要从事土壤微生物环境生态研究.发表论文1篇.

引用本文:

李爽, 李晓敏, 李芳柏. Fe (Ⅱ)对反硝化过程及其功能微生物群落的影响[J]. 中国环境科学, 2018, 38(1): 263-274. LI Shuang, LI Xiao-min, LI Fang-bai. Effect of Fe(Ⅱ) on denitrification and associated functional microbial communities. CHINA ENVIRONMENTAL SCIENCECE, 2018, 38(1): 263-274.

链接本文:

http://manu36.magtech.com.cn/Jweb_zghjkx/CN/ 或 http://manu36.magtech.com.cn/Jweb_zghjkx/CN/Y2018/V38/I1/263

[1]	Zumft W G. Cell biology and molecular basis of denitrification[J]. Microbiology & Molecular Biology Reviews, 1997,61(4):533-616.
[2]	Castresana J, Saraste M. Evolution of energetic metabolism:the respiration-early hypothesis[J]. Trends in Biochemical Sciences, 1995, 20(11):443-448.
[3]	Ducluzeau A L, Lis R V, Duval S, et al. Was nitric oxide the first deep electron sink?[J]. Trends in Biochemical Sciences, 2009, 34(1):9-15.
[4]	Kraft B, Strous M, Tegetmeyer H E. Microbial nitrate respirationgenes, enzymes and environmental distribution[J]. Journal of Biotechnology, 2011,155(1):104-117.
[5]	Palmer K, Biasi C, Horn M A. Contrasting denitrifier communities relate to contrasting N2O emission patterns from acidic peat soils in arctic tundra[J]. ISME Journal, 2012,6(5):1058-1077.
[6]	Deiglmayr K, Philippot L, Kandeler E. Functional stability of the nitrate-reducing community in grassland soils towards high nitrate supply[J]. Soil Biology & Biochemistry, 2006, 38(9):2980-2984.
[7]	鲍林林,王晓燕,陈永娟,等.北运河沉积物中主要脱氮功能微生物的群落特征[J]. 中国环境科学, 2016,36(54):1520-1529.
[8]	Nordhoff M, Tominski C, Halama M, et al. Insights into nitrate-reducing Fe(Ⅱ) oxidation mechanisms by analyzing cellmineral associations, cell encrustation and mineralogy in the chemolithoautotrophic enrichment culture KS[J]. Applied & Environmental Microbiology, 2017,doi:10.1128/AEM.00752-17.
[9]	赵其国.红壤物质循环及其调控[M]. 北京:科学出版社, 2002.
[10]	谢小兰,余光伟,种云霄,等.污染河流底泥亚铁氧化硝酸盐还原菌分离及代谢特性[J]. 环境科学, 2015,35(5):1224-1562.
[11]	Smith R L, Kent D B, Repert D A, et al. Anoxic nitrate reduction coupled with iron oxidation and attenuation of dissolved arsenic and phosphate in a sand and gravel aquifer[J]. Geochimica Et Cosmochimica Acta, 2017,196:102-120.
[12]	Zhang J, Zhao S, Xu Y, et al. Nitrate stimulates anaerobic microbial arsenite oxidation in paddy soils[J]. Environmental Science & Technology, 2017,51(8):4377-4386.
[13]	TongH, Hu M, Li F, et al. Burkholderiales participating in pentachlorophenol biodegradation in iron-reducing paddy soil as identified by stable isotope probing[J]. Environmental Science Processes & Impacts, 2015,17(7):1282-1289.
[14]	Chen X P, Zhu Y G, Hong M N, et al. Effects of different forms of nitrogen fertilizers on arsenic uptake by rice plants[J]. Environmental Toxicology and Chemistry, 2008,27(4):881-887.
[15]	罗希茜,陈哲,胡荣桂,等.长期施用氮肥对水稻土亚硝酸还原酶基因多样性的影响[J]. 环境科学, 2010,31(2):423-430.
[16]	Li X, Zhang W, Liu T, et al. Changes in the composition and diversity of microbial communities during anaerobic nitrate reduction and Fe(Ⅱ) oxidation at circumneutral pH in paddy soil[J]. Soil Biology & Biochemistry, 2016,94:70-79.
[17]	陈鹏程,李晓敏,李芳柏.水稻土Fe(Ⅱ)氧化耦合NO3-还原的微生物变化[J]. 中国环境科学, 2017,37(1):358-366.
[18]	鲁如坤.土壤农业化学分析方法[M]. 北京:中国农业科技出版社, 2000.
[19]	Zhu G, Wang S, Wang Y, et al. Anaerobic ammonia oxidation in a fertilized paddy soil[J]. ISME Journal, 2011,5(12):1905-1912.
[20]	Lovley D R, Phillips E J. Rapid assay for microbially reducible ferric iron in aquatic sediments[J]. Applied & Environmental Microbiology, 1987,53(7):1536-1540.
[21]	Klueglein N, Kappler A. Abiotic oxidation of Fe(Ⅱ) by reactive nitrogen species in cultures of the nitrate-reducing Fe(Ⅱ) oxidizer Acidovorax sp. BoFeN1-questioning the existence of enzymatic Fe(Ⅱ) oxidation[J]. Geobiology, 2013,11(2):180-190.
[22]	Petersen D J, Blazewicz S J, Firestone M, et al. Abundance of microbial genes associated with nitrogen cycling as indices of biogeochemical process rates across a vegetation gradient in Alaska[J]. Environmental Microbiology, 2012,14(4):993-1008.
[23]	Lópezgutiérrez J C, Henry S, Hallet S, et al. Quantification of a novel group of nitrate-reducing bacteria in the environment by real-time PCR[J]. Journal of Microbiological Methods, 2004, 57(57):399-407.
[24]	Bru D, Sarr A, Philippot L. Relative abundances of proteobacterial membrane-bound and periplasmic nitrate reductases in selected environments[J]. Applied & Environmental Microbiology, 2007,73(18):5971-5974.
[25]	Palmer K, Biasi C, Horn M A. Contrasting denitrifier communities relate to contrasting N2O emission patterns from acidic peat soils in arctic tundra[J]. ISME Journal, 2011, 6(5):1058-1077.
[26]	Vilar-Sanz A, Puig S, García-Lledó A, et al. Denitrifying bacterial communities affect current production and nitrous oxide accumulation in a microbial fuel cell[J]. PloS One, 2013, 8(5):e63460.
[27]	Caporaso J G, Lauber C L, Walters W A, et al. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample[J]. Proceedings of the National Academy of Sciences of the United States of America, 2011,108(Suppl 1):4516-4522.
[28]	Throbäck I N, Enwall K, Jarvis A, et al. Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE[J]. FEMS Microbiology Ecology, 2004,49(3):401-417.
[29]	Philippot L, Piutti S, Martin-Laurent F, et al. Molecular analysis of the nitrate-reducing community from unplanted and maizeplanted soils[J]. Applied & Environmental Microbiology, 2002,68(12):6121-6128.
[30]	Walter P B, Knutson M D, Paler-Martinez A, et al. Iron deficiency and iron excess damage mitochondria and mitochondrial DNA in rats[J]. Proceedings of the National Academy of Sciences, 2002,99(4):2264-2269.
[31]	Klausner R D, Rouault T A, Harford J B. Regulating the fate of mRNA:the control of cellular iron metabolism[J]. Cell, 1993,72(1):19-28.
[32]	Ishii S, Yamamoto M, Kikuchi M, et al. Microbial populations responsive to denitrification-inducing conditions in rice paddy soil, as revealed by comparative 16S rRNA gene analysis[J]. Applied & Environmental Microbiology, 2009,75(22):7070-7078.
[33]	Straub K L, Benz M, Schink B, et al. Anaerobic, nitratedependent microbial oxidation of ferrous iron[J]. Applied & Environmental Microbiology, 1996,62(4):1458-1460.
[34]	Livermore J A, Jin Y O, Arnseth R W, et al. Microbial community dynamics during acetate biostimulation of RDX-contaminated groundwater[J]. Environmental Science & Technology, 2013, 47(14):7672-7678.
[35]	Chakraborty A, Picardal F. Neutrophilic, nitrate-dependent, Fe(Ⅱ) oxidation by a Dechloromonas species[J]. World Journal of Microbiology and Biotechnology, 2013,29(4):617-623.
[36]	Grimes D J, Woese C R, Macdonell M T, et al. Systematic study of the genus Vogesella gen. nov. and its type species, Vogesella indigofera comb. Nov[J]. International Journal of Systematic Bacteriology, 1997,47(1):19-27.
[37]	Ishii S, Joikai K, Otsuka S, et al. Denitrification and nitratedependent Fe(Ⅱ) oxidation in various Pseudogulbenkiania strains[J]. Microbes & Environments, 2016,31(3):293-298.
[38]	Sobolev D, Roden, E. E. Characterization of a neutrophilic, chemolithoautotrophic Fe(Ⅱ)-oxidizing β-proteobacterium from freshwater wetland sediments[J]. Geomicrobiology Journal, 2004, 21(1):1-10.
[39]	Philippot L. Denitrifying genes in bacterial and Archaeal genomes[J]. Biochimica Et Biophysica Acta, 2002,1577(3):355-376.
[40]	Chaudhuri S K, Lack J G, Coates J D. Biogenic magnetite formation through anaerobic biooxidation of Fe(Ⅱ)[J]. Applied & Environmental Microbiology, 2001,67(6):2844-8.
[41]	Philippot L, Højberg O. Dissimilatory nitrate reductases in bacteria[J]. Biochimica et Biophysica Acta-Gene Structure and Expression, 1999,1446(1/2):1-23.
[42]	Richardson D J, Berks B C, Russell D A, et al Functional, biochemical and genetic diversity of prokaryotic nitrate reductases[J]. Cellular & Molecular Life Sciences:CMLS, 2001,58(2):165-178.
[43]	Richardson D J, King G F, Kelly D J. The role of auxiliary oxidants in maintaining redox balance during phototrophic growth of Rhodobacter capsulatus on propionate or butyrate[J]. Archives of Microbiology, 1988,150(6):602-602.
[44]	González P J, Correia C, Moura I, et al. Bacterial nitrate reductases:Molecular and biological aspects of nitrate reduction[J]. Journal of Inorganic Biochemistry, 2006,100(6):1015-1023.
[45]	Delgado M J, Tresierra-Ayala A, Bedmar E J, et al. The bradyrhizobium japonicum napEDABC genes encoding the periplasmic nitrate reductase are essential for nitrate respiration[J]. Microbiology, 2003,149(Pt 12):3395-3403.
[46]	Picardal F W, Zaybak Z, Chakraborty A, et al. Microaerophilic, Fe(Ⅱ)-dependent growth and Fe(Ⅱ) oxidation by a Dechlorospirillum species[J]. FEMS Microbiology Letters, 2011,319(1):51-57.
[47]	Baldani J I, Baldani V L D, Seldin L, et al. Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root-associated nitrogen-fixing bacterium[J]. International Journal of Systematic Bacteriology, 1986,36(1):86-93.
[48]	Zhang Z, Lei Z, He X, et al. Nitrate removal by Thiobacillus denitrificans immobilized on poly(vinyl alcohol) carriers[J]. Journal of Hazardous Materials, 2009,163(2/3):1090-1095.