硝化抑制剂对酸性红壤硝化作用及氨氧化微生物丰度和群落结构的影响
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摘要
以一种典型酸性红壤为研究对象,在微宇宙培养条件下,研究硝化抑制剂双氰胺(DCD)和乙炔对酸性土壤硝化过程以及氨氧化微生物丰度和群落结构的影响。结果表明:硫酸铵(铵态氮肥)施加刺激了土壤硝化过程,硝化抑制剂处理显著抑制了硝化作用并且C_2H_2对硝化过程的抑制作用比DCD更强。研究还发现氨氧化细菌(Ammonia-Oxidizing Bacteria,AOB)的丰度与的浓度呈显著正相关关系,而氨氧化古菌(Ammonia-oxidizing Archaea,AOA)的丰度与的浓度呈显著负相关关系。末端限制性片段多态性(T-RFLP)分析和主坐标分析(PCo A)表明单施硫酸铵处理改变了AOB和AOA的群落结构。硝化抑制剂处理对AOA和AOB的群落结构没有显著影响。这些结果表明在高铵氮条件下AOB是土壤硝化过程的主要执行者,硝化抑制剂通过抑制AOB的丰度从而抑制硝化过程。
Effects of two different nitrification inhibitors(NIs, DCD and C_2H_2) on nitrification process, ammonia oxidizers abundance as well as their community structure in an acidic red soil were studied by cultivation experiment. Our results showed that ammonium-fertilizer application significantly increased the net nitrification rates, but were significantly inhibited by both NIs, and the inhibitory effect of C_2H_2 was significantly greater than that of DCD. AOB amoA gene abundance was positively correlated with NO_3~- concentration, while AOA amoA gene abundance was negatively correlated with it. Based on T-RFLP and PCoA, application of ammonium-fertilizer significantly changed the structure of the AOB and AOA community in the acidic soil.However, the application of NIs did not change the structure of ammonia oxidizers. These results indicated that AOB controlled soil nitrification process and the NIs inhibited nitrification by suppressing AOB growth in high ammonium soil.
Effects of two different nitrification inhibitors(NIs, DCD and C_2H_2) on nitrification process, ammonia oxidizers abundance as well as their community structure in an acidic red soil were studied by cultivation experiment. Our results showed that ammonium-fertilizer application significantly increased the net nitrification rates, but were significantly inhibited by both NIs, and the inhibitory effect of C_2H_2 was significantly greater than that of DCD. AOB amoA gene abundance was positively correlated with NO_3~- concentration, while AOA amoA gene abundance was negatively correlated with it. Based on T-RFLP and PCoA, application of ammonium-fertilizer significantly changed the structure of the AOB and AOA community in the acidic soil.However, the application of NIs did not change the structure of ammonia oxidizers. These results indicated that AOB controlled soil nitrification process and the NIs inhibited nitrification by suppressing AOB growth in high ammonium soil.
引文
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[27]CHEN X,ZHANG L M,SHEN J P,et al.Soil type determines the abundance and community structure of ammonia-oxidizing bacteria and archaea in flooded paddy soils[J].Journal of Soils and Sediments,2010,10:1510-1516.
[2]PURKHOLD U,POMMERENING-ROSER A,JURETSCHKO S,et al.Phylogeny of all recognized species of ammonia oxidizers based on comparative 16Sr RNA and amo A sequence analysis:implications for molecular diversity surveys[J].Applied Environment Microbiology,2000,66:5368-5382.
[3]IPCC.Climate change 2013:the physical science basis.Contribution of Working Group I to the Fifth Assessment Report of the Intergovern-mental Panel on Climate Change[M].Cambridge,United Kingdom and New York,NY,USA:Cambridge University Press,2013.
[4]HU H W,CHEN D,HE J Z.Microbial regulation of terrestrial nitrous oxide formation:understanding the biological pathways for prediction of emission rates[J].FEMS Microbiology Review,2015a,39:729-749.
[5]CUI P,FAN F,YIN C,et al.Urea-and nitrapyrin-affected N2O emission is coupled mainly with ammonia oxidizing bacteria growth in microcosms of three typical Chinese arable soils[J].Soil Biology and Biochemistry,2013,66:214-221.
[6]DAI Y,DI H J,CAMERON K C,et al.Effects of nitrogen application rate and a nitrification inhibitor dicyandiamide on ammonia oxidizers and N2O emissions in a grazed pasture soil[J].Science of the Total Environment,2013,465:125-135.
[7]GONG P,ZHANG L L,WU Z J,et al.Responses of Ammonia-Oxidizing Bacteria and Archaea in Two Agricultural Soils to Nitrification Inhibitors DCD and DMPP:A Pot Experiment[J].Pedosphere,2013,23:729-739.
[8]KELLIHER F M,CLOUGH T J,CLARK H,et al.The temperature dependence of dicyandiamide(DCD)degradation in soils:a data synthesis[J].Soil Biology and Biochemistry,2008,40(7):1878-1882.
[9]DI H J,CAMERON K C,SHEN J P,et al.Nitrification driven by bacteria and not archaea in nitrogen-rich grassland soils[J].Nature Geoscience,2009(2):621-624.
[10]DE BORE W,KOWALCHUK G A.Nitrification in acid soils:micro-organisms and mechanisms[J].Soil Biology and Biochemistry,2001,33(7):853-866.
[11]GUBRY-RANGIN C,HAI B,QUINCE C,et al.Niche specialization of terrestrial archaeal ammonia oxidizers[J].Proceedings of the National Academy of Sciences,2011,108(52):21206-21211.
[12]GUBRY-RANGIN C,NICOL G W,PROSSER J I.Archaea rather than bacteria control nitrification in two agricultural acidic soils[J].FEMS Microbiology Ecology,2010,74:566-574.
[13]JIA Z,CONRAD R.Bacteria rather than Archaea dominate microbial ammonia oxidation in an agricultural soil[J].Environ Microbiol,2009,11:1658-1671.
[14]SHEN J P,ZHANG L M,ZHU Y G,et al.Abundance and composition of ammonia-oxidizing bacteria and ammonia-oxidizing archaea communities of an alkaline sandy loam[J].Environmental Microbiology,2008,10:1601-1611.
[15]ZHANG L M,HU H W,SHEN J P,et al.Ammonia-oxidizing archaea have more important role than ammonia-oxidizing bacteria in ammonia oxidation of strongly acidic soils[J].The ISME Journal,2012,6:1032-1045.
[16]PERSSON T,WIREN A.Nitrogen mineralization and potential nitrification at different depths in acid forest soils[J].Plant and Soil,1995,168:55-65.
[17]TOURNA M,FREITAG T E,NICOL G W,et al.Growth,activity and temperature responses of ammonia‐oxidizing archaea and bacteria in soil microcosms[J].Environmental microbiology,2008,10(5):1357-1364.
[18]RAJBANSHI S S,BENCKISRE G,OTTOW J C G.Effects of concentration,incubation temperature,and repeated applications on degradation kinetics of dicyandiamide(DCD)in model experiments with a silt loam soil[J].Biology and Fertility of Soils,1992,13(2):61-64.
[19]HAUSER M,HASELWANDTER K.Degradation of dicyandiamide by soil bacteria[J].Soil Biology and Biochemistry,1990,22:113-114.
[20]CHEN Q H,QI L Y,BI Q F,et al.Comparative effects of3,4-dimethylpyrazole phosphate(DMPP)and dicyandiamide(DCD)on ammonia-oxidizing bacteria and archaea in a vegetable soil[J].Applied Microbiology and Biotechnology,2015,99:477-487.
[21]LEININGER S,URICH T,SCHLOTER M,et al.Archaea predominate among ammonia-oxidizing prokaryotes in soils[J].Nature,2006,442:806-809.
[22]DI H J,CAMERON K C,PODOLYAN A,et al.Effect of soil moisture status and a nitrification inhibitor,dicyandiamide,on ammonia oxidizer and denitrifier growth and nitrous oxide emissions in a grassland soil[J].Soil Biology and Biochemistry,2014,73:59-68.
[23]WANG Q,ZHANG L M,SHEN J P,et al.Nitrogen fertiliser-induced changes in N2O emissions are attributed more to ammonia-oxidising bacteria rather than archaea as revealed using 1-octyne and acetylene inhibitors in two arable soils[J].Biology and Fertility of Soils,2016,52:1163-1171.
[24]SHEN T,STIEGLMEIER M,DAI J,et al.Responses of the terrestrial ammonia-oxidizing archaeon Ca.Nitrososphaera viennensis and the ammonia-oxidizing bacterium Nitrosospira multiformis to nitrification inhibitors[J].FEMS microbiology letters,2013,344(2):121-129.
[25]AVRAHAMI S,CONRAD R,BRAKER G.Effect of soil ammonium concentration on N2O release and on the community structure of ammonia oxidizers and denitrifiers[J].Applied Environment Microbiology,2002,68(11):5685-5692.
[26]OFFRE P,PROSSER J I,NICOL G W.Growth of ammonia-oxidizing archaea in soil microcosms is inhibited by acetylene[J].FEMS Microbiology Ecology,2009,70:99-108.
[27]CHEN X,ZHANG L M,SHEN J P,et al.Soil type determines the abundance and community structure of ammonia-oxidizing bacteria and archaea in flooded paddy soils[J].Journal of Soils and Sediments,2010,10:1510-1516.
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