脲酶抑制剂与硝化抑制剂对稻田土壤硝化、反硝化功能菌的影响
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摘要
【目的】在农业生产中,脲酶抑制剂(urease inhibitor,UI)与硝化抑制剂(nitrification inhibitor,NI)常作为氮肥增效剂来提高肥料利用率。本文研究了在我国南方红壤稻田施用脲酶抑制剂与硝化抑制剂后,土壤中氨氧化细菌(ammonia oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)以及反硝化细菌的丰度以及群落结构的变化特征,旨在揭示抑制剂的作用机理及其对土壤环境的影响。【方法】试验在我国南方红壤稻田进行,共设5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI),3次重复。脲酶抑制剂与硝化抑制剂分别为NBPT[N-(n-butyl) thiophosphrictriamide,N-丁基硫代磷酰三胺]和DMPP (3,4-dimethylpyrazole phosphate,3,4-二甲基吡唑磷酸盐)。通过荧光定量PCR (Real-time PCR)研究水稻分蘖期与孕穗期抑制剂对三类微生物标记基因拷贝数的影响,并分析土壤铵态氮、硝态氮与三种菌群丰度的相关性;利用变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)分析抑制剂对土壤AOB、AOA以及反硝化细菌群落结构的影响,并对优势菌群进行系统发育分析。【结果】1)荧光定量PCR结果表明,施用氮肥对两个时期土壤中AOB的amoA基因与反硝化细菌nirK基因的拷贝数均有显著提高,而对AOA的amoA基因始终没有明显影响;AOB与nirK反硝化细菌的丰度与两个时期的铵态氮含量、分蘖期的硝态氮含量呈极显著正相关,与孕穗期的硝态氮含量相关性不显著;DMPP仅在分蘖期显著减少了AOB的amoA基因拷贝数,表明DMPP主要通过限制AOB的生长来抑制稻田土壤硝化过程;NBPT对三类微生物的丰度无明显影响;2) DGGE图谱表明,在分蘖期与孕穗期,施用氮肥均明显增加了图谱中AOB的条带数,而对AOA却没有明显影响;氮肥明显增加了孕穗期反硝化细菌的条带数;与氮肥的影响相比,抑制剂NBPT与DMPP对AOA、AOB以及反硝化菌的群落结构影响甚微;系统发育分析结果表明,与土壤中AOB的优势菌群序列较为接近的有亚硝化单胞菌和亚硝化螺菌。【结论】在南方红壤稻田中,施入氮肥可显著提高AOB与反硝化细菌的丰度,明显影响两种菌群的群落结构,而AOA较为稳定;NBPT对三类微生物的群落结构丰度无明显影响;硝化抑制剂DMPP可抑制AOB的生长但仅表现在分蘖期,这可能是其缓解硝化反应的主要途径;这也说明二者对土壤生态环境均安全可靠。
【Objectives】Urease inhibitor(UI) and nitrification inhibitor(NI), as nitrogen synergist, are used to improve the efficiency of nitrogen(N) fertilizer in agriculture. The aim of this study was to investigate the effect of UI and NI on the abundance and community structure of ammonia oxidizing bacteria(AOB), ammoniaoxidizing archaea(AOA), and denitrifying bacteria in paddy soil, and reveal the synergy mechanism of the inhibitors and their effects on soil environment.【Methods】A field experiment was conducted in a paddy field of red soil in southern China. A randomized block design with five treatments and three replicates was as follows:1) CK(no N control), 2) U(Urea), 3) U + UI, 4) U + NI, 5) U + UI + NI. The urease inhibitor and nitrification inhibitor were NBPT [N-(n-butyl) thiophosphrictriamide] and DMPP(3,4-dimethylpyrazole phosphate)respectively. The abundance and composition of AOB, AOA and denitrifying bacteria were investigated by fluorescent quantitative real-time PCR and denaturing gradient gel electrophoresis(DGGE) approaches based on amoA and nirK genes, at tillering stage and booting stage, respectively. Then, dominant populations was sequenced and phylogenetic analyzed.【Results】1) Real-time PCR analysis showed that N fertilizer remarkably increased the gene copy numbers of amoA in AOB and nirK in denitrifying bacteria in soil at both stages,respectively. But this effect was not observed in the abundance of AOA all the time. Moreover, the abundance of AOB and denitrifying bacteria was very significantly(P < 0.01) related to the content of NH4+-N at both stages and NO3--N at tillering stage. DMPP mainly decreased the abundance of AOB but not AOA at tillering stage,which revealed that DMPP inhibited the soil nitrification by limiting the growth of AOB. Little changes in the abundance of AOB, AOA and denitrifying bacteria were detected among the treatments added with NBPT. 2) The profile of DGGE fingerprint indicated that, N fertilizer notably increased the band numbers in fingerprint of AOB but not affected that of AOA at both stages. In addition, N fertilizer obviously enhanced the band number in fingerprint of denitrifying bacteria at booting stage. The effects of NBPT and DMPP on the community structures of AOB, AOA and denitrifying bacteria were too little to be observed. Phylogenetic analysis indicated that the sequences of Nitrosomonas and Nitrosospira were more similar to dominant microflora of AOB in paddy field.【Conclusions】The abundance of AOB and denitrifying bacteria was distinctly improved by N fertilizer, and the community structures of both bacteria were sensitive to N fertilizer in paddy field, but AOA was stable, in red soil area of Southern China. The effect of urease inhibitor NBPT on the abundance and community structure of AOA,AOB and denitrifying bacteria was little. The nitrification inhibitor DMPP limited the growth of AOB at tillering stage, while the effect was negligible at booting stage. It may be the main way to alleviate nitrification. It is estimated that both NBPT and DMPP are harmless to the soil environment.
【Objectives】Urease inhibitor(UI) and nitrification inhibitor(NI), as nitrogen synergist, are used to improve the efficiency of nitrogen(N) fertilizer in agriculture. The aim of this study was to investigate the effect of UI and NI on the abundance and community structure of ammonia oxidizing bacteria(AOB), ammoniaoxidizing archaea(AOA), and denitrifying bacteria in paddy soil, and reveal the synergy mechanism of the inhibitors and their effects on soil environment.【Methods】A field experiment was conducted in a paddy field of red soil in southern China. A randomized block design with five treatments and three replicates was as follows:1) CK(no N control), 2) U(Urea), 3) U + UI, 4) U + NI, 5) U + UI + NI. The urease inhibitor and nitrification inhibitor were NBPT [N-(n-butyl) thiophosphrictriamide] and DMPP(3,4-dimethylpyrazole phosphate)respectively. The abundance and composition of AOB, AOA and denitrifying bacteria were investigated by fluorescent quantitative real-time PCR and denaturing gradient gel electrophoresis(DGGE) approaches based on amoA and nirK genes, at tillering stage and booting stage, respectively. Then, dominant populations was sequenced and phylogenetic analyzed.【Results】1) Real-time PCR analysis showed that N fertilizer remarkably increased the gene copy numbers of amoA in AOB and nirK in denitrifying bacteria in soil at both stages,respectively. But this effect was not observed in the abundance of AOA all the time. Moreover, the abundance of AOB and denitrifying bacteria was very significantly(P < 0.01) related to the content of NH4+-N at both stages and NO3--N at tillering stage. DMPP mainly decreased the abundance of AOB but not AOA at tillering stage,which revealed that DMPP inhibited the soil nitrification by limiting the growth of AOB. Little changes in the abundance of AOB, AOA and denitrifying bacteria were detected among the treatments added with NBPT. 2) The profile of DGGE fingerprint indicated that, N fertilizer notably increased the band numbers in fingerprint of AOB but not affected that of AOA at both stages. In addition, N fertilizer obviously enhanced the band number in fingerprint of denitrifying bacteria at booting stage. The effects of NBPT and DMPP on the community structures of AOB, AOA and denitrifying bacteria were too little to be observed. Phylogenetic analysis indicated that the sequences of Nitrosomonas and Nitrosospira were more similar to dominant microflora of AOB in paddy field.【Conclusions】The abundance of AOB and denitrifying bacteria was distinctly improved by N fertilizer, and the community structures of both bacteria were sensitive to N fertilizer in paddy field, but AOA was stable, in red soil area of Southern China. The effect of urease inhibitor NBPT on the abundance and community structure of AOA,AOB and denitrifying bacteria was little. The nitrification inhibitor DMPP limited the growth of AOB at tillering stage, while the effect was negligible at booting stage. It may be the main way to alleviate nitrification. It is estimated that both NBPT and DMPP are harmless to the soil environment.
引文
[1]朱兆良.农田中氮肥的损失与对策[J].土壤与环境,2000,9(1):1-6.Zhu Z L.Loss of fertilizer N from plants-soil system and the strategies and techniques for its reduction[J].Soil and Environmental Sciences,2000,9(1):1-6.
[2]蔡祖聪.尿素和KNO3对水稻土无机氮转化过程和产物的影响Ⅰ.无机氮转化过程[J].土壤学报,2003,40(2):239-245.Cai Z C.Effects of urea and KNO3 on processes and products of inorganic nitrogen transformation in paddy soils I.processes of inorganic nitrogen[J].Acta Pedologica Sinica,2003,40(2):239-245.
[3]贺纪正,张丽梅.土壤氮素转化的关键微生物过程及机制[J].微生物学通报,2013,40(1):98-108.He J Z,Zhang L M.Key processes and microbial mechanisms of soil nitrogen transformation[J].Microbiology China,2013,40(1):98-108.
[4]陈春兰,吴敏娜,魏文学.长期施用氮肥对土壤细菌硝化基因多样性及组成的影响[J].环境科学,2011,32(5):1489-1496.Chen C L,Wu M N,Wei W X.Effect of long-term application of nitrogen fertilizer on the diversity of nitrifying genes(amoA and hao)in paddy soil[J].Environmental Science,2011,32(5):1489-1496.
[5]郝永俊,吴松维,吴伟祥,等.好氧氨氧化菌的种群生态学研究进展[J].生态学报,2007,27(4):1573-1582.Hao Y J,Wu S W,Wu W X,et al.Research progress on the microbial ecology of aerobic ammonia-oxidizing bacteria[J].Acta Ecologica Sinica,2007,27(4):1573-1582.
[6]侯海军,秦红灵,陈春兰,等.土壤氮循环微生物过程的分子生态学研究进展[J].农业现代化研究,2014,35(5):588-594.Hou H J,Qin H L,Chen C L,et al.Research progress of the molecular ecology on microbiological processes in soil nitrogen cycling[J].Research of Agricultural Modernization,2014,35(5):588-594.
[7]刘正辉,李德豪.氨氧化古菌及其对氮循环贡献的研究进展[J].微生物学通报,2015,42(4):774-782.Liu Z H,Li D H.Ammonia-oxidizing archaea and their contribution to global nitrogen cycling:a review[J].Microbiology China,2015,42(4):774-782.
[8]Sayavedra-Soto L A,Hommes N G,Arp D J.Characterization of the gene encoding hydroxylamine oxidoreductase in Nitrosomonas europaea[J].Journal of Bacteriology,1994,176(2):504-510.
[9]Nicol G W,Schleper C.Ammonia-oxidising Crenarchaeota:important players in the nitrogen cycle?[J].Trends in Microbiology,2006,14(5):207-212.
[10]王海涛,郑天凌,杨小茹.土壤反硝化的分子生态学研究进展及其影响因素[J].农业环境科学学报,2013,32(10):1915-1924.Wang H T,Zheng T L,Yang X R.Molecular ecology research progress for soil denitrification and research status for its influencing factors[J].Journal of Agro-Environment Science,2013,32(10):1915-1924.
[11]Hussain Q,Liu Y,Jin Z,et al.Temporal dynamics of ammonia oxidizer(amoA)and denitrifier(nirK)communities in the rhizosphere of a rice ecosystem from Tai Lake region,China[J].Applied Soil Ecology,2011,48(2):210-218.
[12]Chen Z,Luo X,Hu R,et al.Impact of long-term fertilization on the composition of denitrifier communities based on nitrite reductase analyses in a paddy soil[J].Microbial Ecology,2010,60(4):850-861.
[13]李刚,修伟明,王杰,等.不同植被恢复模式下呼伦贝尔沙地土壤反硝化细菌nirK基因组成结构和多样性研究[J].草业学报,2015,24(1):115-123.Li G,Xiu W M,Wang J,et al.Community structure and diversity of soil denitrifying bacteria of the nirK gene type under different vegetation restoration patterns in the Hulun Buir sandy land[J].Acta Prataculturae Sinica,2015,24(1):115-123.
[14]Hou J,Dong Y,Fan Z.Effects of coated urea amended with biological inhibitors on physiological characteristics,yield and quality of peanut[J].Communications in Soil Science and PlantAnalysis,2014,45(7):896-911.
[15]徐星凯,周礼恺.脲酶抑制剂/硝化抑制剂对植稻土壤中尿素N行为的影响[J].生态学报,2001,21(10):1682-1686.Xu X K,Zhou L K.Effect of urease/nitrification inhibitors on the behavior of urea-N in the soil planted to rice[J].Acta Ecologica Sinica,2001,21(10):1682-1686.
[16]Trenkel M E.Slow-and controlled-release and stabilized fertilizers[M].an option for enhancing nutrient efficiency in agriculture(2nd edition)[M].Paris,France:International Fertilizer Industry Association,2010.
[17]张文学,孙刚,何萍,等.脲酶抑制剂与硝化抑制剂对稻田氨挥发的影响[J].植物营养与肥料学报,2013,19(6):1411-1419.Zhang W X,Sun G,He P,et al.Effects of urease and nitrification inhibitors on ammonia volatilization from paddy fields[J].Journal of Plant Nutrition and Fertilizer,2013,19(6):1411-1419.
[18]王慧颖,徐明岗,马想,等.长期施肥下我国农田土壤微生物及氨氧化菌研究进展[J].中国土壤与肥料,2018,(2):1-12.Wang H Y,Xu M G,Ma X,et al.Research advances of microorganism and ammonia oxidizing bacteria under long term fertilization in Chinese typical cropland[J].Soil and Fertilizer Siences in China,2018,(2):1-12.
[19]王军,申田田,车钊,等.有机和无机肥配比对黄褐土硝化和反硝化微生物丰度及功能的影响[J].植物营养与肥料学报,2018,24(3):1-10.Wang J,Shen T T,Che Z,et al.Effects of combination of organic and inorganic fertilizers on abundances of nitrifiers and denitrifiers and their function in yellow-cinnamon soil[J].Journal of Plant Nutrition and Fertilizer,2018,24(3):1-10.
[20]Rotthauwe J H,Witzel K P,Liesack W.The ammonia monooxygenase structural gene amoA as a functional marker:molecular fine-scale analysis of natural ammonia-oxidizing populations[J].Applied and Environmental Microbiology,1997,63(12):4704-4712.
[21]Long X,Chen C,Xu Z,et al.Abundance and community structure of ammonia-oxidizing bacteria and archaea in a temperate forest ecosystem under ten-years elevated CO2[J].Soil Biology and Biochemistry,2012,46:163-171.
[22]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.
[23]Throb?ck I N,Enwall K,Jarvis?,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.
[24]张文学,孙刚,何萍,等.双季稻田添加脲酶抑制剂NBPT氮肥的最高减量潜力研究[J].植物营养与肥料学报,2014,20(4):821-830.Zhang W X,Sun G,He P,et al.Highest potential of subtracting nitrogen fertilizer through addition of urease inhibitor NBPT in double-cropping paddy fields[J].Journal of Plant Nutrition and Fertilizer,2014,20(4):821-830.
[25]陈哲,袁红朝,吴金水,等.长期施肥制度对稻田土壤反硝化细菌群落活性和结构的影响[J].生态学报,2009,29(11):5923-5929.Chen Z,Yuan H C,Wu J S,et al.Activity and com position of the denitrifying bacteria community respond differently to long-term fertilization[J].Acta Ecologica Sinica,2009,29(11):5923-5929.
[26]Wang Y,Ke X,Wu L,et al.Community composition of ammoniaoxidizing bacteria and archaea in rice field soil as affected by nitrogen fertilization[J].Systematic and Applied Microbiology,2009,32(1):27-36.
[27]解开治,徐培智,蒋瑞萍,等.有机无机肥配施提升冷浸田土壤氮转化相关微生物丰度和水稻产量[J].植物营养与肥料学报,2016,22(5):1267-1277.Xie K Z,Xu P Z,Jiang R P,et al.Combined application of inorganic and organic fertilizers improve rice yield and the abundance of soil nitrogen-cycling microbes in cold waterlogged paddy fields[J].Journal of Plant Nutrition and Fertilizer,2016,22(5):1267-1277.
[28]卢圣鄂,王蓥燕,陈勇,等.不同施肥制度对石灰性紫色水稻土中氨氧化古菌群落结构的影响[J].生态学报,2016,36(21):6919-6927.Lu S E,Wang Y Y,Chen Y,et al.Impact of different long-term fertilization systems on ammonia oxidation Archaea community structures in Calcareous Purple Paddy soil[J].Acta Ecologica Sinica,2016,36(21):6919-6927.
[29]Ouyang Y,Norton J M,Stark J M,et al.Ammonia-oxidizing bacteria are more responsive than archaea to nitrogen source in an agricultural soil[J].Soil Biology and Biochemistry,2016,96:4-15.
[30]Huang L,Dong H,Wang S,et al.Diversity and abundance of ammonia-oxidizing archaea and bacteria in diverse Chinese paddy soils[J].Geomicrobiology Journal,2014,31(1):12-22.
[31]Cui P Y,Fan F L,Yin C,et al.Urea-and nitrapyrin-affected N2Oemission 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.
[32]Chen Y,Xu Z,Hu H,et al.Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia[J].Applied Soil Ecology,2013,68:36-45.
[33]Okano Y,Hristova K R,Leutenegger C M,et al.Application of realtime PCR to study effects of ammonium on population size of ammonia-oxidizing bacteria in soil[J].Applied and Environmental Microbiology,2004,70(2):1008-1016.
[34]Di H J,Cameron K C,Shen J P,et al.Ammonia-oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions[J].FEMS Microbiology Ecology,2010,72(3):386-394.
[35]Gong P,Zhang L,Wu Z,et al.Does the nitrification inhibitor dicyandiamide affect the abundance of ammonia-oxidizing bacteria and archaea in a Hap-Udic Luvisol?[J].Journal of Soil Science and Plant Nutrition,2013,13(1):35-42.
[36]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.
[37]Kleineidam K,Ko?mrlj K,Kublik S,et al.Influence of the nitrification inhibitor 3,4-dimethylpyrazole phosphate(DMPP)on ammonia-oxidizing bacteria and archaea in rhizosphere and bulk soil[J].Chemosphere,2011,84(1):182-186.
[38]周卉,田光明,李华.DMPP减少稻田土壤氮素损失的研究进展[J].土壤,2013,45(6):964-969.Zhou H,Tian G M,Li H.Research progress of DMPP in reducing nitrogen losses from paddy fields[J].Soils,2013,45(6):964-969.
[39]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 International Society for Microbial Ecology Journal,2012,6(5):1032-1045.
[40]Gubry-Rangin C,Nicol G W,Prosser J I.Archaea rather than bacteria control nitrification in two agricultural acidic soils[J].FEMSMicrobiology Ecology,2010,74(3):566-574.
[41]Zerulla W,Barth T,Dressel J,et al.3,4-dimethylpyrazole phosphate(DMPP)-a new nitrification inhibitor for agriculture and horticulture[J].Biology and Fertility of Soils,2001,34:79-84.
[42]武志杰,陈利军.缓释/控释肥料:原理与应用[M].北京:科学出版社,2003.Wu Z J,Chen L J.Slow/controlled-release fertilizer:Principle and application[M].Beijing:Science Press,2003.
[43]曾希柏,王亚男,王玉忠,等.施肥对设施菜地nirK型反硝化细菌群落结构和丰度的影响[J].应用生态学报,2014,25(2):505-514.Zeng X B,Wang Y N,Wang Y Z,et al.Effects of different fertilization regimes on abundance and community structure of the nirK-type denitrifying bacteria in greenhouse vegetable soils[J].Chinese Journal of Applied Ecology,2014,25(2):505-514.
[44]Hayatsu M,Tago K,Saito M.Various players in the nitrogen cycle:diversity and functions of the microorganisms involved in nitrification and denitrification[J].Soil Science and Plant Nutrition,2008,54(1):33-45.
[45]Ligi T,Truu M,Truu J,et al.Effects of soil chemical characteristics and water regime on denitrification genes(nirS,nirK,and nosZ)abundances in a created riverine wetland complex[J].Ecological Engineering,2014,72:47-55.
[46]Jin Z,Li L,Liu X,et al.Impact of long-term fertilization on community structure of ammonia oxidizing and denitrifying bacteria based on amoA and nirK genes in a rice paddy from Tai Lake Region,China[J].Journal of Integrative Agriculture,2014,13(10):2286-2298.
[47]宋亚娜,吴明基,林艳.稻田土壤nirS型反硝化细菌群落对氮肥水平的响应[J].中国农业科学,2013,46(9):1818-1826.Song Y N,Wu M J,Lin Y.Response of the denitrifying bacterial nirSgene community to nitrogen fertilizer in paddy field[J].Scientia Agricultura Sinica,2013,46(9):1818-1826.
[48]Barrena I,Menéndez S,Correa-Galeote D,et al.Soil water content modulates the effect of the nitrification inhibitor 3,4-dimethylpyrazole phosphate(DMPP)on nitrifying and denitrifying bacteria[J].Geoderma,2017,303:1-8.
[49]He J Z,C X,Zhang L M,et al.Ammonia-oxidizing bacteria and archaea in different paddy soils of China[A].International Union of Soil Sciences.Proceedings of the 19th World Congress of Soil Science:Soil solutions for a changing world[C].Brisbane,Australia,2010.9-12.
[50]Chen X P,Zhu Y G,Xia Y,et al.Ammonia-oxidizing archaea:important players in paddy rhizosphere soil?[J].Environmental Microbiology,2008,10(8):1978-1987.
[2]蔡祖聪.尿素和KNO3对水稻土无机氮转化过程和产物的影响Ⅰ.无机氮转化过程[J].土壤学报,2003,40(2):239-245.Cai Z C.Effects of urea and KNO3 on processes and products of inorganic nitrogen transformation in paddy soils I.processes of inorganic nitrogen[J].Acta Pedologica Sinica,2003,40(2):239-245.
[3]贺纪正,张丽梅.土壤氮素转化的关键微生物过程及机制[J].微生物学通报,2013,40(1):98-108.He J Z,Zhang L M.Key processes and microbial mechanisms of soil nitrogen transformation[J].Microbiology China,2013,40(1):98-108.
[4]陈春兰,吴敏娜,魏文学.长期施用氮肥对土壤细菌硝化基因多样性及组成的影响[J].环境科学,2011,32(5):1489-1496.Chen C L,Wu M N,Wei W X.Effect of long-term application of nitrogen fertilizer on the diversity of nitrifying genes(amoA and hao)in paddy soil[J].Environmental Science,2011,32(5):1489-1496.
[5]郝永俊,吴松维,吴伟祥,等.好氧氨氧化菌的种群生态学研究进展[J].生态学报,2007,27(4):1573-1582.Hao Y J,Wu S W,Wu W X,et al.Research progress on the microbial ecology of aerobic ammonia-oxidizing bacteria[J].Acta Ecologica Sinica,2007,27(4):1573-1582.
[6]侯海军,秦红灵,陈春兰,等.土壤氮循环微生物过程的分子生态学研究进展[J].农业现代化研究,2014,35(5):588-594.Hou H J,Qin H L,Chen C L,et al.Research progress of the molecular ecology on microbiological processes in soil nitrogen cycling[J].Research of Agricultural Modernization,2014,35(5):588-594.
[7]刘正辉,李德豪.氨氧化古菌及其对氮循环贡献的研究进展[J].微生物学通报,2015,42(4):774-782.Liu Z H,Li D H.Ammonia-oxidizing archaea and their contribution to global nitrogen cycling:a review[J].Microbiology China,2015,42(4):774-782.
[8]Sayavedra-Soto L A,Hommes N G,Arp D J.Characterization of the gene encoding hydroxylamine oxidoreductase in Nitrosomonas europaea[J].Journal of Bacteriology,1994,176(2):504-510.
[9]Nicol G W,Schleper C.Ammonia-oxidising Crenarchaeota:important players in the nitrogen cycle?[J].Trends in Microbiology,2006,14(5):207-212.
[10]王海涛,郑天凌,杨小茹.土壤反硝化的分子生态学研究进展及其影响因素[J].农业环境科学学报,2013,32(10):1915-1924.Wang H T,Zheng T L,Yang X R.Molecular ecology research progress for soil denitrification and research status for its influencing factors[J].Journal of Agro-Environment Science,2013,32(10):1915-1924.
[11]Hussain Q,Liu Y,Jin Z,et al.Temporal dynamics of ammonia oxidizer(amoA)and denitrifier(nirK)communities in the rhizosphere of a rice ecosystem from Tai Lake region,China[J].Applied Soil Ecology,2011,48(2):210-218.
[12]Chen Z,Luo X,Hu R,et al.Impact of long-term fertilization on the composition of denitrifier communities based on nitrite reductase analyses in a paddy soil[J].Microbial Ecology,2010,60(4):850-861.
[13]李刚,修伟明,王杰,等.不同植被恢复模式下呼伦贝尔沙地土壤反硝化细菌nirK基因组成结构和多样性研究[J].草业学报,2015,24(1):115-123.Li G,Xiu W M,Wang J,et al.Community structure and diversity of soil denitrifying bacteria of the nirK gene type under different vegetation restoration patterns in the Hulun Buir sandy land[J].Acta Prataculturae Sinica,2015,24(1):115-123.
[14]Hou J,Dong Y,Fan Z.Effects of coated urea amended with biological inhibitors on physiological characteristics,yield and quality of peanut[J].Communications in Soil Science and PlantAnalysis,2014,45(7):896-911.
[15]徐星凯,周礼恺.脲酶抑制剂/硝化抑制剂对植稻土壤中尿素N行为的影响[J].生态学报,2001,21(10):1682-1686.Xu X K,Zhou L K.Effect of urease/nitrification inhibitors on the behavior of urea-N in the soil planted to rice[J].Acta Ecologica Sinica,2001,21(10):1682-1686.
[16]Trenkel M E.Slow-and controlled-release and stabilized fertilizers[M].an option for enhancing nutrient efficiency in agriculture(2nd edition)[M].Paris,France:International Fertilizer Industry Association,2010.
[17]张文学,孙刚,何萍,等.脲酶抑制剂与硝化抑制剂对稻田氨挥发的影响[J].植物营养与肥料学报,2013,19(6):1411-1419.Zhang W X,Sun G,He P,et al.Effects of urease and nitrification inhibitors on ammonia volatilization from paddy fields[J].Journal of Plant Nutrition and Fertilizer,2013,19(6):1411-1419.
[18]王慧颖,徐明岗,马想,等.长期施肥下我国农田土壤微生物及氨氧化菌研究进展[J].中国土壤与肥料,2018,(2):1-12.Wang H Y,Xu M G,Ma X,et al.Research advances of microorganism and ammonia oxidizing bacteria under long term fertilization in Chinese typical cropland[J].Soil and Fertilizer Siences in China,2018,(2):1-12.
[19]王军,申田田,车钊,等.有机和无机肥配比对黄褐土硝化和反硝化微生物丰度及功能的影响[J].植物营养与肥料学报,2018,24(3):1-10.Wang J,Shen T T,Che Z,et al.Effects of combination of organic and inorganic fertilizers on abundances of nitrifiers and denitrifiers and their function in yellow-cinnamon soil[J].Journal of Plant Nutrition and Fertilizer,2018,24(3):1-10.
[20]Rotthauwe J H,Witzel K P,Liesack W.The ammonia monooxygenase structural gene amoA as a functional marker:molecular fine-scale analysis of natural ammonia-oxidizing populations[J].Applied and Environmental Microbiology,1997,63(12):4704-4712.
[21]Long X,Chen C,Xu Z,et al.Abundance and community structure of ammonia-oxidizing bacteria and archaea in a temperate forest ecosystem under ten-years elevated CO2[J].Soil Biology and Biochemistry,2012,46:163-171.
[22]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.
[23]Throb?ck I N,Enwall K,Jarvis?,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.
[24]张文学,孙刚,何萍,等.双季稻田添加脲酶抑制剂NBPT氮肥的最高减量潜力研究[J].植物营养与肥料学报,2014,20(4):821-830.Zhang W X,Sun G,He P,et al.Highest potential of subtracting nitrogen fertilizer through addition of urease inhibitor NBPT in double-cropping paddy fields[J].Journal of Plant Nutrition and Fertilizer,2014,20(4):821-830.
[25]陈哲,袁红朝,吴金水,等.长期施肥制度对稻田土壤反硝化细菌群落活性和结构的影响[J].生态学报,2009,29(11):5923-5929.Chen Z,Yuan H C,Wu J S,et al.Activity and com position of the denitrifying bacteria community respond differently to long-term fertilization[J].Acta Ecologica Sinica,2009,29(11):5923-5929.
[26]Wang Y,Ke X,Wu L,et al.Community composition of ammoniaoxidizing bacteria and archaea in rice field soil as affected by nitrogen fertilization[J].Systematic and Applied Microbiology,2009,32(1):27-36.
[27]解开治,徐培智,蒋瑞萍,等.有机无机肥配施提升冷浸田土壤氮转化相关微生物丰度和水稻产量[J].植物营养与肥料学报,2016,22(5):1267-1277.Xie K Z,Xu P Z,Jiang R P,et al.Combined application of inorganic and organic fertilizers improve rice yield and the abundance of soil nitrogen-cycling microbes in cold waterlogged paddy fields[J].Journal of Plant Nutrition and Fertilizer,2016,22(5):1267-1277.
[28]卢圣鄂,王蓥燕,陈勇,等.不同施肥制度对石灰性紫色水稻土中氨氧化古菌群落结构的影响[J].生态学报,2016,36(21):6919-6927.Lu S E,Wang Y Y,Chen Y,et al.Impact of different long-term fertilization systems on ammonia oxidation Archaea community structures in Calcareous Purple Paddy soil[J].Acta Ecologica Sinica,2016,36(21):6919-6927.
[29]Ouyang Y,Norton J M,Stark J M,et al.Ammonia-oxidizing bacteria are more responsive than archaea to nitrogen source in an agricultural soil[J].Soil Biology and Biochemistry,2016,96:4-15.
[30]Huang L,Dong H,Wang S,et al.Diversity and abundance of ammonia-oxidizing archaea and bacteria in diverse Chinese paddy soils[J].Geomicrobiology Journal,2014,31(1):12-22.
[31]Cui P Y,Fan F L,Yin C,et al.Urea-and nitrapyrin-affected N2Oemission 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.
[32]Chen Y,Xu Z,Hu H,et al.Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia[J].Applied Soil Ecology,2013,68:36-45.
[33]Okano Y,Hristova K R,Leutenegger C M,et al.Application of realtime PCR to study effects of ammonium on population size of ammonia-oxidizing bacteria in soil[J].Applied and Environmental Microbiology,2004,70(2):1008-1016.
[34]Di H J,Cameron K C,Shen J P,et al.Ammonia-oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions[J].FEMS Microbiology Ecology,2010,72(3):386-394.
[35]Gong P,Zhang L,Wu Z,et al.Does the nitrification inhibitor dicyandiamide affect the abundance of ammonia-oxidizing bacteria and archaea in a Hap-Udic Luvisol?[J].Journal of Soil Science and Plant Nutrition,2013,13(1):35-42.
[36]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.
[37]Kleineidam K,Ko?mrlj K,Kublik S,et al.Influence of the nitrification inhibitor 3,4-dimethylpyrazole phosphate(DMPP)on ammonia-oxidizing bacteria and archaea in rhizosphere and bulk soil[J].Chemosphere,2011,84(1):182-186.
[38]周卉,田光明,李华.DMPP减少稻田土壤氮素损失的研究进展[J].土壤,2013,45(6):964-969.Zhou H,Tian G M,Li H.Research progress of DMPP in reducing nitrogen losses from paddy fields[J].Soils,2013,45(6):964-969.
[39]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 International Society for Microbial Ecology Journal,2012,6(5):1032-1045.
[40]Gubry-Rangin C,Nicol G W,Prosser J I.Archaea rather than bacteria control nitrification in two agricultural acidic soils[J].FEMSMicrobiology Ecology,2010,74(3):566-574.
[41]Zerulla W,Barth T,Dressel J,et al.3,4-dimethylpyrazole phosphate(DMPP)-a new nitrification inhibitor for agriculture and horticulture[J].Biology and Fertility of Soils,2001,34:79-84.
[42]武志杰,陈利军.缓释/控释肥料:原理与应用[M].北京:科学出版社,2003.Wu Z J,Chen L J.Slow/controlled-release fertilizer:Principle and application[M].Beijing:Science Press,2003.
[43]曾希柏,王亚男,王玉忠,等.施肥对设施菜地nirK型反硝化细菌群落结构和丰度的影响[J].应用生态学报,2014,25(2):505-514.Zeng X B,Wang Y N,Wang Y Z,et al.Effects of different fertilization regimes on abundance and community structure of the nirK-type denitrifying bacteria in greenhouse vegetable soils[J].Chinese Journal of Applied Ecology,2014,25(2):505-514.
[44]Hayatsu M,Tago K,Saito M.Various players in the nitrogen cycle:diversity and functions of the microorganisms involved in nitrification and denitrification[J].Soil Science and Plant Nutrition,2008,54(1):33-45.
[45]Ligi T,Truu M,Truu J,et al.Effects of soil chemical characteristics and water regime on denitrification genes(nirS,nirK,and nosZ)abundances in a created riverine wetland complex[J].Ecological Engineering,2014,72:47-55.
[46]Jin Z,Li L,Liu X,et al.Impact of long-term fertilization on community structure of ammonia oxidizing and denitrifying bacteria based on amoA and nirK genes in a rice paddy from Tai Lake Region,China[J].Journal of Integrative Agriculture,2014,13(10):2286-2298.
[47]宋亚娜,吴明基,林艳.稻田土壤nirS型反硝化细菌群落对氮肥水平的响应[J].中国农业科学,2013,46(9):1818-1826.Song Y N,Wu M J,Lin Y.Response of the denitrifying bacterial nirSgene community to nitrogen fertilizer in paddy field[J].Scientia Agricultura Sinica,2013,46(9):1818-1826.
[48]Barrena I,Menéndez S,Correa-Galeote D,et al.Soil water content modulates the effect of the nitrification inhibitor 3,4-dimethylpyrazole phosphate(DMPP)on nitrifying and denitrifying bacteria[J].Geoderma,2017,303:1-8.
[49]He J Z,C X,Zhang L M,et al.Ammonia-oxidizing bacteria and archaea in different paddy soils of China[A].International Union of Soil Sciences.Proceedings of the 19th World Congress of Soil Science:Soil solutions for a changing world[C].Brisbane,Australia,2010.9-12.
[50]Chen X P,Zhu Y G,Xia Y,et al.Ammonia-oxidizing archaea:important players in paddy rhizosphere soil?[J].Environmental Microbiology,2008,10(8):1978-1987.