厌氧条件下土壤反硝化气体(N2、N2O、NO)和CO2排放——氦环境培养—气体同步直接测定法的应用初探

冯琪; 王睿; 郑循华; 张伟; 邹建文

doi:10.3878/j.issn.1006-9585.2013.11014

厌氧条件下土壤反硝化气体(N₂、N₂O、NO)和CO₂排放——氦环境培养—气体同步直接测定法的应用初探

doi: 10.3878/j.issn.1006-9585.2013.11014

1.
南京农业大学资源与环境科学学院, 南京 210095;中国科学院大气物理研究所大气边界层物理和大气化学国家重点实验室, 北京 100029
2.
中国科学院大气物理研究所大气边界层物理和大气化学国家重点实验室, 北京 100029
3.
南京农业大学资源与环境科学学院, 南京 210095

基金项目: 公益性行业(农业)科研专项项目200803036;国家自然科学基金国际合作项目40711130636

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出版历程
- 收稿日期: 2011-01-27
- 修回日期: 2013-02-25

Direct Measurements of Denitrification Gas (N₂, N₂O, NO) and CO₂ Emissions Using the Gas-Flow-Soil-Core Technique with Helium Environment Incubation

1.
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095;State Key Laboratory of Atmospheric Boundary Layer Physics and Atmospheric Chemistry, Institute of Atmospheric Physics, Chinese Academy of Sciences, Beijing 100029
2.
State Key Laboratory of Atmospheric Boundary Layer Physics and Atmospheric Chemistry, Institute of Atmospheric Physics, Chinese Academy of Sciences, Beijing 100029
3.
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095

摘要

摘要: 反硝化过程是维系闭合氮循环所必需的氮素形态转化环节。土壤反硝化过程速率及产物比的直接测定是研究氮循环过程机理的基础,但却是一个难题。为解决此难题,德国卡尔斯鲁厄技术研究所与中国科学院大气物理研究所最近合作新建了一套通过氦环境培养—气体同步直接测定土壤反硝化气体——氮气(N₂)、氧化亚氮(N₂O)、一氧化氮(NO)和二氧化碳(CO₂)排放的系统和与之配套的三阶段培养方法。为检验该新建系统和配套方法测定土壤反硝化过程的准确性和可靠性,以华北地区广泛分布的盐碱地农田土壤(采自山西运城)为研究对象开展实验室培养试验,在初始可溶性有机碳(DOC)供应比较充足约300 mgC kg^-1 干土(d.s.)的条件下,测试了不同初始土壤硝态氮含量水平(10、100 mgN kg^-1d.s.左右,分别表示为10N和100N)的反硝化气体和CO₂排放过程。结果显示:100N的反硝化速率(定义为N₂、N₂O和NO排放速率之和)显著高于10N处理(统计检验显著水平p<0.01);两个处理的反硝化产物均以N₂为主(质量比分别占77%和75%),产物的NO/N₂O摩尔比分别为1.2和1.5,N₂O/N₂摩尔比均为0.19;土壤反硝化气体动态排放速率及相关指标的测定结果表明,培养土壤中消失的硝态氮被回收81%～87%,培养前后的氮平衡率达92%～95%。因此,该新建方法测定土壤反硝化速率和产物比的结果具有很好的可靠性,为定量研究土壤反硝化过程提供了有效的直接测定手段。研究中检测到的土壤反硝化产物NO/N₂O摩尔比大于1,不同于以往用液体培养基纯培养反硝化细菌得出的NO/N₂O摩尔比远小于1的结论。这意味着,不能用NO/N₂O摩尔比小于1与否来推断土壤排放的N₂O和NO是主要来源于反硝化作用还是硝化作用。
- 氦环境培养—气体同步直接测定法 /
- 反硝化作用 /
- N₂、N₂O、NO和CO₂排放 /
- NO/N₂O比 /
- N₂O/N₂比 /
- 回收率
Abstract: Denitrification is the key process of nitrogen transformation to close the global nitrogen cycle. Quantification of microbial denitrification rate and its ratios of products is the key step for obtaining insights into nitrogen cycling processes, though it is very difficult. To enable precise quantification of the rates of the entire process, as well as individual products, a system of gas-flow-soil-core technique with helium-environment incubation was recently established and a three-period incubation method was set up by the Karlsruhe Institute of Technology and the Institute of Atmospheric Physics, Chinese Academy of Sciences. Using this system, dynamic emission rates of dinitrogen (N₂), nitrous oxide (N₂O), nitric oxide (NO) and carbon dioxide (CO₂), which are the gaseous products of microbial denitrification, can be simultaneously and directly measured. In this study, we conducted the first soil incubation experiment to test the reliability of the measurements by this system in association with the newly proposed incubation method. Our experiment included two levels of initial soil nitrate (NO₃^-) content, which were around 10 and 100 mgN kg^-1d.s. (dry soil), respectively (hereinafter referred to as 10N and 100N). For either nitrate level, sufficient dissolved organic carbon (DOC) was initially supplied (glucose was added to establish an initial DOC content of around 300 mgC kg^-1d.s.). The incubated fresh soil was a silty clay loam of the northern China. It was sampled from a typical cropland rotationally cultivated with a double cropping system of summer maize and winter wheat and a single cropping system of cotton. Our results show that the microbial denitrification rate was significantly higher for the 100N than 10N treatments (p<0.01), and N₂ was the main product in both treatments (with mass fractions of 77% and 75%, respectively). The molar ratios were 1.2 (10N) to 1.5 (100N) for NO/N₂O and were 0.19 (both treatments) for N₂O/N₂. The measurements of individual denitrification gases recovered 81%-87% of disappeared nitrate during incubation. The direct dynamic detection of individual denitrification gases and the measurements of DOC, ammonium, nitrate, microbial biomass carbon, and microbial biomass nitrogen at the beginning and end of incubation gave mass balance rates of 92%-95% for nitrogen. These results suggest that the tested system, in association with the proposed incubation method, could precisely determine the dynamical rate of microbial denitrification. The molar ratios of NO/N₂O given by our data were greater than 1. This differs from previous knowledge of much lower ratios yielded by denitrifiers. This difference implicates that the NO/N₂O ratio of 1 may not be used as the threshold to indicate nitrification or denitrification processes.
- Gas-flow-soil-core technique with helium-environment incubation /
- Denitrification /
- Dynamics of N₂ /
- N₂O /
- NO /
- and CO₂emissions

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参考文献(27)

[1]	Anderson I C, Levine J S. 1986. Relative rates of nitric oxide and nitrous oxide production by nitrifiers, denitrifiers, and nitrate respirers[J]. Applied and Environmental Microbiology, 51 (5): 938-945.
[2]	鲍士旦. 2000. 土壤农化分析[M]. 北京: 中国农业出版社, 49-56. Bao Shidan. 2000. Soil Agricultural Chemistry Analysis (in Chinese)[M]. Beijing: China Agriculture Press, 49-56.
[3]	Bollmann A, Conrad R. 1997. Acetylene blockage technique leads to underestimation of denitrification rates in oxic soils due to scavenging of intermediate nitric oxide[J]. Soil Biology and Biochemistry, 29 (7): 1067-1077.
[4]	Bridgham S D, Updegraff K, Pastor J. 1998. Carbon, nitrogen, and phosphorus mineralization in northern wetlands[J]. Ecology, 79 (5): 1545-1561.
[5]	Butterbach-Bahl K, Willibald G, Papen H. 2002. Soil core method for direct simultaneous determination of N₂and N₂O emissions from forest soils[J]. Plant and Soil, 240 (1): 105-116.
[6]	Cárdenas L M, Hawkins J M B, Chadwick D, et al. 2003. Biogenic gas emissions from soils measured using a new automated laboratory incubation system[J]. Soil Biology and Biochemistry, 35 (6): 867-870.
[7]	Caskey M H, Tiedje J M. 1979. Evidence for clostridia as agents of dissimilatory reduction of NO₃^- to H₄⁺ in soils[J]. Soil Science Society of America Journal, 43 (5): 931-936.
[8]	Cuhel J, Simek M, Laughlin R J, et al. 2010. Insights into the effect of soil pH on N₂O and N₂ emissions and denitrifier community size and activity[J]. Applied and Environmental Microbiology, 76 (6): 1870-1878.
[9]	Dannenmann M, Butterbach-Bahl K, Gasche R, et al. 2008. Dinitrogen emissions and the N₂: N₂O emission ratio of a Rendzic Leptosol as influenced by pH and forest thinning[J]. Soil Biology and Biochemistry, 40 (9): 2317-2323.
[10]	Davidson E A, Seitzinger S. 2006. The enigma of progress in denitrification research[J]. Ecological Applications, 16 (6): 2057-2063.
[11]	范晓晖, 朱兆良. 2002. 旱地土壤中的硝化—反硝化作用[J]. 土壤通报, 33 (5): 385-391. Fan Xiaohui, Zhu Zhaoliang. 2002. Nitrification and denitrification in upland soils[J]. Chinese Journal of Soil Science (in Chinese), 33 (5): 385-391.
[12]	Focht D D. 1985. Differences in nitrogen-15 enrichments of evolvednitrous oxide and dinitrogen and the question of auniform nitrate-15 pool[J]. Soil Science Society of America Journal, 49 (3): 786-790.
[13]	Groffman P M, Altabet M A, Bohlke J K, et al. 2006. Methods for measuring denitrification: diverse approaches to a difficult problem[J]. Ecological Applications, 16 (6): 2091-2122.
[14]	Hutchinson G L, Davidson E A. 1993. Processes for production and consumption of gaseous nitrogen oxides in soil[M]//Harper L A, Mosier A R, Duxbury J M, et al. Agricultural Ecosystem Effects on Trace Gases and Global Climate Change. Madison, WI: ASA Special Publication, 79-93.
[15]	IPCC. 2007. Climate Change 2007: The Physical Scientific Basis[M]//Contribution of Working Group 1 to the Fourth Assessment Report of the Intergovernmental Panel on Climate Chang. Solomon S, Qin D, Manning M, et al., Eds. Cambridge, United Kingdom and New York, NY, USA: Cambridge University Press, 996.
[16]	Jordan T E, Weller D E, Correll D L. 1998. Denitrification in surface soils of a riparian forest: Effects of water, nitrate, and sucrose[J]. Soil Biology and Biochemistry, 30 (7): 833-843.
[17]	Klemedtsson L, Hansson G, Mosier A. 1990. The use of acetylene for the quantification of N₂ production from biological processes in soil[M]//Revsbech J P, Sorensen J. Denitrification in Soil and Sediment. New York: Plenum Press, 167-180.
[18]	Knowles R. 1982. Denitrification[J]. Microbiological Reviews, 46 (1): 43-70.
[19]	李明, 梁旺国, 郑循华, 等. 2009. 晋南地区典型盐碱地棉田的NO排放特征[J]. 气候与环境研究, 14 (3): 318-328. Li Ming, Liang Wangguo, Zheng Xunhua, et al. 2009. Characteristics of NO emission from typical saline soil of southern Shanxi cotton land[J]. Climatic and Environmental Research (in Chinese), 14 (3): 318-328.
[20]	李振高, 潘映华, 伍期途, 等. 1989. 太湖地区水稻土优势反硝化细菌的数量、组成与酶活性[J]. 土壤学报, 26 (1): 79-86. Li Zhengao, Pan Yinghua, Wu Qitu, et al. 1989. Numbers, compositions and enzyme activities of denitrifiers in paddy soils of Taihu Lake district[J]. Acta Pedologica Sinica (in Chinese), 26 (1): 79-86.
[21]	McCready R G L, Gould W D, Brendregt R W. 1983. Nitrogen isotope fractionation during the reduction of NO₃^-to NH₄⁺ by Desulfovibrio sp.[J]. Canadian Journal of Microbiology, 29 (2): 231-234.
[22]	Molstad L, Dörsch P, Bakken L R. 2007. Robotized incubation system for monitoring gases (O₂, NO, N₂O, N₂) in denitrifying cultures[J]. Journal Microbiological Methods, 71 (3): 202-211.
[23]	Nömmik H. 1956. Investigations on denitrification in soil[J]. Acta Agriculturae Scandinavica, 6 (2): 195-228.
[24]	Reddy K R, DeLaune R D. 2008. Biogeochemistry of Wetlands: Science and Applications[M]. CRC Press: Boca Raton: 136-151.
[25]	Rice W A, Paul E A. 1972. The organisms and biological processes involved in asymbiotic nitrogen fixation in waterlogged soil amended with straw[J]. Canadian Journal of Microbiology, 18 (6): 715-723.
[26]	Robertson G P, Groffman P M. 2007. Nitrogen Transformations[M]//Paul E 摁??楓潯捩桬攠浍楩獣瑲牯祢???????????????水???扤爠?卩睯散牨瑥獭?????攮爠挳歲硤?剥??嘠汁慭獳獴慥歲????ㄠ???捴???渠晈汥畩敤湥捬敢?潲晧?挠慌牯扮潤湯?愬瘠慎楥汷愠扙楯汲楫琬礠?潸湦?瑲桤攬?灐牡潲摩畳挬琠楓潡湮?潄晩?乧佯??乓?獮甠扆????獩畳扣?伬??乩?獧畡扰????猠畓批????伬?獔畯扫????獅畬扳??扩祥?猬漠椳水?挭漳父攴献?摢畲爾楓湡杨?慡湷慡整爠潋戠楌挬?楋湥捥畮扥慹琠楄漠湒嬮?崱??倶氮愠湎瑩?慲湯摵?匠潯楸汩??ㄠ????????ㄠ????????扬牳?半灝愮爠汁楤湶条???倠??圠敓獯瑩???坣?????????漠搱椰昳椭挱愴琸椮漼湢獲￣瑓潣?瑥桥敲?晃甬洠楗条慳瑳業潡湮?攠硒琬爠慂捵瑴楴潥湲?瑡散捨栭湂楡煨畬攠?琬漠?灴攠牡浬椮琠′猰椰洹甮氠瑔慨湥攠潲略獬?整硩瑯牮慳捨瑩楰漠湢?慴湷摥?敮猠瑎椼浳慵瑢椾漲渼?潳晵?猾潏椬氠?浏椬挠牡潮扤椠慎氼???愾渲搼?浳極换爾漠扦楬慵汸?乳嬠?嵲???潦浥浲畴湩楬捩慺瑥楤漠湡獮?椠湩?卲潩楧污?卥捤椠敤湲捹敬?慮湤搠?偯汩慬湳琠??渠慴汨祥猠楁獲??ㄠ????????????????扩牳?呡楮敛摊橝攮??????ㄠ??????挳漱水漠木礱?漲昩?搠攲渷椳琭爲椸昳椮挼慢瑲椾潓湣?慯湬摥?摩楥獬獤椠浄椬氠慈瑡潷牫祩?湳椠瑊爠慍琠敂?爠敊摡畣捫瑳楯潮渠?琠潍?愠洱洹漹渷楡甮洠孄?嵶??婯数桭湥摮整爠?????????極潭氠潡杴祭?潳晰??湲慥攠牳潯扩楬挠??楣捵牢潡潴物杯慮渠楴獥浣獨??乱敵睥?奦潯牲欠??坲楥汣整礠??扡牳?坲慥湭来?剴??坦椠汮汩楴扲慯汵摳?????敥渠条?兤??敩瑮?慴汲???のㄠ????敥慳猠畤牵敲浩敮湧琠獤?潮晩?乲?獦畩扣????獮畛扊???乓?獩畬戠????獯畧批?佡??丠佂?慯湣摨??佩?獴畲批?㈠??猠用戹??攰洩椺猠猱椳漴渵猭?昳爵漲洮?獢潲椾汓獣?睯楬瑥桦?瑥桬敤?杄愬猠?晡汷潫睩?獳漠楊氠?挠潂爬攠?瑡散捫桳湯楮焠畓攠孍?崠???渷癢椮爠潕湳浥攠湯瑦愠污?卦捬楯敷湩据敧?慨湥摬?呵敭挠桡湴潭汯潳杰票????????????は????っ????扵牥?坴敯椠敭牥???????潨牥愠湥???坣??倠潯睦攠牤??????敦瑩?慡汴?????????敯湬楳琠牡楰晰楬捩慥瑤椠潴湯?慩湮摴?瑣桴攠?摯楲湥楳琠牯潦朠敡渠?湬楡瑹爠潳畯獩?潛硊楝搮攠?牯慩瑬椠潂?慯獬?慧晹映敡据瑤攠摂?扯祣?獥潭楩汳?睲慹琬攠爲??愨瘹愭椱氰愩戺氠攱″挳愷爭戱漳渴??愼湢摲￣湓楩瑭牡慲瑭敡孴?崠??匠潂楥汮?卩捳楥敲渠捇攬?协潴捴楯敷琠祊?潃映??洠攱爹椹挳愮??潦畦牥湣慴氠?????????????????扣牡?奢慯潮?????潲湡牴慥搭?删??坴慩獯猠浯慮渠湴?剥??敥瑬?慡汢?????????晡晣敥捴瑹?潥普?猠潩楮氠?捬桯慣牫慩据瑧攠牴楨獥琠楎挼獳?潢渾′猼支煳畵敢渾瑏椭慲汥?牵散摴畡捳瑥椠潡湣?慩湶摩?浹攠瑯桦愠湤敥?灩牴潲摩畦捹瑩楮潧渠?楡湣?獥楲硩瑡攠敩湮?牳楯捩敬?灊慝搮搠祂?獯潬楯汧獹?晡牮潤洠??桲楴湩慬??瑹栠敯?倠桓楯汩楬灳瀬椠渱攵猠?′愩渺搠??琷愭氱礱嬲?崼???楓潴来敦潡据桳敯浮椠獒琠牃礬??????????????㈠????戮爠?奥潡獳桵楲湥慭牥楮?吠???湮潩睴汲敯獧?剮?????????捯敵瑳礠汯數湩敤?椠湥桶楯扬極瑴楩潯湮?潦晲?湭椠瑳牯潩畬猭?潬硡楮摴攠?特敳摴略捭瑳椠潵湳?扮祧?摳敥湡楬瑥牤椠晧祲楯湷杴?戠慣捨瑡敭牢楥慲孳?嵊???楓潯捩桬攠浓楣捩慥汮?慥測搠??椹漠瀨栳礩猺椠挲愰氳?刲攰猶攮愼牢捲栾??潥浲浴畳渠楍挬愠瑕楹潴湴獥???????????き????へ??戬爠?婴栠敡湬朮?報?????敓楥??????坩慮湵杯?女????敳瑵?慥汭????は???兯畩慬渠瑡楴晭楯捳慰瑨楥潲湥?潧晡?乥?猠畷扩????獡畳戭?佬?晷氠畳硯敩獬?晣牯潲浥?獭潥楴汨?灤汛慊湝琮?獓祯獩瑬攠浓獣?浥慮祣?戠敓?扣楩慥獴敹搠?扦礠?瑭桥敲?慣灡瀠汊楯敵摲?条慬猬?挵根爠漨洵愩琺漠朱爳愳瀶栭?洳攴琲栮漼摢潲氾潓杷祥孲?嵳??倬氠慍湥瑲?慫湸搠?匬漠楖汬?????????㈱?????ㄠ?????rification, N₂-fixation and fermentation during anaerobic incubation of soils amended with glucose and nitrate[J]. Biology and Fertility of Soils, 23 (3): 229-235.
[27]	Swerts M, Merckx R, Vlassak K. 1996b. Denitrification followed by N₂ fixation during anaerobic incubation[J]. Soil Biology an