Nitrogen isotope fractionation mechanism, analysis measurement tracer technology and its application in ecological environment
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摘要:
研究目的 氮（N）是地球陆地生态系统中的关键营养元素，也是引起水体富营养化的污染元素之一。随着分析测试技术的不断提高，氮稳定同位素技术已经发展成为一种常用的研究方法和分析手段，在氮的生物地球化学循环、水体富营养化和地下水污染来源识别等方面广泛应用。
研究方法 本文查阅近年来国内外生态环境领域氮稳定同位素的相关文献，综述了氮同位素分馏机制、氮同位素分析技术以及氮同位素在生态环境中应用的研究现状。
研究结果 （1）氮同位素质谱分析技术与氮同位素示踪技术目前已经建立了成熟体系。（2）硝化作用与反硝化作用是土壤氮转化循环的主要机制，生物固氮实现氮的输入，而植物或微生物产生含氮气体或矿化作用是氮输出的主要途径，并伴随着不同程度的氮同位素分馏效应。（3）氮同位素可以用来测定土壤氮素周转速率、N2O排放途径与生物固氮量、指示大气氮沉降变化、探究植物与土壤的相互作用及确定植物对氮素的吸收利用、识别农作物产地与水体、大气的氮污染来源。
结论 未来应将研究重点放在提升氮循环过程中不确定性来源的定量检测能力，确定未被发现的氮输入、积累和损失途径，完善并发展生态系统氮循环模型。
Abstract:This paper is the result of environmental geological survey engineering.
Objective Nitrogen (N) is a key nutrient across Earth's terrestrial ecosystems and one of the pollution elements that cause water eutrophication. Owing to the continuous improvements of analysis and testing techniques, nitrogen stable isotope technology has developed into a common research method and analysis mean, and has been widely used in nitrogen biogeochemical cycle, water eutrophication and groundwater pollution source identification.
Methods In this paper, the relevant literatures on nitrogen stable isotope in the field of ecological environment domestic and overseas in recent years were reviewed, and the research status of nitrogen isotope fractionation mechanism, nitrogen stable isotope analysis technology and nitrogen isotope applications in ecological environment were summarized, the development of remediation technologys of nitrate pollution in groundwater were briefly described.
Results (1) A mature system of nitrogen isotope mass spectrometry and nitrogen isotope tracer technology has been established. (2) Nitrification and denitrification are the main mechanisms of soil nitrogen conversion cycle. Nitrogen input is realized by biological nitrogen fixation, and nitrogen output is mainly through nitrogen gas or ammoniation produced by plants or microorganisms, which is accompanied by different degrees of nitrogen isotope fractionation. (3) Nitrogen isotopes can be used to measure soil nitrogen turnover rates and N2O emission rates, improve biological nitrogen fixation, indicate changes in atmospheric nitrogen deposition, investigate the interaction between plant and soil and determine nitrogen uptake and utilization by plants, and identify crop area sources and pollution in groundwater and atmosphere.
Conclusions Future researches should focus on improving the ability of quantitative detection of uncertainty sources in the nitrogen cycle, identifying undiscovered nitrogen input, accumulation and loss pathways, and perfecting and developing ecosystem nitrogen cycle model.
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图 3 温度（a，据Yun et al., 2011）和湿度影响下土壤硝化过程氮同位素分馏系数变化（b，据Yun and Ro, 2014）
Figure 3.
图 4 δ15N−NH4+随NH4+受硝化作用和同化作用影响的变化（据Choi et al., 2017）
Figure 4.
图 10 不同生态系统氮矿化速率（N为观测值个数；据Elrys et al., 2021）
Figure 10.
图 11 土壤N2O产生途径（据Baggs, 2008）
Figure 11.
图 12 土壤孔隙水含量（a，据Wang et al., 2023）与pH（b，据Zhang et al., 2021）对硝化和反硝化作用对N2O生成贡献的影响
Figure 12.
图 13 土壤15N−N2O（a）与土壤排放气体N2O/N2（b）随温度变化（据Lai et al., 2019）
Figure 13.
图 14 不同土壤类型产生N2O的15N丰度（据Zhang et al., 2017）
Figure 14.
图 15 不同土壤类型N2O产生途径nh、na和nd分别是异养硝化、自养硝化和反硝化对N2O生成的贡献（据Zhang et al., 2018）
Figure 15.
图 17 作物生物固氮过程（据Guo et al., 2023）
Figure 17.
图 18 氮沉降与不同地区森林叶片δ15N的相关性（据Choi et al., 2023）
Figure 18.
图 20 温带地区（a）和冷温带白桦和泥炭藓（b）吸收氮偏好（据Gao et al., 2020）
Figure 20.
图 21 不同污染来源δ15N值分布（据Xue et al., 2009）
Figure 21.
图 22 不同污染来源δ15N与δ18O值分布（据Kendall, 1998）
Figure 22.
图 23 典型污染来源δ15N与δ11B值分布（据Sankoh et al., 2022）
Figure 23.
表 1 不同细菌在不同实验条件下反硝化作用的氮同位素分馏效应（ε，δ 15N/‰）
Table 1. Nitrogen isotope fractionation effect of denitrification by different bacteria under different experimental conditions (ε, δ 15N/‰)
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表 2 同种细菌在不同实验条件下反硝化作用的氮同位素分馏效应（ε，δ 15N/‰）（据Frey et al., 2014；Treibergs and Granger, 2017）
Table 2. Nitrogen isotope fractionation effect of denitrification by the same bacteria under different experimental conditions (ε, δ 15N/‰) (after Frey et al., 2014; Treibergs and Granger, 2017）
细菌 温度/
℃NO3−浓度/
mM细菌数量/
个氮同位素分馏效应
（ε，δ 15N/‰）脱氮副球菌 20 0.2 4 22.90 20 0.2 4 25.80 20 1 3 33.00 20 1 5 31.50 硫单胞菌 15 5 5 24.29 15 5 4 21.17 15 1 5 17.69 10 1 5 13.08 10 1 5 13.08
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表 3 不同生态系统反硝化作用氮同位素分馏效应
Table 3. Denitrification effect of nitrogen isotope fractionation in different ecosystems
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表 4 不同生态系统平均硝化速率（据Li et al., 2020）
Table 4. Average nitrification rate in different ecosystems (after Li et al., 2020)
生态系统 平均硝化速率/（mg/kg/d） 观测数据/个 农田 3.82 1423 森林 2.58 841 草地 1.70 368 湿地 3.29 80
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表 5 不同N2O产生过程中δ15N与SP值（据Decock and Six, 2013）
Table 5. Different N2O production processes of δ15N and SP values (after Decock and Six, 2013)
N2O产生过程 δ15N/‰ SP 氨氧化细菌羟氨氧化 −0.3±3.4 33.0±1.6 甲烷氧化菌羟氨氧化 1.9±1.9 32.7±5.0 氨氧化细菌氨氧化 −45.5±2.3 31.4±4.2 非生物氧化 −5.8±9.2 29.9±1.5 真菌反硝化作用 −9.9±6.7 35.2±4.3 反硝化作用 −13.8±4.9 −2.2±3.2 硝化细菌反硝化作用 −29.0±6.0 −1.0±4.3
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表 6 全球范围内不同气候带和海拔范围内的土壤δ15N统计特征值（据昝麒麟等，2022）
Table 6. Number of samples and statistical descriptions of soil δ15N on global scale in different climate zones and elevation ranges (after Zan Qilin et al., 2022)
土壤环境 土壤δ15N值/‰ 气候带 赤道气候带 5.34 干旱气候带 5.78 暖温气候带 3.91 冷温气候带 3.98 极地气候带 3.98 海拔 <500 m 4.39 500~1000 m 4.70 1000~2000 m 4.72 2000~3500 m 4.78 >3500 m 4.18
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表 7 不同种类肥料δ15N值（据Choi et al., 2017）
Table 7. δ15N values of different kinds of fertilizers (after Choi et al., 2017)
肥料种类 δ15N/‰ 平均δ15N/‰ 合成肥料 尿素 −5.9~1.9 −1.2 ± 0.4 硫酸铵 −3.9~6.6 −0.5 ± 0.7 硝酸铵 −1.7~2.6 0.1 ± 0.5 牲畜粪便 牛粪 3.5~16.5 8.4 ± 0.9 猪粪 4.4 ~11.3 6.7 ± 1.0 马粪 7.2~11.8 9.5 ± 2.3 家禽粪便 2.7~14.6 7.1 ± 1.5
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表 8 利用氮同位素技术提高作物相关微生物固氮的研究现状
Table 8. Research status of nitrogen isotope technology to improve nitrogen fixation by crop−related microorganisms
作物种类 研究现状 参考文献 水稻 利用15N直接标记测定稻田固氮量，发现水稻种植极大地促进了固氮菌nifH基因的表达，提高了固氮菌的固氮活性，增加了稻田各层土壤的固氮量 张燕辉等(2021) 玉米 利用15N自然丰度法进行实验证明，墨西哥玉米品种ierra Mixe根部产生的黏液中具有活性nif基因的细菌菌株，可以满足该玉米品种氮需求的29%~82% Van Deynze et al. (2018) 将氮肥替换为30% atom15N标记的(NH4)2SO4，测定γ−变形杆菌通过生物固氮贡献了玉米茎中积累的11.8%的总氮 Zhang et al. (2022) 甘蔗 利用15N自然丰度法确定接种重氮营养菌使RB92579和RB2003两种甘蔗品种的生物固氮量分别增加了50和17 kg/hm2 Martins et al. (2020) 通过对植株组织、植株产物和叶绿素的15N同位素稀释，证实了克莱伯菌与甘蔗共生，能缓解氮素缺乏症的发生 Luo et al. (2015a) 豆类 利用15N贫化法确定接种根瘤菌有利于提高籽粒固氮和籽粒重。与Apore品种相比，Ouro Negro品种的根瘤数量和重量以及从大气中获得的氮量更高 Silva et al. (2017)
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表 9 不同大陆和气候带森林的大气氮沉降速率（据Schwede et al., 2018）
Table 9. Atmospheric N deposition rates for forests in different continents and climate zones (after Schwede et al., 2018)
气候带 地区 氮沉降速率/（kg/hm2/a） 亚洲 7.8 温带 美洲 5.4 欧洲 10.8 亚洲 29.3 北美洲 7.2 亚热带 南美洲 10.5 欧洲 10.5 非洲 7.9 热带 亚洲 11.8 北美洲 7.4 南美洲 5.8 欧洲 7.1
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表 10 不同地区植物叶片的δ15N和氮含量（据Choi et al., 2023）
Table 10. δ15N and N concentration of foliage in different zones (after Choi et al., 2023)
气候带 叶片δ15N/‰ 叶片氮含量/（g/kg） 温带 −2.4±2.2 16.4±5.2 亚热带 −2.5±2.0 14.6±5.0 热带 1.6±4.3 18.5±7.3
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表 11 入侵物种对土壤有效氮的影响
Table 11. Effects of invasive species on soil available nitrogen
地区 入侵物种 对土壤有效氮影响 参考文献 北美 矢车菊 根系分泌儿茶素显著抑制土壤硝化速率，导致入侵地土壤NO3−浓度降低 Thorpe and Callaway，2011 北美 虎杖 分泌单宁抑制土壤的氮矿化作用，导致土壤无机氮浓度较低，总游离氨基酸和可溶性有机氮的浓度升高 Tharayil et al.，2013 中国北方 火炬树 提高土壤的硝化速率，土壤NO3−含量升高，土壤NH4+含量降低 黄乔乔等，2013 海南 金钟藤 土壤NO3−、NH4+含量升高 纵熠等，2015 澳大利亚 须芒草 提高土壤氨化作用，抑制土壤氮硝化，土壤NH4+含量增加 Rossiter−Rachor et al.，2009
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表 12 不同地区小麦样品δ15N值（据Luo et al., 2015b）
Table 12. δ15N values of wheat samples from different regions (after Luo et al., 2015b)
来源 δ15N/‰ 美国 3.04±0.79 加拿大 2.13±0.70 澳大利亚 7.25±1.18 中国 2.45±0.5
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表 13 不同地区谷物δ15N与δ13C值（据Wu et al., 2015）
Table 13. δ15N and δ13Cvalues of different cereal grains from different origins (after Wu et al., 2015)
谷物 平均δ15N/‰ 平均δ13C/‰ 黑龙江 山东 江苏 黑龙江 山东 江苏 水稻 3.070 3.434 3.221 −26.285 −26.946 −27.578 大豆 1.365 1.462 1.388 −23.679 −25.642 −26.102 小米 1.098 1.251 1.112 −11.985 −12.346 −12.897 小麦 1.827 1.934 1.850 −22.843 −25.694 −25.162 玉米 0.237 0.432 0.292 −10.965 −10.444 −11.292
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表 14 不同排放源大气氮δ15N值（据Choi et al., 2023）
Table 14. Atmospheric nitrogen δ15N values of different emission sources (after Choi et al., 2023)
来源 δ15N−NOx/‰ δ15N−NH3/‰ 生物质燃烧 0.2 化石燃料燃烧 11.7 −13 施肥土壤 −32.5 −40.5 牲畜粪便 −19.3 −27.9 车辆 −8.2 −21.7 天然气燃烧 −16.7 有机废物 −37.8
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