Comparison of natural 15N abundance technique and 15N dilution technique in the determination of plant nitrogen fixation

doi:10.7523/j.ucas.2021.0008

Abstract

Abstract: Biological nitrogen fixation (BNF) is an important source of nitrogen in ecosystem, therefore, several experimental techniques and calculating methods have been developed to quantify the contribution of BNF to plants and vegetations. In this study, percentage of plant N derived from N₂-fixation (%Ndfa) was evaluated for a leguminous species, Astragalus arnoldii, in an alpine steppe in Qinghai-Tibet Plateau in order to compare two dominant in situ experimental techniques with different calculating approaches, reference plants, and sampling times. The δ¹⁵N values of four reference plants, i.e., Stipa purpurea, Kobresia pygmaea, Leontopodium nanum, and Carex moorcroftii, were significantly lower in late August than in late July (P<0.05). However, they were significantly higher in later August, 30 days after application of (¹⁵NH₄)₂SO₄ to soil, than late July which was 24h after labeling (P<0.05). In either site with or without (¹⁵NH₄)₂SO₄ application, δ¹⁵N of A. arnoldii did not differ in the two sampling times. Besides,%Ndfa of A. arnoldii did not differ between calculation based on ¹⁵N excess relative to atmospheric N₂ and that relative to unlabeled plants. Nevertheless,%Ndfa measured by natural abundance technique was significantly higher than that by isotope dilution technique (P<0.05) except for L. nanum as the reference species. The sampling time had a significant effect on estimated%Ndfa values in both the naturalabundance technique (F=89.906, P<0.01), and isotope dilution technique (F=496.712, P<0.01).

Key words: biological nitrogen fixation, natural abundance technique, isotope dilution technique, Qinghai-Tibet plateau, alpine steppe, ¹⁵N

CLC Number:

Q948.1

LI Runfu, NIU Haishan, KONG Qian, LIU Qiang. Comparison of natural ¹⁵N abundance technique and ¹⁵N dilution technique in the determination of plant nitrogen fixation[J]. Journal of University of Chinese Academy of Sciences, 2022, 39(1): 34-42.

References

[1] Vicente E J, Dean D R. Keeping the nitrogen-fixation dream alive[J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(12):3009-3011.
[2] 何道文, 孙辉, 黄雪菊. 利用N-15自然丰度法研究固氮植物生物固氮量[J]. 干旱地区农业研究, 2004, 22(1):132-137.
[3] 陈朝勋, 席琳乔, 姚拓, 等. 生物固氮测定方法研究进展[J]. 草原与草坪, 2005, 25(2):24-26.
[4] Hong J T, Ma X X, Zhang X K, et al. Nitrogen uptake pattern of herbaceous plants:coping strategies in altered neighbor species[J]. Biology and Fertility of Soils, 2017, 53(7):729-735.
[5] Xu X L, Ouyang H, Cao G M, et al. Dominant plant species shift their nitrogen uptake patterns in response to nutrient enrichment caused by a fungal fairy in an alpine meadow[J]. Plant and Soil, 2011, 341(1/2):495-504.
[6] Schleuss P M, Heitkamp F, Sun Y, et al. Nitrogen uptake in an alpine Kobresia pasture on the Tibetan Plateau:localization by ¹⁵N labeling and implications for a vulnerable ecosystem[J]. Ecosystems, 2015, 18(6):946-957.
[7] Carranca C, de Varennes A, Rolston D E. Biological nitrogen fixation estimated by ¹⁵N dilution, natural ¹⁵N abundance, and N difference techniques in a subterranean clover-grass sward under Mediterranean conditions[J]. European Journal of Agronomy, 1999, 10(2):81-89.
[8] Lonati M, Probo M, Gorlier A, et al. Nitrogen fixation assessment in a legume-dominant alpine community:comparison of different reference species using the ¹⁵N isotope dilution technique[J]. Alpine Botany, 2015, 125(1):51-58.
[9] Yang B J, Qiao N, Xu X L, et al. Symbiotic nitrogen fixation by legumes in two Chinese grasslands estimated with the ¹⁵N dilution technique[J]. Nutrient Cycling in Agroecosystems, 2011, 91(1):91-98.
[10] McAuliffe C, Chamblee D S, Uribe-Arango H, et al. Influence of inorganic nitrogen on nitrogen fixation by legumes as revealed by N¹⁵[J]. Agronomy Journal, 1958, 50(6):334-337.
[11] Arnone Ⅲ J A. Symbiotic N₂ fixation in a high Alpine grassland:effects of four growing seasons of elevated CO₂[J]. Functional Ecology, 1999, 13(3):383-387.
[12] Che R X, Deng Y C, Wang F, et al. Autotrophic and symbiotic diazotrophs dominate nitrogen-fixing communities in Tibetan grassland soils[J]. Science of the Total Environment, 2018, 639:997-1006.
[13] Jacot K A, Lüscher A, Nösberger J, et al. The relative contribution of symbiotic N₂ fixation and other nitrogen sources to grassland ecosystems along an altitudinal gradient in the Alps[J]. Plant and Soil, 2000, 225(1/2):201-211.
[14] 代冬雪. 西藏高寒草原豆科与非豆科优势植物群落生物固氮功能比较研究[D]. 北京:中国科学院大学, 2015.
[15] Yang K, He J, Tang W J, et al. On downward shortwave and longwave radiations over high altitude regions:observation and modeling in the Tibetan Plateau[J]. Agricultural and Forest Meteorology, 2010, 150(1):38-46.
[16] Ledgard S F, Simpson J R, Freney J R, et al. Field evaluation of ¹⁵N techniques for estimating nitrogen fixation in legume-grass associations[J]. Australian Journal of Agricultural Research, 1985, 36(2):247.
[17] Rennie R J, Dubetz S. Nitrogen-15-determined nitrogen fixation in field-grown chickpea, lentil, fababean, and field pea[J]. Agronomy Journal, 1986, 78(4):654-660.
[18] Sun Y, Schleuss P M, Pausch J, et al. Nitrogen pools and cycles in Tibetan Kobresia pastures depending on grazing[J]. Biology and Fertility of Soils, 2018, 54(5):569-581.
[19] Ti C P, Pan J J, Xia Y Q, et al. A nitrogen budget of mainland China with spatial and temporal variation[J]. Biogeochemistry, 2012, 108(1/2/3):381-394.
[20] Bowman W D, Schardt J C, Schmidt S K. Symbiotic N₂-fixation in alpine tundra:ecosystem input and variation in fixation rates among communities[J]. Oecologia, 1996, 108(2):345-350.
[21] Jacot K A, Lüscher A, Nösberger J, et al. Symbiotic N₂ fixation of various legume species along an altitudinal gradient in the Swiss Alps[J]. Soil Biology and Biochemistry, 2000, 32(8/9):1043-1052.
[22] Boddey R M, Urquiaga S, Neves M C P, et al. Quantification of the contribution of N₂ fixation to field-grown grain legumes:a strategy for the practical application of the ¹⁵N isotope dilution technique[J]. Soil Biology and Biochemistry, 1990, 22(5):649-655.
[23] 罗绪强, 张桂玲, 杨鸿雁, 等. 喀斯特山地不同退化植被下烟管荚蒾氮同位素组成的季节变化[J]. 科学技术与工程, 2020, 20(11):4243-4249.
[24] 苏波, 韩兴国, 黄建辉. ¹⁵N自然丰度法在生态系统氮素循环研究中的应用[J]. 生态学报, 1999, 19(3):408-416.
[25] Chalk P M, Ladha J K. Estimation of legume symbiotic dependence:an evaluation of techniques based on ¹⁵N dilution[J]. Soil Biology and Biochemistry, 1999, 31(14):1901-1917.
[26] Yang L J, Zhang L L, Geisseler D, et al. Available C and N affect the utilization of glycine by soil microorganisms[J]. Geoderma, 2016, 283:32-38.
[27] Hossain S A, Waring S A, Strong W M, et al. Estimates of nitrogen fixations by legumes in alternate cropping systems at Warra, Queensland, using enriched-¹⁵N dilution and natural ¹⁵N abundance techniques[J]. Australian Journal of Agricultural Research, 1995, 46(3):493-505.
[28] Burchill W, James E K, Li D, et al. Comparisons of biological nitrogen fixation in association with white clover (Trifolium repens L.) under four fertiliser nitrogen inputs as measured using two ¹⁵N techniques[J]. Plant and Soil, 2014, 385(1/2):287-302.
[29] Høgh-Jensen H, Schjoerring J K. Measurement of biological dinitrogen fixation in grassland:comparison of the enriched ¹⁵N dilution and the natural ¹⁵N abundance methods at different nitrogen application rates and defoliation frequencies[J]. Plant and Soil, 1994, 166(2):153-163.

Comparison of natural ¹⁵N abundance technique and ¹⁵N dilution technique in the determination of plant nitrogen fixation

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