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Appraisal of 15N enrichment and 15N natural abundance methods for estimating N2 fixation by understorey Acacia leiocalyx and A. disparimma in a native forest of subtropical Australia

  • SOILS, SEC 1 · SOIL ORGANIC MATTER DYNAMICS AND NUTRIENT CYCLING · RESEARCH ARTICLE
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Abstract

Purpose

It is anticipated that global climate change will increase the frequency of wildfires in native forests of eastern Australia. Understorey legumes such as Acacia species play an important role in maintaining ecosystem nitrogen (N) balance through biological N fixation (BNF). This is particularly important in Australian native forests with soils of low nutrient status and frequent disturbance of the nutrient cycles by fires. This study aimed to examine 15N enrichment and 15N natural abundance techniques in terms of their utilisation for evaluation of N2 fixation of understorey acacias and determine the relationship between species ecophysiological traits and N2 fixation.

Materials and methods

A trial was established at sites 1 and 2 located at Toohey Forest, Queensland, Australia, a eucalypt-dominated native forest, to examine the determination of BNF using 15N enrichment and 15N natural abundance methods. Toohey Forest is an urban forest and subjected to frequent fuel reduction burns to protect the adjacent properties. Plant physiological status was measured to determine the relationship between physiological and N2 fixation activities.

Results and discussion

Both 15N enrichment and 15N natural abundance techniques may be used to estimate N2 fixation of acacia tree species. The estimation of BNF using 15N enrichment was higher than those of the 15N natural abundance method. A grass reference plant, Themeda triandra, as well as tree reference plants provided an appropriate δ15N signal. Potential B values for Acacia spp. between −0.3‰ and 1.0‰ provided an acceptable BNF estimation. This suburban forest is located nearby a busy highway leading to N deposition over time with consequent negative δ15N signal. This N deposition may explain the separation between the δ15N signal of the acacias and that of the reference plants which led to the successful use of the 15N natural abundance technique. Acacia leiocalyx demonstrated greater N2 fixation as well as photosynthesis and instantaneous water use efficiency than Acacia disparimma. However, no strong relationship between plant photosynthesis and N2 fixation was observed in this study. A high within-treatment variation may have masked the relationships between plant BNF activities and photosynthesis

Conclusions

The 15N natural abundance technique is preferred to be used for future studies as it is simple and inexpensive compared with 15N enrichment method. The dependence of both species on BNF at site 2, where fuel reduction burning had not taken place for 8 years, suggests that the frequent burning impoverished the soil, and this has wider implications as higher fire frequencies are to be expected in other Australian ecosystems as a result of global climate change.

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References

  • Adams MA, Simon J, Pfautsch S (2010) Woody legumes: a (re)view from the South. Tree Physiol 30:1072–1082

    Article  Google Scholar 

  • Androsoff GL, Vankessel C, Pennock DJ (1995) Landscape-scale estimates of dinitrogen fixation by Pisum sativum by 15N natural abundance and enriched isotope dilution. Biol Fertil Soils 20:33–40

    Article  Google Scholar 

  • Aranjuelo I, Irigoyen JJ, Sánchez-Díaz M (2007) Effect of elevated temperature and water availability on CO2 exchange and nitrogen fixation of nodulated alfalfa plants. Environ Exp Bot 59:99–108

    Article  CAS  Google Scholar 

  • Blumfield TJ, Xu ZH (2003) Impact of harvest residues on soil mineral nitrogen dynamics following clearfall harvesting of a hoop pine plantation in subtropical Australia. For Ecol Manag 179:55–67

    Article  Google Scholar 

  • Blumfield TJ, Xu ZH, Prasolova NV, Mathers NJ (2006) Effect of overlying windrowed harvest residues on soil carbon and nitrogen in hoop pine plantations of subtropical Australia. J Soil Sediment 6:243–248

    Article  Google Scholar 

  • Boddey RM, Peoples MB, Palmer B, Dart PJ (2000) Use of the 15N natural abundance technique to quantify biological nitrogen fixation by woody perennials. Nutr Cycl Agroecosyts 57:235–270

    Article  Google Scholar 

  • Bouillet JP, Laclau JP, Goncalves JLM, Moreira MZ, Trivelin PCO, Jourdan C, Silva EV, Piccolo MC, Tsai SM, Galiana A (2008) Mixed-species plantations of Acacia mangium and Eucalyptus grandis in Brazil 2: nitrogen accumulation in the stands and biological N2 fixation. For Ecol Manag 255:3918–3930

    Article  Google Scholar 

  • Burton J, Chen CR, Xu ZH, Ghadiri H (2007) Gross nitrogen transformations in adjacent native and plantation forest’s of subtropical Australia. Soil Biol Biochem 39:426–433

    Article  CAS  Google Scholar 

  • Catterall CP, Wallace CP, Griffith University, Institute of Applied Environmental Research (1987) An island in suburbia: the natural and social history of toohey forest. Institute of Applied Environmental Research, Griffith University, Nathan

    Google Scholar 

  • Choi W-J, Lee S-M, Chang SX, Ro H-M (2005) Variations of δ13C and δ15N in Pinus densiflora tree-rings and their relationship to environmental changes in eastern Korea. Water Air Soil Pollut 164:173–187

    Article  CAS  Google Scholar 

  • Crutzen PJ, Andreae MO (1990) Biomass burning in the tropics—impact on atmospheric chemistry and biogeochemical cycles. Science 250:1669–1678

    Article  CAS  Google Scholar 

  • Danso SKA, Bowen GD, Sanginga N (1992) Biological nitrogen fixation in trees in agroecosystems. Plant Soil 141:177–196

    Article  CAS  Google Scholar 

  • Evans JR (1989) Photosynthesis and nitrogen relationships in leaves of C-3 plants. Oecologia 78:9–19

    Article  Google Scholar 

  • Evans RD (2001) Physiological mechanisms influencing plant nitrogen isotope composition. Trends Plant Sci 6:121–126

    Article  CAS  Google Scholar 

  • Farquhar GD, Richards RA (1984) Isotopic composition of plant carbon correlates with water-use efficiency in wheat genotypes. Aust J Plant Physiol 11:539–552

    Article  CAS  Google Scholar 

  • Forrester DI, Bauhus J, Cowie AL, Vanclay JK (2006) Mixed-species plantations of Eucalyptus with nitrogen-fixing trees: a review. For Ecol Manag 233:211–230

    Article  Google Scholar 

  • Fried M, Middelboe V (1977) Measurement of amount of nitrogen fixed by legume crop. Plant Soil 43:713–715

    Article  Google Scholar 

  • Fujita K, Masuda T, Ogata S (1988) Dinitrogen fixation, ureide concentration in xylem exudate and translocation of photosynthates in soybean as influenced by ood removal and defoliation. Soil Sci Plant Nutr 34:265–275

    Article  CAS  Google Scholar 

  • Galiana A, Balle P, Kanga AN, Domenach AM (2002) Nitrogen fixation estimated by the 15N natural abundance method in Acacia mangium Willd. inoculated with Bradyrhizobium sp. and grown in silvicultural conditions. Soil Biol Biochem 34:251–262

    Article  CAS  Google Scholar 

  • Gehring C, Vlek PLG (2004) Limitations of the 15N natural abundance method for estimating biological nitrogen fixation in Amazonian forest legumes. Basic Appl Ecol 5:567–580

    Article  CAS  Google Scholar 

  • Guerrieri MR, Siegwolf RTW, Saurer M, Jäggi M, Cherubini P, Ripullone F, Borghetti M (2009) Impact of different nitrogen emission sources on tree physiology as assessed by a triple stable isotope approach. Atmos Environ 43:410–418

    Article  CAS  Google Scholar 

  • Guinto DF, Xu ZH, House APN, Saffigna PG (2000) Assessment of N2 fixation by understorey acacias in recurrently burnt eucalypt forests of subtropical Australia using 15N isotope dilution techniques. Can J For Res 30:112–121

    CAS  Google Scholar 

  • Hamilton SD, Hopmans P, Chalk PM, Smith CJ (1993) Field estimation of N2 fixation by Acacia Spp. using 15N isotope dilution and labeling with S35. For Ecol Manag 56:297–313

    Article  Google Scholar 

  • Hansen AP, Pate JS (1987) Evaluation of the 15N natural abundance method and xylem sap analysis for assessing N2 fixation of understorey legumes in jarrah (Eucalyptus marginata Donn-Ex-Sm) forest in SW Australia. J Exp Bot 38:1446–1458

    Article  CAS  Google Scholar 

  • Heaton THE (1990) 15N/14N ratios of NOx from vehicle engines and coal-fired power stations. Tellus B 42:304–307

    Google Scholar 

  • Hogberg P (1997) Tansley review no 95–15N natural abundance in soil-plant systems. New Phytol 137:179–203

    Article  Google Scholar 

  • Høgh-Jensen H, Schjoerring JK (1994) 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. Plant Soil 166:153–163

    Article  Google Scholar 

  • Hopmans P, Stewart HTL, Flinn DW (1993) Impacts of harvesting on nutrients in a eucalypt ecosystem in Southeastern Australia. For Ecol Manag 59:29–51

    Article  Google Scholar 

  • Houngnandan P, Yemadje R, Oikeh S, Djidohokpin C, Boeckx P, Van Cleemput O (2008) Improved estimation of biological nitrogen fixation of soybean cultivars (Glycine max L. Merril) using 15N natural abundance technique. Biol Fertil Soils 45:175–183

    Article  CAS  Google Scholar 

  • Imsande J (1988) Enhanced nitrogen fixation increases net photosynthetic output and seed yield of hydroponically grown soybean. J Exp Bot 39:1313–1321

    Article  Google Scholar 

  • Jung M, Reichstein M, Ciais P, Seneviratne SI, Sheffield J, Goulden ML, Bonan G, Cescatti A, Chen JQ, de Jeu R, Dolman AJ, Eugster W, Gerten D, Gianelle D, Gobron N, Heinke J, Kimball J, Law BE, Montagnani L, Mu QZ, Mueller B, Oleson K, Papale D, Richardson AD, Roupsard O, Running S, Tomelleri E, Viovy N, Weber U, Williams C, Wood E, Zaehle S, Zhang K (2010) Recent decline in the global land evapotranspiration trend due to limited moisture supply. Nature 467:951–954

    Article  CAS  Google Scholar 

  • Kaschuk G, Kuyper TW, Leffelaar PA, Hungria M, Giller KE (2009) Are the rates of photosynthesis stimulated by the carbon sink strength of rhizobial and arbuscular mycorrhizal symbioses? Soil Biol Biochem 41:1233–1244

    Article  CAS  Google Scholar 

  • Koopmans CJ, Dam DV, Tietema A, Verstraten JM (1997) Natural 15N abundance in two nitrogen saturated forest ecosystems. Oecologia 111:470–480

    Article  Google Scholar 

  • Kwak J-H, Lim S-S, Chang S, Lee K-H, Choi W-J (2011) Potential use of δ13C, δ15N, N concentration, and Ca/Al of Pinus densiflora; tree rings in estimating historical precipitation pH. J Soil Sediment 11:709–721

    Article  CAS  Google Scholar 

  • Ladha JK, Peoples MB, Garrity DP, Capuno VT, Dart PJ (1993) Estimating dinitrogen fixation of hedgerow vegetation using the 15N natural abundance method. Soil Sci Soc Am J 57:732–737

    Article  CAS  Google Scholar 

  • Lambers H, Chapin FS, Pons TL (2008) Plant physiological ecology. Springer, New York

    Book  Google Scholar 

  • May BM, Attiwill PM (2003) Nitrogen fixation by Acacia dealbata and changes in soil properties 5 years after mechanical disturbance or slash burning following timber harvest. For Ecol Manag 181:339–355

    Article  Google Scholar 

  • McKeon GM, Stone GS, Syktus JI, Carter JO, Flood NR, Ahrens DG, Bruget DN, Chilcott CR, Cobon DH, Cowley RA, Crimp SJ, Fraser GW, Howden SM, Johnston PW, Ryan JG, Stokes CJ, Day KA (2009) Climate change impacts on northern Australian rangeland livestock carrying capacity: a review of issues. Rangeland J 31:1–29

    Article  Google Scholar 

  • Naab J, Chimphango S, Dakora F (2009) N2 fixation in cowpea plants grown in farmers’ fields in the Upper West Region of Ghana, measured using15N natural abundance. Symbiosis 48:37–46

    Article  CAS  Google Scholar 

  • Okito A, Alves B, Urquiaga S, Boddey RM (2004) Isotopic fractionation during N2 fixation by four tropical legumes. Soil Biol Biochem 36:1179–1190

    Article  CAS  Google Scholar 

  • Otieno DO, Schmidt MWT, Kinyamario JI, Tenhunen J (2005) Responses of Acacia tortilis and Acacia xanthophloea to seasonal changes in soil water availability in the savanna region of Kenya. J Arid Environ 62:377–400

    Article  Google Scholar 

  • Pareek RP, Ladha JK, Watanabe I (1990) Estimating N2 fixation by Sesbania rostrata and S. cannabina (Syn S. aculeata) in lowland rice soil by the 15N dilution method. Biol Fertil Soils 10:77–88

    Google Scholar 

  • Parrotta JA, Baker DD, Fried M (1994) Application of 15N enrichment methodologies to estimate nitrogen fixation in Casuarina equisetifolia. Can J For Res 24:201–207

    Article  Google Scholar 

  • Peoples MB, Faizah AW, Rerkasem B, Herridge DF (eds) (1989) Methods for evaluating nitrogen fixation by nodulated legumes in the field. ACIAR monograph series; 11. Australian Centre for International Agricultural Research, Canberra

    Google Scholar 

  • Peoples MB, Palmer B, Lilley DM, Duc LM, Herridge DF (1996) Application of 15N and xylem ureide methods for assessing N2 fixation of three shrub legumes periodically pruned for forage. Plant Soil 182:125–137

    Article  CAS  Google Scholar 

  • Prasolova NV, Xu ZH, Farquhar GD, Saffigna PG, Dieters MJ (2000) Variation in canopy C-13 of 8-year-old hoop pine families (Araucaria cunninghamii) in relation to canopy nitrogen concentration and tree growth in subtropical Australia. Tree Physiol 20:1049–1055

    Article  CAS  Google Scholar 

  • Raddad EAY, Luukkanen O (2006) Adaptive genetic variation in water use efficiency and gum yield in Acacia senegal provenances grown on clay soil in the Blue Nile region, Sudan. For Ecol Manag 226:219–229

    Article  Google Scholar 

  • Reverchon F, Xu ZH, Blumfield TJ, Chen CR, Abdullah KM (2011) Impact of global climate change and prescribed burning on understorey legumes and associated belowground communities: implications for biogeochemical cycles in forest ecosystems. J Soil Sediment 12:150–160

    Article  Google Scholar 

  • Sanford P, Pate JS, Unkovich MJ (1994) A survey of proportional dependence of subterranean clover and other pasture legumes on N2 fixation in South-West Australia utilizing 15N natural-abundance. Aust J Agr Res 45:165–181

    Article  Google Scholar 

  • Savard M, Bégin C, Smirnoff A (2009) Tree-ring nitrogen isotopes reflect anthropogenic NOX emissions and climatic effects. Environ Sci Tech 43:604–609

    Article  CAS  Google Scholar 

  • Shearer G, Kohl DH (1986) N2 fixation in field settings: estimations based on natural 15N abundance. Aust J Plant Physiol 13:699–756

    CAS  Google Scholar 

  • Somado EA, Kuehne RF (2006) Appraisal of the 15N isotope dilution and 15N natural abundance methods for quantifying nitrogen fixation by flood-tolerant green manure legumes. Afr J Biotechnol 5:1210–1214

    CAS  Google Scholar 

  • Stevenson FC, Knight JD, Vankessel C (1995) Dinitrogen fixation in pea—controls at the landscape-scale and microscale. Soil Sci Soc Am J 59:1603–1611

    Article  CAS  Google Scholar 

  • Thonicke K, Spessa A, Prentice IC, Harrison SP, Dong L, Carmona-Moreno C (2010) The influence of vegetation, fire spread and fire behaviour on biomass burning and trace gas emissions: results from a process-based model. Biogeosciences 7:1991–2011

    Article  CAS  Google Scholar 

  • Unkovich MJ, Pate JS (2000) An appraisal of recent field measurements of symbiotic N2 fixation by annual legumes. Field Crop Res 65:211–228

    Article  Google Scholar 

  • Unkovich MJ, Pate JS, Lefroy EC, Arthur DJ (2000) Nitrogen isotope fractionation in the fodder tree legume tagasaste (Chamaecytisus proliferus) and assessment of N2 fixation inputs in deep sandy soils of Western Australia. Aust J Plant Physiol 27:921–929

    Google Scholar 

  • van Kessel C, Farrel RE, Roskoski JP, Keane KM (1994) Recycling of the naturally-occurring 15N in an established stand of Leucaena leucocephala. Soil Biol Biochem 26:757–762

    Article  Google Scholar 

  • Warembourg FR (1993) Nitrogen fixation in soil and plant systems. In: Knowles R, Blackburn TH (eds) Nitrogen isotope techniques. Academic Press, INC, London, pp 127–156

    Google Scholar 

  • Xiao HY, Liu CQ (2002) Sources of nitrogen and sulfur in wet deposition at Guiyang, southwest China. Atmos Environ 36:5121–5130

    Article  CAS  Google Scholar 

  • Xiao H-Y, Tang C-G, Xiao H-W, Liu X-Y, Liu C-Q (2010) Stable sulphur and nitrogen isotopes of the moss haplocladium microphyllum at urban, rural and forested sites. Atmos Environ 44:4312–4317

    Article  CAS  Google Scholar 

  • Xu ZH, Saffigna PG, Farquhar GD, Simpson JA, Haines RJ, Walker S, Osborne DO, Guinto D (2000) Carbon isotope discrimination and oxygen isotope composition in clones of the F-1 hybrid between slash pine and Caribbean pine in relation to tree growth, water-use efficiency and foliar nutrient concentration. Tree Physiol 20:1209–1217

    Article  CAS  Google Scholar 

  • Xu ZH, Prasolova NV, Lundkvist K, Beadle C, Leaman T (2003) Genetic variation in branchlet carbon and nitrogen isotope composition and nutrient concentration of 11-year-old hoop pine families in relation to tree growth in subtropical Australia. For Ecol Manag 186:359–371

    Article  Google Scholar 

  • Xu ZH, Chen CR, He JZ, Liu JX (2009) Trends and challenges in soil research (2009) linking global climate change to local long-term forest productivity. J Soil Sediment 9:83–88

    Article  Google Scholar 

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Acknowledgments

The authors would like to gratefully acknowledge the financial support and assistance of Powerlink QLD. The authors are particularly grateful to Ms. Elizabeth Gordon and Marijke Heenan for their technical supports and Mr. Bob Coutts for identifying plant species at the experimental sites.

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Authors and Affiliations

  1. Environmental Futures Centre, School of Biomolecular and Physical Sciences, Griffith University, Nathan, Brisbane, Queensland, 4111, Australia

    Shahla Hosseini Bai, Zhihong Xu & Timothy J. Blumfield

  2. Research Centre for Quality, Safety and Standard of Agricultural Products, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China

    Fangfang Sun

  3. Environmental Futures Centre, School of Environment, Griffith University, Nathan, Brisbane, Queensland, 4111, Australia

    Chengrong Chen

  4. Environmental Futures Centre, School of Environment, Griffith University, Gold Coast, Queensland, 9726, Australia

    Clyde Wild

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  1. Shahla Hosseini Bai
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  2. Fangfang Sun
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Correspondence to Shahla Hosseini Bai.

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Hosseini Bai, S., Sun, F., Xu, Z. et al. Appraisal of 15N enrichment and 15N natural abundance methods for estimating N2 fixation by understorey Acacia leiocalyx and A. disparimma in a native forest of subtropical Australia. J Soils Sediments 12, 653–662 (2012). https://doi.org/10.1007/s11368-012-0492-2

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