Water source partitioning and nitrogen facilitation promote coexistence of nitrogen-fixing and neighbor species in mixed plantations in the semiarid Loess Plateau

  • Yakun Tang
  • Xu Wu
  • Chen Chen
  • Chang Jia
  • Yunming ChenEmail author
Regular Article



Effects of water source relations on the facilitation of nitrogen (N) acquisition between N2-fixing species (NFS) and neighbor species require further investigation, especially in water- and N-limited conditions.


Plant water sources and leaf physiological parameters of Hippophae rhamnoides and two neighbor species were investigated in pure and mixed plantations. Then, N stable isotope and N content of leaves were analyzed to examine the N facilitation of H. rhamnoides to neighbor species. Finally, interactions between plant water sources and N acquisition were detected using the dissimilarity analysis.


H. rhamnoides and Pinus tabuliformis obtained water from similar soil depths, whereas, water sources were partitioned between H. rhamnoides and Ulmus pumila. Compared with pure plantations, P. tabuliformis and H. rhamnoides could not maintain a stable net photosynthetic rate (Pn) during the dry period in mixed plantation. However, H. rhamnoides and U. pumila in mixed plantation could alleviate drought effect on Pn through stomatal adjustments, comapred with pure plantations. Furthermore, H. rhamnoides facilitated N to neighbor species regardless of their water source relations, however, this facilitation only significantly improved the leaf N content of U. pumila.


Water source partitioning, stomatal adjustment, and N facilitation promoted stable coexistence of NFS and neighbor species in water- and N-limited environments.


Loess Plateau Nitrogen facilitation Nitrogen stable isotope Stomatal adjustment Water stable isotope 





N2-fixing species


Stable N isotope


Potential evapotranspiration


Stable hydrogen isotope


Stable oxygen isotope


Soil gravimetric water content


Stomatal conductance


Net photosynthesis rate


Midday leaf water potential


General linear model


Coefficient of variation


Pinus tabuliformisHippophae rhamnoides plantation


Ulmus pumilaH. rhamnoides plantation


Ectomycorrhizal fungi


Arbuscular mycorrhizal fungi


Common mycorrhizal networks



We are grateful to the constructive and insightful suggestions of section editor Rafael S. Oliveira and two anonymous reviewers. This work was supported by the National Natural Science Foundation of China (41977425), the National Key R&D Program of China (2017YFA0604801), and the Fundamental Research Funds for the Central Universities (2452016105).

Supplementary material

11104_2019_4301_MOESM1_ESM.docx (550 kb)
ESM 1 (DOCX 549 kb)


  1. Bouillet JP, Laclau JP, Goncalves JLM, Moreira M, 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 N(2) fixation. Forest Ecol Manag 255:3918–3930CrossRefGoogle Scholar
  2. Bremner JM, Mulvaney CS (1982) Nitrogen-total. Methods of soil analysis, Part 2-Chemical and microbiological properties. Madison Press, Wisconsin, USA. pp 595–624Google Scholar
  3. Chalk PM, Ladha JK (1999) Estimation of legume symbiotic dependence: an evaluation techniques based on N-15 dilution. Soil Biol Biochem 31:1901–1917CrossRefGoogle Scholar
  4. Chalk PM, Peoples MB, McNeill AM, Boddey RM, Unkovich MJ, Gardener MJ, Silva CF, Chen DL (2014) Methodologies for estimating nitrogen transfer between legumes and companion species in agro-ecosystems: a review of 15N-enriched techniques. Soil Biol Biochem 73:10–21CrossRefGoogle Scholar
  5. Chen YJ, Cao KF, Schnitzer SA, Fan ZX, Zhang JL, Bongers F (2015) Water-use advantage for lianas over trees in tropical seasonal forests. New Phytol 205:128–136PubMedCrossRefGoogle Scholar
  6. Chimner RA, Cooper DJ (2004) Using stable oxygen isotopes to quantify the water source used for transpiration by native shrubs in the San Luis Valley, Colorado USA. Plant Soil 260:225–236CrossRefGoogle Scholar
  7. Collino DJ, Salvagiotti F, Perticari A, Piccinetti C, Ovando G, Urquiaga S, Racca RW (2015) Biological nitrogen fixation in soybean in Argentina: relationships with crop, soil, and meteorological factors. Plant Soil 392:239–252CrossRefGoogle Scholar
  8. Craine JM, Elmore AJ, Aidar MPM, Bustamante M, Dawson TE, Hobbie EA, Kahmen A, Mack MC, McLauchlan KK, Michelsen A, Nardoto GB, Pardo LH, Penuelas J, Reich PB, Schuur EAG, Stock WD, Templer PH, Virginia RA, Welker JM, Wright IJ (2009) Global patterns of foliar nitrogen isotopes and their relationships with climate, mycorrhizal fungi, foliar nutrient concentrations, and nitrogen availability. New Phytol 183:980–992CrossRefGoogle Scholar
  9. Ellsworth PZ, Sternberg LSL (2016) Strategies in nitrogen uptake and use by deciduous and evergreen woody species in a seasonally dry sandhill community. Plant Soil 400:165–175CrossRefGoogle Scholar
  10. Emmerton KS, Callaghan TV, Jones HE, Leake JR, Michelsen A, Read DJ (2001) Assimilation and isotopic fractionation of nitrogen by mycorrhizal fungi. New Phytol 151:503–511CrossRefGoogle Scholar
  11. Evans JR (1989) Photosynthesis and nitrogen relationships in levels of C3 plants. Oecologia 78:9–19CrossRefGoogle Scholar
  12. February EC, Allsopp N, Shabane T, Hattas D (2011) Coexistence of a C-4 grass and a leaf succulent shrub in an arid ecosystem. The relationship between rooting depth, water and nitrogen. Plant Soil 349:253–260CrossRefGoogle Scholar
  13. Feng X, Ackerly DD, Dawson TE, Manzoni S, McLaughlin B, Skelton RP, Vico G, Weitz AP, Thompson SE (2019) Beyond isohydricity: the role of environmental variability in determining plant drought responses. Plant Cell Environ 42:1104–1111PubMedCrossRefGoogle Scholar
  14. Forrester DI (2014) The spatial and temporal dynamics of species interactions in mixed-species forests: from pattern to process. Forest Ecol Manag 312:282–292CrossRefGoogle Scholar
  15. Forrester DI, Bauhus J, Cowie AL, Mitchell PA, Brockwell J (2007) Productivity of three young mixed-species plantations containing N-2-fixing Acacia and non-N2-fixing Eucalyptus and Pinus trees in southeastern Australia. For Sci 53:426–434Google Scholar
  16. Fu BJ, Meng QH, Qiu Y, Zhao WW, Zhang QJ, Davidson DA (2004) Effects of land use on soil erosion and nitrogen loss in the hilly area of the Loess Plateau, China. Land Degrad Dev 15:87–96CrossRefGoogle Scholar
  17. Gao XD, Li HC, Zhao XN, Ma W, Wu PT (2018) Identifying a suitable revegetation technique for soil restoration on water-limited and degraded land: considering both deep soil moisture deficit and soil organic carbon sequestration. Geoderma 319:61–69CrossRefGoogle Scholar
  18. Gebauer RLE, Ehleringer JR (2000) Water and nitrogen uptake patterns following moisture pulses in a cold desert community. Ecology 81:1415–1424CrossRefGoogle Scholar
  19. Gentili F (2006) Phosphorus, nitrogen and their interactions affect N-2 fixation, N isotope fractionation and N partitioning in Hippophae rhamnoides. Symbiosis 41:39–45Google Scholar
  20. Griesmann M, Chang Y, Liu X, Song Y, Haberer G, Crook MB, Billault-Penneteau B, Lauressergues D, Keller J, Imanishi L, Roswanjaya YP, Kohlen W, Pujic P, Battenberg K, Alloisio N, Liang YH, Hilhorst H, Salgado MG, Hocher V, Gherbi H, Svistoonoff S, Doyle JJ, He SX, Xu Y, Xu SY, Qu J, Gao Q, Fang XD, Fu Y, Normand P, Berry AM, Wall LG, Ane JM, Pawlowski K, Xu X, Yang HM, Spannagl M, Mayer KFX, Wong GKS, Parniske M, Delaux PM, Cheng SF (2018) Phylogenomics reveals multiple losses of nitrogen-fixing root nodule symbiosis. Science 361:144Google Scholar
  21. Guinet M, Nicolardot B, Revellin C, Durey V, Carlsson G, Voisin AS (2018) Comparative effect of inorganic N on plant growth and N-2 fixation of ten legume crops: towards a better understanding of the differential response among species. Plant Soil 432:207–227CrossRefGoogle Scholar
  22. Guo DL, Xia MX, Wei X, Chang WJ, Liu Y, Wang ZQ (2008) Anatomical traits associated with absorption and mycorrhizal colonization are linked to root branch order in twenty-three Chinese temperate tree species. New Phytol 180:673–683PubMedCrossRefPubMedCentralGoogle Scholar
  23. He XH, Critchley C, Ng H, Bledsoe C (2005) Nodulated N-2-fixing Casuarina cunninghamiana is the sink for net N transfer from non-N-2-fixing Eucalyptus maculata via an ectomycorrhizal fungus Pisolithus sp using (NH4+)-N-15 or (NO3-)-N-15 supplied as ammonium nitrate. New Phytol 167:897–912PubMedCrossRefPubMedCentralGoogle Scholar
  24. He XH, Xu MG, Qiu GY, Zhou JB (2009) Use of N-15 stable isotope to quantify nitrogen transfer between mycorrhizal plants. J Plant Ecol 2:107–118CrossRefGoogle Scholar
  25. Hernandez P, Picon-Cochard C (2016) Presence of trifolium repens promotes complementarity of water use and N facilitation in diverse grass mixtures. Front Plant Sci 7.
  26. Herridge DF, Peoples MB, Boddey RM (2008) Global inputs of biological nitrogen fixation in agricultural systems. Plant Soil 311:1–18CrossRefGoogle Scholar
  27. Hobbie EA, Högberg P (2012) Nitrogen isotopes link mycorrhizal fungi and plants to nitrogen dynamics. New Phytol 196:367–382CrossRefGoogle Scholar
  28. Hobbie EA, Macko SA, Williams M (2000) Correlations between foliar delta N-15 and nitrogen concentrations may indicate plant-mycorrhizal interactions. Oecologia 122:273–283PubMedCrossRefPubMedCentralGoogle Scholar
  29. Husse S, Luscher A, Buchmann N, Hoekstra NJ, Huguenin-Elie O (2017) Effects of mixing forage species contrasting in vertical and temporal nutrient capture on nutrient yields and fertilizer recovery in productive grasslands. Plant Soil 420:505–521CrossRefGoogle Scholar
  30. Levitt J (1980) Responses of plants to environmental stresses. Volume II water, radiation, salt, and other stresses. Academic Press, New YorkGoogle Scholar
  31. Li YY, Chen WY, Chen JC, Shi H (2016) Vulnerability to drought-induced cavitation in shoots of two typical shrubs in the southern Mu Us Sandy Land, China. J Arid Land 8:125–137CrossRefGoogle Scholar
  32. Liu WJ, Wang PY, Li JT, Liu WY, Li HM (2014) Plasticity of source-water acquisition in epiphytic, transitional and terrestrial growth phases of Ficus tinctoria. Ecohydrology 7:1524–1533CrossRefGoogle Scholar
  33. Liu GB, Shangguan ZP, Yao WY, Yang QK, Zhao MJ, Dang XH, Guo MH, Wang GL, Wang B (2017) Ecological effects of soil conservation in Loess Plateau. Bull Chin Acad Sci 32:11–19Google Scholar
  34. Louarn G, Pereira-Lopes E, Fustec J, Mary B, Voisin AS, Carvalho PCD, Gastal F (2015) The amounts and dynamics of nitrogen transfer to grasses differ in alfalfa and white clover-based grass-legume mixtures as a result of rooting strategies and rhizodeposit quality. Plant Soil 389:289–305CrossRefGoogle Scholar
  35. Marino D, Frendo P, Ladrera R, Zabalza A, Puppo A, Arrese-Igor C, Gonzalez EM (2007) Nitrogen fixation control under drought stress. Localized or systemic? Plant Physiol 143:1968–1974PubMedPubMedCentralCrossRefGoogle Scholar
  36. Martinez-Vilalta J, Garcia-Forner N (2017) Water potential regulation, stomatal behaviour and hydraulic transport under drought: deconstructing the iso/anisohydric concept. Plant Cell Environ 40:962–976PubMedCrossRefPubMedCentralGoogle Scholar
  37. Meng QH, Fu BJ, Tang XP, Ren HC (2008) Effects of land use on phosphorus loss in the hilly area of the loess plateau, China. Environ Monit Assess 139:195–204PubMedCrossRefPubMedCentralGoogle Scholar
  38. Montesinos-Navarro A, Verdu M, Querejetac JI, Sortibran L, Valiente-Banuet A (2016) Soil fungi promote nitrogen transfer among plants involved in long-lasting facilitative interactions. Perspect Plant Ecol 18:45–51CrossRefGoogle Scholar
  39. Montesinos-Navarro A, Verdu M, Querejeta JI, Valiente-Banuet A (2017) Nurse plants transfer more nitrogen to distantly related species. Ecology 98:1300–1310PubMedCrossRefPubMedCentralGoogle Scholar
  40. Moore JW, Semmens BX (2008) Incorporating uncertainty and prior information into stable isotope mixing models. Ecol Lett 11: 470–480Google Scholar
  41. Moreno-Gutierrez C, Dawson TE, Nicolas E, Querejeta JI (2012) Isotopes reveal contrasting water use strategies among coexisting plant species in a Mediterranean ecosystem. New Phytol 196:489–496PubMedCrossRefGoogle Scholar
  42. Paynel F, Murray PJ, Cliquet JB (2001) Root exudates: a pathway for short-term N transfer from clover and ryegrass. Plant Soil 229:235–243CrossRefGoogle Scholar
  43. Peoples MB, Chalk PM, Unkovich MJ, Boddey RM (2015) Can differences in N-15 natural abundance be used to quantify the transfer of nitrogen from legumes to neighbouring non-legume plant species? Soil Biol Biochem 87:97–109CrossRefGoogle Scholar
  44. Phillips DL, Newsome SD, Gregg JW (2005) Combining sources in stable isotope mixing models: alternative methods. Oecologia 144:520–527PubMedPubMedCentralCrossRefGoogle Scholar
  45. Pivovaroff AL, Pasquini SC, De Guzman ME, Alstad KP, Stemke JS, Santiago LS (2016) Multiple strategies for drought survival among woody plant species. Funct Ecol 30:517–526CrossRefGoogle Scholar
  46. Priestley CHB, Taylor RJ (1972) On the assessment of surface heat flux and evaporation using large-scale parameters. Mon Weather Rev 100:81–92CrossRefGoogle Scholar
  47. Renninger HJ, Carlo N, Clark KL, Schafer KVR (2014) Physiological strategies of co-occurring oaks in a water- and nutrient-limited ecosystem. Tree Physiol 34:159–173PubMedCrossRefGoogle Scholar
  48. Richards AE, Forrester DI, Bauhus J, Scherer-Lorenzen M (2010) The influence of mixed tree plantations on the nutrition of individual species: a review. Tree Physiol 30:1192–1208PubMedCrossRefGoogle Scholar
  49. Roggy JC, Prevost MF, Gourbiere F, Casabianca H, Garbaye J, Domenach AM (1999) Leaf natural N-15 abundance and total N concentration as potential indicators of plant N nutrition in legumes and pioneer species in a rain forest of French Guiana. Oecologia 120:171–182PubMedCrossRefGoogle Scholar
  50. Saha AK, Sternberg LDO, Ross MS, Miralles-Wilhelm F (2010) Water source utilization and foliar nutrient status differs between upland and flooded plant communities in wetland tree islands. Wetl Ecol Manag 18:343–355CrossRefGoogle Scholar
  51. Schultz NM, Griffis TJ, Lee XH, Baker JM (2011) Identification and correction of spectral contamination in H-2/H-1 and O-18/O-16 measured in leaf, stem, and soil water. Rapid Commun Mass Sp 25:3360–3368CrossRefGoogle Scholar
  52. Schwinning S, Kelly CK (2013) Plant competition, temporal niches and implications for productivity and adaptability to climate change in water-limited environments. Funct Ecol 27:886–897CrossRefGoogle Scholar
  53. Silvertown J, Araya Y, Gowing D (2015) Hydrological niches in terrestrial plant communities: a review. J Ecol 103:93–108CrossRefGoogle Scholar
  54. Tang YK, Chen YM, Wen XF, Sun XM, Wu X, Wang HM (2016) Variation of carbon use efficiency over ten years in a subtropical coniferous plantation in Southeast China. Ecol Eng 97:196–206CrossRefGoogle Scholar
  55. Tang YK, Wu X, Chen YM, Wen J, Xie YL, Lu SB (2018) Water use strategies for two dominant tree species in pure and mixed plantations of the semiarid Chinese loess plateau. Ecohydrology 11.
  56. Tardieu F, Simonneau T (1998) Variability among species of stomatal control under fluctuating soil water status and evaporative demand: modelling isohydric and anisohydric behaviours. J Exp Bot 49:419–432CrossRefGoogle Scholar
  57. Taylor BN, Chazdon RL, Bachelot B, Menge DNL (2017) Nitrogen-fixing trees inhibit growth of regenerating Costa Rican rainforests. P Natl Acad Sci USA 114:8817–8822CrossRefGoogle Scholar
  58. Tegeder M, Masclaux-Daubresse C (2018) Source and sink mechanisms of nitrogen transport and use. New Phytol 217:35–53. CrossRefPubMedGoogle Scholar
  59. Unkovich M (2013) Isotope discrimination provides new insight into biological nitrogen fixation. New Phytol 198:643–646PubMedCrossRefGoogle Scholar
  60. Wang X, Gao Y, Zhang H, Shao Z, Sun B, Gao Q (2019) Enhancement of rhizosphere citric acid and decrease of NO3−/NH4+ ratio by root interactions facilitate N fixation and transfer. Plant Soil.
  61. Wei LL, Lockington DA, Yu S, Lovelock CE (2015) Nitrogen sharing and water source partitioning co-occur in estuarine wetlands. Funct Plant Biol 42:410–417CrossRefGoogle Scholar
  62. West AG, Hultine KR, Burtch KG, Ehleringer JR (2007) Seasonal variations in moisture use in a pinon-juniper woodland. Oecologia 153:787–798PubMedCrossRefPubMedCentralGoogle Scholar
  63. West AG, Dawson TE, February EC, Midgley GF, Bond WJ, Aston TL (2012) Diverse functional responses to drought in a Mediterranean-type shrubland in South Africa. New Phytol 195:396–407PubMedCrossRefPubMedCentralGoogle Scholar
  64. Williams DG, Ehleringer JR (2000) Intra- and interspecific variation for summer precipitation use in pinyon-juniper woodlands. Ecol Monogr 70: 517–537Google Scholar
  65. Wu HW, Li XY, Jiang ZY, Chen HY, Zhang CC, Xiao X (2016a) Contrasting water use pattern of introduced and native plants in an alpine desert ecosystem, Northeast Qinghai-Tibet Plateau, China. Sci Total Environ 542:182–191PubMedCrossRefPubMedCentralGoogle Scholar
  66. Wu JN, Liu WJ, Chen CF (2016b) Can intercropping with the world's three major beverage plants help improve the water use of rubber trees? J Appl Ecol 53:1787–1799CrossRefGoogle Scholar
  67. Wurzburger N, Miniat CF (2014) Drought enhances symbiotic dinitrogen fixation and competitive ability of a temperate forest tree. Oecologia 174:1117–1126PubMedCrossRefPubMedCentralGoogle Scholar
  68. Xu GQ, Li Y, Xu H (2011) Seasonal variation in plant hydraulic traits of two co-occurring desert shrubs, Tamarix ramosissima and Haloxylon ammodendron, with different rooting patterns. Ecol Res 26:1071–1080CrossRefGoogle Scholar
  69. Yang B, Wen XF, Sun XM (2015) Seasonal variations in depth of water uptake for a subtropical coniferous plantation subjected to drought in an east Asian monsoon region. Agric For Meteorol 201:218–228CrossRefGoogle Scholar
  70. Zhang HD, Wei W, Chen LD, Yang L (2017) Evaluating canopy transpiration and water use of two typical planted tree species in the dryland loess plateau of China. Ecohydrology 10. CrossRefGoogle Scholar
  71. Zhou GY, Yin GC, Tang XL, Wen ZD, Liu GP, Kuang YW, Wang WT (2018) Carbon reserves in forest ecosystems of China: biomass allometric equation. Science Press, Beijing, pp 44–54 (In Chinese)Google Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Soil Erosion and Dry-land Farming on the Loess PlateauNorthwest A&F UniversityYanglingChina
  2. 2.State Key Laboratory of Soil Erosion and Dry-land Farming on the Loess Plateau, Institute of Soil and Water ConservationChinese Academy of Sciences and the Ministry of Water ResourcesYanglingChina
  3. 3.University of Chinese Academy of ScienceBeijingChina

Personalised recommendations