, Volume 27, Issue 4, pp 311–319 | Cite as

Nutrient enrichment effects on mycorrhizal fungi in an Andean tropical montane Forest

  • Camille S. Delavaux
  • Tessa Camenzind
  • Jürgen Homeier
  • Rosa Jiménez-Paz
  • Mark Ashton
  • Simon A. Queenborough
Original Article


Nitrogen (N) and phosphorus (P) deposition are increasing worldwide largely due to increased fertilizer use and fossil fuel combustion. Most work with N and P deposition in natural ecosystems has focused on temperate, highly industrialized, regions. Tropical regions are becoming more developed, releasing large amounts of these nutrients into the atmosphere. Nutrient enrichment in nutrient-poor systems such as tropical montane forest can represent a relatively large shift in nutrient availability, especially for sensitive microorganisms such as arbuscular mycorrhizal fungi (AMF). These symbiotic fungi are particularly critical, given their key role in ecosystem processes affecting plant community structure and function.

To better understand the consequences of nutrient deposition in plant communities, a long-term nutrient addition experiment was set up in a tropical montane forest in the Andes of southern Ecuador. In this study, we investigated the impacts of 7 years of elevated N and P on AMF root colonization potential (AMF-RCP) through a greenhouse bait plant method in which we quantified root colonization. We also examined the relationship between AMF-RCP and rarefied tree diversity.

After 7 years of nutrient addition, AMF-RCP was negatively correlated with soil P, positively correlated with soil N, and positively correlated with rarefied tree diversity. Our results show that AMF in this tropical montane forest are directly affected by soil N and P concentrations, but may also be indirectly impacted by shifts in rarefied tree diversity. Our research also highlights the need to fully understand the benefits and drawbacks of using different sampling methods (e.g., AMF-RCP versus direct root sampling) to robustly examine AMF-plant interactions in the future.


Tropical Arbuscular mycorrhizas Ecuador Nitrogen fertilization Phosphorus fertilization 



We would like to acknowledge the Ministry of the Environment of Zamora, Ecuador, for allowing us to carry out our research. Sample collection was authorized under permit 0012-IC-FAU/FLO-DPZCH-MA from the Ministry of the Environment of Zamora, Ecuador; sample export was authorized under permit no. 3075376 from the Ministry of Agriculture, Ranching, Aquaculture, and Fisheries. The Ecuadorian Nutrient Manipulation Experiment was established and is maintained by funds from the German Research Foundation DFG (current project Ho3296/4).

In addition, we greatly thank Joseph Morton and the International Culture Collection of (Vesicular) Arbuscular Mycorrhizal Fungi (INVAM) for research input throughout this study. We recognize contribution from Craig Brodersen for lab resources, Dan Maynard for statistical support, and Marlyse Duguid for fruitful discussion and editing. We are grateful for comments on this manuscript by the Ashton, Comita, and Klironomos labs. We would also like to thank two anonymous reviewers and the editor for exceptional comments on this manuscript. Funding for this project was made available by the Yale Tropical Resources Institute, the Jubitz Family Endowment for Research Internships Fund, the F&ES Carpenter-Sperry Internship and Research Fund, and the Tinker Foundation through the Council on Latin American & Iberian Studies.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

572_2016_749_MOESM1_ESM.docx (73 kb)
Table S1 (DOCX 72 kb)


  1. Azcón-Aguillar C, Barea JM (1996) Arbuscular mycorrhizal and biological control of soil-borne plant pathogens—an overview of the mechanisms involved. Mycorrhiza 6:457–464CrossRefGoogle Scholar
  2. Augé RM (2001) Water relations, drought and vesicular–arbuscular mycorrhizal symbiosis. Mycorrhiza 11:3–42CrossRefGoogle Scholar
  3. Bates D, Maechler M, Bolker B and Walker S (2014). lme4: Linear mixed-effects models using Eigen and S4.R package version 1.1–7.
  4. Bobbink R et al (2010) Global assessment of nitrogen deposition effects on terrestrial plant diversity: a synthesis. Ecol Appl 20:30–59CrossRefPubMedGoogle Scholar
  5. Camenzind T, Hempel S, Homeier J, Horn S, Velescu A, Wilcke W, Rillig MC (2014) Nitrogen and phosphorous additions impact arbuscular mycorrhizal abundance and molecular diversity in a tropical montane forest. Glob Chang Biol 20:3646–3659CrossRefPubMedGoogle Scholar
  6. Camenzind T, Homeier J, Dietrich K, Hempel S, Hertiel D, John A, Leuschner C, Oelmann Y, Olsson PA, Suárez JP, Rillig MC (2016) Opposing effects of nitrogen versus phosphorus additions on mycorrhizal fungal abundance along an elevational gradient in tropical montane forests. Soil Biol Biochem 94:37–47CrossRefGoogle Scholar
  7. Camenzind T, Rillig MC (2013) Extraradical arbuscular mycorrhizal fungal hyphae in an organic tropical montane forest soil. Soil Biol Biochem 64:96–102CrossRefGoogle Scholar
  8. Chagnon PL, Bradley RL, Maherali H, Klironomos JN (2013) A trait-based framework to understand life history of mycorrhizal fungi. Trends Plant Sci 18:484–491CrossRefPubMedGoogle Scholar
  9. Chaudhary VB, Bowker MA, O’Dell TE, Grace TE, Redman AE, Rillig MC, Johnson NC (2009) Untangling the biological contributions to soil stability in semiarid shrublands. Ecol Appl 19:110–202CrossRefPubMedGoogle Scholar
  10. Chaudhary B, O’dell T, Rillig MC, Johnson N (2014) Multiscale patterns of arbuscular mycorrhizal fungal abundance and diversity in semiarid shrublands. Fungal Ecol 12:32–43CrossRefGoogle Scholar
  11. Cipollini D, Rigsby CM, Barto EK (2012) Microbes as targets and mediators of allelopathy in plants. J Chem Ecol 38:714–727CrossRefPubMedGoogle Scholar
  12. Egerton-Warburton LM, Querejeta JI, Allen MF (2007) Common mycorrhizal networks provide a potential pathway for the transfer of hydraulically lifted water between plants. J Exp Bot 58:1473–1483. doi: 10.1093/jxb/erm009 CrossRefPubMedGoogle Scholar
  13. Eom A, Hartnett D, Wilson G, Figge D (1999) The effect of fire, mowing and fertilizer amendment on arbusuclar mycorrhizas in tallgrass prairie. Am Midl Nat 142:55–70CrossRefGoogle Scholar
  14. Eriksson A (2001) Arbusuclar mycorrhiza in relation to management history, soil nutrients and plant diversity. Plant Ecol 155:129–137CrossRefGoogle Scholar
  15. Fangmeier A, Hadwiger-Fangmeier A, Van der Eerden L, Jäger HJ (1993) Effects of atmospheric ammonia on vegetation—a review. Environ Pollut 86:43–82CrossRefGoogle Scholar
  16. Gai J, Gao W, Liu L, Chen Q, Feng G, Zhang J, Christie P, Li X (2015) Infectivity and community composition of arbuscular mycorrhizal fungi fro different soil depths in intensively managed agricultural ecosystem. Journal of Soil Sediments 15:1200–1211CrossRefGoogle Scholar
  17. Garcia MO, Ovasapyan T, Greas M, Treseder KK (2008) Mycorrhizal dynamics under elevated CO2 and nitrogen fertilization in a warm temperate forest. Plant Soil 303:301–310CrossRefGoogle Scholar
  18. Giauque H, Hawkes CV (2013) Climate affects symbiotic fungal endophyte diversity and performance. Am J Bot 100:1435–1444CrossRefPubMedGoogle Scholar
  19. Giovannetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular–arbuscular mycorrhizal infection in roots. New Phytol 84:489–500CrossRefGoogle Scholar
  20. Gotelli NJ, Colwell RK (2001) Quantifying biodiversity: procedures and pitfalls in the measurement and comparison of species richness. Ecol Lett 4:379–391CrossRefGoogle Scholar
  21. Grman E (2012) Plant species differ in their ability to reduce allocation to non-beneficial arbusular mycorrhizal fungi. Ecology 93(4):711–718CrossRefPubMedGoogle Scholar
  22. Grman E, Robinson TMP (2013) Resource availability and imbalance affect plant–mycorrhizal interactions: a field test of three hypotheses. Ecology 94(1):62–71CrossRefPubMedGoogle Scholar
  23. Hart MM, Reader RJ, Klironomos JN (2003) Plant coexistence mediated by arbuscular mycorrhizal fungi. Trends Ecol Evol 18(8):418–423CrossRefGoogle Scholar
  24. Hoeksema JD, Chaudhary VB, Gehring CA, Johnson NC, Karst J, Koide RT, Pringle A, Zabinski C, Bever JD, Moore JC, Wilson GWT, Klironomos JN, Unbanhowar J (2010) A meta-analysis of context-dependency in plant response to inoculation with mycorrhizal fungi. Ecol Lett 13:394–407CrossRefPubMedGoogle Scholar
  25. Homeier J, Breckle SW, Günter S, Rollenbeck RT, Leuschner C (2010) Tree diversity, forest structure and productivity along altitudinal and topographic gradients in a species-rich Ecuadorian montane rain forest. Biotropica 42:140–148CrossRefGoogle Scholar
  26. Homeier J et al (2012) Tropical Andean forests are highly susceptible to nutrient inputs—rapid effects of experimental N and P addition to an Ecuadorian montane forest. PLoS One 7:e47128CrossRefPubMedPubMedCentralGoogle Scholar
  27. Homeier J, Werner FA, Gradstein SR, Breckle SW, Richter M (2008) Gradients in a tropical mountain ecosystem of Ecuador, ecological studies 198. Chapter 10.1, potential vegetarion and floristic composition of Andean forests in southern Ecuador, with a focus on RBSF. Springer-Verlag, Berlin Heidelberg, pp. 87–100Google Scholar
  28. Janos D (1996) Mycorrhizas, succession, and the rehabilitation of deforested lands in the humid tropics. In: Cambridge University Press, for British Mycological SocietyGoogle Scholar
  29. Johnson NC (2010) Resource stoichiometry elucidates the structure and function of arbuscular mycorrhizas across scales. New Phytol 185:631–647CrossRefPubMedGoogle Scholar
  30. Johnson D, Leake JR, Read DJ (2001) Novel in-growth core system enables functional studies of grassland mycorrhizal mycelial networks. New Phytol 152:555–562CrossRefGoogle Scholar
  31. Johnson D, Gilbert L (2015) Interplant signaling through hyphal networks. New Phytol 205:1448–1453CrossRefPubMedGoogle Scholar
  32. Johnson NC, Graham JH, Smith FA (1997) Functioning of mycorrhizal associations along the mutualism–parasitism continuum. New Phytol 135:575–585CrossRefGoogle Scholar
  33. Johnson NC, Wilson GWT, Wilson JA, Miller RM, Bowker MA (2015) Mycorrhizal phenotypes and the law of the minimum. New Phytol 205:1473–1484CrossRefPubMedGoogle Scholar
  34. Jung SC, Martinez-Medina A, Lopez-Raez J, Pozo MJ (2012) Mycorrhiza-induced resistance and priming of plant defences. J Chem Ecol 38:651–664CrossRefPubMedGoogle Scholar
  35. Klironomos J (2000) Host-specificity and functional diversity among arbuscular mycorrhizal fungi Microbial Biosystems: New. Frontiers:845–851Google Scholar
  36. Koide RT (1991) Nutrient supply, nutrient demand and plant response to mycorrhizal infection. New Phyologist 117:365–386CrossRefGoogle Scholar
  37. Koide RT, Mosse B (2004) A history of research on arbuscular mycorrhiza. Mycorrhiza 14:145–163CrossRefPubMedGoogle Scholar
  38. Kottke I, Beck A, Oberwinkler F, Homeier J, Neill D (2004) Arbuscular endomycorrhizas are dominant in the organic soil of a neotropical montane cloud forest. J Trop Ecol 20:125–129CrossRefGoogle Scholar
  39. Krashevska V, Sandmann D, Maraun M, Scheu S (2014) Moderate changes in nutrient input alter tropical microbial and protist communities and belowground linkages. The ISME Journal 8:1126–1134CrossRefPubMedGoogle Scholar
  40. Krupa SV (2003) Effects of atmospheric ammonia (NH3) on terrestrial vegetation: a review. Environ Pollut 124:179–221CrossRefPubMedGoogle Scholar
  41. Lambers H, Martinoia E, Renton M (2015) Plant adaptations to severely phosphorus-impoverished soils. Curr Opin Plant Biol 25:23–31CrossRefPubMedGoogle Scholar
  42. Maherali H, Klironomos JN (2012) Phyogenetic and trait-based assemly of arbuscular mycorrhizal fungal communities. PLoS One 7:1–9CrossRefGoogle Scholar
  43. Mahowald NM, Artaxo P, Baker AR, Jickells TD, Okin GS, Randerson JT, Townsend AR (2005) Impacts of biomass burning emissions and land use change on Amazonian atmospheric phosphorus cycling and deposition. Glob Biogeochem Cycles 19(4)Google Scholar
  44. Manning P, Saunders M, Bardgett RD, Bonkowski M, Bradford MA, Ellis RJ, Kandeler E, Marhan S, Tscherko D (2008) Direct and indirect effects of nitrogen deposition on litter decomposition. Soil Biol Biochem 40:688–698CrossRefGoogle Scholar
  45. Matson PA, McDowell WH, Townsend AR, Vitousek PM (1999) The globalization of N deposition: ecosystem consequences in tropical environments. Biochemistry 46:67–83Google Scholar
  46. McGonigle TP, Miller MH, Evans DG, Fairchild GL, Swarn JA (1990) A new method which gives an objective measure of colonization of roots by vesicular–arbuscular mycorrhizal fungi. New Phytol 115:495–501CrossRefGoogle Scholar
  47. Moorman T, Reeves FB (1979) The role of endomycorrhizae in revegetation practices in the semi-arid west. II. A bioassay to determine the effect of lang disturbance on endomycorrhizal populations. Am J Bot 66:14–18CrossRefGoogle Scholar
  48. Oades JM (1984) Soil organic matter and structural stability: mechanisms and implications for management. Plant Soil 76:319–337CrossRefGoogle Scholar
  49. Pagano M (2012) Mycorrhiza: Occurrence in Natural and Restored Environments. Chapter 4. Arbuscular Mycorrhiza in the Amazon Region, Nova Science pp 75–86Google Scholar
  50. Pozo MJ, Azcon-Aguilar C (2007) Unraveling mycorrhiza-induced resistance. Curr Opin Plant Biol 10:393–398. doi: 10.1016/j.pbi.2007.05.004 CrossRefPubMedGoogle Scholar
  51. R core team 2014: R Core Team (2014). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL
  52. Read DJ, Perez-Moreno J (2003) Mycorrhizas and nutrient cycling in ecosystems—a journey towards relevance? New Phytol 157:475–492CrossRefGoogle Scholar
  53. Rillig MC, Aguilar-Trigueros CA, Bergmann J, Verbruggen E, Veresoglou SD, Lehmann A (2015) Plant root and mycorrhizal fungal traits for understanding soil aggregation. New Phytol 205:1385–1388CrossRefPubMedGoogle Scholar
  54. Rillig MC, Mummey DL (2006) Mycorrhizas and soil structure. New Phytol 171:41–53CrossRefPubMedGoogle Scholar
  55. Schindelin J et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682CrossRefPubMedGoogle Scholar
  56. Schnitzer SA, Klironomos JN, HilleRisLambers J, Kinkel LL, Reich PB, Xiao K, Rillig MC, Sikes BA, Callaway RM, Mangan SA, van Nes EG, Scheffer M (2011) Soil microbes drive the classic plant diversity-productivity pattern. Ecology 92:296–303CrossRefPubMedGoogle Scholar
  57. Simard SW, Durall DM (2004) Mycorrhizal networks: a review of their extent, function, and importance. Can J Bot 82(8):1140–1165CrossRefGoogle Scholar
  58. Smith SE, Read DJ (2008) Mycorrhizal Symbiosis. Growth and carbon economy of arbuscular mycorrhizal symbionts, 3rd edn. Elsevier, Oxford, pp. 117–144Google Scholar
  59. Shantz AA, Lemoine NP, Burkepile DE (2016) Nutrient loading alters the performance of key nutrient exchanges mutualisms. Ecol Lett 19:20–28CrossRefPubMedGoogle Scholar
  60. Sykorova Z, Ineichen K, Wiemken A, Redecker D (2007) The cultivation bias: different communities of arbuscular mycorrhizal fungi detected in roots from the field, from bait plants transplanted to the field, and from a greenhouse trap experiment. Mycorrhiza 18:1–14CrossRefPubMedGoogle Scholar
  61. Tilman D (1999) The ecological consequences of changes in biodiversity: a search for general principles. Ecology 80:1455–1474Google Scholar
  62. Torti SD, Coley P, Janos DP (1997) Vesicular–arbuscular mycorrhizae in two tropical monodominant trees. J Trop Ecol 13:623–629CrossRefGoogle Scholar
  63. Treseder KK (2004) A meta-analysis of mycorrhizal responses to nitrogen, phosphorous, and atmospheric CO2 in field studies. New Phytol 164:347–355CrossRefGoogle Scholar
  64. Treseder KK, Allen MF (2002) Direct nitrogen and phosphorus limitation of arbuscular mycorrhizal fungi: a model and field test. New Phytol 155:507–515CrossRefGoogle Scholar
  65. Urgiles N, Loján P, Aguirre N, Blaschke H, Günter S, Stimm B, Kottke I (2009) Application of mycorrhizal roots improves growth of tropical tree seedlings in the nursery: a step towards reforestation with native species in the Andes of Ecuador. New For 38:229–239CrossRefGoogle Scholar
  66. Urgiles N, Strauß A, Loján P, Shüßler A (2014) Cultured arbuscular mycorrhizal fungi and native soil inocula improve seedling development of two pioneer trees in the Andean region. New For 45:859–874CrossRefGoogle Scholar
  67. van der Heijden MGA, Horton TA (2009) Socialism in soil? The important of mycorrhizal fungal networks for facilitation in natural ecosystems. J Ecol 97:1139–1150CrossRefGoogle Scholar
  68. van der Heijden MGA, Klironomos JN, Ursic M, Moutoglis P, Streitwolf-Engel R, Boller T, Wiemken A, Sanders IR (1998) Mycorrhizal fungal diversity determines plant biodiversity, ecosystem variability and productivity. Nature 396:69–72CrossRefGoogle Scholar
  69. van der Heijden MGA, Scheublin TR (2007) Functional traits in mycorrhizal ecology: their use for predicting the impact of arbuscular mycorhizal fungal communities on plant growth and ecosystem functioning. New Phytol 174:244–250CrossRefPubMedGoogle Scholar
  70. Vierheilig H, Schweiger P, Brundrett M (2005) An overview of methods for detection and observation of arbuscular mycorrhizal fungi in roots. Physiol Plant 125:393–404Google Scholar
  71. Vitousek PM (1994) Beyond global warming: ecology and global change. Ecological Society of America 75:1861–1876Google Scholar
  72. Wagg C, Jansa J, Stadler M, Schmid B, van der Heijden MGA (2014) Mycorrhizal fungal identitiy and diversity relaxes plant-plant competition. Ecological Society of America 92:1303–1313Google Scholar
  73. Wehner JP, Antunes M, Powell JR, Mazukatow J, Rillig MC (2010) Plant pathogen protection by arbsucular mycorrhizas: a role for fungal diversity? Pedobiologia 53:197–201CrossRefGoogle Scholar
  74. Weremijewicz J, Janos DP (2012) Common mycorrhizal networks amplify size inequality in Andropogon gerardii monocultures. New Phytol 198:203–213CrossRefGoogle Scholar
  75. Werner GDA, Kiers ET (2015) Partner selection in the mycorrhizal mutualism. New Phytol 205:1437–1442CrossRefPubMedGoogle Scholar
  76. Wilke W, Yasin S, Abramowski U, Valarezo C, Zech W (2002) Nutrient storage and turnover in organic layers under tropical montane rain forest in Ecuador. Eur J Soil Sci 53:15–27CrossRefGoogle Scholar
  77. Wolf K, Veldkamp E, Homeier J, Martinson GO (2011) Nitrogen availability links forests productivity, soil nitrous oxide and nitric oxide fluxes of a tropical montane forest in southern Ecuador. Glob Biogeochem Cycles 25:GB4009Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Camille S. Delavaux
    • 1
    • 2
  • Tessa Camenzind
    • 3
    • 4
  • Jürgen Homeier
    • 5
  • Rosa Jiménez-Paz
    • 6
  • Mark Ashton
    • 1
  • Simon A. Queenborough
    • 1
  1. 1.Yale School of Forestry and Environmental Studies, Yale UniversityNew HavenUSA
  2. 2.Ecology and Evolutionary Biology, University of KansasLawrenceUSA
  3. 3.Institute of Biology, Freie Universität BerlinBerlinGermany
  4. 4.Berlin- Brandenburg Institute of Advanced Biodiversity Research, Plant EcologyBerlinGermany
  5. 5.Albrecht von Haller Institute of Plant Sciences, University of GöttingenGöttingenGermany
  6. 6.Laboratorio de Ecología de Plantas, Escuela de Ciencias Biológicas, Pontificia Universidad Católica del EcuadorQuitoEcuador

Personalised recommendations