Plant and Soil

, Volume 195, Issue 2, pp 329–338 | Cite as

Uptake of 15N-labelled alanine, ammonium and nitrate in Pinus sylvestris L. ectomycorrhiza growing in forest soil treated with nitrogen, sulphur or lime

  • Håkan Wallander
  • Kristina Arnebrant
  • Fredrik Östrand
  • Ola Kårén


The uptake of 15N-labelled alanine, ammonium and nitrate was studied in ectomycorrhizal morphotypes of intact Pinus sylvestris seedlings. PCR-RFLP analysis of the ITS-region of fungal rDNA was used to identify the morphotypes. Seedlings were grown in forest soil collected at an experimental site in southern Sweden. The treatments compared were a control, N fertilisation (600 kg N ha-1 as urea), sulfur application (1200 kg S ha-1) and lime application (6000 kg CaCO3 ha-1). The forest, which had been dominated by Picea abies, was clear-cut two years before the forest soil was sampled. Soil was also collected from an adjacent standing forest. The aim of the present study was to detect changes in the ectomycorrhizal communities in forest soils and relate these changes to the functional parameter of uptake of nitrogen from organic (alanine and protein) and inorganic (ammonium and nitrate) sources.

Liming resulted in the detection of a morphotype not found in other samples, and one morphotype was only found in samples from the standing forest (the fungi in these two morphotypes could not be identified). All mycorrhizal root tips showed a higher 15N concentration after exposure to different nitrogen forms than non-mycorrhizal long roots. Uptake of15 N from a labelled solution of alanine or ammonium was higher (about tenfold) than uptake from a 15N-labelled solution of nitrate. Uptake of ammonium and alanine varied between 0.2 and 0.5 mg N g-1 h-1 and between 0.1 and 0.33 mg N g-1 h-1, respectively, among the different morphotypes.

In seedlings grown in the control soil and in soil from standing forest, alanine and ammonium were taken up to a similar degree from a supply solution by all morphotypes, whereas ammonium uptake was higher than alanine uptake in seedlings grown in lime-treated soil (about twofold) and, to a lesser extent, in the nitrogen- and sulfur-treated soils. The higher ammonium uptake by morphotypes from the limed soil was confirmed in pure culture studies. In cases where ammonium was used as the N source, an isolate of the S. variegatus morphotype collected in the limed soil produced more biomass compared with isolates of S. variegatus collected in nitrogen- or sulphur-treated soil. One isolate of a silvery white morphotype produced about equal amounts of biomass on alanine and ammonium, whereas all S. variegatus isolated performed better with ammonium as their N source. Based on the results it is hypothesised that liming can induce a shift in the ectomycorrhizal community, favouring individuals that mainly utilise inorganic nitrogen over those that primarily utilise organic nitrogen.

ectomycorrhizal community nitrogen uptake organic nitrogen PCR Suillus 


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  1. Abuzinadah R A and Read D J 1986a The role of proteins in the nitrogen nutrition of ectomycorrhizal plants. II. Utilization of protein by mycorrhizal plants of Pinus contorta. New Phytol. 103, 495–506.Google Scholar
  2. Abuzinadah R A and Read D J 1986b The role of proteins in the nitrogen nutrition of ectomycorrhizal plants. III. Protein utilization by Betula, Picea and Pinus in mycorrhizal association with Hebeloma crustuliniforme. New Phytol. 103, 481–493.Google Scholar
  3. Andersson S and Söderström B 1995 Effects of lime (CaCO3) on ectomycorrhizal colonization of Picea abies (L.) Karst. seedlings planted in a spruce forest. Scand. J. For. Res. 10, 149–154. Agerer R 1996 Colour Atlas of Ectomycorrhizae. Einhorn-Verlag, Munich.Google Scholar
  4. Andersson F Bergholm J Hallbäcken Popovic B Möller G and Pettersson F 1996 Farabolförsöket, försurning, kalkning och kvävegödsling av en sydöstsvensk granskog. Swedish University of Agricultural Sciences, Department of Ecology and Environmmetal Sciences. Box 7072, 750 07 Uppsala, Sweden. Rapport 70.Google Scholar
  5. Arnebrant K 1994 Nitrogen amendments reduce the growth of extramatrical ectomycorrhizal mycelium. Mycorrhiza 5, 7–15.Google Scholar
  6. Arnebrant K and Bå åth E 1991 Measurements of ATP in forest humus. Soil Biol. Biochem. 23, 501–506.Google Scholar
  7. Arnebrant K and Söderström B 1992 Effects of different fertilizer treatments on ectomycorrhizal colonization potential in two Scots pine forests in Sweden. For. Ecol. Man. 53, 77–89.Google Scholar
  8. Buchmann N, Schulze E D and Gebauer G 1995 N-15-ammonium and N-15-nitrate uptake of a 15 year old Picea abies plantation. Oecologia 102, 361–370.Google Scholar
  9. Bå åth E and Arnebrant K 1994 Growth rate and response of bacterial communities to pH in limed and ash treated forest soil. Soil Biol. Biochem. 26, 995–1001. 338Google Scholar
  10. Dahlberg A and Stenström E 1991 Dynamic changes in nursery and indigenous mycorrhiza of Pinus sylvestris seedlings planted out in forest and clearcuts. Plant Soil 136, 73–86.Google Scholar
  11. Dighton J and Mason P A 1988 Mycorrhizal dynamics during forest tree development. In Developmental Biology of Higher Fungi. Eds D. Moore, L A Casselton, D A Wood and J C Frankland. Cambridge University Press, New York, USA.Google Scholar
  12. Erland S and Söderström B 1990 Effects of liming on ectomycorrhizal fungi infecting Pinus sylvestris L. I. Mycorrhizal infection in lined humus in the laboratory and isolation of fungi from mycorrhizal roots. New Phytol. 115, 675–682.Google Scholar
  13. Finlay R D, FrostegårdÅ A and Sonnerfeldt A-M 1992 Utilization of organic and inorganic nitrogen sources by ectomycorrhizal fungi in pure culture and in symbiosis with Pinus contorta Dougl. ex Loud. New Phytol. 120, 105–115.Google Scholar
  14. Gardes M White T J Fortin J A Bruns T D and Taylor J W 1991 Identification of indigenous and introduced symbiotic fungi in ectomycorrhizae by amplification of nuclear and mitocondrial ribosomal DNA. Can. J. Bot. 69, 180–190.Google Scholar
  15. Jacobsson S 1993 Inventering av svampfloran i kalkade provrutor på hallandsås. Jordstjärnan 14, 13–23.Google Scholar
  16. Jones HE Quarmby C and Harrison AF 1991 Aroot bioassay test for nitrogen deficiency in forest trees. For. Ecol. Man. 42, 267–282.Google Scholar
  17. Kårén O Högberg N Dahlberg A Jonsson L and Nylund J-E 1997 Inter-and intraspecific variation in the ITS region of rDNA of ectomycorrhizal fungi in Fennoscandia as detected by endonuclease analysis. New Phytol. 136.Google Scholar
  18. Nohrstedt HO Sikstrom U Ring E Näsholm T Högberg P Persson T 1996 Nitrate in soil water in three Norway spruce stands in southwest Sweden as related to N-deposition and soil, stand, and foliage properties. Can. J. For. Res. 26, 836–848.Google Scholar
  19. Persson T and Wirén A 1995 Nitrogen mineralization and potential nitrification at different depths in acid forest soils. Plant Soil 168–169, 55–66.Google Scholar
  20. Plassard C Scheromm P Mousain D and Salsac L 1991 Assimilation of mineral nitrogen and ion balance in the two partners of ectomycorrhizal symbiosis: Data and hypothesis. Experientia 47, 340–349.Google Scholar
  21. Read DJ 1991 Mycorrhizas in ecosystems. Experientia 47, 376–391. Schaffer H E and Sederoff R R 1981 Improved estimation of DNA fragment lengths from agarose gels. Anal. Biochem. 115, 113– 122.Google Scholar
  22. Sneath P H A and Sokal R R 1973 Numerical Taxonomy. W.H. Freeman, San Fransisco.Google Scholar
  23. Sokal R R and Rohlf F J 1995 Biometry, The Principles and Practice of Statistics in Biological Research.W.H. Freeman and Company, New York.Google Scholar
  24. Stevenson FJ 1982 Organic forms of soil nitrogen. In Humus Chemistry. Genesis, Composition, Reactions. Ed. F J Stevenson. pp 55–92. John Wiley and Sons, New York.Google Scholar
  25. Wallander H 1994 A new hypothesis to explain allocation of dry matter between mycorrhizal fungi and pine seedlings in relation to nutrient supply. Plant Soil 168–169, 243–248.Google Scholar
  26. White T J, Bruns T, Lee S and Taylor J 1990 Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR protocols: A Guide to Methods and Applications. Eds. M A Innis, D H Gelfand, J J Sninsky, T J White. pp 315–322. Academic Press, San Diego.Google Scholar

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© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Håkan Wallander
  • Kristina Arnebrant
  • Fredrik Östrand
  • Ola Kårén

There are no affiliations available

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