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Detection of extracellular protease activity in different species and genera of ectomycorrhizal fungi

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Abstract

In northern forest ecosystems, most soil nitrogen (N) is in organic form and forest trees are largely dependent on ectomycorrhizal (ECM) fungi and their degradative abilities for N uptake. The ability of ECM fungi to acquire N from organic substrates should, therefore, be a widespread trait given its ecological importance. However, little is known about the degradative abilities of most ECM fungi as they remain untested due to problems of isolation or extremely slow growth in pure culture. In this paper, we present data on extracellular protease activity of 32 species of ECM fungi, most of which have not previously been cultured. Milk powder plates and zymograms were compared for detecting protease activity in these intractable species. In total, 29/32 of the species produced extracellular protease activity, but detection was method dependent. Growth on milk powder plates detected protease activity in 28 of 32 species, while zymograms only detected proteases in Amanita muscaria, Russula chloroides, Lactarius deterrimus and Lactarius quieticolor. The study supports the hypothesis that protease excretion is a widespread physiological trait in ECM fungi and that this ability is of considerable significance for nitrogen uptake in forest ecosystems.

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References

  • Abuzinadah RA, Finlay RD, Read DJ (1986) 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

    Article  CAS  Google Scholar 

  • Agerer R (2001) Exploration types of ectomycorrhizae: a proposal to classify ectomycorrhizal mycelial systems according to their patterns of differentiation and putative ecological importance. Mycorrhiza 11:107–114

    Article  Google Scholar 

  • Barbour MG, Burk JH, Pitts WD (1987) Terrestrial plant ecology. Benjamin/Cummings, Menlo Park, CA

    Google Scholar 

  • Benson DA, Karsch-Mizrachi I, Lipman DJ, Ostell J, Wheeler DL (2005) GenBank. Nucleic Acids Res 33:D34–D38, Database issue

    Article  CAS  PubMed  Google Scholar 

  • Cai YJ, Chapman SJ, Buswell JA, Chang ST (1999) Production and distribution of endoglucanase, cellobiohydrolysate and β-glucosidase components of the cellulolytic system of Volvariella volvacea, the edible straw mushroom. Appl Environ Microbiol 65:553–559

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cohen BL (1981) Regulation of protease production in Aspergillus. Trans Br Mycol Soc 76:447–450

    Article  CAS  Google Scholar 

  • Finlay RD, Frostegård Å, Sonnerfeldt AM (1992) Utilization of organic and inorganic nitrogen sources by ectomycorrhizal fungi in pure culture and in symbiosis with Pinus contorta. New Phytol 120:105–115

    Article  Google Scholar 

  • Gomi K, Arikawa K, Kamiya N, Kitamoto K, Kumagai C (1993) Cloning and nucleotide sequence of the acid protease encoding gene (pep A) from Aspergillus oryzae. Biosci Biotechnol Biochem 57:592–621

    Article  Google Scholar 

  • Horton TR, Bruns TD (2001) The molecular revolution in ectomycorrhizal ecology: peeking into the black box. Mol Ecol 10:1855–1871

    Article  CAS  PubMed  Google Scholar 

  • Kõljalg U, Larsson K-H, Abarenkov K et al (2005) UNITE—a database providing web-based methods for the molecular identification of ectomycorrhizal fungi. New Phytol 166:1063–1068

    Article  PubMed  Google Scholar 

  • Leake JR, Read DJ (1989) The biology of mycorrhiza in the Ericaceae XIII. Some characteristics of the extracellular proteinase activity of the ericoid endophyte Hymenoscyphus ericae. New Phytol 112:69–76

    Article  CAS  Google Scholar 

  • Leake JR, Read DJ (1990a) Proteinase activity in mycorrhizal fungi I. The effect of extracellular pH on the production and activity of proteinase by ericoid endophytes from soils of contrasted pH. New Phytol 115:243–250

    Article  CAS  Google Scholar 

  • Leake JR, Read DJ (1990b) Proteinase activity in mycorrhizal fungi II. The effects of mineral and organic nitrogen sources on induction of extracellular proteinase in Hymenoscyphus ericae (Read) Korf and Kernan. New Phytol 116:123–128

    Article  CAS  Google Scholar 

  • Leake JR, Read DJ (1991) Proteinase activity in mycorrhizal fungi III. Effects on protein, protein hydrolysate, glucose and ammonium on production of extracellular proteinase by Hymenoscyphus ericae (Read) Korf and Kernan. New Phytol 117:309–317

    Article  CAS  Google Scholar 

  • Leake JR, Read DJ (1997) Mycorrhizal fungi in terrestrial habitats. The mycota IV. In: Wicklow D, Söderström B (eds) Environmental and microbial relationships. Springer, Berlin Heidelberg New York, pp 281–301

    Google Scholar 

  • Lilleskov EA, Hobbie EA, Fahey TJ (2002) Ectomycorrhizal fungal taxa differing in response to nitrogen deposition also differ in pure culture organic nitrogen use and natural abundance of nitrogen isotopes. New Phytol 154:219–231

    Article  CAS  Google Scholar 

  • Lindahl BD, Taylor AFS (2004) Occurrence of N-acetylhexosaminidase-encoding genes in ectomycorrhizal basidiomycetes. New Phytol 164:193–199

    Article  CAS  Google Scholar 

  • Lindahl BD, Finaly RD, Cairney JWG (2005) Enzymatic activities of mycelia in mycorrhizal fungal communities. In: Dighton J, White JF, Oudemans P (eds) The fungal community—its organisation and role in the ecosystem. Taylor and Francis, Boca Raton, FL, pp 331–348

    Google Scholar 

  • Lopez-Llorca LV, Carbonell T, Gomez-Vidal S (2002) Degradation of insect cuticle by Paecilomyces farinosus proteases. Mycol Prog 3:249–256

    Article  Google Scholar 

  • Maijala P, Fagerstedt KV, Raudaskoski M (1991) Detection of extracellular cellulolytic and proteolytic activity in ectomycorrhizal fungi and Heterobasidion annosum (Fr.) Bref. New Phytol 117:643–648

    Article  CAS  Google Scholar 

  • Martino E, Franco B, Piccoli G, Stocchi V, Perotto S (2002) Influence of zinc ions on protein secretion in heavy metal tolerant strain of the ericoid mycorrhizal fungus Oidiodendron maius. Mol Cell Biochem 231:179–185

    Article  CAS  PubMed  Google Scholar 

  • Marx DH (1969) The influence of ectotrophic fungi on the resistance of pine roots to pathogenic infection. Phytopathology 59:153–163

    Google Scholar 

  • Nehls U, Bock A, Einig W, Hampp R (2001) Excretion of two proteases by the ectomycorrhizal fungus Amanita muscaria. Plant Cell Environ 24:741–747

    Article  CAS  Google Scholar 

  • Norkrans B (1949) Some mycorrhiza forming Tricholoma species. Svensk Botanisk Tidskrift 43:485–490

    Google Scholar 

  • Näsholm T, Persson J (2001) Plant acquisition of organic nitrogen in boreal forests. Physiol Plant 111:419–426

    Article  PubMed  Google Scholar 

  • Ogawa Y, Tatsumi H, Murakami S et al (1990) Secretion of Aspergillus oryzae alkaline protease in an osmophilic yeast, Zygosaccharomyces rouxii. Agric Biol Chem 54:2521–2529

    CAS  PubMed  Google Scholar 

  • Plassard C, Bonafos B, Touraine B (2000) Differential effects of mineral and organic N sources, and of ectomycorrhizal infection by Hebeloma cylindrosporum, on growth and N utilization in Pinus pinaster. Plant Cell Environ 23:1195–1205

    Article  Google Scholar 

  • Raidl S (1997) Studien zur Ontogenie an Rhizomorphen von Ektomykorrhizen. Bibl Mycol 169:184

    Google Scholar 

  • Rao MB, Tanksale AM, Ghatge MS, Deshpande VV (1998) Molecular and biotechnological aspects of microbial proteases. Microb Mol Biol Rev 62:597–635

    CAS  Google Scholar 

  • Read DJ, Perez-Moreno J (2003) Mycorrhizas and nutrient cycling in ecosystems—a journey towards relevance? New Phytol 157:475–492

    Article  Google Scholar 

  • Roche PC, Ryan RJ (1986) Electrophoretic analysis of membrane proteases in the luteinized rat ovary. Endocrinology 119:495–501

    Article  CAS  PubMed  Google Scholar 

  • Sandhya C, Sumantha A, Szakacs G, Pandey A (2005) Comparative evaluation of neutral protease production by Aspergillus oryzae in submerged and solid-state fermentation. Process Biochem 40:2689–2694

    Article  CAS  Google Scholar 

  • Sinsabaugh RL (2005) Fungal enzymes at the community scale. In: Dighton J, White JF, Oudemans P (eds) The fungal community—its organisation and role in the ecosystem. Taylor and Francis, Boca Raton, FL, pp 349–360

    Chapter  Google Scholar 

  • Smith S, Read DJ (1997) Mycorrhizal symbiosis. Academic, London

    Google Scholar 

  • Tamm CO (1991) Nitrogen in terrestrial ecosystems. Springer, Berlin Heidelberg New York

    Book  Google Scholar 

  • Taylor AFS, Martin F, Read DJ (2000) Fungal diversity in ectomycorrhizal communities of Norway spruce (Picea abies (L.) Karst.) and Beech (Fagus sylvatica L.) along north–south transects in Europe. In: Schulze E-D (ed) Carbon and nitrogen cycling in European forest ecosystems. Ecol Stud 142:343–365

  • Taylor AFS, Gebauer G, Read DJ (2004) Uptake of nitrogen and carbon from double-labelled 15N and 13C glycine by mycorrhizal pine seedlings. New Phytol 164:383–388

    Article  CAS  Google Scholar 

  • Vrålstad T, Schumacher T, Taylor AFS (2002) Mycorrhizal synthesis between fungal strains of the Hymenoscyphus ericae aggregate and potential ectomycorrhizal and ericoid hosts. New Phytol 153:143–152

    Article  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Ursula Eberhardt and Rasmus Kjøller for the use of their private sequence databases. This work was financially supported by the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (FORMAS-21.4/2003-0894).

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Correspondence to Cajsa M. R. Nygren.

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Nygren, C.M.R., Edqvist, J., Elfstrand, M. et al. Detection of extracellular protease activity in different species and genera of ectomycorrhizal fungi. Mycorrhiza 17, 241–248 (2007). https://doi.org/10.1007/s00572-006-0100-7

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