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Appl Environ Microbiol, February 1998, p. 613-617, Vol. 64, No. 2
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Use of Fluorogenic Substrates To Measure Fungal Presence and Activity in Soil

Morten Miller,1,* Ansa Palojärvi,2 Andrea Rangger,3 Morten Reeslev,1 and Annelise Kjøller1

Department of General Microbiology, University of Copenhagen, DK-1307 Copenhagen K, Denmark,1 GSF-Forschungszentrum für Umwelt- und Gesundheit GmbH, Institut für Bodenökologie, D-85764 Oberschleißheim, Germany,2 and Institute of Microbiology, University of Innsbruck, A-6020 Innsbruck, Austria3

Received 9 June 1997/Accepted 24 November 1997

Our objective was to determine if 4-methylumbelliferyl-labelled enzyme substrates could be used to detect and quantify specific components of chitinase and cellulase activities as specific indicators of the presence and activity of fungal biomass. The fluorogenic substrates 4-methylumbelliferyl (MUF) N-acetyl-beta -D-glucosaminide and MUF beta -D-lactoside were used for the detection and quantification of beta -N-acetylglucosaminidase (EC 3.2.1.30) (NAGase) and endo 1,4-beta -glucanase (EC 3.2.1.4)/cellobiohydrolase (EC 3.2.1.91) (CELase), respectively. Culture screenings on solid media showed a widespread ability to produce NAGase among a taxonomically diverse selection of fungi on media with and without added chitin. NAGase activity was expressed only in a limited number of bacteria and on media supplemented with chitin. The CELase activity was observed only in a limited number of fungi and bacteria. Bacterial CELase activity was expressed on agar media containing a cellulose-derived substrate. In soil samples, NAGase activity was significantly correlated with estimates of fungal biomass, based on the content of two fungus-specific indicator molecules, 18:2omega 6 phospholipid fatty acid (PLFA) and ergosterol. CELase activity was significantly correlated with the PLFA-based estimate of fungal biomass in the soil, but no correlation was found with ergosterol-based estimates of fungal biomass.

* Corresponding author. Mailing address: Department of General Microbiology, University of Copenhagen, Sølvgade 83 H, DK-1307 Copenhagen K, Denmark. Phone: 45 35 32 20 54. Fax: 45 35 32 20 40. E-mail: mmiller{at}mermaid.molbio.ku.dk.




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