title: Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae.
abstract: The aim of a joint effort by different research teams was to provide an improved procedure for enzyme activity profiling of field-sampled ectomycorrhizae, including recommendations on the best conditions and maximum duration for storage of ectomycorrhizal samples. A more simplified and efficient protocol compared to formerly published procedures was achieved by using manufactured 96-filter plates in combination with a vacuum manifold and by optimizing incubation times. Major improvements were achieved by performing the series of eight enzyme assays with a single series of root samples instead of two series, reducing the time needed for sample preparation, minimizing error-prone steps such as pipetting and morphotyping, and facilitating subsequent DNA analyses due to the reduced sequencing effort. The best preservation of samples proved to be storage in soil at 4-6 °C in the form of undisturbed soil cores containing roots. Enzyme activities were maintained for up to 4 weeks under these conditions. Short-term storage of washed roots and ectomycorrhizal tips overnight in water did not cause substantial changes in enzyme activity profiles. No optimal means for longer-term storage by freezing at -20 °C or storage in 100% ethanol were recommended.
authors.affiliation: Institute of Soil Ecology, German Research Center for Environmental Health (GmbH), 85764, Neuherberg, Germany. pritsch@helmholtz-muenchen.de.
authors.forename: Karin
authors.initials: K
authors.lastname: Pritsch
authors.affiliation: UMR 1136, Interactions Arbres Micro-organismes, Centre INRA de Nancy, 54280, Champenoux, France.|Botanical Institute, University of Basel, 4056, Basel, Switzerland.
authors.forename: Pierre Emanuel
authors.initials: PE
authors.lastname: Courty
authors.affiliation: UMR 1136, Interactions Arbres Micro-organismes, Centre INRA de Nancy, 54280, Champenoux, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Churin
authors.affiliation: UMR 1222, Ecologie Fonctionelle et Biogéochimie des Sols, INRA/IRD/SupAgro, 34060, Montpellier Cedex 01, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Cloutier-Hurteau
authors.affiliation: UMR 1222, Ecologie Fonctionelle et Biogéochimie des Sols, INRA/IRD/SupAgro, 34060, Montpellier Cedex 01, France.
authors.forename: Muhammad Arif
authors.initials: MA
authors.lastname: Ali
authors.affiliation: UMR CNRS 5557 d'Ecologie Microbienne, Université de Lyon, Université Lyon 1, 69622, Villeurbanne Cedex, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Damon
authors.affiliation: UMR 1222, Ecologie Fonctionelle et Biogéochimie des Sols, INRA/IRD/SupAgro, 34060, Montpellier Cedex 01, France.
authors.forename: Myriam
authors.initials: M
authors.lastname: Duchemin
authors.affiliation: Swiss Federal Research Institute WSL, 8903, Birmensdorf, Switzerland.
authors.forename: Simon
authors.initials: S
authors.lastname: Egli
authors.affiliation: Institute of Soil Ecology, Terrestrial Ecogenetics, German Research Center for Environmental Health (GmbH), 85764, Neuherberg, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Ernst
authors.affiliation: UMR CNRS 5557 d'Ecologie Microbienne, Université de Lyon, Université Lyon 1, 69622, Villeurbanne Cedex, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Fraissinet-Tachet
authors.affiliation: Biodiversidad y Biología Experimental, CABA, Universidad de Buenos Aires, Buenos Aires, 1428, Argentina.|Institute of Soil Ecology, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), 85764, Neuherberg, Germany.
authors.forename: Francisco
authors.initials: F
authors.lastname: Kuhar
authors.affiliation: UMR 1222, Ecologie Fonctionelle et Biogéochimie des Sols, INRA/IRD/SupAgro, 34060, Montpellier Cedex 01, France.
authors.forename: Elvira
authors.initials: E
authors.lastname: Legname
authors.affiliation: UMR CNRS 5557 d'Ecologie Microbienne, Université de Lyon, Université Lyon 1, 69622, Villeurbanne Cedex, France.
authors.forename: Roland
authors.initials: R
authors.lastname: Marmeisse
authors.affiliation: Swiss Federal Research Institute WSL, 8903, Birmensdorf, Switzerland.
authors.forename: Alex
authors.initials: A
authors.lastname: Müller
authors.affiliation: Institute of Soil Ecology, German Research Center for Environmental Health (GmbH), 85764, Neuherberg, Germany.
authors.forename: Petia
authors.initials: P
authors.lastname: Nikolova
authors.affiliation: Swiss Federal Research Institute WSL, 8903, Birmensdorf, Switzerland.
authors.forename: Martina
authors.initials: M
authors.lastname: Peter
authors.affiliation: UMR 1222, Ecologie Fonctionelle et Biogéochimie des Sols, INRA/IRD/SupAgro, 34060, Montpellier Cedex 01, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Plassard
authors.affiliation: UMR 5175, Centre d'Ecologie Fonctionnelle et Evolutive, 34293, Montpellier 5, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Richard
authors.affiliation: Institute of Soil Ecology, Terrestrial Ecogenetics, German Research Center for Environmental Health (GmbH), 85764, Neuherberg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schloter
authors.affiliation: UMR 5175, Centre d'Ecologie Fonctionnelle et Evolutive, 34293, Montpellier 5, France.
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Selosse
authors.affiliation: UMR Biodivers Genes & Communautes, INRA Pierroton, 33612, Cestas, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Franc
authors.affiliation: UMR 1136, Interactions Arbres Micro-organismes, Centre INRA de Nancy, 54280, Champenoux, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Garbaye
mesh_terms: D003080:Cold Temperature; D004798:Enzymes / Q000032:analysis*; D008828:Microbiological Techniques / Q000379:methods; D009172:Mycology / Q000379:methods*; D038821:Mycorrhizae / Q000201:enzymology*; D018517:Plant Roots / Q000382:microbiology*; D011309:Preservation, Biological / Q000379:methods*; D013997:Time Factors
chemical_list: D004798:Enzymes