Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

Pritsch, K. and Courty, Pierre-Emmanuel and Churin, J. L. and Cloutier-Hurteau, B. and Ali, M. A. and Damon, C. and Duchemin, M. and Egli, S. and Ernst, J. and Fraissinet-Tachet, L. and Kuhar, F. and Legname, E. and Marmeisse, R. and Muller, A. and Nikolova, P. and Peter, M. and Plassard, C. and Richard, F. and Schloter, M. and Selosse, M. A. and Franc, A. and Garbaye, J.. (2011) Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae. Mycorrhiza, 21 (7). pp. 589-600.

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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 4-6A degrees 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 -20A degrees C or storage in 100% ethanol were recommended.
Faculties and Departments:05 Faculty of Science > Departement Umweltwissenschaften > Ehemalige Einheiten Umweltwissenschaften > Pflanzenphysiologie Zuckermetabolismus (Wiemken)
UniBasel Contributors:Courty, Pierre-Emmanuel
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:30 Nov 2017 10:17
Deposited On:30 Nov 2017 10:17

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