Optimization of cryopreservation protocols of ectomycorrhizal fungi : effects on plant root colonization, growth and nutrition and on fungal genetic stability
(2017)
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- Abstract
- The objective of the thesis was to develop a cryopreservation protocol adapted to different ectomycorrhizal (ECM) fungi. Those fungi form symbiotic associations with the roots of woody and shrub plant species. They provide nutrients (in particular nitrogen and phosphorus) to the plant, in exchange for carbohydrates. As a result, ECM fungi usually improve plant growth and productivity. Cryopreservation of ECM fungi would secure their viability and biological stability over long periods of storage in culture collections. Different cryopreservation protocols were evaluated to identify which are the most appropriate factors for ECM fungi. Over the experiments, a cryopreservation protocol was developed. In this, so named cryovial protocol (CP), ECM fungal mycelium was directly grown in cryovials then incubated in glycerol. The cultures were then cooled at a freezing rate of -1 °C min-1 and stored at -130 °C. For revival, the samples were rapidly thawed in a water bath at 38 °C and transferred on agar plates. The CP was efficient for nearly 100 ECM fungal isolates belonging to 11 species. Phenotypic and genetic stability were maintained following cryopreservation using the CP. Our results provide culture collections an efficient and reliable method for the maintenance of ECM fungi - usually considered recalcitrant to cryopreservation - as an alternative to the widely-used continuous sub-cultivation method.
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Citations
Crahay, C. (2017). Optimization of cryopreservation protocols of ectomycorrhizal fungi : effects on plant root colonization, growth and nutrition and on fungal genetic stability. https://hdl.handle.net/2078.5/172972