CONTROLLED RATE COOLING OF FUNGI USING A STIRLING CYCLE FREEZER

摘要

BACKGROUND: The use of a Stirling cycle freezer for cryopreservation is considered to have significant advantages over traditional methodologies including N2 free operation, application of low cooling rates, reduction of sample contamination risks and control of ice nucleation. OBJECTIVE: The study assesses the suitability of an 'N_2-free' Stirling Cycle controlled rate freezer for fungi cryopreservation. METHODS: In total, 77 fungi representing a broad taxonomic coverage were cooled using the N_2 free cooler following a cooling rate of -1°C min~(-1). Of these, 15 strains were also cryopreserved using a traditional 'N_2 gas chamber' controlled rate cooler and a comparison of culture morphology and genomic stability against non-cryopreserved starter cultures was undertaken. RESULTS: In total of 75 fungi survived cryopreservation, only a recalcitrant Basidiomycete and filamentous Chromist failed to survive. No changes were detected in genomic profile after preservation, suggesting that genomic function is not adversely compromised as a result of using 'N_2 free' cooling. CONCLUSION: The results demonstrate the potential of 'N_2-free' cooling for the routine cryopreservation of fungi in Biological Resource Centres.