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DEVELOPMENT OF IMPROVED TECHNIQUES FOR THE PRESERVATION AND MAINTENANCE OF CULTURES OF FUNGAL STRAINS OF BIOTECHNOLOGICAL IMPORTANCE

Objectif

IMPROVED PRESERVATION TECHNIQUES FOR FUNGAL STRAINS ENSURING THE LONG TERM STABILITY OF THEIR BIOCHEMICAL AND PHYSIOLOGICAL ACTIVITIES.

THUS, CULTURE COLLECTIONS WILL HAVE THE POSSIBILITY TO PROVIDE STRAINS WITH MORE STABLE AND DEFINED PROPERTIES TO RESEARCH LABORATORIES AND INDUSTRY.
Our research programme has resulted in the development of improved long-term preservation procedures to retain purity, viability and stability of fungi. Cryomicroscopy has been used for the first time on fungi and the observed responses to cooling used to predict their survival following freezing techniques.

The techniques of freeze drying and cryopreservation have been used for many years but were not specifically developed for fungi. No fundametnal research had been carried out to determine the effect of cooling on fungal cells; this was needed for a better understanding of preservation in order to develop improved methods.
To optimize the techniques of freezing and freeze drying for the perservation of viability and stability of properties of fungi of biotechnological importance. The primary approach was to determine the effects of cooling and subsequent rewarming on selected strains of fungi, and employing the findings to devise optimal methods of preservation. The viability, morphology, physiology, biochemistry and other aspects of genetic stability were monitored before and after perservation and during storage under a variety of conditions. The results of preservation by the newly developed techniques were compared with those by traditional methods of freeze drying and cryopreservation. This enabled recommendations to be made on optimum techniques to adopt for different fungi.

25 of 26 fungi were shown to have optimum cooling rates for recovery. Death was related to shrinkage at slow cooling rates and, in Lentinus edodes, membrane integrity was lost during fast cooling. Many fungi gradually lost viability during storage at temperatures of -40 C or above. Most strains of fungi belonging to the Mastigomycotina failed to recover after 1 year of storage at -20 and -40 C but remained viable in liquid nitrogen vapour (approximately -175 C). 45 (60%) fungi failed to survive or died in storage when freeze dried using methods involving accelerated drying. The properties of the survivors did not deteriorate or change with loss of viability. Change seemed to relate more to storage time. Recovery time preserved strains became longer with storage time. In general, the properties of the fungi and yeasts tested were retained (3 y) but must be reassessed in the long term.
CULTURE COLLECTIONS OF FUNGI PROVIDE A VITAL GENETIC RESOURCE FOR BIOTECHNOLOGY, THE BIOCHEMICAL AND PHYSIOLOGICAL ACTIVITIES OF FUNGAL STRAINS BEING THE MAJOR ELEMENT OF INTEREST.

PRESERVATION METHODS OF FUNGI ARE EMPIRICALLY DEFINED AND MOSTLY NONSPECIFIC.

THE PRESENT PROJECT AIMS AT DEFINING PRESERVATION TECHNIQUES TO ENSURE LONG TERM OPTIMAL STABILITY OF FUNGAL STRAINS.

TEST STRAINS OF FUNGI WILL BE PRESERVED IN COLD STORAGE AND BY FREEZE DRYING. SELECTED PROPERTIES OF THESE STRAINS WILL BE TESTED BOTH BEFORE AND AFTER PRESERVATION AND AT INTERVALS DURING STORAGE.

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Coordinateur

CAB International Mycological Institute
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Adresse
Ferry Lane Kew
TW9 3AF Richmond
Royaume-Uni

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