Forschungs- & Entwicklungsinformationsdienst der Gemeinschaft - CORDIS

Development of new cryopreservation protocols

New / adapted cryopreservation protocols have been established. The protocols are not only applicable to the mandate plant species, which were initially foreseen in the project, but also are readily applicable other plant species. Key developed protocols: Vitrification droplet method for apical meristems and highly proliferating meristem clumps: With this method, cryopreservation of banana became a routine procedure since all banana accessions can now be cryopreserved (up to date, more than 400 banana accessions are stored in liquid nitrogen for long term at K.U.Leuven).

Moreover, this method can be efficiently applied to a wide variety of plant species like potato, chicory, strawberry, taro, pelargonium and date palm. Other plant species are currently being tested. Simple freezing method: A very unique method only based on a preculture phase on high concentrations of sucrose. The application of specific sterols, polyamines or aromatic amines as well as some mono-unsaturated fatty acids into the sucrose preculture medium could enhance the post thaw shoot regeneration of proliferating meristems of different banana cultivars.

Encapsulation / dehydration cryopreservation method: The encapsulation/dehydration method as applied to Ribes spp. utilized DSC thermal analysis studies to evaluate and optimise the loading of sucrose into the alginate matrix and the critical desiccation times required for silica gel treatment as compared to evaporative airflow. This method is also applicable to garlic, apple and almond, hawthorn, service tree and strawberry tree, etc. The encapsulation method was combined with vitrification for several woody plant species in vitro germplasm types demonstrating that this method is particularly useful in aiding post storage survival in desiccation sensitive systems.

Controlled rate freezing method: This method has been applied to olive.

Seed cryopreservation method: Five key factors of non-orthodox seed cryopreservation procedures have been investigated for the first time (seed-lot quality, effect of slow drying, rewarming procedure) or re-examined (optimal hydration status, cooling rate). For each of these five factors, important findings were obtained, allowing immediate modifications of standard cryopreservation procedure for coffee seeds and formulation of important new recommendations for the setting up of a cryopreservation protocol in any new given species producing non-orthodox oily seeds.

Droplet freezing method: Compared with other methods, the ultra-rapid droplet freezing method is the simplest approach for the cryopreservation of potato germplasm. Innovative benefits: bespoke techniques for application by end user practitioners: germplasm repositories, seed banks, culture collections, botanic gardens and biotechnology companies. Helping to meet their needs to hold germplasm in cryogenic storage provides the only long-term means of securing non-orthodox germplasm ex situ. The major strength of the result is the integration of technological and fundamental approaches to assist end users to cryopreserve their germplasm collections.

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Reported by

Katholieke Universiteit Leuven
Laboratory of Tropical Crop Improvement, Kasteelpark Arenberg 13
3001 Heverlee
Belgium
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