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Cryopreservation of Dendrobium heterocarpum Lindl. via encapsulation-dehydration method

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Author(s): Witaya Pimda | Sumontip Bunnag

Journal: Extreme Life, Biospeology & Astrobiology
ISSN 2066-7671

Volume: 2;
Issue: 1;
Start page: 7;
Date: 2010;
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Keywords: Dendrobium heterocarpum Lindl. | encapsulation-dehydration | protocorms | cryopreservation | cold acclimation | regrowth | preculture

ABSTRACT
The species Dendrobium heterocarpum Lindl., a wild orchid native to north and northeastThailand, creates an aura of mystique and grandeur with their intricate flower appearance which enthralsmany collectors within the country. An increase in collection for trade or any orther purposes may lead toa dramatic decrase in the population of this species, thus becoming rare or endangered species in thenear future. Cryopreservation is an alternative means for conservation of plant genetic resourceswhich can be applied to this species. This study, therefore, was aimed to optimize the conditions forencapsulation-dehydration method in D. heterocarpum. The protocorms were successfully cryopreservedas the following protocol. The protocorms were encapsulated with Ca-alginate, precultured on solid NDmedium supplemented with 0.25 M sucrose for 1 week in the dark at 0°C and subsequently preculturedin liquid ND medium supplemented with 0.25 M sucrose for 2 days in the dark at 0°C. Then, they weredehydrated by air-drying for 12 hours and plunged into LN for 1 day. Recovery percentage was 8.33%after 2 weeks of regrowth. However, preculture with ABA was found to insufficiently enhance freezingand thawing tolerance. TTC staining was negative and further regrowth on solid ND medium confirmedno survival of the cryopreserved protocorms.
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