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AOBPreview published online on November 13, 2002
Annals of Botany, doi:10.1093/aob/mcg009
© 2002 by Annals of Botany Company
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Submitted on January 3, 2002
Revised on March 19, 2002
Accepted on October 6, 2002

Suitability of Cryopreservation for the Long-term Storage of Rare and Endangered Plant Species: a Case History for Cosmos atrosanguineus

TIM WILKINSON1, ANDREW WETTEN2, CHRISSIE PRYCHID1, and MICHAEL F. FAY1*

Affiliation of the authors: 1 Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK; 2 School of Plant Sciences, University of Reading, Department of Agricultural Botany, Whiteknights, PO Box 221, Reading RG6 6AS, UK

* To whom correspondence should be addressed. E-mail: m.fay{at}rbgkew.org.uk.

The suitability of cryopreservation for the secure, long-term storage of the rare and endangered species Cosmos atrosanguineus was investigated. Using encapsulation/dehydration of shoot tips in alginate strips, survival rates of up to 100 % and shoot regeneration of up to 35 % were achieved. Light and electron microscopy studies indicated that cellular damage to some regions of the shoot tip during the freeze/thaw procedure was high, although cell survival in and around the meristematic region allowed shoot tip regeneration. The genetic fingerprinting technique, amplified fragment length polymorphisms (AFLPs), showed that no detectable genetic variation was present between material of C. atrosanguineus at the time of initiation into tissue culture and that which had been cryopreserved, stored in liquid nitrogen for 12 months and regenerated. Weaned plantlets that were grown under glasshouse conditions exhibited no morphological variation from non-frozen controls.
Key words: Cosmos atrosanguineus, cryopreservation, shoot tips, alginate encapsulation, somaclonal variation, AFLP, TEM.


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