Journal Article

Cryopreservation of Shoot Tips from Six Endangered Australian Species using a Modified Vitrification Protocol

S. R. Turner, T. Senaratna, E. Bunn, B. Tan, K. W. Dixon and D. H. Touchell

in Annals of Botany

Published on behalf of The Annals of Botany Company

Volume 87, issue 3, pages 371-378
Published in print March 2001 | ISSN: 0305-7364
Published online March 2001 | e-ISSN: 1095-8290 | DOI: https://dx.doi.org/10.1006/anbo.2000.1346
Cryopreservation of Shoot Tips from Six Endangered Australian Species using a Modified Vitrification Protocol

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An efficient vitrification procedure was developed and successfully applied to cryopreserve six endangered West Australian species (family Haemodoraceae: Anigozanthos humilis ssp. chrysanthus Hopper;A. kalbarriensis Hopper;A. viridis ssp. terraspectans Hopper;Conostylis dielsia ssp.teres Hopper;C. micrantha Hopper and C. wonganensis Hopper). Species were initially evaluated for cryostorage using a basic vitrification protocol involving: culturing plantlets in vitro for 21 d; excision of shoot apices; preculture of apical tips on 0.4 M sorbitol for 2 d, followed by incubation in PVS2 (plant vitrification solution 2) for 25 min at 0 °C, then direct immersion in liquid nitrogen (LN). Warming of retrieved material was for 1 min in a 40 °C water bath. Using this protocol five of the six species exhibited low post-storage survival, while the sixth species, A. viridis ssp. terraspectans posted higher survival (61.1%). Using A. viridis ssp. terraspectans as an indicator species, the initial protocol was modified to include: 3 d preculture on 0.80 M glycerol, loading treatment with 2.0 M glycerol plus 0.4 M sucrose solution for 20 min, followed by 25 min exposure to a modified PVS2. Survival was significantly improved in the test species, and in further experiments three other species also showed significant improvements with the new protocol. Key findings include: effectiveness of glycerol in the preculture medium; the effect of preculture duration; the importance of a loading stage for these species; and the successful use of modified PVS2 solutions with reduced or zero dimethyl sulfoxide (DMSO). Copyright 2001 Annals of Botany Company

Keywords: A. humilis ssp. chrysanthus, A. kalbarriensis, A. viridis ssp.terraspectans , Conostylis dielsia ssp. teres, C. micrantha, C. wonganensis, kangaroo paws, Haemodoraceae, vitrification, cryopreservation, rare and endangered, conservation

Journal Article.  5 words. 

Subjects: Ecology and Conservation ; Evolutionary Biology ; Plant Sciences and Forestry

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