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Title: CRYOPRESERVATION OF ARABIDOPSIS THALIANA SHOOT TIPS.

Author
item Towill, Leigh
item Bonnart, Remi
item Volk, Gayle

Submitted to: CryoLetters
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/6/2006
Publication Date: 12/31/2006
Citation: Towill, L.E., R. Bonnart and G.M. Volk. 2006. Cryopreservation of Arabidopsis thaliana shoot tips. CryoLetters 27:353-360.

Interpretive Summary: We are developing a model to evaluate mechanisms of injury and of protection from the diverse stresses that occur during cryopreservation. Shoot tips from Arabidopsis thaliana 'Columbia' were exposed to liquid nitrogen using established techniques that provide exogenous cryoprotectants and controlled cooling rates to cryogenic temperatures. High levels of survival and regrowth occurred after LN exposure. The abundance of mutants available in this species will aid us in assessing biochemical, metabolic and developmental processes that affect injury and protection during cryopreservation.

Technical Abstract: Arabidopsis thaliana shoot tips provide a model to study processes important for cryopreservation. Cryopreservation was accomplished using both so-called vitrification and two-step cooling methods. With vitrification methods, shoot formation after liquid nitrogen exposure was as high as 100% and 95%, respectively, for shoot tips exposed to PVS2 at 0°C and to PVS3 at 23°C. A two-step cooling method also gave greater than 90% survival if shoot tips were cooled at 0.3°C min-1 to below -30°C before immersing the samples into liquid nitrogen. The high levels of shoot formation after LN exposure in Arabidopsis thaliana shoot tips will allow use of mutants to examine how alterations in biochemical, metabolic and developmental paths affect survival and growth.