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Title: Gene expression in response to cryoprotectant and liquid nitrogen exposure in Arabidopsis shoot tips

Author
item Volk, Gayle
item Henk, Adam
item CHHANDAK, BASU - University Of Northern Colorado

Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/19/2009
Publication Date: 9/1/2011
Citation: Volk, G.M., Henk, A.D., Chhandak, B. 2011. Gene expression in response to cryoprotectant and liquid nitrogen exposure in Arabidopsis shoot tips. Acta Horticulturae. 908: 55-66.

Interpretive Summary: Arabidopsis thaliana is an ideal model system to study plant cryopreservation at the molecular level. Arabidopsis shoot tips can be successfully cryopreserved when treated with cryoprotectant solutions that desiccate hydrated tissues. We extracted RNA from excised Arabidopsis shoot tips (Trt. A), treated with cryoprotectants (Trt. B), treated with cryoprotectants, liquid nitrogen, warmed, and grown for one day (Trt. C), and shoot tips treated with cryoprotectants and then grown for one day (Trt. D). We have made use of the fully sequenced Arabidopsis genome and readily available microarray slides with 29,000 genes to compare the gene expression patterns and we identified genes for which the transcript abundance was at least two-fold higher or lower in Trt. B, C, and D relative to Trt. A. Genes upregulated in Trt. C encode proteins involved in response to stimuli such as cold, heat, water deprivation, and oxidative stress. Similarly classified proteins were also upregulated in Trt. C and downregulated in Trt. D. Further investigation of the identified genes may reveal common genetic responses to cryoprotectants as well as suites of genes whose expression correlates with enhanced survival after cryoexposure.

Technical Abstract: Arabidopsis thaliana is an ideal model system to study plant cryopreservation at the molecular level. We have developed reliable cryopreservation methods for Arabidopsis shoot tips using Plant Vitrification Solution 2 and Plant Vitrification Solution 3 (PVS3) cryoprotectants. We have made use of the fully sequenced Arabidopsis genome and readily available microarray slides with 29,000 genes to compare the gene expression patterns among dissected control shoot tips, cryoprotectant-treated shoot tips, liquid nitrogen-treated shoot tips, and post-exposure recovering shoot tips. Genes upregulated during recovery after PVS3 and liquid nitrogen exposure encode proteins involved in response to stimuli such as cold, heat, water deprivation, and oxidative stress. Further investigation of the identified genes may reveal common genetic responses to cryoprotectants as well as suites of genes whose expression correlates with enhanced survival after cryoexposure.