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United States Department of Agriculture

Agricultural Research Service

Research Project: RESEARCH TO DEVELOP STRATEGIES AND TECHNOLOGIES FOR PRESERVING PLANT GENETIC DIVERSITY IN EX SITU GENEBANKS

Location: Plant Germplasm Preservation Research Unit

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

Authors
item Volk, Gayle
item Henk, Adam
item Chhandak, Basu - UNIVERSITY OF NO. CO

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: November 14, 2008
Publication Date: April 13, 2009
Citation: Volk, G.M., Henk, A.D., Chhandak, B. Gene expression in response to cryoprotectant and liquid nitrogen exposure in Arabidopsis shoot tips. I International Symposium on Cryopreservation in Horticultural Species, April 5-8, 2009. Leuven, Belgium. pp. 24. Meeting abstract.

Interpretive Summary: Arabidopsis thaliana serves as an ideal model system to study cryopreservation at the molecular level. We have developed reliable cryopreservation methods for Arabidopsis shoot tips using Plant Vitrification Solution 2, Plant Vitrification Solution 3 and polyethylene glycol-glucose-dimethylsulfoxide cryoprotectants. We have made use of the fully sequenced Arabidopsis genome and readily available microarray slides with 26,000 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. We have identified key gene families that show changes in gene expression in fully replicated experiments and have confirmed gene expression patterns using real-time PCR. Key gene families encode proteins involved in lipid transport, dehydration responses, seed maturation, and detoxification. Some genes are specifically upregulated in response to cryoprotectant treatment while others have a more general response expressed during shoot tip recovery. Our results reveal that cryoprotectant treatments induce gene expression and critical pathways may include those involved in lipid transport and osmoregulation.

Technical Abstract: Arabidopsis thaliana serves as an ideal model system to study cryopreservation at the molecular level. We have developed reliable cryopreservation methods for Arabidopsis shoot tips using Plant Vitrification Solution 2, Plant Vitrification Solution 3 and polyethylene glycol-glucose-dimethylsulfoxide cryoprotectants. We have made use of the fully sequenced Arabidopsis genome and readily available microarray slides with 26,000 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. We have identified key gene families that show changes in gene expression in fully replicated experiments and have confirmed gene expression patterns using real-time PCR. Key gene families encode proteins involved in lipid transport, dehydration responses, seed maturation, and detoxification. Some genes are specifically upregulated in response to cryoprotectant treatment while others have a more general response expressed during shoot tip recovery. Our results reveal that cryoprotectant treatments induce gene expression and critical pathways may include those involved in lipid transport and osmoregulation.

Last Modified: 4/23/2014
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