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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 arabidopsis shoot tips after liquid nitrogen exposure

Authors
item VOLK, GAYLE
item HENK, ADAM
item BONNART, REMI
item Gross, Briana -

Submitted to: International Society for Horticultural Science Meeting
Publication Type: Abstract Only
Publication Acceptance Date: August 13, 2013
Publication Date: August 13, 2013
Citation: Volk, G.M., Henk, A.D., Bonnart, R.M., Gross, B. 2013. Gene expression in arabidopsis shoot tips after liquid nitrogen exposure. Meeting Abstract. 2nd International Society for Horticultural Science Meeting. p.17.

Technical Abstract: Arabidopsis thaliana shoot tips can be successfully cryopreserved using either Plant Vitrification Solution 2 (PVS2) or Plant Vitrification Solution 3 (PVS3) as the cryoprotectant. We used this model system to identify suites of genes that were either upregulated or downregulated as shoot tips recovered from cryoprotective treatments. Treatments included: 0.3 M sucrose overnight (SUC), 0.3 M sucrose + 16 h recovery (SUC-LN), PVS2 + 16 h recovery (PVS2-LN), PVS2 + LN + 16 h recovery (PVS2+LN), PVS3 + 16 h recovery (PVS3-LN) and PVS3 + LN + 16 h recovery (PVS3+LN). RNA was extracted and expression patterns were compared using Affymetrix Arabidopsis ATH1 Genechips. In total, 180 genes showed significant 2-fold changes in expression level after treatment with either the cryoprotectant or cryopreservation followed by recovery. Of these 180 genes, 67 were related to stress, defense, wounding, lipid, carbohydrate, abscisic acid, oxidation, temperature (cold/heat) or osmoregulation. Five genes (At4g33030, At1g66180, At3g24500, At4g012500, and At1g75040) were selected for use in qPCR to confirm expression results that were observed in the microarray experiments. These genes are believed to be expressed in response to various abiotic and biotic stresses. These experiments have allowed us to better understand which sets of genes are expressed as shoot tips begin the process of recovering after cryoprotectant and liquid nitrogen exposure.

Last Modified: 8/19/2014
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