title: Changes in transcript expression patterns as a result of cryoprotectant treatment and liquid nitrogen exposure in Arabidopsis shoot tips.
abstract: Transcripts related to abiotic stress, oxidation, and wounding were differentially expressed in Arabidopsis shoot tips in response to cryoprotectant and liquid nitrogen treatment. Cryopreservation methods have been implemented in genebanks as a strategy to back-up plant genetic resource collections that are vegetatively propagated. Cryopreservation is frequently performed using vitrification methods, whereby shoot tips are treated with cryoprotectant solutions, such as Plant Vitrification Solution 2 (PVS2) or Plant Vitrification Solution 3 (PVS3); these solutions remove and/or replace freezable water within the meristem cells. We used the model system Arabidopsis thaliana to identify suites of transcripts that are up- or downregulated in response to PVS2 and PVS3 treatment and liquid nitrogen (LN) exposure. Our results suggest that there are many changes in transcript expression in shoot tips as a result of cryoprotection and that these changes exceed the number detected as a result of LN exposure. In total, 180 transcripts showed significant changes in expression level unique to treatment with either the cryoprotectant or cryopreservation followed by recovery. Of these 180 transcripts, 67 were related to stress, defense, wounding, lipid, carbohydrate, abscisic acid, oxidation, temperature (cold/heat), or osmoregulation. The responses of five transcripts were confirmed using qPCR methods. The transcripts responding to PVS2 + LN suggest an oxidative response to this treatment, whereas the PVS3 + LN treatment invoked a more general metabolic response. This work shows that the choice of cryoprotectant can have a major influence on the patterns of transcript expression, presumably due to the level and extent of stress experienced by the shoot tip. As a result, there may be divergent responses of study systems to PVS2 and PVS3 treatments.
authors.affiliation: University of Minnesota Duluth, 207 Swenson Science Building, 1035 Kirby Drive, Duluth, MN, 55812, USA.
authors.forename: Briana L
authors.initials: BL
authors.lastname: Gross
authors.affiliation: USDA-ARS National Laboratory for Genetic Resources Preservation, 1111 S. Mason St., Fort Collins, CO, 80521, USA.
authors.forename: Adam D
authors.initials: AD
authors.lastname: Henk
authors.affiliation: USDA-ARS National Laboratory for Genetic Resources Preservation, 1111 S. Mason St., Fort Collins, CO, 80521, USA.
authors.forename: Remi
authors.initials: R
authors.lastname: Bonnart
authors.affiliation: USDA-ARS National Laboratory for Genetic Resources Preservation, 1111 S. Mason St., Fort Collins, CO, 80521, USA. Gayle.Volk@ars.usda.gov.
authors.forename: Gayle M
authors.initials: GM
authors.lastname: Volk
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D063990:Gene Ontology; D018519:Meristem / Q000187:drug effects / Q000235:genetics*; D009584:Nitrogen / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D003451:Cryoprotective Agents; D012333:RNA, Messenger; D009584:Nitrogen
keywords: Cryopreservation; Cryoprotection; Liquid nitrogen; Microarray; Transcript expression; Vitrification