DIFFERENTIAL GENE EXPRESSION OCCURS DURING PROCESSING FOR AND RECOVERY FROM CRYOPRESERVATION IN ARABIDOPSIS THALIANA SHOOT TIPS

Authors: BASU, CHHANDAK - UNIV. OF NORTHERN CO; Volk, Gayle; BENNETT, AUDREY - UNIV. OF NORTHERN CO

Submitted to: American Society of Plant Biologists
Publication Type: Abstract Only
Publication Acceptance Date: 6/16/2006
Publication Date: 8/5/2006
Citation: Basu, C., Volk, G.M., Bennett, A.A. 2006. Differential gene expression occurs during processing for and recovery from cryopreservation in arabidopsis thaliana shoot tips. American Society of Plant Biologists. August 5-9, 2006, Boston, Massachusetts. Meeting Abstract.

Interpretive Summary: Cryopreservation is a specialized technique where plant materials are stored at liquid nitrogen temperatures (-196 C) for extended lengths of time. Cryopreservation is a way to preserve germplasm of agriculturally important crops. Most cellular and biochemical activities in plant cells are essentially stopped during cryostorage. Cryogenically stored plant tissues need to be dehydrated by cryoprotectants to prevent intracellular ice crystal formation. Understanding the molecular and genomic basis of plant responses to cryostress will open doors for further research into many aspects of plant abiotic stress physiology, especially dehydration stress. To understand differential gene expression patterns in Arabidopsis thaliana shoot tips, we treated excised shoot tips with cryoprotectants and then flash froze them in liquid nitrogen for RNA extraction. A second treatment included Arabidopsis shoot tips that were sampled 1 day after samples were warmed and had begun to recover. The third treatment included untreated Arabidopsis shoot tips that served as 'controls' in our experiments. Total RNA was extracted from all three treatments of Arabidopsis shoot tips and cDNA was synthesized using reverse transcriptase. Differential display experiments were performed using the Gene Fishing kit from Seegene, USA. Polymerase chain reaction (PCR) was performed with 10 sets of random primer combinations to amplify cDNAs from different treatments. We obtained 10 differentially expressed genes from our experiments. These genes will be sequenced and Real-Time Reverse Transcriptase PCR will be performed to confirm gene expression. The goal of the project is to acquire knowledge and understanding of roles of genes in plant abiotic stress response.

Technical Abstract: Cryopreservation is a specialized technique where plant materials are stored at liquid nitrogen temperatures (-196 C) for extended lengths of time. Cryopreservation is a way to preserve germplasm of agriculturally important crops. Most cellular and biochemical activities in plant cells are essentially stopped during cryostorage. Cryogenically stored plant tissues need to be dehydrated by cryoprotectants to prevent intracellular ice crystal formation. Understanding the molecular and genomic basis of plant responses to cryostress will open doors for further research into many aspects of plant abiotic stress physiology, especially dehydration stress. To understand differential gene expression patterns in Arabidopsis thaliana shoot tips, we treated excised shoot tips with cryoprotectants and then flash froze them in liquid nitrogen for RNA extraction. A second treatment included Arabidopsis shoot tips that were sampled 1 day after samples were warmed and had begun to recover. The third treatment included untreated Arabidopsis shoot tips that served as 'controls' in our experiments. Total RNA was extracted from all three treatments of Arabidopsis shoot tips and cDNA was synthesized using reverse transcriptase. Differential display experiments were performed using the Gene Fishing kit from Seegene, USA. Polymerase chain reaction (PCR) was performed with 10 sets of random primer combinations to amplify cDNAs from different treatments. We obtained 10 differentially expressed genes from our experiments. These genes will be sequenced and Real-Time Reverse Transcriptase PCR will be performed to confirm gene expression. The goal of the project is to acquire knowledge and understanding of roles of genes in plant abiotic stress response.