ADAPTABILITY Project Description
Workpackage 1 Workpackage 1.1 Workpackage 1.2 Workpackage 1.3 Workpackage 1.4 Workpackage 1.5 Workpackage 2 Workpackage 2.1 Workpackage 2.2 Workpackage 2.3 Workpackage 3 Workpackage 3.1 Workpackage 3.2 Workpackage 4 Workpackage 4.1	Workpackage 3.2. Differential display of newly identified genes
	Start date: 1st month		Partners responsible: UNI GOE
	Completion date: 36th month		Other partners involved: TUM
	Objectives
	Identification of common genes involved in general stress response Identification of differential expressed mRNAs deriving from identical genotypes exposed to different stress treatments Identification of newly synthesised proteins involved in stress response
	Description on the work
	mRNA will be isolated from the seedlings of different full-sib families produced in different crossing environments and grown under different conditions (control, drought and freezing stress). In plants exit some general mechanisms in stress response, therefore these mRNAs will be first characterised by Dot-Blot-hybridisation with 140 newly identified cDNA clones from an elicitor induced cDNA library from spruce (provided by P2). This will allow the identification of genes which are generally expressed in stress situations. Using RT-PCR the mRNAs of the seedlings will be reverse transcribed in cDNA strands which will be fingerprinted with arbitrary primers. This DDRT-PCR allows the amplification of differentially expressed mRNA deriving from identical genotypes exposed to drought and freezing stress in order to identify and characterise the specific genes involved in each of these stress situations. Known that stress situations caused the increase of typical sets of proteins, these mRNAs will be translated in vitro using a rabbit reticulosyte lysate to give a survey of newly synthesised proteins according to drought and freezing stress. Additionally the reaction mixture will be used for immunoprecipitation with antibodies of candidate proteins (see WP 1.5.) like SOD
	Deliverables
	Manuscript on differentially expressed mRNA
	Milestones
	1° year: Characterisation of generally expressed genes in stress response 3° year: Characterisation of differentially expressed mRNA according to drought and freezing stress
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