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 1.5. Investigations of physiological traits of metabolic pathways, which are involved in drought and frost tolerance
	Start date: Start date: 1st month		Partners responsible: Text
	Completion date: 36th month		Other partners involved
	Objectives
	Grow spruce seedlings under semi-controlled conditions and perform drought stress experiments, [optional: supply stressed materials to other partners] Characterise stress levels by determining the water potentials, electrolyte leakage and total osmolyte concentrations [optional: Prolin] Identify progenies with high antioxidative capacities and responsiveness to drought stress, provide functional molecular and physiological markers Characterise antioxidative protection in progenies after freezing stress (supplied by other partners) and supply functional markers
	Description on the work
	Seedlings (or seeds) obtained from partners will be preconditioned by growth under semi-controlled greenhouse conditions. The seedlings will then be exposed gradually to drought stress by withholding water for several weeks. The development of drought stress will be controlled by the determination of soil water content, by chlorophyll fluorescence measurements and by the pre-dawn water potential of the seedlings. Additionally the osmolyte concentration and electrolyte leakage of roots will be determined regularly on aliquots. Above and below-ground biomass will be determined (Optional: Seedlings in defined stages of stress will be supplied to partners for further analyses). Tolerant and sensitive seedlings harvested in well-defined stress conditions will be used for antioxidants analyses. Based on previous experience, SOD and GR will be analysed as the most likely candidates. mRNA will be isolated after established protocols and used for Northern blots with heterologous cDNAs for cytosolic and chloroplastic SOD. SOD-protein will be quantified by Western blot analysis. If these studies do not give conclusive results, other antioxidative systems will also be screened using HPLC and spectrophotometric methods well-established in our laboratory. Frozen plant materials, collected under well-defined conditions, obtained from other partners, will be used for the screening for physiological traits as described above
	Deliverables
	Supply of stressed material to other partners 0-12 Report about stress sensitivity within and between full-sib families 9-24 Report about the role of antioxidants as functional, physiological and molecular markers 18-36
	Milestones
	1° year Grow spruce seedlings and drought stress experiments 2° year Characterisation of drought stress sensitivity in different progenies 3° year Characterisation of antioxidants as functional, physiological and molecular markers
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