The main objectives are to analyse how the environment during sexual reproduction affects the climatic adaptation of Norway spruce , with respect to:

• Selection processes, acting on gametophytes during male and female meiosis (meiotic drive), pollen tube growth, megaspore degeneration, fertilisation and embryo competition
• Environmentally induced gene expression (genomic imprinting) during the reproductive process specifies the production of particular gene products (proteins, enzymes or regulatory molecules) shaping the expression of adaptive traits in the progenies

Specific objects are:

1. Crossing experiments with the same genotypes under different environments (climates) in a phytotron. Seedlings from these seeds will be tested to separate the impacts of photoperiod and temperature given during sexual reproduction of the mother clones on the progeny performance in adaptive traits

2. Characterisation of cold resistance and bud-set phenology differences between full-sib families from crossings under different environments to test the hypothesis that either photoperiod or temperature or a combination of the two perceived by mother clones during sexual reproduction affect the progeny performance in adaptive traits, measured by terminal bud-set and cold hardiness

3. A short term field trial to investigate how fast can Norway spruce adapt to different climatic conditions to test how fast landraces develop and to describe the effects of early selection for cold hardiness in the autumn on traits related to the synchronisation of growth and dormancy with seasonal changes in temperature

4. Characterisation of drought resistance differences between full-sib families from crossings under different environments to study to what extent adaptability to drought-stress conditions in Picea abies is influenced by the environment in which the seed is produced and matured

5. Investigations of physiological traits of metabolic pathways, which are involved in drought and frost tolerance to characterise stress levels by determining the water potentials, electrolyte leakage and total osmolyte concentrations, to identify progenies with high antioxidative capacities and responsiveness to drought stress, provide functional molecular and physiological markers and characterisation of antioxidative protection in progenies after freezing stress and supply functional markers

7. Investigations on the basis of highly variable DNA-markers (microsatellites). Analyses of seed samples (megagametophyt/embryo). Inheritance analyses of the investigated markers. To compare the potential of different types of molecular markers for evaluating adaptive properties of the progenies

8. Investigations on the basis of EST-markers. Full-sib families which were produced from the same set of parents in different crossing environments will be characterized by means of stress related EST markers in order to study the existence of genetic selection and „after effects“

9. Environmental effects on metylation of DNA and transcription and translation of phytochrome genes, cycline and other identified genes. To determine the general methylation pattern of DNA in vegetative buds, the levels of mRNA encoding different phytochromes during the process of cold acclimation and the localisation pattern and pattern of accumulation in vegetative buds of the two known phytochromes

10. Differential display of newly identified expressed genes. Identification of common genes involved in general stress response, of differential expressed mRNAs deriving from identical genotypes exposed to different stress treatments and of newly synthesised proteins involved in stress response

11. Development of practical recommendations for seed producers, seed centers and tree nurseries. Development of practical recommendations for tree breeding, seed transfer, silviculture and gene resource conservation based on the project results. Establishment of a web site which provides the project results

The results of this proposal will significantly contribute to our understanding of the naturally occurring adaptation processes in Norway spruce (and other tree species) and will have far-reaching consequences for all practical aspects of tree breeding, seed transfer, silviculture and gene resource conservation within the Community. Furthermore the results of this project might have a bearing on the Council Directive on the marketing of forest reproductive Material.