Work package 2 (WP2) Topic: Cyanobacterial Dormant Forms in an Aquatic environment Objective: To reveal conditions and processes involved in germination TaskDeliverable #month 1. Determine environmental variables/stimuli responsible for the arousal of akinetes from the dormant stage D1012 (M12) 2. Metabolic pathways associated with the germination of akinetes D1030 (M30) 3. Identification of compounds and metabolites involved in germination D1330 (M30) 4. Gene expression during akinete germination D1135 (M35) 5. Quantitative analyses of the expression of specific genes by QPCR D1235 (M35) 6. Summary and ReportD1436 (M36)

# D10 and # D13 Studies on conditions and mechanisms involved in akinetes germination: Environmental stimuli: Light (intensity, duration, spectrum), Temperature, Oxygen Nutrients: Phosphorus, Nitrogen [inhibitory effects (ammonium), depletion- heterocysts formation) Phosphate uptake: Is there an increase in alkaline phosphatase activity? Nitrogen metabolism: enzymes involved - nitrate reductase, glutamine synthetase Protein synthesis – enzymes induced (protein kinase) Metabolic pathways: Resumption of photosynthetic apparatus (synthesis of pigments), CO 2 uptake, O 2 evolution/consumption (respiration), energetic budget (ATP) Degradation of metabolites: [carbohydrates, cyanophycin (N)] – enzymes involved, e.g cyanophycinase DNA, RNA synthesis

# D11, # D12 Identification of genes associated with germination in cyanobacteria Genes that are expressed Gene expression/repression studies – regulated up/down during akinete germination Microarrays using samples collected from germinating akinetes and vegetative cells Proteome profiling to identify specific proteins associated with germinating akinetes