Work package 2 (WP2)

Topic: Cyanobacterial Dormant Forms in an Aquatic environment

Objective: To reveal conditions and processes involved in germination

Task Deliverable # month

1. Determine environmental variables/stimuli responsible for the arousal of akinetes from the dormant stage

D10 12 (M12)

2. Metabolic pathways associated with the

germination of akinetes

D10 30 (M30)

3. Identification of compounds and metabolites involved in germination

D13 30 (M30)

4. Gene expression during akinete germination

D11 35 (M35)

5. Quantitative analyses of the expression of specific genes by QPCR

D12 35 (M35)

6. Summary and Report D14 36 (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), CO2 uptake, O2 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