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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