1. Work package 1 (WP1) Topic: Cyanobacterial Dormant Forms in an Aquatic environment Objective: Establishment of dormant stage/ Induction of akinetes in cyanobacteria TaskDeliverable #month 1. Determine conditions for the induction of akinetes D49 (M9) 2. Physiological processes involved in the induction of the dormant stage D830 (M30) 3. Compounds and metabolites associated with the dormant stage D830 (M30) 4. Development of molecular tools: DNA libraries. Gene expression during akinete formation (avaK, hetR) D5 D6 18 (M18) 27 (M27) 5. Quantitative analyses of the expression of specific genes by QPCR D735 (M35) 6. Summary and Report(M36)

2. # D4 # D8 Basic studies to reveal conditions and mechanisms involved in induction of akinetes formation, dormancy and desiccation: Nutrient Depletion (P), Light, Temp, O 2 (hypoxia), K Storage of metabolites [carbohydrates, cyanophycin (N)] – enzymes involved, e.g cyanophycine synthase Mature akinetes Respiration and Photosynthetic apparatus (is there pigments production?) O 2 evolution/consumption To collect adequate material for further genomic, proteomic and metabolomic studies.

3. 1.Akinete formation K > HL > P 2. Storage metabolites in akinetes Cyanophycin the storage protein found in cyanobacteria is found in akinetes of A. ovalisporum and is enriched in aspartic and arginine amino acids. 3. Photosynthesis apparatus Young akinetes perform photosynthetic capabilities as found for vegetative cells grown in akinete-induced media. Mature akinetes maintain residual photosynthetic activity even though some components of the photosynthetic apparatus appear to remain intact.

4. # D5 4. Molecular approach Two genomic libraries were cloned into pCC1Fos. The number of clones per culture: 13,056 fosmids 15,360 fosmids A total of 3481 sequencing reads were made from both library fosmids, using primers T7 and M13(-28) Comparison between Anabaena genome and Aphanizomenon contigs led to the selection of nine fosmids that may contain candidate genes loci.

5. GeneFunctionFosmid AvaKAkinete markersbfos01-3p12, sbfos02-6e7 HetRprotease with DNA binding activitysbfos02-9m21,sbfos02-7e10 DevRtwo-component system, regulatory proteinsbfos01-3d6, sbfos02-8h10 HepAABC transportersbfos01-9d6 CphACyanophycin synthetasesbfos01-4l16 CphBCyanophycinasesbfos01-4l16 Kdp operonK+ transporting ATPasesbfos01-3f13