1. PICODIV Aims: Establish diversity of picoplankton Measure abundance of key picoplanktonic taxa with molecular methods

2. Workpackage 1 - Cultures Achievements Novel cultures established from Med Sea (PROSOPE), Roscoff and Blanes sites Pigement, ultrastructural and molecular data accumulated

3. Novel cultures Dictyochophyceae Bolidophyceae ? Eikrem unpublished

4. Workpackage 1 - Cultures Year 2 challenges and key directions Culture establishment is much slower than anticipated. It is difficult to get rid of heterotrophic contaminants (but...). Focus on successful strategies We need more coastal cultures (in particular from Helgoland) Describe formerly novel taxa

5. Workpackage 2 - Clone libraries Achievements Eukaryotic clone libraries for all three sites DGGE analysis (Blanes)

6. Autotrophs: 24%

7. Autotrophs: 48 %

8. Mamiellales

9. Workpackage 2 - Clone libraries Year 2 challenges and key directions Obtain cyanobacteria clone libraries Should we do more clone libraries and which ones? Synthesize and publish results already obtained from partial sequences Select subset of clones for full sequencing

10. Workpackage 3 - Probe design Achievements Probes designed and under testing –Prochlorococcus and Synechococcus (U of Warwick) –Prasinophytes (Roscoff) –Cryptophytes (AWI) –Stramenopiles (Barcelona)

11. Workpackage 3 - Probe design Year 2 challenges and key directions Design probes for some of the uncultivated groups (Alveolates) Select minimum set of probes for annual monitoring

12. Workpackage 4 - Probe measurement Achievements FISH-TSA with microscopy operational FISH-TSA with flow cytometry promising

13. Probes 50 µm ABC E 10 µm D Euk 1209RCHLO01NCHLO01 BOLIDO01 PELA01 Not et al. submitted

14. Probes West et al. submitted

15. Workpackage 4 - Probe measurement Year 2 challenges and key directions Start quantitative PCR DNA arrays ???

16. Workpackage 5 - Monitoring Achievements Protocol finalized Monthly sampling started at all sites

17. Workpackage 5 - Monitoring Year 2 challenges and key directions Verify sample quality ( TEM, FISH, pigments ) Set up databases

18. To be done during this meeting Wk 1: Select cultures for focus Wk 2: Define strategy for clone libraries Wk 2: Select clones for full sequencing Wk 3: Select probes to be developed and used Wk 4: Define strategy for DNA chips Update list of deliverables with partner responsability Define structure of year 1 report Discuss strategy for publication of papers

19. Publications Papers –2 published –5 submitted Presentations at meetings –12

20. Report Wk 1: Cultures –Status of RCC (0.5 p)DV –Prokaryotes (1 p)DS –Eukaryotes TEM (2 p)WE Pigments (1 p)ML Sequences (0.5 p)DV –Plans for year 2 (0.25 p)DV Wk 2: Clone librairies –Synechococcus (1 p)DS –Roscoff (1 p)KR –Helgoland (1 p)KV –Blanes (1 p)RM –Plans for year 2 (0.25 p)DS

21. Report Wk 3: Probes –Overview (1 p)LM –Syn/Pro (1 p)DS –Pras (1 p) FN –Crypto (0.5 p) LM –Prym (0.5 p) LM –Stram (1 p) LM –Plans for year 2 (0.25 p)LM Wk 4: Probe technology –Overview (1 p)LM –FISH-TSA (1 p)FN –FISH-TSA cyto (1 p)IB –Quantitative PCR (0.5 p)IB –DNA chips (0.5 p)LM –Plans for year 2 (0.25 p)LM

22. Report Wk 5: Sampling sites –Protocol detailedDV –Plans for year 2 (0.25 p)CP

23. All contributions synthetic (stick to pages max) Use Word styles; no tab; Times roman 10; single space All figures are “special” pasted as image (not object). One fig max per contribution. Files should be as small as possible (< 1 Mo, if necessary lower resolution...) Sent to Roscoff by May 17 at most