PICODIV

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 Eikrem unpublished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 andSynechococcus (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. A B C D 50 µm 50 µm 50 µm E 50 µm 10 µm Probes Euk 1209R CHLO01 NCHLO01 PELA01 BOLIDO01 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. 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

20. 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

21. Report • Wk 5: Sampling sites • Protocol detailed DV • Plans for year 2 (0.25 p) CP

22. 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