1. Christophe Six UMR UPMC-CNRS 7144 “Adaptation & Diversité en Milieu Marin” Equipe “Procaryotes Photosynthétiques Marins" Station Biologique de Roscoff Université Pierre et Marie Curie (Paris VI). Bureau 354, 3 ème étage (Batiment GT) six@sb-roscoff.fr UE Evolution of marine Phytoplankton & Biogeochemistry Research Worshop 2 : Functional diversity among Marine Synechococcus

2. Research workshop 2. Personal work : a short report in the form of a research article  Introduction, Methods, Results, Discussion  Mini-article due by January 29 th on pdf Format at six@sb-roscoff.fr. Methods : culturing phytoplankton, flow cytometry and biophysics of photosynthesis, phylogeny. Bringing out a scientific question from a scientific context. Setting up a experiment plan. Carring out the experiments. Synthetising and presenting the results. Interpreting the results. Communicating the results and their interpretation properly. A scientific approach. Results and interpretation : together

3. 0.1 0.5 10 µg/L Chlorophyll concentration in the World Ocean

4. 0.1 0.5 10 µg/L Latitudinal environmental gradients in the World Ocean ?

5. Latitudinal temperature gradient in the World Ocean Average temperature for year 2005

6. Latitudinal Light irradiance gradient in the World Ocean

7. Latitudinal environmental gradients in the World Ocean

8. 0.01 Chl µg/L) 1 60 UV Exposure Nutrients Enviro. stability Cell size Offshore-coast gradients Turbidity Light colour in water « Longitudinal » Environmental gradients In the World Ocean

9. Research Workshop 2 Functional diversity among marine Synechococcus Global scientific questions : How Synechococcus cells can adapt to such contrasted environmental conditions ? Did the evolution shape different cyanobacterial « ecotypes » ? More focused questions :.  How to start giving elements of answers to these questions ? What may influence the distribution of different pigment types ? What may inflence the distribution of the different phylogenetic clades ? What may inflence the pigment cell content ? What may inflence the photosynthetic activity ?

10. Research Workshop 2 Functional diversity among marine Synechococcus. To take advantage of the Roscoff Culture Collection >3000 phytoplankton strains from all aroud the world, including  350 Synechococcus

11. Research Workshop 2 Functional diversity among marine Synechococcus  Panel of  16 Synechococcus strains from the North Atlantic Ocean Post 1: Characterise the strains and their isolation sites and build a preliminary phylogeny. Gathering information on your strains : Strain denominations, coordinates, depth, ocean, sea, etc.. Build a preliminary phylogeny : Retrieve 16S sequences from NCBI and other databases, align and truncate the sequences on the BIOEDIT software. Then build a neighbor joining phylogeny with bootstrap values on the MEGA5 software  Use the RCC website to build a table on an Excel sheet  Install Google Earth (http://www.google.fr/earth/download/ge/agree.html)  2 sessions to acquire all the data (Monday-Tuesday)  Install Bioedit (http://www.mbio.ncsu.edu/bioedit/bioedit.html)http://www.mbio.ncsu.edu/bioedit/bioedit.html  Install MEGA5 (http://www.megasoftware.net/mega.php)  At least one laptop by student triplet  3-4 strains for 2-3 students (triplicate cultures)

12. Research Workshop 2 Functional diversity among marine Synechococcus Post 2: Determine the pigment type (photosynthetic antenna) of each strain by spectrofluorimetry. Determine phycobiliprotein composition. Calculate phycobilin fluorescence ratios. Deduce the pigment type  Justine Pittera (PhD student), spectrofluorimeter at the plankton lab

13. Research Workshop 2 Functional diversity among marine Synechococcus Post 3 : Determine the chlorophyll cell content for each strain (spectrophotometry) Determine the cell concentration and the strain cytometry parameters. Compare the chl a cell content with fluorescence parameters. Compare to cell size proxy  Cytometry analyses on Wednesday 16th

14. Research Workshop 2 Functional diversity among marine Synechococcus Post 4 :Study photosynthetic capacities by Pulse Amplitude Modulated Fluorimetry. Using the Junior PAM, draw a fluorescence light response curve. Determine photosynthetic parameters

15. Research Workshop 2 Functional diversity among marine Synechococcus  Synthetise all the data for all the strains : - Table with all information. TD sessions - Whole phylogenic tree with bootstrap values - All fluorescence spectra with pigment types determined  Try to understand whether some features seem to be associated to isolation sites Considering environmental gradients (latitude, longitude, depth, etc.) - Pigment contents and cytometric parameters - Light response curves  Compare the different strains