picodiv aims: establish diversity of picoplankton measure abundance of key picoplanktonic taxa with...
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PICODIVPICODIV
Aims:• Establish diversity of picoplankton
• Measure abundance of key picoplanktonic
taxa with molecular methods
Workpackage 1 - Cultures
AchievementsAchievements
• Novel cultures established from Med Sea
(PROSOPE), Roscoff and Blanes sites
• Pigement, ultrastructural and molecular data
accumulated
Novel cultures
Dictyochophyceae
Bolidophyceae ?
Eikrem unpublished
Workpackage 1 - Cultures
Year 2 challenges and key directionsYear 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
Workpackage 2 - Clone libraries
AchievementsAchievements
• Eukaryotic clone libraries for all three sites
• DGGE analysis (Blanes)
Coastal: Helgoland March 2000
Group I OLI11001Alveolata
He0003xx.26
Alveolata
Rhizopoda
Group II AmoebophryaAlveolata
CryptophyceaeCryptophyta
Stramenopiles
PrasinophyceaeChlorophyta
DinophyceaeAlveolata
CiliophoraAlveolata
Autotrophs: 24%Autotrophs: 24%
Coastal: Roscoff April 2000
CiliophoraAlveolata
DinophyceaeAlveolata
MamiellalesPrasinophyceae
ChlorophytaChoanoflagellida
ThraustochytriidaeLabyrinthulidaStramenopiles
SlopalinidaStramenopiles
CryptophyceaeCryptophyta
PrymnesiophyceaeHaptophyta
Stramenopiles
DictyochophyceaeStramenopiles
Rhizopoda
RA000412.151
AmoebophryaDinophyceae
Alveolata
Autotrophs: 48 %Autotrophs: 48 %
Mamiellales
Workpackage 2 - Clone libraries
Year 2 challenges and key directionsYear 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
Workpackage 3 - Probe design
AchievementsAchievements
• Probes designed and under testing
– Prochlorococcus and
Synechococcus (U of Warwick)
– Prasinophytes (Roscoff)
– Cryptophytes (AWI)
– Stramenopiles (Barcelona)
Workpackage 3 - Probe design
Year 2 challenges and key directionsYear 2 challenges and key directions
• Design probes for some of the
uncultivated groups (Alveolates)
• Select minimum set of probes for annual
monitoring
Workpackage 4 - Probe measurement
AchievementsAchievements
• FISH-TSA with microscopy operational
• FISH-TSA with flow cytometry promising
Probes
50 µm50 µm
50 µm
A B C
50 µm
E
10 µm
D
Euk 1209R CHLO01 NCHLO01
BOLIDO01 PELA01
Not et al. submittedNot et al. submitted
Probes
West et al. submittedWest et al. submitted
Workpackage 4 - Probe measurement
Year 2 challenges and key directionsYear 2 challenges and key directions
• Start quantitative PCR
• DNA arrays ???
Workpackage 5 - Monitoring
AchievementsAchievements
• Protocol finalized
• Monthly sampling started at all sites
Workpackage 5 - Monitoring
Year 2 challenges and key directionsYear 2 challenges and key directions
• Verify sample quality (TEM, FISH, pigments)
• Set up databases
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
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
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
Report• Wk 5: Sampling sites
– Protocol detailed DV– Plans for year 2 (0.25 p) CP
• 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