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Initial results from the screening of PROSOPE and Roscoff cultures
Florence LE GALL, Héloïse FELMAN, Frédéric PARTENSKY,Nathalie SIMON, Dominique MARIE, Daniel VAULOT
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The first steps of isolation
Surface water of ASTAN
2 filtrations on 3µm
Enumeration by flow cytometry
Culture inliquid mediumK/10, K/100, K/10+Si,F/20
Serial dilutionsK/10
Culture on plateK medium (0,6%agar)
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Isolation and purification of microalgae
Flow cytometry sortingSerial dilutions
160 precultures (PROSOPE)
38 strains isolated
•24 eukaryotes
•14 Synechococcus
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Synechococcus spectra
NOUM 97010
0
0.2
0.4
0.6
0.8
1
400 450 500 550
Longueur d'ondewavelength
PROSOPE 157-13
PUBPEB
NOUM 97012
0
0.2
0.4
0.6
0.8
1
400 450 500 550
Longueur d'ondewavelength
PROSOPE 53-4
PEB
PUB
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HPLC analysis and presence of caracteristic pigments
2 4 6 8 10 12 14 16 18
2 4 6 8 10 12 14 16 18
• PROSOPE culture 3
• PROSOPE culture 18
MgDVP Pras ViolaNeo
Diadchr
Chl b
Chl a
Neo Viola
Lut Chl b
Chl a
ß-car
MgDVP : Mg-2,4-divinyl pheoporphyrin a5Pras : PrasinoxanthinDiadchr : DiadinochromeNeo : NeoxanthinViola : ViolaxanthinChl b : Chlorophyll bChl a : Chlorophyll aLut : Luteinß-car : ß-carotene
Prasinophyceae
Chlorophyceae
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2 4 6 8 10 12 14 16 18
•PROSOPE culture 96
•ASTAN culture R 000412-4
Chl c2
Chl c3
19’HF
FucoDiadino
Chl a
2 4 6 8 10 12 14 16 18
Chl c3
Chl c2
Fuco
DiadinoDiato
Chl a
ß-car
19’HF :19’-HexanoyloxyfucoxanthinFuco : FucoxanthinDiadino : DiadinoxanthinChl c2 : Chlorophyll c2Chl c3 : Chlorophyll c3Chl a : Chlorophyll aDiato : Diatoxanthinß-car : ß-carotene
Prymnesiophyceae
Bacillariophyceae
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Morphological characterization
Optical microscopy Transmission electronic microscopy
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0.1 1.0 10.0
Scatter
0.1
1
10
100
1000C
hl f
luo
res
ce
nc
e
Ostreo. ocea
Bolid.medBolid.pacifi
Ostreo. taur
Micro pusi
Iso. Galb.E. huxleyi
Phaeocystis
PRO_3
PRO_22
PRO_5Peuk Astan
PRO_36PRO_63
Euk Astan (K)
PRO_114-2PRO_18-1
PRO_96
PRO_118-1
PRO_90
Euk Astan (F/2)
PRO_140-3
PRO_1
PRO_64
PRO_32
PRO_54
Prymnesiophyceae
Bolidophyceae
Prasinophyceae
Chlorophyceae
Prymnesiophyceae
Cultures from - Med Sea (PRO)- Roscoff
Diatom
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Summary
Prymnesiophyceae : 11 strains
Prasinophyceae : 3 strains
Chlorophyceae : 1 strain
Bacillariophyceae : 1 strain