Initial results from the screening of PROSOPE and Roscoff cultures Florence LE GALL, Héloïse...
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Transcript of Initial results from the screening of PROSOPE and Roscoff cultures Florence LE GALL, Héloïse...
Initial results from the screening of PROSOPE and Roscoff cultures
Florence LE GALL, Héloïse FELMAN, Frédéric PARTENSKY,Nathalie SIMON, Dominique MARIE, Daniel VAULOT
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)
Isolation and purification of microalgae
Flow cytometry sortingSerial dilutions
160 precultures (PROSOPE)
38 strains isolated
•24 eukaryotes
•14 Synechococcus
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
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
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
Morphological characterization
Optical microscopy Transmission electronic microscopy
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
Summary
Prymnesiophyceae : 11 strains
Prasinophyceae : 3 strains
Chlorophyceae : 1 strain
Bacillariophyceae : 1 strain