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