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Assessement of picoeukaryote diversity of the Roscoff Culture Collection (RCC)

Assessement of picoeukaryote diversity of the Roscoff Culture Collection (RCC). Khadidja Romari, Florence Le Gall & Daniel Vaulot Station Biologique de Roscoff. Marine picoeukaryote species known to-date. Year Name Size (µm)

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Assessement of picoeukaryote diversity of the Roscoff Culture Collection (RCC)

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  1. Assessement of picoeukaryote diversity of the Roscoff Culture Collection (RCC) Khadidja Romari, Florence Le Gall & Daniel Vaulot Station Biologique de Roscoff

  2. Marine picoeukaryote species known to-date • Year Name Size (µm) • 1952 Chromulina pusilla Butcher (renamed Micromonas) 1-1.5 • Nannochloris atomus, N. maculata (renamed Nannochloropsis) 1.5-4 • 1955 Nannochloris oculata (renamed Nannochloropsis)1.5-4 • 1957 Monallantus salina (renamed Nannochloropsis)1.5-4 • 1967 Hillea marina 2-2.5 • 1969 Pedinomonas mikron (renamed Resultor) 1.5-2.5 • Scourfieldia marina (renamed Pseudoscourfieldia) 2-3 • 1974 Imantonia rotunda 2-4 • 1977 Pelagococcus subviridis 2.5-5.5 • 1978 Chlorella nana 1.8-2.6 • 1982 Nannochloropsis gaditana Lubian2.5-5 • Nanochlorum eucaryotum (renamed Nannochloris) • 1987 Triparma spp, Tetraparma pelagica 2.2-4.7 • 1988 Aureococcus anophagefferens 2-4 • 1990 Bathycoccus prasinos 1.5-2.5 • Pycnococcus provasolii 1- 4 • 1993 Pelagomonas calceolata 1.3-3 • 1995 Ostreococcus tauri 0.8-1.1 • 1996 Prasinoderma coloniale 2.5-5.5 • Nannochloropsis granulata 2- 4 • 1997 Aureoumbra lagunensis 2.5-5 • Bolidomonas pacifica and mediterranea 1.5 • Picophagusflagellatus 1.5 • 2000 Symbiomonasscintillans 1.5 • 2000 Ostreococcus oceanica1.2

  3. Morphology SSU rRNA gene Pigment signature Strategy

  4. Metodology DNA extraction : CTAB method 328f 18S rDNA HpaII & HaeIII           329r Clonning 18S rDNA Sequence Identification by RFLP Program 

  5. M’ M’ M’ M’ RCC 365 RCC 305 RCC 366 RCC 305.1 RCC 305.2 RCC 305.1 RCC 305.2 RCC 82 RCC 82.1 RCC 82.2 RCC 305 RCC 82.1 RCC 82.2 RCC 366 RCC 366 RCC 305 RCC 82 RCC 82 M M 600bp 2000bp A HpaII B HaeIII Fig.1: Agarose gel electrophoresis showing A : 18S rDNA PCR product of culture (RCC). B: restriction profiles of 18S rDNA fragment (culture and clones) obtained using the enzyme HpaII and HaeIII. M: Smart Ladder, M’: molecular mass marker 100bp

  6. 0.1 Fig.2: Parcimony ML dendogram showing the position of the two clones of RCC 82

  7. M’ M’ M’ M’ RCC 365 RCC 305 RCC 366 RCC 305.1 RCC 305.2 RCC 305.1 RCC 305.2 RCC 82 RCC 82.1 RCC 82.2 RCC 305 RCC 82.1 RCC 82.2 RCC 366 RCC 366 RCC 305 RCC 82 RCC 82 M M 600bp 2000bp A HpaII B HaeIII Fig.1: Agarose gel electrophoresis showing A : 18S rDNA PCR product of culture (RCC). B: restriction profiles of 18S rDNA fragment (culture and clones) obtained using the enzyme HpaII and HaeIII. M: Smart Ladder, M’: molecular mass marker 100bp

  8. 0.10 Fig. 3 : Phylogenetic position of the RCC 366. Part of tree extracted from ARB Database (Euk algae 98)

  9. M’ M’ M’ M’ RCC 365 RCC 305 RCC 366 RCC 305.1 RCC 305.2 RCC 305.1 RCC 305.2 RCC 82 RCC 82.1 RCC 82.2 RCC 305 RCC 82.1 RCC 82.2 RCC 366 RCC 366 RCC 305 RCC 82 RCC 82 M M 600bp 2000bp A HpaII B HaeIII Fig.1: Agarose gel electrophoresis showing A : 18S rDNA PCR product of culture (RCC). B: restriction profiles of 18S rDNA fragment (culture and clones) obtained using the enzyme HpaII and HaeIII. M: Smart Ladder, M’: molecular mass marker 100bp

  10. Conclusion • Problem of double sequences : increase [ ] of the antibiotic • Competitive amplification : Two or three PCR product mixed • Roscoff Culture Collection hide a very interesting species

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