Presentation on theme: "A spatio-temporal comparison of picoeukaryotes in the Alborán Sea (SW Mediterranean) by denaturing gradient gel electrophoresis and HPLC pigment analysis."— Presentation transcript:
A spatio-temporal comparison of picoeukaryotes in the Alborán Sea (SW Mediterranean) by denaturing gradient gel electrophoresis and HPLC pigment analysis Beatriz Díez Mikel Latasa Carlos Pedrós-Alió Ramon Massana Institut de Ciències del Mar, CSIC Passeig Joan de Borbó, s/n, 08039 Barcelona, Catalunya, Spain.
Location of stations along the studied Alborán Sea area (SW Mediterranean Sea)
- Study of temporal (short and long-term) variability of picoeukaryotes in Alborán Sea - Study of spatial variability of picoeukaryotes in Alborán Sea - Identification of the dominant groups Objetives
The DGGE protocol follows the protocol of Díez et al. (2001) 0.75 mm-thick vertical gel containing 6% (w/v) polyacrylamide (37:5:1) acrylamide:bisacrylamide) Linear gradient of denaturing agents, increasing from 45% at the top to 65% at the bottom of the gel (100% denaturing agents is defined as 7M urea and 40% deoinized formamide) 600-800 ng PCR product Electrophoresis performed at 100v/cm 16h Stain: 1 x TAE buffer + Syber Gold for 45 min and visualized with a FluorS MultiImager and the MultiAnalyst Imaging software (Biorad Laboratories) EUK516 R EUK1F
The HPLC method follows the protocol of Zapata et al. (2000) adapted to our system
In general, % of Chl a <5µm was smaller at station A than at B and C
Picoeukaryotes are ubiquitous in this area Distribution of picoeukaryotes was dependent on the physical conditions. Both DGGE and HLPC data showed that differences in picoplankton composition were larger between years than among stations. Both techniques agreed in presenting chlorophyll b-containing algae and prymnesiophytes as the main components of the picoplankton during our cruises in the Alborán Sea. Conclusions