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.
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A spatio-temporal comparison of picoeukaryotes in the Alborán Sea (SW Mediterranean) by denaturing gradient gel electrophoresis and HPLC pigment analysis.
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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-AlióRamon Massana
Institut de Ciències del Mar, CSIC
Passeig Joan de Borbó, s/n, 08039 Barcelona, Catalunya, Spain.
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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
ABC
November 1997
May 1998
September 1999
Table 3. During the MTP-III-MATER/HESP/97, MTP-II-MATER/HESP/04-98 andMTP-I-MATER/HESP/99 cruises, on board the R/V Hesperides, stations (A, B, C, in acoastal-offshore gradient) located across the Western Alborán Sea Gyre (SW
Mediterranean) were sampled in November 1997, May 1998 and September 1999(fraction between 0,2-5 µm in size).
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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
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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
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 andprymnesiophytes as the main components of the picoplankton during our cruises in the Alborán Sea.