Aquatic Sciences Meeting, Albuquerque 2001

SS14 Microbial Diversity (Disciplinary Connections)

Date: Monday, February 12, 2001, Time: 2:45:00 PM

Location: Brazos

Countway, P, D, University of Southern California, Los Angeles, USA, countway@usc.edu

Caron, D, A, University of Southern California, Los Angeles, USA, dcaron@usc.edu

Gast, R, J, Woods Hole Oceanographic Institution, Woods Hole, USA, rgast@whoi.edu

Dennett, M, R, Woods Hole Oceanographic Institution, Woods Hole, USA, mdennett@whoi.edu

TERMINAL RESTRICTION FRAGMENT LENGTH POLYMORPHISM (TRFLP) ANALYSIS OF PROTISTAN ASSEMBLAGES OF THE NORTH ATLANTIC

TRFLP analysis was conducted on 18S rDNA, extracted and amplified from naturally occurring (<200 um) marine eukaryote assemblages with a 6-FAM labeled forward primer. Products of appropriate length were digested with restriction endonucleases recognizing 4-bp sequences. Enzymes used for the digestion of DNA were chosen based on 'theoretical' fragment patterns predicted for a wide variety of protistan taxa using publicly available 18S rDNA sequence data (NCBI), and recognition sites using MacVector 7.0. In addition, analyses of TRFLP patterns for a large protistan culture collection were conducted to provide real fragment patterns with which to compare and interpret environmental samples. Patterns of protistan diversity were examined throughout the water-column at sites in the Western North Atlantic (Sargasso Sea, Gulf Stream, Shelf water) during August 2000. Samples were collected at water-column features including the surface mixed-layer, deep chlorophyll maximum, base of the euphotic zone, and >500m at deep sites. The resulting TRFLP patterns provide a novel, culture-independent approach for examining and dissecting protistan community structure that can be compared directly with information from sequence databases and existing culture collections.

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Aquatic Sciences Meeting, Albuquerque 2001

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