Development of High Throughput Analysis of Plankton Communities

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Over the past decade, research employing molecular approaches and other new technologies has permitted increased resolution of plankton diversity and led to the discovery of important new taxa throughout the world's oceans. Progress in understanding the significance of this newfound diversity has been hampered by a lack of high throughput technologies for the simultaneous identification and quantification of the many taxa comprising such communities. Bead arrays offer a promising approach to the problem of high throughput analysis of multiple taxa. The goal of this project is to test the applicability of bead array technology to the quantitative analysis of diverse marine microbial communities. As applied here, a bead array consists of a set of taxon-specific oligonucleotides attached to 5 micron beads. Up to 100 different oligonucleotide probes, each attached to beads of a different color, are used to quantitatively probe DNA extracted from planktonic organisms filtered from bulk seawater. Probes with different levels of resolution can be mixed in a single multiplex assay. Two test systems for the development of this technology will be employed. An array will be developed to assay seasonal changes in picophytoplankton diversity in a coastal environment. A second array will be developed to distinguish among the 25 species of the diatom genus Pseudo-nitzschia, ten of which are known to produce the neurotoxin domoic acid. Multiplex bead arrays will provide a new methodological tool for the analysis of marine microbial communities. The planned investigations also will provide excellent training opportunities for undergraduate, graduate and postdoctoral researchers seeking to apply new molecular technologies to important questions in marine ecology.