Experimental studies to understand and evaluate acclimation of marine plankton assemblages to increased CO2 and temperature.

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Intellectual Merit. Progressing ocean acidification and increasing sea surface temperature may significantly impact marine plankton community structure and community-level processes. Yet, our ability to predict specific responses is still limited because of the tremendous taxonomic complexity of microbial assemblages and the limitations of the methodological and experimental tools presently available to test specific hypotheses. Research to study community level effects due to a changing CO2/temperature regime often involve short-term field incubations that subject organisms to simulated 'greenhouse' conditions. A central question for understanding global climate change is whether the trends and patterns that are observed in communities during short-term manipulations can be extrapolated to the responses of fully acclimated plankton communities over decadal or longer timescales. The specific objectives of this research program are: 1) to examine how protistan communities restructure in response to increased seawater CO2 concentrations and temperature in semi-continuous field incubation experiments, and 2) to evaluate if the dominant algal species that are isolated from either ambient or increased CO2 and temperature treatments in field experiments will re-establish dominance under the same conditions in acclimated laboratory culture competition studies. Changes in community structure of natural protistan assemblages in our experimental treatments will be followed using image-based methods (flow cytometry, FlowCAM and microscopy) in combination with state-of the art molecular tools (DNA fingerprinting). Molecular approaches have begun to reveal an incredible high diversity for marine microbes and stimulate debate in regard to the ubiquitous presence of a microbial 'Rare Biosphere'; that is, the presence of a huge number of species that are present at extremely small percentages of the total abundance of microbes, among a much smaller percentage of dominant ones. Little is known about the ecological significance of these rare species, and the investigators hypothesize that change in CO2 and temperature will select for some of these members that are inconspicuous under ambient conditions. The unique aspect of this experimental approach is the combined use of field incubations that encompass entire natural microbial assemblages, with a series of laboratory culture competition trials that focus on the same groups of algae after extended acclimation, to evaluate the validity of short-term experiments that examine changing CO2 and temperature. First, field incubation experiments will be conducted to characterize changes in protistan community structure under ambient and future CO2/temperature regimes. Second, clonal algal strains will be isolated from dominant taxa in present day and greenhouse treatments, and cultivated for extended periods under their 'preferred' CO2/temperature conditions. Finally, mixtures of these acclimated strains will be competed against each other, to re-examine their responses to ambient and greenhouse conditions and compare them to the responses observed in the unacclimated field incubation experiments.Broader Impacts. Two graduate students will make this project the focus of their Ph.D. research at USC, and undergraduate students will be involved in the field and laboratory work. Results from this research will be incorporated in lesson plans on microbial diversity and global climate change. Dissemination of data and results is planned on a project website. The PIs in this project also participate in an on-going, innovative, NSF-funded program (Centers for Ocean Science Education Excellence; COSEE-West) which focuses on personal involvement of faculty in a custom framework to allow an effective connection with K-12 teachers, thus improving math and science education in disadvantaged parts of Southern California.