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Qualitative Determination of Exopolymeric Substances in Particle-Associated Bacteria from the Sargasso Sea

Description

The biological carbon pump acts as part of the global carbon cycle through the photosynthetic fixation of inorganic carbon into dissolved and particulate organic carbon by phytoplankton. Previously, the biological carbon pump was attributed to large aggregates and zooplankton fecal pellets since their size and density results in faster sinking…

The biological carbon pump acts as part of the global carbon cycle through the photosynthetic fixation of inorganic carbon into dissolved and particulate organic carbon by phytoplankton. Previously, the biological carbon pump was attributed to large aggregates and zooplankton fecal pellets since their size and density results in faster sinking rates, efficiently exporting organic carbon to deeper depths in the ocean. However, recent studies have indicated that small cells, known as picoplankton, contribute significantly to the formation of sinking particles. The presence of exopolymeric substances (EPS), among them sticky transparent exopolymeric particles (TEP) and proteinaceous coomassie stainable particles (CSP), serve as influential factors of export flux and aggregation. The presence of heterotrophic bacteria can also affect aggregation and sinking velocity, as seen in previous studies, and is likely attributed to their EPS and TEP production. The staining and visualization of TEP and CSP allow for the qualitative determination of these types of EPS from bacteria isolated from sinking particles collected with particle interceptor traps at various depths in the Sargasso Sea. I study the presence of TEP and CSP in particle-associated bacteria. Cultures of picocyanobacteria, consisting of xenic Synechococcus and axenic Prochlorococcus, were used to establish positive and negative controls for stained isolate analysis. Marinobacter adhaerens served as a tertiary control for an axenic culture that stains positive for TEP. I chose six isolates of bacteria isolated from sinking particles to be stained and visualized to test for the secretion of TEP and CSP. Four of the isolates stained positive for both TEP and CSP, including Pseudoalteromonas sp., Erythrobacter sp., and Marinobacter sp., while one isolate, Micrococcus sp., stained positive only for TEP, and the last isolate, another Marinobacter sp., stained positive for only CSP. These results are important in understanding the role of plankton organisms in the formation of sinking particles.
ContributorsLivar, Britni (Author) / Neuer, Susanne (Thesis director) / Cadillo-Quiroz, Hinsby (Committee member) / Cruz, Bianca (Committee member) / School of Life Sciences (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Impact of Nutrient Limitation on Growth, Aggregation, and TEP Production of Marine Nano- and Picophytoplankton

Description
The oceanic biological carbon pump is a key component of the global carbon cycle in which dissolved carbon dioxide is taken up by phytoplankton during photosynthesis, a fraction of which then sinks to depth and contributes to oceanic carbon storage. The small-celled phytoplankton (<5 µm) that dominate the phytoplankton community…

The oceanic biological carbon pump is a key component of the global carbon cycle in which dissolved carbon dioxide is taken up by phytoplankton during photosynthesis, a fraction of which then sinks to depth and contributes to oceanic carbon storage. The small-celled phytoplankton (<5 µm) that dominate the phytoplankton community in oligotrophic oceans have traditionally been viewed as contributing little to export production due to their small size. However, recent studies have shown that the picocyanobacterium Synechococcus produces transparent exopolymer particles (TEP), the sticky matrix of marine aggregates, and forms abundant microaggregates (5-60 µm), which is enhanced under nutrient limited growth conditions. Whether other small phytoplankton species exude TEP and form microaggregates, and if these are enhanced under growth-limiting conditions remains to be investigated. This study aims to analyze how nutrient limitation affects TEP production and microaggregate formation of species that are found to be associated with sinking particles in the Sargasso Sea. The pico-cyanobacterium Prochlorococcus marinus (0.8 µm), the nano-diatom Minutocellus polymorphus (2 µm), and the pico-prasinophyte Ostreococcus lucimarinus (0.6 µm) were grown in axenic batch culture experiments under nutrient replete and limited conditions. It was hypothesized that phytoplankton subject to nutrient limitation will aggregate more than those under replete conditions due to an increased exudation of TEP and that Minutocellus would produce the most TEP and microaggregates while Prochlorococcus would produce the least TEP and microaggregates of the three phytoplankton groups. As hypothesized, nutrient limitation increased TEP concentration in all three species, however they were only significant in nitrogen-limited treatments of Prochlorococcus as well as nitrogen- and phosphorus-limited treatments of Minutocellus. Formation of microaggregates was significantly enhanced in Minutocellus and Ostreococcus cultures in distinct microaggregate size ranges. Minutocellus produced the most TEP per cell and aggregated at higher volume concentrations compared to Prochlorococcus and Ostreococcus. Surprisingly, Ostreococcus produced more TEP than Prochlorococcus and Minutocellus per unit cell volume. These findings show for the first time how nutrient limited conditions enhance TEP production and microaggregation of Prochlorococcus, Minutocellus, and Ostreococcus, providing a mechanism for their incorporation into larger, sinking particles and contribution to export production in oligotrophic oceans.
ContributorsShurtleff, Catrina (Author) / Neuer, Susanne (Thesis advisor) / Lomas, Michael W. (Committee member) / Garcia-Pichel, Ferran (Committee member) / Arizona State University (Publisher)
Created2022