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Aspects of Nitrogen Metabolism in Symbiotic Cnidarians

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Title: Aspects of Nitrogen Metabolism in Symbiotic Cnidarians
Author: Boutilier, Ryan Michael
Department: Department of Integrative Biology
Program: Integrative Biology
Advisor: Ballantyne, James Stuart
Abstract: The pathway of seawater ammonium assimilation and influence of light on amino acid synthesis remain unresolved in cnidarian symbioses. Labeled ammonium (10 μM 15NH4Cl) in seawater was used to trace the pathway of the incorporation into amino acids in host tissue, Zoanthus sp., and zooxanthellae, Symbiodinium microadriaticum. Freshly isolated zooxanthellae were exposed to 20 μM 15NH4Cl with coral homogenate to evaluate the role of host factors on amino acid synthesis. High performance liquid chromatography and mass spectrometry was used to measure percent labeling and concentrations of amino acids. In zooxanthellae, ammonium was assimilated into glutamine likely via glutamine synthetase and into glutamate via glutamine 2-oxoglutarate amidotransferase. Interrupting photosynthesis with DCMU did not inhibit glutamine and tryptophan synthesis however reduced the 15N-enrichment of glutamate, aspartate, and ornithine in zooxanthellae, as well as arginine, ornithine, and lysine in host tissue. Coral homogenate had little effect on the 15N-enrichment of glutamine, aspartate, and alanine in freshly isolated zooxanthellae. Evidence is presented to support the uptake of ammonium ions and data shows that glutamine and not glutamate is translocated to the coral host.
Date: 2012-05

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