A microsampling method for genotyping coral symbionts
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Genotypic characterization of Symbiodinium symbionts in hard corals has routinely involved coring, or the removal of branches or a piece of the coral colony. These methods can potentially underestimate the complexity of the Symbiodinium community structure and may produce lesions. This study demonstrates that microscale sampling of individual coral polyps provided sufficient DNA for identifying zooxanthellae clades by RFLP analyses, and subclades through the use of PCR amplification of the ITS-2 region of rDNA and denaturing-gradient gel electrophoresis. Using this technique it was possible to detect distinct ITS-2 types of Symbiodinium from two or three adjacent coral polyps. These methods can be used to intensely sample coral-symbiont population/communities while causing minimal damage. The effectiveness and fine scale capabilities of these methods were demonstrated by sampling and identifying phylotypes of Symbiodinium clades A, B, and C that co-reside within a single Montastraea faveolata colony.
KeywordsSymbiodinium Coral sampling Zooxanthellae Microhabitat Symbiosis Genotyping
We would like to thank Brigitte Bruns, Daniel Thornhill, Jennifer McCabe Reynolds, Robin Smith, Randi Rotjan, Meredith Meyers, Xavier Hernandez-Pech, John Ware and Gordon Hendler for their laboratory, diving and field assistance. Special thanks to Scott Santos for discussions inspiring development of these techniques. This work was supported by the World Bank Center of Excellence Coral Bleaching Working Group and National Science Foundation grant NSF-0137007 to W.K. Fitt and G.W.Schmidt.
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