AME 80:1-13 (2017) - DOI: https://doi.org/10.3354/ame01833
Development of a protocol for specific detection and quantification of free-living and endosymbiotic Symbiodinium communities in coral reefs
Zhenyue Lin1,2, Mingliang Chen2, Jianming Chen2,*
ABSTRACT: This study focused on the evaluation of a novel specific Taqman PCR assay to accurately determine the intercolony cell density of Symbiodinium in surrounding seawaters and intracolony cell density in coral tissue, which can be used to constantly monitor health status of the coral in field studies. Our Taqman method using ITS-rDNA-based specific primer-probe sets can differentiate Symbiodinium genotypes at the clade level. An additional primer-probe set based on the coral paired-box (Pax) gene was used as a universal internal reference to estimate the relative abundance of coral host cells. This assay was highly reproducible and reliable, which allowed accurate quantification of extremely low Symbiodinium DNA (up to 2.0 pg) in different coral DNA backgrounds with high specificity and efficiency. The Taqman PCR assay was further successfully applied for the detection and quantification of Symbiodinium in complex samples from multiple origins, including free-living individuals and endosymbionts within the coral Galaxea fascicularis.
KEY WORDS: Taqman · Coral bleaching · Primer-probe design
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Full text in pdf format Supplementary material | Cite this article as: Lin Z, Chen M, Chen J
(2017) Development of a protocol for specific detection and quantification of free-living and endosymbiotic Symbiodinium communities in coral reefs. Aquat Microb Ecol 80:1-13. https://doi.org/10.3354/ame01833
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