Development of a protocol for specific detection and quantification of free-living and endosymbiotic Symbiodinium communities in coral reefs

作者：Lin, Zhenyue; Chen, Mingliang; Chen, Jianming^*

来源：Aquatic Microbial Ecology, 2018, 80(1): 1-13.

DOI：10.3354/ame01833

• Taqman
• Coral bleaching
• Primer-probe design

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.

• 出版日期2018
• 单位厦门大学; 国家海洋局第三海洋研究所