Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens

doi:10.1002/lom3.10344

	Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens
	Zhang, Qing-Chun 1,2,3; Chen, Zhen-Fan 1,4; Zhao, Jia-Yu1,4 ; Yu, Ren-Cheng1,2,3,4 ; Qiu, Li-Mei 3,5,6; Kong, Fan-Zhou1,2,3 ; Wang, Yun-Feng 1; Yan, Tian 1,2,3; Zhou, Ming-Jiang 1
	2019-12-20
发表期刊	LIMNOLOGY AND OCEANOGRAPHY-METHODS
ISSN	1541-5856
页码	11
摘要	Brown tides of pelagophyte Aureococcus anophagefferens have been recorded in the coastal waters of Qinhuangdao in the Bohai Sea (BS) since 2009. Current understandings on the dynamics and mechanisms of brown tides in the BS, however, are still quite limited due to the lack of accurate methods for detecting low-abundance of A. anophagefferens cells. In this study, a Taqman qPCR method was developed, which targeted 28S rDNA D1-D2 domain of A. anophagefferens. The specificity of the qPCR method was tested using 25 strains of microalgae commonly present in the coastal waters, and the targeted species A. anophagefferens was the only species that had reliable positive signal. The method was highly sensitive and reproducible, and the lower detection limit could reach 0.3 cells of A. anophagefferens. A calibration curve was established for A. anophagefferens using the DNA template extracted from different number of cells, and the lower and upper limits of detection for the qPCR assay is 0.3 cells and 1.2 x 10(5) cells, respectively. A high recovery rate ranged from 96.5% to 107% was achieved for A. anophagefferens at different cell densities. The qPCR assay has been successfully applied to the study on spatial and temporal variations of A. anophagefferens in the coastal waters of Qinhuangdao in 2014.
DOI	10.1002/lom3.10344
收录类别	SCI
语种	英语
资助项目	S&T Basic Resources Investigation Program of China[2018YF100206]###3592; National Key Research and Development Program Sino-Australian Centre for Healthy Coasts[2016YFE0101500]###3591; Program of the Qingdao National Laboratory for Marine Science and Technology[2016ASKJ02]###2760; National Natural Science Foundation of China[41276117]###2759; National Natural Science Foundation of China[41576121]###2758; National Natural Science Foundation of China[U1606404]###798; National key R&D Program of China[2017YFC1404304]###190; National key R&D Program of China[2017YFC1404304]; National Natural Science Foundation of China[U1606404]; National Natural Science Foundation of China[41576121]; National Natural Science Foundation of China[41276117]; Program of the Qingdao National Laboratory for Marine Science and Technology[2016ASKJ02]; National Key Research and Development Program Sino-Australian Centre for Healthy Coasts[2016YFE0101500]; S&T Basic Resources Investigation Program of China[2018YF100206]
WOS研究方向	Marine & Freshwater Biology ; Oceanography
WOS类目	Limnology ; Oceanography
WOS记录号	WOS:000503554100001
出版者	WILEY
引用统计	被引频次：8[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
版本	出版稿
条目标识符	http://ir.qdio.ac.cn/handle/337002/164179
专题	海洋生态与环境科学重点实验室实验海洋生物学重点实验室
通讯作者	Yu, Ren-Cheng
作者单位	1.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Marine Ecol & Environm Sci, Qingdao, Shandong, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Ecol & Environm Sci, Qingdao, Shandong, Peoples R China 3.Chinese Acad Sci, Ctr Ocean Megasci, Qingdao, Shandong, Peoples R China 4.Univ Chinese Acad Sci, Beijing, Peoples R China 5.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Expt Marine Biol, Qingdao, Shandong, Peoples R China 6.Qingdao Natl Lab Marine Sci & Technol, Marine Biol & Biotechnol Lab, Qingdao, Shandong, Peoples R China
第一作者单位	中国科学院海洋研究所; 中国科学院海洋大科学研究中心
通讯作者单位	中国科学院海洋研究所; 中国科学院海洋大科学研究中心
推荐引用方式 GB/T 7714	Zhang, Qing-Chun,Chen, Zhen-Fan,Zhao, Jia-Yu,et al. Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens[J]. LIMNOLOGY AND OCEANOGRAPHY-METHODS,2019:11.
APA	Zhang, Qing-Chun.,Chen, Zhen-Fan.,Zhao, Jia-Yu.,Yu, Ren-Cheng.,Qiu, Li-Mei.,...&Zhou, Ming-Jiang.(2019).Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens.LIMNOLOGY AND OCEANOGRAPHY-METHODS,11.
MLA	Zhang, Qing-Chun,et al."Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens".LIMNOLOGY AND OCEANOGRAPHY-METHODS (2019):11.

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