Institutional Repository of Key Laboratory of Marine Ecology & Environmental Sciences, CAS
| Development of a sensitive qPCR method for the detection of pelagophyte Aureococcus anophagefferens | |
Zhang, Qing-Chun1,2,3; Chen, Zhen-Fan1,4; Zhao, Jia-Yu1,4; Yu, Ren-Cheng1,2,3,4; Qiu, Li-Mei3,5,6; Kong, Fan-Zhou1,2,3 ; Wang, Yun-Feng1; Yan, Tian1,2,3; Zhou, Ming-Jiang1
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| 2019-12-20 | |
| 发表期刊 | LIMNOLOGY AND OCEANOGRAPHY-METHODS
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| 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] ; National Key Research and Development Program Sino-Australian Centre for Healthy Coasts[2016YFE0101500] ; Program of the Qingdao National Laboratory for Marine Science and Technology[2016ASKJ02] ; National Natural Science Foundation of China[41276117] ; National Natural Science Foundation of China[41576121] ; National Natural Science Foundation of China[U1606404] ; National key R&D Program of China[2017YFC1404304] ; 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 | |
| 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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| Development of a sen（666KB） | 期刊论文 | 作者接受稿 | 限制开放 | CC BY-NC-SA | 浏览 | |
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