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 | |
| Source Publication | LIMNOLOGY AND OCEANOGRAPHY-METHODS
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| ISSN | 1541-5856 |
| Pages | 11 |
| Corresponding Author | Yu, Ren-Cheng(rcyu@qdio.ac.cn) |
| Abstract | 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 |
| Indexed By | SCI |
| Language | 英语 |
| Funding Project | National Natural Science Foundation of China[41576121] ; National Natural Science Foundation of China[41276117] ; National Natural Science Foundation of China[U1606404] ; National Key Research and Development Program Sino-Australian Centre for Healthy Coasts[2016YFE0101500] ; S&T Basic Resources Investigation Program of China[2018YF100206] ; National Key R&D Program of China[2017YFC1404304] ; Program of the Qingdao National Laboratory for Marine Science and Technology[2016ASKJ02] |
| WOS Research Area | Marine & Freshwater Biology ; Oceanography |
| WOS Subject | Limnology ; Oceanography |
| WOS ID | WOS:000503554100001 |
| Publisher | WILEY |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.qdio.ac.cn/handle/337002/164179 |
| Collection | 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 |
| Corresponding Author | Yu, Ren-Cheng |
| Affiliation | 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 |
| First Author Affilication | Institute of Oceanology, Chinese Academy of Sciences |
| Corresponding Author Affilication | Institute of Oceanology, Chinese Academy of Sciences |
| Recommended Citation 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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