title: Rapid Identification and Quantification ofAureococcus anophagefferensby qPCR Method (Taqman) in the Qinhuangdao Coastal Area: A Region for Recurrent Brown Tide Breakout in China.
abstract: Since 2009,Aureococcus anophagefferenshas caused brown tide to occur recurrently in Qinhuangdao coastal area, China. Because the algal cells ofA. anophagefferensare so tiny (~3 µm) that it is very hard to identify exactly under a microscope for natural water samples, it is very urgent to develop a method for efficient and continuous monitoring. Here specific primers and Taqman probe are designed to develop a real-time quantitative PCR (qPCR) method for identification and quantification continually. The algal community and cell abundance ofA. anophagefferensin the study area (E 119°20'-119°50' and N 39°30'-39°50') from April to October in 2013 are detected by pyrosequencing, and are used to validate the specification and precision of qPCR method for natural samples. Both pyrosequencing and qPCR shows that the targeted cells are present only in May, June and July, and the cell abundance are July > June > May. Although there are various algal species including dinoflagellata, diatom, Cryptomonadales, Chrysophyceae and Chlorophyta living in the natural seawater simultaneously, no disturbance happens to qPCR method. This qPCR method could detect as few as 10 targeted cells, indicating it is able to detect the algal cells at pre-bloom levels. Therefore, qPCR with Taqman probe provides a powerful and sensitive method to monitor the brown tide continually in Qinhuangdao coastal area, China. The results provide a necessary technology support for forecasting the brown tide initiation, in China.
authors: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing, 100012 People's Republic of China ; State Environmental Protection Key Laboratory of Estuarine and Coastal Research, Chinese Research Academy of Environmental Sciences, Beijing, 100012 People's Republic of China.
authors: Li-Ping
authors: LP
authors: Wang
authors: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing, 100012 People's Republic of China ; State Environmental Protection Key Laboratory of Estuarine and Coastal Research, Chinese Research Academy of Environmental Sciences, Beijing, 100012 People's Republic of China.
authors: Kun
authors: K
authors: Lei
keywords: Aureococcus anophagefferens; Brown tide bloom; Harmful algal bloom (HAM); Pyrosequencing; Real-time quantitative PCR (qPCR)