| 应用实时定量PCR技术检测秦皇岛近岸海域褐潮原因种——抑食金球藻(Aureococcus anophagefferens) |
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| 作者姓名： | GUO Hao LIU Yongjian ZHANG Qi YUAN Xiutang ZHANG Weiwei ZHANG Zhifeng |
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| 作者单位： | Dalian Maritime University, Dalian 116026, China;Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China,Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China,Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China;College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China,Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China,Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China,Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China |
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| 基金项目： | The National Natural Science Foundation of China under contract No.40906081; the Public Science and Technology Research Funds Projects of Ocean under contract Nos 2011418021 and 201305030; Integrated Environmental Governances and Restoration in Beidaihe Coast of Hebei Province. |
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| 摘 要： | Aureococcus anophagefferens, a small pelagophyte algae, has caused brown tide blooms in coastal waters of Qinhuangdao in recent years, presenting significant negative impacts on the shellfish mariculture industry. Under standard light microscopy, it is visually indistinguishable from other small algae in field samples due to its extremely small size. In this study, quantitative polymerase chain reaction(q PCR) based on 18 S r DNA sequences was developed and used to detect and enumerate A. anophagefferens. A linear regression(R2 = 0.91) was generated based on cycle thresholds value(Ct) versus known concentrations of A. anophagefferens. Twenty-two field samples collected in coastal waters of Qinhuangdao were subjected to DNA extraction and then analyzed using q PCR. Results showed that A. anophagefferens had a wide distribution in coastal waters along Qinhuangdao. Elevated A. anophagefferens abundance, category 3 brown tide blooms(200 000 cells/m L) occurred at Dongshan Beach and Tiger-stone Beach in August in 2013. In shellfish mariculture areas along coastal waters of Qinhuangdao, 4 stations had category 3 blooms, and 6 stations had category 2 blooms(35 000–200 000 cells/m L) in August and all stations had category 1 blooms(0 to ≤35 000 cells/m L) in October. Quantitative PCR allows for detection of A. anophagefferens cells at low levels in filed samples, which is essential to effective management and prediction of brown tide blooms.
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| 关 键 词： | 褐潮 抑食金球藻 实时定量PCR 环境样品 |
| 收稿时间： | 1/6/2014 12:00:00 AM |
| 修稿时间： | 2014/6/20 0:00:00 |
A quantitative polymerase chain reaction assay for the enumeration of brown tide algae Aureococcus anophagefferens in coastal waters of Qinhuangdao |
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| GUO Hao,LIU Yongjian,ZHANG Qi,YUAN Xiutang,ZHANG Weiwei,ZHANG Zhifeng.A quantitative polymerase chain reaction assay for the enumeration of brown tide algae Aureococcus anophagefferens in coastal waters of Qinhuangdao[J].Acta Oceanologica Sinica,2015,34(2):132-136. |
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| Authors: | GUO Hao LIU Yongjian ZHANG Qi YUAN Xiutang ZHANG Weiwei and ZHANG Zhifeng |
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| Institution: | 1. Dalian Maritime University, Dalian 116026, China
2. Key Laboratory of Coastal Ecology and Environment of State 0ceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China |
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| Abstract: | Aureococcus anophagefferens, a small pelagophyte algae, has caused brown tide blooms in coastal waters of Qinhua-ngdao in recent years, presenting significant negative impacts on the shellfish mariculture industry. Under standard light microscopy, it is visually indistinguishable from other small algae in field samples due to its extremely small size. In this study, quantitative polymerase chain reaction (qPCR) based on 18S rDNA sequences was developed and used to detect and enumerate A. anophagefferens. A linear regression (R2= 0.91) was generated based on cycle thr-esholds value (Ct) versus known concentrations of A. anophagefferens. Twenty-two field samples collected in coastal waters of Qinhuangdao were subjected to DNA extraction and then analyzed using qPCR. Results showed that A. anophagefferens had a wide distribution in coastal waters along Qinhuangdao. Elevated A. anophagefferens abun-dance, category 3 brown tide blooms (>200 000 cells/mL) occurred at Dongshan Beach and Tiger-stone Beach in August in 2013. In shellfish mariculture areas along coastal waters of Qinhuangdao, 4 stations had category 3 blooms, and 6 stations had category 2 blooms (35 000-200 000 cells/mL) in August and all stations had category 1 blooms (>0 to ≤35 000 cells/mL) in October. Quantitative PCR allows for detection of A. anophagefferens cells at low levels in filed samples, which is essential to effective management and prediction of brown tide blooms. |
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| Keywords: | Aureococcus anophagefferens quantitative polymerase chain reaction (qPCR) field samples |
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