| GUO Hao,LIU Yongjian,ZHANG Qi,YUAN Xiutang,ZHANG Weiwei,ZHANG Zhifeng. 2015. A quantitative polymerase chain reaction assay for the enumeration of brown tide algae Aureococcus anophagefferens in coastal waters of Qinhuangdao. Acta Oceanologica Sinica, 34(2):132－136 |
| A quantitative polymerase chain reaction assay for the enumeration of brown tide algae Aureococcus anophagefferens in coastal waters of Qinhuangdao |
| 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) |
| Received:January 06, 2014 Revised:June 20, 2014 |
| DOI：10.1007/s13131-015-0615-x |
| Key words:Aureococcus anophagefferens quantitative polymerase chain reaction (qPCR) field samples |
| 中文关键词: 褐潮 抑食金球藻 实时定量PCR 环境样品 |
| 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. |
| Author Name | Affiliation | E-mail | | GUO Hao | 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 | hguo@nmemc.org.cn | | LIU Yongjian | Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China | | | ZHANG Qi | 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 | | | YUAN Xiutang | Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China | | | ZHANG Weiwei | Key Laboratory of Coastal Ecology and Environment of State Oceanic Administration, National Marine Environmental Monitoring Center, Dalian 116023, China | | | ZHANG Zhifeng | Key Laboratory of Coastal Ecology and Environment of State Oceanic 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. |
| 中文摘要: |
| 近年来,秦皇岛近岸海域连续发生抑食金球藻褐潮,对当地贝类水产养殖带来负面影响.由于其个体及其微小,在光学显微镜下很难将抑食金球藻和水体中其他微型藻类准确区分.基于18S rDNA 序列,本研究应用实时定量PCR技术检测和计数环境中的抑食金球藻.实验结果显示:循环阈值和抑食金球藻细胞丰度之间存在良好线性关系(R2= 0.91).应用实时定量PCR技术对采自秦皇岛近岸海域的24份海水样品进行了检测分析,发现抑食金球藻在秦皇岛近岸海域广泛分布.根据抑食金球藻细胞丰度及褐潮分级标准,2013年8月份,东山海水浴场和老虎石海水浴场发生3级抑食金球藻褐潮;在贝类养殖区,4个站位发生3级褐潮,6个站位发生2级抑食金球藻褐潮.而在2013年10月份,贝类养殖区所有站位均发生1级抑食金球藻褐潮.实时定量PCR技术可以检测到海水样品中的微量抑食金球藻,这对褐潮灾害的提前预警及灾害管理至关重要. |
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