A quantitative polymerase chain reaction assay for the enumeration of brown tide algae Aureococcus anophagefferens in coastal waters of Qinhuangdao

作者：Guo Hao^*; Liu Yongjian; Zhang Qi; Yuan Xiutang; Zhang Weiwei; Zhang Zhifeng

来源：Acta Oceanologica Sinica, 2015, 34(2): 132-136.

DOI：10.1007/s13131-015-0615-x

• Aureococcus anophagefferens
• quantitative polymerase chain reaction (qPCR)
• field samples

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 (qPCR) based on 18S rDNA sequences was developed and used to detect and enumerate A. anophagefferens. A linear regression (R-2 = 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 qPCR. 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/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.

• 出版日期2015-2
• 单位国家海洋环境监测中心; 大连海事大学; 大连海洋大学