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刘洋,胡佩茹,马思三,叶金云.实时荧光定量PCR方法检测南太湖入湖口产毒微囊藻.湖泊科学,2016,28(2):246-252. DOI:10.18307/2016.0202
LIU Yang,HU Peiru,MA Sisan,YE Jinyun.Detection of microcystin-producing Microcystis cells at the entrance of rivers to southern Lake Taihu by real-time fluorescence quantitative PCR. J. Lake Sci.2016,28(2):246-252. DOI:10.18307/2016.0202
实时荧光定量PCR方法检测南太湖入湖口产毒微囊藻
Detection of microcystin-producing Microcystis cells at the entrance of rivers to southern Lake Taihu by real-time fluorescence quantitative PCR
投稿时间:2015-04-21  修订日期:2015-06-19
DOI:10.18307/2016.0202
中文关键词: 实时荧光定量PCR  产毒微囊藻  入湖口  南太湖  铜绿微囊藻
Keywords: Real-time fluorescence quantitative PCR  microcystin producing cells  the entrance of lake  southern Lake Taihu  Microcystis aeruginosa
基金项目:浙江省科技计划项目-分析测试科技计划项目(2014C37091)资助.
作者单位E-mail
刘洋 湖州师范学院生命科学院, 湖州 313000;浙江省水生生物资源养护与开发技术研究重点实验室, 湖州 313000;中国水产科学研究院水生动物繁育与营养重点实验室, 湖州 313000  
胡佩茹 湖州师范学院生命科学院, 湖州 313000  
马思三 湖州师范学院生命科学院, 湖州 313000  
叶金云 湖州师范学院生命科学院, 湖州 313000;浙江省水生生物资源养护与开发技术研究重点实验室, 湖州 313000;中国水产科学研究院水生动物繁育与营养重点实验室, 湖州 313000 ziff2006@163.com. 
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中文摘要:
      南太湖入湖口产毒微囊藻的丰度对于周边县市取水口的水质安全和太湖水质有着重要影响.以藻毒素合成酶基因mcyE/ndaF为靶基因,建立实时荧光定量PCR检测产毒微囊藻的方法,并对南太湖入湖口7个监测点水样中产毒微囊藻的丰度进行检测,结果表明:该实时荧光定量PCR方法的特异性强及准确性、重复性较好.建立铜绿微囊藻(Microcystis aeruginosa)的标准曲线方程为y=-3.454x+49.88,斜率为-3.454,R2=0.991,扩增效率E为94.6%,定量检测区间为1.689×104~1.689×108拷贝数/μl.对南太湖入湖口7个监测点检测表明,夹浦和合溪2个监测点的产毒微囊藻数量最高,预测产毒微囊藻浓度分别为(1.99±0.35)×105和(1.47±0.23)×105 cells/ml.7个监测点的产毒微囊藻的种类较为一致,均为铜绿微囊藻.该方法可以快速准确地检测水体中微囊藻毒素产毒藻种种类和数量,为蓝藻水华监测、预警提供技术依据.
Abstract:
      The amount of Microsystis cells at the entrance of rivers to southern Lake Taihu has important influence on the drinking water security and water quality to county nearby and Lake Taihu.A real-time fluorescence quantitative PCR method was established using the microcystin synthetase gene mcyE/ndaF as target in this study.The quantity of Microsystis cells at the entrance of rivers to southern Lake Taihu was assayed.Results showed that the specificity, accuracy and repetitiveness of the method were good.The standard curve to detect Microcystis aeruginosa was y=-3.454x+49.88.The slope, correlation coefficient and amplication efficiency were -3.454, 0.991 and 94.6%, respectively.It was revealed that the amount of microcystin producing blue-green algae at site of Jiapu and Hexi was the highest among the 7 sampling sites, reaching (1.99±0.35)×105 and (1.47±0.23)×105 cells/ml, respectively.The main microcystin producing species was Microcystis aeruginosa in all the sampling sites.This method could be used rapidly and accurately to detect the quantity and type of the microcystin producing blue-green algae, and provide evidence for algal bloom and early warning.
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