应用实时荧光定量PCR方法检测水库水体中微囊藻毒素

Detection of Micocystin of Reservoir by Real-Time Fluorescence Quantitative PCR

水库微囊藻毒素对供水水质安全有着重要影响.以微囊藻毒素合成酶基因mcyA、mcyB、mcyC、mcyD、mcyE、mcyF、mcyG、mcyH、mcyI和mcyJ为靶基因,应用实时荧光定量PCR方法,检测水库进水口、库内和输水口3个监测点,6月-9月水体中微囊藻毒素丰度.结果表明,水库输水口微囊藻毒素合成基因拷贝数低于进水口,特别是7月和8月温度较高时,这种趋势比较明显.微囊藻特异性16S rDNA基因拷贝数比MC合成基因拷贝数高3个数量级,这说明水库水体中除了产毒微囊藻外,还存在着不产生毒素的微囊藻.相关性分析表明,mcyA、mcyB、mcyD、mcyE、mcyF、mcyH、mcyI和mcyJ具有一定的相关性.综上所述,该实时荧光定量PCR方法可用于水库水体中微囊藻毒素检测,为蓝藻水华监测、预警提供技术支撑.

The content of microcystin in reservoir has important effect on drinking water quality.A real-time fluorescence quantitative PCR method was established using microcystin synthetase(mcyA,mcyB,mcyC,mcyD,mcyE,mcyF,mcyG,mcyH,mcyI and mcyJ)as target gene in the study.The content of microcystin in the reservoir from June to September was detected.The sampling site included raw water from river,water inside reservoir,water piping to drinking water plant.Results found that target genes were detected in each sample.Compared to inlet sluice,copy number of microcystin biosynthetic genes was lower in pump station,especially in July and August when the temperature was higher,that the trend was more obvious.In addition to toxic microcystis,there were a large number of nontoxic microcystis in the reservoir.Correlation analysis showed that mcyA,mcyB,mcyD,mcyE,mcyF,mcyH,mcyI and mcyJ had significant correlation.The method could be used rapidly and accurately to detect microcystin synthetase,and provide technical support for monitoring and early warning of cyanobacteria bloom.