郑州市主要生活饮用水源中微囊藻细胞的分离培养与毒性鉴定

Isolation and identification of microcystis in Zhengzhou main drinking water source

目的:对郑州市主要生活饮用水源中微囊藻细胞进行分离培养和毒性鉴定。方法:采用改进的96孔板分离技术对郑州市2个主要生活饮用水源西流湖和黄河花园口段某调蓄池中采集的浮游藻类进行分离培养;全细胞PCR测定藻青蛋白基因中间序列(PC-IGS)和微囊藻毒素合成酶基因(mcyB);ELISA检测毒素浓度。结果:应用96孔板结合极限稀释法成功得到了所要分离微囊藻细胞的单克隆;所分离3株微囊藻细胞PC-IGS基因序列和mcyB基因序列扩增结果均为阳性;ELISA检测3株微囊藻细胞干粉的产毒量分别为1.07mg/g、4.70mg/g、4.71mg/g。结论:改进的96孔板分离技术能够简便、快速地分离各种藻细胞;所分离的3株微囊藻细胞均为蓝藻种属,且能够产毒。

Aim ： To isolate the microcystis in the main source water in Zhengzhou city and identify the toxicity. Methods：The algae cell was isolated by the 96-microbiologlcal assay and the phycocyanin intergenie spacer region （PC-IGS） and the microcystin synthetase gene B （McyB） of the isolated algae cell were detected by the whole-cell PCR, and the toxicity was identified by ELISA. Results： Single cell clones were acquired successfully, and the whole-cell PCR results were positive of PC-IGS and McyB,which suggested that the 3 microcystis were cyanobaeteria and toxic. The toxin contents of the three microcystis powder were 1.07 mg/g,4.70 mg/g, and 4.71 mg/g. Conclusion ： The 96-microbiological assay is a simple and fast method which can isolate the algae cells from the water successfully. The microcystis isolated from the main source water in Zhengzhou city are blue-green algae and they can produce microcystin.