摘要
目的建立微囊藻的分离纯化培养方法,并鉴定其生长和产毒特性。方法采用96孔板结合极限稀释法,对郑州市西流湖微囊藻进行分离纯化培养;用倒置显微镜观察分离微囊藻株的细胞学和生长特点;用可见分光光度法测定分离微囊藻培养液的吸光度,并绘制分离藻株的生长曲线,计算其生长速率常数和倍增时间;用全细胞PCR和ELISA鉴定分离微囊藻株的产毒性,用ELISA测定其微囊藻毒素粗提物的浓度,并计算其产毒量。结果成功从郑州市西流湖分离出2株微囊藻Microcystis XLH6和Microcystis XLH10,其生长曲线均呈“S”型,生长速率常数均呈先迅速升高然后逐渐降低的趋势;一个生长周期内Microcystis XLH6和Microcystis XLH10的平均生长速率常数分别为0.294和0.345,平均倍增时间分别为82h和70h。全细胞PCR和ELISA结果均为阳性,2株微囊藻均为产毒株,冻干藻细胞Microcystis XLH6和Microcystis XLH10的微囊藻毒素产量分别为1.0μg/mg和2.4μg/mg。结论郑州市西流湖有产毒微囊藻污染,96孔细胞培养板可用于分离纯化微囊藻。
Objective To establish methods of isolation and purification of microcystis, and to identify it＇s biological property. Methods By the means of 96 well microplates and utmost dilution, microcystis was isolated and purified from Xiliu Lake of Zhengzhou. Absorbency of isolated microcystis was determined by visble light spectrophotometric method, and it＇ s growth curve was ploted, calculated growth rate constant and double time. Toxigenicity of isolated microcystis was identified by means of the Whole cell PCR and ELISA, and the concentration of microcystin in crude extractive solution from freeze-dried microcystis cells was determined by ELISA. Results Two strains of microcystis, Microcystis XLH6 and Microcystis XLH10 were isolated successfully from Xiliu Lake, Their growth curves were ＂S＂ style, their growth rate constants rapidly ascended firstly, and then slowly descended, Average growth rate constants of the two strains were 0. 294 and 0.345, average double times of the two strains were 82h and 70h, respectively. The results of the Whole cell PCR and ELISA of the two strains were positive, and both of them were toxigenic, Every milligram of freeze-dried Microcystis XLH6 and Microcystis XLH10 produced 1,4,ttg and 2,4μg microcystin. Conclusion Xiliu Lake was polluted by toxigenic microcystis, 96 well microplates can be used to isolate microcystis.
出处
《卫生研究》
CAS
CSCD
北大核心
2007年第4期424-426,共3页
Journal of Hygiene Research
基金
河南省医学创新人才工程资助项目(No.200311205)
关键词
微囊藻
微囊藻毒素
分离纯化培养
产毒特性
microcystis, microcystin, isolation and purification culture, toxigenic characteristic
