产毒微囊藻PCR检测方法的建立

目的]建立产毒微囊藻的PCR检测方法。方法]根据GenBank上发表的mcyA基因序列保守区域设计一对引物,建立和优化检测产毒微囊藻的PCR方法,并以此方法检测产毒和非产毒微囊藻参考株系。结果]经PCR检测,产毒藻株出现特异性扩增条带,而不产毒株未出现特异性条带;以10倍系列稀释纯培养的蓝藻细胞作为PCR反应的模板,检测限均为2.46×103细胞/ml;运用建立的方法从天然水体中检测到了产毒微囊藻,对PCR产物序列进行比对,发现PCR扩增片段与铜绿微囊藻(PCC7806)核苷酸序列的同源性为98%。结论]该研究为水华产毒微囊藻的监测提供了一种新方法。

Development of PCR Method for Detection of Potential Microcystin-producing Microcystis

Objective ] The aim was to develop a PCR method for detection of potential microcystin-producing microcystis. Method ] Accord- ing to conserved regions of mcyA gene sequences in GenBank, a pair of primers was designed, and a PCR method for the detection of microcys- tin-producing microcystis was developed and optimized. Then the referenced microcystin-producing and non-microcystin-producing microcystis strains were detected by this method. Result] The microcystin-producing microcystis stains appeared specific DNA band and the non-micro- cystin-producing microcystis strains didn＇t show specific band by the PCR test; the intact cells were 10 times diluted and used as template, and the results indicated that the detection limit of the method was up to 2.46 × 103 cells/ml ; the microcystin-producing microcystis was de- tected from natural water by using the established method and the Blast N result showed that this PCR fragments had 98% homology to that of Microcystis aeruginosa （PCC7806）. Conclusion ] This study provided a new method for the monitoring of microcystin-producing microcystis in bloom.