水体中蓝绿藻分子标志物——mcyD基因的检测

Detection of Potential Microcystin-Producing Cyanobacteria in Waters With a mcyD Molecular Biomarker

目的建立一种筛选自然水体中产微囊藻毒素藻株的方法。方法应用编码微囊藻毒素合成酶mcyD基因的特异引物结合藻类保守序列16SrRNA对所培养的纯种蓝绿藻(铜绿微囊藻FACHB469、铜绿微囊藻PCC7806、微囊藻属FACHB569、浮游蓝丝藻属小颤藻FACHB247、泡沫节球藻FACHB377)和采集的水样进行聚合酶链反应,观察扩增结果,并以酶联免疫吸附方法检测其微囊藻毒素浓度。结果在产微囊藻毒素(铜绿微囊藻PCC7806、微囊藻属FACHB569)的藻株和水样均可以扩增出1条约870bp的单一条带,而不产微囊藻毒素的藻株(铜绿微囊藻FACHB469、浮游蓝丝藻属小颤藻FACHB247、泡沫节球藻FACHB377)和水样未扩增出此条带。结论以mcyD基因为微囊藻毒素分子标志物的分子生物学方法鉴定自然水体中的产微囊藻毒素和非产微囊藻毒素蓝绿藻是可行和实用的。

Objective To develop a new method to screen the potential microeystin-producing eyanobacteria in natural waters. Methods Cyanobaeteria were cultured and waters samples were analyzed by polymerase chain reaction with special primers for gene of the operon mcyD which encodes a microcystin synthetase combining conserved gene 16S rRNA of alga, subsequently, whose microcystins were detected by ELISA. Results There was a unique amplified product of approximately 870 bp in mieroeystin-produeing blue-green alga and water samples, but no target band in non-toxic strains and samples. Conclusion It is feasible to use the mol^ular biological method used meyD as the microcystin molecular biomarker to screen mierocystin-producing cyanobacteria in natural water.