采用抗生素与倍比稀释相结合的分离方法,对太湖梅梁湾水域水华的优势藻进行分离培养;结合全细胞聚合酶链反应(PCR)、高效液相色谱法(HPLC)和实时荧光定量PCR(RTQ-PCR)法进行藻属与产毒特性分析.结果显示:自太湖梅梁湾水域成功分离获得2株藻株TH1和TH2,2株藻株藻蓝蛋白基因中间序列(PC-IGS)、微囊藻16S rDNA保守序列(Micr16s rDNA)扩增均为阳性,TH1的微囊藻毒素合成酶基因B(mcyB)扩增为阳性,而TH2的mcyB扩增为阴性.培养15d的TH1藻株每108个藻细胞产生的总微囊藻毒素-LR(TMC-LR)为0.594μg,胞外微囊藻毒素-LR(EMC-LR)为0.085μg,分别为铜绿微囊藻产毒株的61.93%和86.09%;TH2藻株未检出MC-LR.TH1藻株mcyB的mRNA相对表达水平为铜绿微囊藻产毒株的5.9%.结果表明:分离自太湖梅梁湾的2株藻细胞均为蓝藻门中的微囊藻属,其中1株产毒微囊藻具有较强的产毒能力,太湖梅梁湾水域有产毒微囊藻污染.
Algae were isolated from Meiliang Bay of Lake Taihu by dilution culture method and the antibiotics were applied to inhibit the growth of bacteria.The genus and toxigenicity of algae were analyzed by whole-cell polymerase chain reaction(PCR),high performance liquid chromatography(HPLC) and real-time quantitative PCR(RTQ-PCR) method.The results show that two algae strains are successfully isolated from Meiliang Bay of Lake Taihu bloom named as TH1 and TH2.The Phycocyanin intergenic spacer(PC-IGS) gene and 16S ribosoma deoxyribonucleic acid of Microcystis(Micr 16S rDNA) gene are amplified as positive in both algae strains and the Microcystin Synthetase B gene(mcyB) is only found in TH1 strain in whole-cell PCR.Microcystins-LR produced by TH1 strain is detected as 0.594μg in total and 0.085μg in extracellular per 108 cells in 15 d culture,which is 61.39% and 86.09% of MC-LR produced by Microcystis aeruginosa strain,respectively.A negative result of MC-LR is obtained in TH2 strain.The express intensity of mcyB in mRNA level of TH1 is 5.9% of Microcystis aeruginosa.It is concluded that two isolated algae strains from the Meiliang Bay of Lake Taihu are Microcystis and one of them has stronger ability of MC-LR production.Water pollution of toxic Microcystis exists in the area.