实时荧光定量PCR技术在太湖蓝藻监测和评估中的应用
Application of Quantitative PCR Technology in Biomonitoring and Assessment of Harmful Cyanobacteria in Lake Tai
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摘要: 我国东部地区的湖泊深受蓝藻水华的影响，监测可产生毒素的藻种类及其浓度对于水质安全预警与管理有重要意义。然而传统形态学方法无法鉴别藻类产生毒素及嗅味物质的潜在趋势。为综合评估湖泊及其水源水体的藻类有毒代谢物发生风险，采用实时定量聚合酶链式反应（quantitative real-time polymerase chain reaction，qPCR）方法对太湖产毒微囊藻基因、产毒柱孢藻基因和产2-甲基异莰醇基因进行监测，同时采用盘式酶联免疫法（enzyme-linked immuno-sorbent assay，ELISA）以及气相色谱-质谱法分析微囊藻毒素、柱孢藻毒素和2-甲基异莰醇浓度，并进行相关性分析。结果表明，太湖水体各项蓝藻产毒基因水平与相关藻毒素浓度具有良好相关性。由蓝藻产毒基因/藻种基因丰度的比值可知，高丰度的微囊藻菌株不具有产藻毒素的潜力，而绝大多数柱孢藻都含有产毒素的基因，表明采用产毒基因的拷贝数预测藻种的产毒能力更准确。不同类型的产毒藻基因在太湖具有显著的时空分布差异，产毒微囊藻基因在9月达到浓度峰值，在太湖西部浓度更高，而产毒柱孢藻和产2-甲基异莰醇基因则在7-8月达到峰值，在中部和东部维持较高浓度。太湖的饮用水源地2-甲基异莰醇风险显著高于微囊藻毒素和柱孢藻毒素。因此，分子生物监测和评估可为蓝藻预警与管理提供重要支撑。Abstract: The lakes in eastern China have been heavily affected by cyanobacterial blooms. Monitoring the toxic algae species and concentrations of their corresponding toxic metabolites are pre-requirement for water quality management. However, traditional morphological methods are limited in the identification of the potential trend of algae producing toxins and odorants. To comprehensively assess the risk of toxic metabolites of algae in Lake Tai, the real-time quantitative polymerase chain reaction (qPCR) assays were developed for quantifying potentially toxigenic Microcystis, cylindrospermopsin-producing and 2-methylisoborneol (2-MIB)-synthesis cyanobacteria. Furthermore, the concentrations of corresponding toxins were measured by enzyme-linked immuno-sorbent assay and gas chromatography-mass spectrometry method. The results showed statistically significant correlations between the levels of toxigenic genes and the concentrations of corresponding toxins in Lake Tai. The low ratio of the gene copies from toxic Microcystis to that from total Microcystis showed that the high abundance of Microcystis strains weren't potential to produce toxins. However, most of Cylindrospermopsis cells were potentially toxigenic as showed by the data. It is indicated that the copy numbers of toxin producing genes are accurate enough to diagnose the presence of harmful cyanobacteria and the potential presence of cyanotoxins in Lake Tai. Significant differences in the temporal and spatial distribution of toxigenic genes were observed for different types of toxic algae. The gene copies of toxigenic Microcystis reached the peak in the west of Lake Tai and showed higher abundance in September, while the copies of cylindrospermopsin-producing gene and 2-MIB synthesis gene of cyanobacteria remained high in the middle and east of Lake Tai and reached peaks in July and August. The risk of 2-MIB in drinking water sources of Lake Tai was significantly higher than that of microcystin and cylindrospermopsin. In conclusion, molecular biological monitoring and assessment can provide important powerful tools for cyanobacteria early warning and management.
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Key words:
- cyanotoxin /
- odor compound /
- Lake Tai /
- Microcystis /
- cylindrospermopsin /
- qPCR
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