水体富营养化,由于工业和农业废水的排放,导致生态退化。然而,到目前为止,如何评估和管理水污染的风险,尤其是氮污染,仍然是一个特别值得注意的问题。硝酸盐,最重要的含氮化合物,已成为制约全氮管理的瓶颈。监测硝酸盐污染的生物记者的发展有助于估计水质,尤其是营养的可用性。在这项研究中,我们从基于不同宿主构建的40种蓝藻衍生物中获得了9种生物报告基因,副本编号,和传感元件,并评估了生物报道分子的性能。结果表明,单细胞集胞藻对硝酸盐的敏感性高于丝状鱼,在大多数生物报告基因中,报道基因luxABCDE的反应比sfgfp快,相对中等拷贝的质粒提高了传感元件的性能。九个生物报告基因在生物利用硝酸盐检测中表现良好,其中AD-AS-X和AR-NI-X,通过硝酸盐填充活化,具有最短的响应时间(2小时)和最宽的响应范围(20-800μM),分别。此外,SR-GLN-SG,硝酸盐缺乏激活,表现出最佳的线性响应(R2=0.998)。参数优化后,指数生长期生物记者,培养温度为30°C,将200μL的样品体积确定为最佳监测条件。我们发现常见的水污染物(铜,镉,和磷)对生物记者的性能没有影响,表明生物记者的稳定性。9名生物记者中有6名,尤其是SR-NB-X,在检测废水样品中的生物可利用硝酸盐方面非常有效。该研究为开发更多蓝藻生物分子及其在硝酸盐检测中的实际应用提供了有价值的参考。
The water eutrophication, resulting from the discharge of industrial and agricultural wastewater, leads to ecological degradation. However, to date, how to assess and manage the risks of water pollution, especially nitrogen pollution, remains a particularly noteworthy issue. Nitrate, the most important nitrogen compound, has become a bottleneck restricting total nitrogen management. The development of bioreporters monitoring nitrate pollution contributes to the estimation of water quality, especially the availability of nutrients. In this study, we obtained 9 bioreporters from 40 cyanobacterial derivatives which were constructed based on different hosts, copy numbers, and sensing elements and evaluated the performance of bioreporters. The results showed that single-celled Synechocystis was more sensitive to nitrate than filamentous Anabaena, that the reporter gene luxABCDE responded faster than sfgfp in most bioreporters, and that relatively medium-copy plasmid improved the performance of sensing elements. Nine bioreporters performed well in bioavailable nitrate detection, of which AD-AS-X and AR-NI-X, activated by nitrate repletion, had the shortest response time (2 h) and the widest response range (20-800 μM), respectively. Moreover, SR-GLN-SG, activated by nitrate deficiency, exhibited the best linear response (R2 = 0.998). After parameter optimization, exponential growth phase bioreporters, culture temperature of 30 °C, sample volume of 200 μL were determined as optimal monitoring conditions. We found that common water contaminants (copper, cadmium, and phosphorus) had no impact on the performance of bioreporters, indicating the stability of bioreporters. Six out of 9 bioreporters, especially the SR-NB-X, were highly effective in detecting the bioavailable nitrate in wastewater sample. This study provides valuable references for developing more cyanobacterial bioreporters and their practical application in nitrate detection.