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朝向新一代全细胞生物报告器的发展,以感测海洋系统中的铁生物利用率——从聚球藻 PCC7002 铁生物报告器的案例中学习。

Towards the development of a new generation of whole-cell bioreporters to sense iron bioavailability in oceanic systems-learning from the case of Synechococcus sp. PCC7002 iron bioreporter.

机构信息

Department F.-A. Forel for Environmental and Aquatic Sciences, Faculty of Science, University of Geneva, Geneva, Switzerland.

出版信息

J Appl Microbiol. 2019 Nov;127(5):1291-1304. doi: 10.1111/jam.14277. Epub 2019 Jun 26.

Abstract

Whole-cell bioreporters are genetically modified micro-organisms designed to sense bioavailable forms of nutrients or toxic compounds in aquatic systems. As they represent the most promising cost-efficient tools available for such purpose, engineering and use of bioreporters is rapidly growing in association with wide applicability. Bioreporters are urgently needed to determine phytoplankton iron (Fe) limitation, which has been reported in up to 30% of the ocean, with consequences affecting Earth's global carbon cycle and climate. This study presents a critical evaluation and optimization of the only Cyanobacteria bioreporter available to sense Fe limitation in marine systems (Synechococcus sp. PCC7002). The nonmonotonic biphasic dose-response curve between the bioreporters' signal and Fe bioavailability impairs an appropriate data interpretation, highlighting the need for new carefully designed bioreporters. Here, limitations under low Fe concentrations were related to cellular energy stress, nonlinear expression of the targeted promoter and siderophore expression. Furthermore, we provide critical standard criteria for the development of new Fe bioreporters. Finally, based on gene expression data under a range of marine Fe concentrations, we propose novel sensor genes for the development of new Cyanobacteria Fe bioreporters for distinct marine regions.

摘要

整细胞生物报告器是经过基因改造的微生物,旨在感知水生系统中生物可利用形式的营养物质或有毒化合物。由于它们是目前用于此类目的最有前途的具有成本效益的工具,因此生物报告器的工程设计和使用与广泛的适用性迅速发展。迫切需要生物报告器来确定浮游植物铁(Fe)限制,据报道,海洋中高达 30%的区域存在这种限制,其后果会影响地球的全球碳循环和气候。本研究对目前唯一可用于感测海洋系统中 Fe 限制的蓝细菌生物报告器(聚球藻 PCC7002)进行了批判性评估和优化。生物报告器的信号与 Fe 生物利用度之间的非单调双相剂量反应曲线会影响到对数据的适当解释,这突出了需要新的精心设计的生物报告器。在这里,低 Fe 浓度下的限制与细胞能量应激、靶向启动子的非线性表达和铁载体表达有关。此外,我们还为新的 Fe 生物报告器的开发提供了关键的标准标准。最后,根据一系列海洋 Fe 浓度下的基因表达数据,我们提出了用于开发新的蓝细菌 Fe 生物报告器的新型传感器基因,以用于不同的海洋区域。

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