Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200
Dai Jie1,2,3; Tang Hongzhi1,2,3; Li Xuegong1,2; Santini, Claire-Lise1,5; Cui Wenpeng1,2,3; Liu Na1,2,3; Qi Xiaoqing1,2; Cui Xuehua1,2; Grossi, Alain4; Notareschi, Philippe5
刊名JOURNAL OF OCEANOLOGY AND LIMNOLOGY
2020-04-20
页码9
关键词oxic-reduction indicator absorption spectra bacterial growth marine microorganism
ISSN号2096-5508
DOI10.1007/s00343-020-9301-8
英文摘要

Resazurin (RZ) is a weakly fluorescent blue dye and can be reduced irreversibly to highly fluorescent pink resorufin (RF) that is reduced reversibly to colorless dihydroresorufin (hRF) by photo, deoxygenation, chemical reaction and reductive organic compounds produced through cell metabolism. Because of the reliable and sensitive fluorescence-color change and noninvasive features, RZ has been used widely as a redox indicator in cell viability/proliferation assays for bacteria, yeast, and mammalian cells. However, RZ is used rarely for physiological characterization of marine microorganisms. Here, we developed a custom-made irradiation and absorption-analysis device to assess the reducing capacity and physiologic status of marine bacterial cultures. We measured the absorption spectra of RZ, RF, and hRF in the presence of the reducing compound Na2S and under visible-light irradiation. After establishing appropriate parameters, we monitored the color changes of RZ and its reduced derivatives to evaluate the coherence between reducing capacity, bioluminescence and growth of the deep-sea bacterium Photobacterium phosphoreum strain ANT-2200 under various conditions. Emission of bioluminescence is an oxidation process dependent upon cellular reducing capacity. Growth and bioluminescence of ANT-2200 cell cultures were impeded progressively with increasing concentrations of RZ, which suggested competition for reducing molecules between RZ at high concentration with reductive metabolism. Therefore, caution should be applied upon direct addition of RZ to growth media to monitor redox reactions in cell cultures. Analyses of the instantaneous reduction velocity of RZ in ANT-2200 cell cultures showed a detrimental effect of high hydrostatic pressure and high coherence between the reducing capacity and bioluminescence of cultures. These data clearly demonstrate the potential of using RZ to characterize the microbial metabolism and physiology of marine bacteria.

资助项目National Key R&D Program of China[2016YFC0302502] ; National Key R&D Program of China[2018YFC0309904] ; National Key R&D Program of China[2016YFC0304905] ; NSFC of China[91751202] ; NSFC of China[91751108] ; NSFC of China[41806174] ; Sanya Municipality[2018YD01] ; Sanya Municipality[2018YD02] ; CNRS
WOS关键词ALAMAR BLUE ; REDUCTION ; RESORUFIN
WOS研究方向Marine & Freshwater Biology ; Oceanography
语种英语
出版者SCIENCE PRESS
WOS记录号WOS:000527509100006
资助机构National Key R&D Program of China ; NSFC of China ; Sanya Municipality ; CNRS
内容类型期刊论文
版本出版稿
源URL[http://ir.idsse.ac.cn/handle/183446/7610]  
专题深海科学研究部_深海生物学研究室_深海微生物细胞生物学研究组
通讯作者Zhang Wei-Jia; Wu Long-Fei
作者单位1.Chinese Acad Sci, CNRS, Int Associated Lab Evolut & Dev Magnetotact Multi, Beijing, Peoples R China
2.Chinese Acad Sci, Inst Deep Sea Sci & Engn, Lab Deep Sea Microbial Cell Biol, Sanya 572000, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100864, Peoples R China
4.Aix Marseille Univ, CNRS, IMM, FR3479, F-13402 Marseille, France
5.Aix Marseille Univ, CNRS, LCB, IMM,FR3479,IM2B, F-13402 Marseille, France
推荐引用方式
GB/T 7714
Dai Jie,Tang Hongzhi,Li Xuegong,et al. Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200[J]. JOURNAL OF OCEANOLOGY AND LIMNOLOGY,2020:9.
APA Dai Jie.,Tang Hongzhi.,Li Xuegong.,Santini, Claire-Lise.,Cui Wenpeng.,...&Wu Long-Fei.(2020).Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200.JOURNAL OF OCEANOLOGY AND LIMNOLOGY,9.
MLA Dai Jie,et al."Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200".JOURNAL OF OCEANOLOGY AND LIMNOLOGY (2020):9.
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