Cite this paper:
Jie DAI, Hongzhi TANG, Xuegong LI, Claire-Lise SANTIN, Wenpeng CUI, Na LIU, Xiaoqing QI, Xuehua CUI, Alain GROSSI, Philippe NOTARESCHI, Wei-Jia ZHANG, Long-Fei WU. 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, 2021, 39(1): 297-305

Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200

Jie DAI1,2,3, Hongzhi TANG1,2,3, Xuegong LI1,3, Claire-Lise SANTIN3,4, Wenpeng CUI1,2,3, Na LIU1,2,3, Xiaoqing QI1,3, Xuehua CUI1,3, Alain GROSSI5, Philippe NOTARESCHI4, Wei-Jia ZHANG1,3, Long-Fei WU3,4
1 Laboratory of Deep Sea Microbial Cell Biology, Institute of Deep Sea Science and Engineering, Chinese Academy of Sciences, Sanya 572000, China;
2 University of Chinese Academy of Sciences, Beijing 100049, China;
3 International Associated Laboratory of Evolution and Development of Magnetotactic Multicellular Organisms (LIA-MagMC), CNRS-CAS;
4 Aix Marseille University, CNRS, LCB, IMM, FR3479, IM2B, Marseille 13402, France;
5 Aix Marseille University, CNRS, IMM, FR3479, Marseille 13402, France
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 photodeoxygenation, 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.
Key words:    oxic-reduction indicator|absorption spectra|bacterial growth|marine microorganism   
Received: 2019-11-13   Revised: 2019-12-17
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Articles by Jie DAI
Articles by Hongzhi TANG
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Articles by Xiaoqing QI
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Articles by Philippe NOTARESCHI
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