Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium <i>Photobacterium</i> <i>phosphoreum</i> ANT-2200 -- Journal of Oceanology and Limnology,2021,39(1):297-305

	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 DAI^1,2,3, Hongzhi TANG^1,2,3, Xuegong LI^1,3, Claire-Lise SANTIN^3,4, Wenpeng CUI^1,2,3, Na LIU^1,2,3, Xiaoqing QI^1,3, Xuehua CUI^1,3, Alain GROSSI⁵, Philippe NOTARESCHI⁴, Wei-Jia ZHANG^1,3, Long-Fei WU^3,4

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

Abstract:

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

Articles by Xuegong LI

Articles by Claire-Lise SANTIN

Articles by Wenpeng CUI

Articles by Na LIU

Articles by Xiaoqing QI

Articles by Xuehua CUI

Articles by Alain GROSSI

Articles by Philippe NOTARESCHI

Articles by Wei-Jia ZHANG

Articles by Long-Fei WU

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