Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium <italic style="font-style: italic">Photobacterium</italic> <italic style="font-style: italic">phosphoreum</italic> ANT-2200

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

doi:10.1007/s00343-020-9301-8

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

Biology | Updated：2023-05-20

- 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 Vol. 39, Issue 1, Pages: 297-305(2021)
- Affiliations：
  
  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
- Author bio：
  
  Wei-Jia ZHANG, E-mail:wzhang@idsse.ac.cn
  Long-Fei WU, E-mail:wu@imm.cnrs.fr
- Funds：
- DOI：10.1007/s00343-020-9301-8
  CLC：
- Received：13 November 2019，
  
  Accepted：17 December 2019，
  
  Online First：12 February 2020，
  
  Published：2021-01
- Accepted：
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Jie DAI, Hongzhi TANG, Xuegong LI, 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, 2021, 39(1): 297-305.
DOI：

Jie DAI, Hongzhi TANG, Xuegong LI, 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, 2021, 39(1): 297-305. DOI： 10.1007/s00343-020-9301-8.

摘要

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

and hRF in the presence of the reducing compound Na

S 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.

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references

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

DOI：10.1007/s00343-020-9301-8

摘要

Abstract

关键词

Keywords

references