Abstract
A method for estimating the oxygen availability in plant cell cultures grown in stationary liquid media (e.g. many protoplast cultures) was developed. The method is based on short-term measurements of respiration rate versus oxygen concentration on a sample of cells, suspended in liquid media. From such data it is possible to estimate the oxygen concentration at the bottom of a stagnant liquid culture, by calculating the amount of oxygen reaching the cells by diffusion. As an example, rape (Brassica napus L. cv. Omega) hypocotyl protoplasts were grown with different oxygen concentrations at the site of the cells, obtained by varying the cell density, the height of the liquid layer and the oxygen content of the gas phase. The number of surviving calli was positively correlated with the estimated oxygen availability in the range between 60 and 350 μM O2, below 60 μM all cells died. This indicates that oxygen availability can be a limiting factor in the range usually encountered in protoplast cultures, and that the method can be useful when designing optimal growth conditions for stationary cultures of plant cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- C1 :
-
bulk oxygen concentration in agitated medium
- Co :
-
oxygen concentration in medium at the gas-liquid interface, in equilibrium with the gas
- Cx :
-
oxygen concentration at cell level
- D:
-
diffusion constant of oxygen in water
- KLa:
-
oxygen transfer rate
- l:
-
height of liquid above cells
- n:
-
number of cells per ml
- Rx :
-
respiration rate per cell
References
Christen AA & Gibson DM (1987) A method for growing stationary plant suspension cell cultures. In Vitro Cell & Dev. Bio. 23: 315–316
Delieu T & Walker DA (1972) An improved cathode for the measurement of photosynthetic oxygen evolution by isolated chloroplasts. New Phytol. 71: 201–225
Ducos JP & Pareilleux A (1986) Effect of aeration rate and influence of pCO2 in large-scale cultures of Catharanthus roseus cells. Appl. Microbiol. Biotechnol. 25: 101–105
Edwards DK, Denny VE & Mills AF (1979) Transfer Processes: An Introduction to Diffusion, Convection and Radiation (pp 67) (equation 3.32. Hemisphere Publishing Corporation, Washington, New York, London
Elmsheuser HA (1977) Untersuchungen über die vegetative Vermehrung von Brassicaceen mittels Gewebekulturen-insbesondere von Raps (Brassica napus L. ssp. oleifera Metzg. Sinsk.). Thesis, Justus Liebig-Univ. Giessen, FRG, pp 28
Glimelius K (1984) High growth rate and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol. Plant 61: 38–44
Hegarty PK, Smart NJ, Scragg AH & Fowler MW (1986) The aeration of Catharanthus roseus L.G. Don suspension cultures in airlift bioreactors: The inhibitory effect at high aeration rates on culture growth. J. Exp. Bot. 37: 1911–1920
Journet E-P, Bligny R & Douce R (1986) Is the availability of substrate for the tricarboxylic acid cycle a limiting factor for uncoupled respiration in sycamore (Acer pseudoplatanus) cells? Biochem. J. 233: 571–576
Kato A, Shimizu Y & Nagai S (1975) Effect of initial kL a on the growth of tobacco cells in batch culture. J. Ferment. Technol. 53: 744–751
Kessell RHJ & Carr AH (1972) The effect of dissolved oxygen concentration on growth and differentiation of carrot (Daucus carota) tissue. J. Exp. Bot. 23: 996–1007
Millington RJ (1955) Diffusion constant and diffusion coefficient. Science 122: 1090–1091
Öpik H (1980) The Respiration of Higher Plants. Edward Arnold Publishers, London
Power JB & Chapman JV (1985) Isolation, culture and genetic manipulation of protoplasts. In: Dixon RA (Ed) Plant Cell Culture: A Practical Approach. IRL Press, Oxford
Smart NJ & Fowler MW (1981) Effect of aeration on largescale cultures of plant cells. Biotechnol. Lett. 3: 171–176
Snape JB & Thomas NH (1989) How suspension cultures of Catharanthus roseus respond to oxygen limitation: Small-scale tests with applications to large-scale cultures. Biotechnol. Bioeng. 34: 1058–1062
van der Plas LHW & Wagner MJ (1986) Effect of oxygen on growth and respiration of potato tuber callus. Plant Cell Tiss. Org. Cult. 7: 217–225
Verma DPS & Marcus A (1974) Oxygen availability as a control factor in the density-dependent regulation of protein synthesis in cell culture. J. Cell Sci. 41: 331–337
Wimpenny JWT & Cooms JP (1983) Penetration of oxygen into bacterial colonies. J. Gen. Microbiol. 129: 1239–1242
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Brandt, K. A method for estimating oxygen availability of stationary plant cell cultures in liquid media. Plant Cell Tiss Organ Cult 26, 195–201 (1991). https://doi.org/10.1007/BF00039944
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00039944