A 'fragile cell' sub-population revealed during cytometric assessment of Saccharomyces cerevisiae viability in lipid-limited alcoholic fermentation.
OBJECTIVE
To show that in anaerobic fermentation with limiting lipid nutrients, cell preparation impacts the viability assessment of yeast cells, and to identify the factors involved.


RESULTS
Saccharomyces cerevisiae viability was determined using propidium iodide staining and the flow cytometry. Analyses identified intact cells, dead cells and, under certain conditions, the presence of a third subpopulation of apparently damaged cells. This intermediate population could account for up to 40% of the entire cell population. We describe, analyse and discuss the effects of different solutions for cell resuspension on the respective proportion of these three populations, in particular that of the intermediate population. We show that this intermediate cell population forms in the absence of Ca(2+)/Mg(2+).


CONCLUSIONS
Cell preparation significantly impacts population viability assessment by FCM. The intermediate population, revealed under certain conditions, could be renamed as 'fragile cells'. For these cells, Ca(2+) and Mg(2+) reduce cell membrane permeability to PI.


CONCLUSIONS
This is the first study that analyses and discusses the factors influencing the formation of an intermediate population when studying viability in yeast alcoholic fermentation. With a wider application in biological research, this study provides important support to the relatively new questioning of propidium iodide staining as a universal cell death indicator.
INRA, UMR1083, Sciences pour l'Oenologie, Montpellier, France SupAgro, UMR1083, Sciences pour l'Oenologie, Montpellier, France.
P
P
Delobel
INRA, UMR1083, Sciences pour l'Oenologie, Montpellier, France SupAgro, UMR1083, Sciences pour l'Oenologie, Montpellier, France.
M
M
Pradal
INRA, UMR1083, Sciences pour l'Oenologie, Montpellier, France SupAgro, UMR1083, Sciences pour l'Oenologie, Montpellier, France.
B
B
Blondin
INRA, UMR1083, Sciences pour l'Oenologie, Montpellier, France SupAgro, UMR1083, Sciences pour l'Oenologie, Montpellier, France.
C
C
Tesniere
D000693:Anaerobiosis / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods; D050356:Lipid Metabolism; D008274:Magnesium / Q000378:metabolism; D011419:Propidium / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000254:growth & development* / Q000378:metabolism*
D011419:Propidium; D000431:Ethanol; D008274:Magnesium; D002118:Calcium