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Journal of Bacteriology, January 1999, p. 270-283, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Adaptation of Mycobacterium smegmatis to
Stationary Phase
Marjan J.
Smeulders,
Jacquie
Keer,
Richard A.
Speight,
and
Huw D.
Williams*
Department of Biology, Imperial College of
Science, Technology and Medicine, London SW7 2AZ, United Kingdom
Received 25 August 1998/Accepted 28 October 1998
Mycobacterium tuberculosis can persist for many years
within host lung tissue without causing clinical disease. Little is known about the state in which the bacilli survive, although it is
frequently referred to as dormancy. Some evidence suggests that cells
survive in nutrient-deprived stationary phase. Therefore, we are
studying stationary-phase survival of Mycobacterium
smegmatis as a model for mycobacterial persistence. M. smegmatis cultures could survive 650 days of either carbon,
nitrogen, or phosphorus starvation. In carbon-limited medium, cells
entered stationary phase before the carbon source (glycerol) had been
completely depleted and glycerol uptake from the medium continued
during the early stages of stationary phase. These results suggest that the cells are able to sense when the glycerol is approaching limiting concentrations and initiate a shutdown into stationary phase, which
involves the uptake of the remaining glycerol from the medium. During
early stationary phase, cells underwent reductive cell division and
became more resistant to osmotic and acid stress and pool mRNA
stabilized. Stationary-phase cells were also more resistant to
oxidative stress, but this resistance was induced during late
exponential phase in a cell-density-dependent manner. Upon recovery in
fresh medium, stationary-phase cultures showed an immediate increase in
protein synthesis irrespective of culture age. Colony morphology
variants accumulated in stationary-phase cultures. A flat colony
variant was seen in 75% of all long-term-stationary-phase cultures and
frequently took over the whole population. Cryo scanning electron
microscopy showed that the colony organization was different in flat
colony strains, flat colonies appearing less well organized than
wild-type colonies. Competition experiments with an
exponential-phase-adapted wild-type strain showed that the flat strain
had a competitive advantage in stationary phase, as well a providing
evidence that growth and cell division occur in stationary-phase
cultures of M. smegmatis. These results argue against
stationary-phase M. smegmatis cultures entering a quiescent
state akin to dormancy but support the idea that they are a dynamic
population of cells.
*
Corresponding author. Mailing address: Department of
Biology, Imperial College of Science, Technology and Medicine, Sir
Alexander Fleming Building, Imperial College Rd., London SW7 2AZ,
United Kingdom. Phone: 44 (171) 5945383. Fax: 44 (171) 5842056. E-mail: h.d.williams{at}ic.ac.uk.

Present address: National Institute for Medical Research, London
NW7 1AA, United
Kingdom.
Journal of Bacteriology, January 1999, p. 270-283, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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