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Applied and Environmental Microbiology, December 2008, p. 7750-7758, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00982-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Population Heterogeneity of Lactobacillus plantarum WCFS1 Microcolonies in Response to and Recovery from Acid Stress{triangledown}

Colin J. Ingham,1,2* Marke Beerthuyzen,1,3 and Johan van Hylckama Vlieg1,3

Top Institute Food and Nutrition, Nieuwe Kanaal 9A, 6709 PA Wageningen, The Netherlands,1 Wageningen University and Research Centre, Laboratory of Microbiology, P.O. Box 8033, 6700 EJ Wageningen, The Netherlands,2 NIZO food research, P.O. Box 20, 6710 BA Ede, The Netherlands3

Received 30 April 2008/ Accepted 15 October 2008

Within an isogenic microbial population in a homogenous environment, individual bacteria can still exhibit differences in phenotype. Phenotypic heterogeneity can facilitate the survival of subpopulations under stress. As the gram-positive bacterium Lactobacillus plantarum grows, it acidifies the growth medium to a low pH. We have examined the growth of L. plantarum microcolonies after rapid pH downshift (pH 2 to 4), which prevents growth in liquid culture. This acidification was achieved by transferring cells from liquid broth onto a porous ceramic support, placed on a base of low-pH MRS medium solidified using Gelrite. We found a subpopulation of cells that displayed phenotypic heterogeneity and continued to grow at pH 3, which resulted in microcolonies dominated by viable but elongated (filamentous) cells lacking septation, as determined by scanning electron microscopy and staining cell membranes with the lipophilic dye FM4-64. Recovery of pH-stressed cells from these colonies was studied by inoculation onto MRS-Gelrite-covered slides at pH 6.5, and outgrowth was monitored by microscopy. The heterogeneity of the population, calculated from the microcolony areas, decreased with recovery from pH 3 over a period of a few hours. Filamentous cells did not have an advantage in outgrowth during recovery. Specific regions within single filamentous cells were more able to form rapidly dividing cells, i.e., there was heterogeneity even within single recovering cells.

* Corresponding author. Mailing address: Wageningen University and Research Centre, Laboratory of Microbiology, P.O. Box 8129, 6700 EV Wageningen, The Netherlands. Phone: 31 317 482105. Fax: 31 317 484978. E-mail: colin.ingham{at}wur.nl

{triangledown} Published ahead of print on 24 October 2008.

Applied and Environmental Microbiology, December 2008, p. 7750-7758, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00982-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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