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Journal of Bacteriology, April 2003, p. 2131-2142, Vol. 185, No. 7
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.7.2131-2142.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genomic Diversification among Archival Strains of Salmonella enterica Serovar Typhimurium LT7

Gui-Rong Liu,1,2 Kelly Edwards,3 Abraham Eisenstark,3 Ying-Mei Fu,4 Wei-Qiao Liu,5 Kenneth E. Sanderson,5 Randal N. Johnston,6 and Shu-Lin Liu1,2*

Departments of Microbiology and Infectious Diseases,1 Biological Sciences,5 Biochemistry and Molecular Biology, University of Calgary, Calgary, T2N 4N1 Alberta, Canada,6 Cancer Research Center and University of Missouri, Columbia, Missouri 65201,3 Department of Microbiology, Harbin Medical University, Harbin 150086 ,4 Department of Microbiology, Peking University School of Basic Medical Sciences, Beijing 100083, China2

Received 6 May 2002/ Accepted 7 January 2003

To document genomic changes during long periods of storage, we analyzed Salmonella enterica serovar Typhimurium LT7, a mutator strain that was previously reported to have higher rates of mutations compared to other serovar Typhimurium strains such as LT2. Upon plating directly from sealed agar stabs that had been stocked at room temperature for up to four decades, many auxotrophic mutants derived from LT7 gave rise to colonies of different sizes. Restreaking from single colonies consistently yielded colonies of diverse sizes even when we repeated single-colony isolation nine times. Colonies from the first plating had diverse genomic changes among and even within individual vials, including translocations, inversions, duplications, and point mutations, which were detected by rare-cutting endonuclease analysis with pulsed-field gel electrophoresis. Interestingly, even though the colony size kept diversifying, all descendents of the same single colonies from the first plating had the same sets of detected genomic changes. We did not detect any colony size or genome structure diversification in serovar Typhimurium LT7 stocked at -70°C or in serovar Typhimurium LT2 stocked either at -70°C or at room temperature. These results suggest that, although colony size diversification occurred during rapid growth, all detected genomic changes took place during the storage at room temperature and were carried over to their descendents without further changes during rapid growth in rich medium. We constructed a genomic cleavage map on the LT7 strain that had been stocked at -70°C and located all of the detected genomic changes on the map. We speculated on the significance of mutators for survival and evolution under environmentally stressed conditions.

* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, University of Calgary, 3330 Hospital Dr., NW, Calgary, Alberta T2N 4N1, Canada. Phone: (403) 220-3799. Fax: (403) 270-0834. E-mail: slliu{at}ucalgary.ca.

Journal of Bacteriology, April 2003, p. 2131-2142, Vol. 185, No. 7
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.7.2131-2142.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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