Luria-Delbruck experiment

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Luria-Delbruck experiment (1943) (also called the Fluctuation Test) demonstrates that in bacteria, genetic mutations arise in the absence of selection, rather than being a response to selection. Therefore, Darwin's theory of natural selection acting on random mutations applies to bacteria as well as to higher organisms. These experiments also introduced the microbiological method of replica plating. Max Delbruck and Salvador Luria won the 1969 Nobel Prize in part for this work.

In their experiment, Luria and Delbruck inoculated a small number of bacteria into separate culture tubes. After a period of growth, they plated equal volumes of these separate cultures onto phage (virus) containing agar. If virus resistance in bacteria were caused by a spontaneous activation in bacteria, then each plate should contain the same number of resistant colonies. This, however was not what Delbruck and Luria found. Instead, the number of resistant colonies on each plate varied drastically.

Luria and Delbruck proposed that these results could be explained by the occurrence of a constant rate of random mutations in each generation of bacteria growing in the initial culture tubes. Delbruck developed a sophisticated mathematical model based on this hypothesis that was entirely consistent with these results. The conclusion was that mutations in bacteria, like higher organisms, are random rather than directed.

The results of Luria and Delbruck were confirmed in more graphical, but less quantitative, way by Newcombe (1949). Newcombe incubated bacteria in a Petri dish for a few hours, then replica plated it onto two new Petri dishes treated with phage. The first plate is left unspread, and the second plate was then respread. Both plates were incubated for growth. There were as much as 50 times greater number of bacterial colonies on the respread dish. This showed that bacterial mutations to virus resistance had randomly occurred during the first incubation. Once again, the mutations occurred before selection was applied.

In recent years, the results of Luria and Delbruck were questioned by Cairns and others (Cairns et al. 1988), who studied mutations in sugar metabolism as a form of environmental stress. However, eventually this result was found to have been caused by selection for gene amplification and/or a higher mutation rate in cells unable to divide. Nevertheless, there is no directed mutagenesis: only those mutations that allow the cells to respond to the environmental stress accumulate in a growing population (Slechta et al. 2002).

References

Cairns, J. Overbaugh, J. & Miller, S. (1988) The Origin of Mutants. Nature, 335: 142-145.
Foster, P. 2000. Sorting out mutation rates. Proc. Natl. Acad. Sci. USA 96: 7617-7618.
Luria, SE, Delbruck, M. (1943) "Mutations of Bacteria from Virus Sensitivity to Virus Resistance" Genetics 28:491-511.
Newcombe, H.B. (1949) "Origin of Bacterial Variants." Nature. 164:150-151.
Slechta ES, Liu J, Andersson DI, Roth JR. "Evidence that selected amplification of a bacterial lac frameshift allele stimulates Lac(+) reversion (adaptive mutation) with or without general hypermutability." Genetics. 2002 Jul;161(3):945-56.

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