Confounders of mutation-rate estimators: Selection and phenotypic lag in Thermus thermophilus

• The measurement of microbial mutation rates is peppered with hazards.
• These include diverse kinds of phenotypic lag and differential growth.
• Ways are described to counter such problems.
• These methods are applied to the case of Thermus thermophilus.
• The methods are generally applicable.

In a recent description of the rate and character of spontaneous mutation in the hyperthermophilic bacterium Thermus thermophilus, the mutation rate was observed to be substantially lower than seen in several mesophiles. Subsequently, a report appeared indicating that this bacterium maintains an average of about 4.5 genomes per cell. This number of genomes might result in a segregation lag for the expression of a recessive mutation and might therefore lead to an underestimate of the rate of mutation. Here we describe some kinds of problems that may arise when estimating mutation rates and outline ways to adjust the rates accordingly. The emphasis is mainly on differential rates of growth of mutants versus their parents and on various kinds of phenotypic lag. We then apply these methods to the T. thermophilus data and conclude that there is as yet no reliable impact on a previously described rate.