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Evolutionary physiology merges the disciplines of evolution and physiology, and it is a research approach that has not received much attention for studying the development of herbicide resistance. This paper makes a case for using evolutionary physiology more frequently when studying herbicide resistance, and illustrates this using three areas where more work would be useful: (i) the interaction among major and minor alleles over many generations during the evolution of physiological responses that lead to specific mechanisms of resistance; (ii) the role of epigenetic factors, especially at an early stage of evolution, on the physiological modifications that result in phenotypes that become insensitive to herbicides; and (iii) the interaction between fitness and physiological performance over time, with emphasis on understanding mechanisms that improve the fitness of herbicide-resistant phenotypes during selection.