Genomic information and tools are beginning to be used to increase our understanding of how organisms of all types interact with their environment. The study of the expression of all genes, at the genome, transcriptome, proteome and metabolome level, in response to exposure to a toxicant, is known as toxicogenomics. Here, we show how this new field of environmental genomics has enhanced the development of fundamental knowledge on the mechanisms behind the toxicity of and resistance to the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). Although 2,4-D is one of the most successfully and widely used herbicides, its intensive use has led to the emergence of resistant weeds and might give rise to several toxicological problems when present in concentrations above those recommended. This review summarizes recent mechanistic insights into 2,4-D toxicity and the corresponding adaptive responses based on studies carried out using Saccharomyces cerevisiae and Arabidopsis thaliana as model organisms.