To increase both the yield potential and stability of crops, integrated breeding strategies are used that have mostly a direct genetic basis, but the utility of epigenetics to improve complex traits is unclear. A better understanding of the status of the epigenome and its contribution to agronomic performance would help in developing approaches to incorporate the epigenetic component of complex traits into breeding programs. Starting from isogenic canola (Brassica napus) lines, epilines were generated by selecting, repeatedly for three generations, for increased energy use efficiency and drought tolerance. These epilines had an enhanced energy use efficiency, drought tolerance, and nitrogen use efficiency. Transcriptome analysis of the epilines and a line selected for its energy use efficiency solely revealed common differentially expressed genes related to the onset of stress tolerance-regulating signaling events. Genes related to responses to salt, osmotic, abscisic acid, and drought treatments were specifically differentially expressed in the droughttolerant epilines. The status of the epigenome, scored as differential trimethylation of lysine-4 of histone 3, further supported the phenotype by targeting drought-responsive genes and facilitating the transcription of the differentially expressed genes. From these results, we conclude that the canola epigenome can be shaped by selection to increase energy use efficiency and stress tolerance. Hence, these findings warrant the further development of strategies to incorporate epigenetics into breeding.The need to improve crop yield in both quantity (yield potential) and stability (actual yield) to meet the increasing demand for food, feed, and plant-derived materials is a major challenge. Very different complementary technologies are used to optimize yield and to develop crops with increased resilience against adverse environmental growth conditions (Botella et al., 2008;Cattivelli et al., 2008; Jhaet al., 2014). Plant breeding programs are a basic component of this improvement process encompassing a wide range of technologies, such as exploration of the genetic potential by intraspecific and interspecific crosses, combination of genetic pools in hybrid breeding, mutational breeding, molecular breeding, and transgene technologies.Rather recently, epigenetics has been investigated as a potential breeding platform (Springer, 2013). Epigenetic variations or heritable changes in gene expression that are not linked to changes in the DNA sequence, but associated with differences in DNA methylation or histone modifications, provide an alternative source of phenotypic variability. A vast and growing number of studies implicate DNA methylation and histone modification in the modulation of gene expression in general, control of developmental transitions, and plant responses to biotic 1 This work was supported by the Agency for Innovation by Science and Technology (grant no. IWT100268), the Ghent University Special Research Fund (grant no. 01J11311), the European Training and...