Regions traditionally destined to agriculture report an ever increasing exposure to cold and drought conditions. This is especially important in countries like Chile where crop management options are limited. The development of new cultivars with better yields under adverse conditions is fundamental if the ever increasing demand for food is to be matched; however, improving tolerance to abiotic stresses has proved to be a complex task. In this regard, development in plant physiology and genomics in the last 20 years has led to a deeper understanding of how plants respond to stress and mechanisms responsible for different ranges of tolerance observed in nature. This review discusses the techniques currently most in use in gene expression analysis, together with some important experimental design variables, such as the developmental stage of the plant, stress intensity and duration, and how different stresses may interact when performing assays. On the other hand, it is fundamental to properly select gene expression techniques according to the available information on the genome, the crop and the final objective of the research. All these points must be considered to ease transition from genomics to practical applications to crop species in order to increase their tolerance to stress. In this regard, the rapid development of new techniques in gene expression analysis with lower costs will determine a new revolution in crop research in coming decades. Therefore, Chile needs to be prepared in this area to continue its development as a major food producer worldwide.