Luciferase activity assays to evaluate the regulation of specific promoters from tomato and Arabidopsis

Abstract

Fundamental questions in plant biology including how cells and tissues maintain their identity over time and how they switch from one stable state to the next. Understanding such basic questions may help us to better predict how plants modulate their shoot architecture in response to endogenous cues and the environment. Shoot architecture is one of the main developmental factors affecting plant productivity. Predictable, plants have evolved intricate regulatory networks to module their shoot architecture, which include phytohormones, transcription factor-regulated transcriptional programming, and epigenetic factors like microRNAs. However, how these different factors integrate to control shoot architecture at molecular and cellular levels is still unclear in most species. Following our findings that microRNAs (miRNAs) can regulate different aspects of sugarcane and tomato development by interacting with phytohormones, the characterization of miRNA-based patterning mechanisms and their association with phytohormones is a central aspect of our work. For example, we are studying the interaction between the gibberellin-negative regulator DELLA with miRNA-controlled pathways during flower and shoot development. Moreover, we are evaluating the interactions between phytohormones and microRNA modules in the control of axillary shoot branching, which directly influences shoot architecture. To that end, it is fundamental to study the interaction between transcription factors and target genes. Thus, the main objective of this project is to establish luciferase activity assays to evaluate the activity of specific promoters (fused to luciferase) regulated by selected transcription factors. (AU)