Phosphate (Pi) limitation is one of the major factors negatively impacting crop yield worldwide. Next generation sequencing (NGS) was used to profile the transcriptomes of rice (Oryza sativa) roots and shoots after phosphate starvation and recovery, shedding further light on the complex and dynamic mechanisms involved in Pi homeostasis. The use of NGS also enabled the identification of previously not annotated loci and novel isoforms of genes that are specifically induced by Pi starvation. Furthermore, phosphate re-feeding was observed to have a unique response with a variety of transcription factors and kinases induced in a transient manner. Expression profiles of miRNAs were also assessed upon longterm Pi starvation in roots and shoots revealing several novel miRNAs associated with Pi starvation. Altogether, this study provides key findings regarding Pi homeostasis in plants that will provide a valuable resource for research aimed at generating crops with increased Pi acquisition/ use efficiency. © 2014 Landes Bioscience.