Transcriptome sequencing of non-model organisms is valuable resource for the genetic basis of ecological-meaningful traits. The Royal Irises, Iris section Oncocyclus, are a Middle-East group of species in the course of speciation. The species are characterized with extremely large flowers, a huge range of flower colors and a unique pollination system. The Royal Irises serve as a model for evolutionary processes of speciation and plant ecology. However, there are no transcriptomic and genomic data for molecular characterization. Here we describe the de novo transcriptome sequencing and assembly of Iris atropurpurea of the Royal Irises. We employed RNA-seq to analyze the transcriptomes of root, leaf and three stages of flower development. We generated over 195 million paired-end sequencing reads. De novo assembly yielded 184,341 transcripts with an average length of 535 bp. At the protein level, a total of 28,709 Iris transcripts showed significant similarity with known proteins from UniProt database. Orthologues of key flowering genes and genes related to pigment synthesis were identified, showing differential expression in different tissues. In addition, we identified 1,503 short sequence repeats that can be developed for molecular markers for population genetics in irises. In the era of large genetic datasets, the Iris transcriptome sequencing provides valuable resource for studying adaptation-associated traits in this non-model plant. Although intensive eco-evolutionary studies, this is the first reported transcriptome for the Royal Irises. The data available from this study will facilitate gene discovery, functional genomic studies and development of molecular markers in irises, and will provide genetic tools for their conservation.