Identification of abiotic stress responsive genes: A genome wide analysis of the cytokinin response regulator gene family in rice.

Response regulators (RRs), which are implicated in various developmental processes as well as environmental responses by acting as either positive or negative regulators, are crucial components of cytokinin signaling in plants. We characterized 36 RRs using in silico and computational analyses of publicly available data. A comprehensive analysis of OsRR family members was performed covering their physicochemical properties, chromosomal distribution, subcellular localization, phylogeny, gene structure, distribution of conserved motifs and domains, and gene duplication events. Gene Ontology analysis results indicate that 22 OsRR genes contribute mainly to the cytokinin-response and signal transduction. Predicted cis-elements in RRs promoter sequences related to phytohormones and abiotic stresses indicate that RRs are involved in hormonal and environmental responses as described in previous studies. MicroRNA (miRNA) target analysis showed that 148 miRNAs target 29 OsRR genes. In some cases, those RRs are targets of the same miRNA group, and may be controlled by common stimulus responses. Based on the analysis of publicly available gene expression data, OsRR4, OsRR6, OsRR9, OsRR10, OsRR22, OsPRR73, and OsPRR95 were found to be involved in response to abiotic stresses. Using quantitative reverse transcription polymerase chain reaction (qPCR) we confirmed that those RRs, namely OsRR4, OsRR6, OsRR9, OsRR10, OsRR22, and OsPRR73, are involved in the response to salinity, osmotic, alkaline and wounding stresses, and can potentially be used as models to understand molecular mechanisms underlying stress responsiveness.