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In vitro , a new protocol of plant regeneration in rose was achieved via protocorm-like bodies (PLBs) induced from the root-like organs named rhizoids that developed from leaf explants. The development of rhizoids is a critical stage for efficient regeneration, which is triggered by exogenous auxin. However, the role of cytokinin in the control of organogenesis in rose is as yet uncharacterized. The aim of this study was to elucidate the molecular mechanism of cytokinin-modulated rhizoid formation in  Rosa canina.  Here, we found that cytokinin is a key regulator in the formation of rhizoids. Treatment with cytokinin reduced callus activity and significantly inhibited rhizoid formation in  Rosa canina . We further isolated the full-length cDNA of a type-A response regulator gene of cytokinin signaling,  RcRR1 , from which the deduced amino acid sequence contained the conserved DDK motif. Gene expression analysis revealed that  RcRR1  was differentially expressed during rhizoid formation and its expression level was rapidly up-regulated by cytokinin. In addition, the functionality of  RcRR1  was tested in  Arabidopsis . RcRR1 was found to be localized to the nucleus in  GFP-RcRR1  transgenic plants and overexpression of  RcRR1  resulted in increased primary root length and lateral root density. More importantly,  RcRR1  overexpression transgenic plants also showed reduced sensitivity to cytokinin during root growth; auxin distribution and the expression of auxin efflux carriers  PIN  genes were altered in  RcRR1  overexpression plants. Taken together, these results demonstrate that RcRR1 is a functional type-A response regulator which is involved in cytokinin-regulated rhizoid formation in  Rosa canina .

RcRR1, a Rosa canina Type-A Response Regulator Gene, Is Involved in Cytokinin-Modulated Rhizoid Organogenesis