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Genes encoding pseudo-response regulator (PRR) proteins play significant roles in plant circadian clocks. In this study, four genes related to flowering time were isolated from  Chrysanthemum morifolium . Phylogenetic analysis showed that they are highly homologous to the counterparts of PRRs of  Helianthus annuus  and named as  CmPRR2 ,  CmPRR7 ,  CmPRR37 , and  CmPRR73 . Conserved motifs prediction indicated that most of the closely related members in the phylogenetic tree share common protein sequence motifs, suggesting functional similarities among the PRR proteins within the same subtree. In order to explore functions of the genes, we selected two  Chrysanthemum  varieties for comparison; that is, a short-day sensitive Zijiao and a short-day insensitive Aoyunbaixue. Compared to Aoyunbaixue, Zijiao needs 13 more days to complete the flower bud differentiation. Evidence from spatio-temporal gene expression patterns demonstrated that the  CmPRRs  are highly expressed in flower and stem tissues, with a growing trend across the  Chrysanthemum  developmental process. In addition, we also characterized the  CmPRRs  expression patterns and found that  CmPRRs  can maintain their circadian oscillation features to some extent under different photoperiod treatment conditions. These lines of evidence indicated that the four  CmPRRs  undergo circadian oscillation and possibly play roles in regulating the flowering time of  C. morifolium .

Expression analysis of four pseudo-response regulator (PRR) genes in Chrysanthemum morifolium under different photoperiods