The genetic network resulting in the production of an inflorescence is complex, involving one or more pathways including the photoperiod, maturity, gibberellin and autonomous pathways, and induction and repression of genes along the pathways. Understanding the cyclic expression profile of genes involved with photoperiod perception and floral pathway induction in sugarcane, an intermediate–short day plant (ISD), is crucial for identifying key genes and understanding how the profile changes in response to floral induction signals under decreasing daylengths. Homologues of 21 genes, and some gene alleles, associated with photoperiod perception and the flower induction pathway were examined in sugarcane variety Q174 over a 24-h light-dark cycle. The strongest expression of these genes was seen in the immature spindle leaves and levels of expression generally decreased with increasing leaf age. Significant changes in gene expression levels during a 24-h cycle were observed for 16 of the 21 genes tested. We have now defined an important baseline for expression patterns over a 24-h cycle in non-inductive conditions in sugarcane. These results can be utilised to select the optimal time for detecting changes during floral induction, differences between varieties that are responsive/non-responsive to photoperiod induction, and to identify genes that may be manipulated to enhance or inhibit flowering.