Plant nuclear factor (NF-Y) is a transcriptional activating factor composed of three subfamilies: NF-YA, NF-YB, and NF-YC. These transcriptional factors are reported to function as activators, suppressors, and regulators under different developmental and stress conditions in plants. However, there is a lack of systematic research on theNF-Ygene subfamily in sugarcane. In this study, 51NF-Ygenes (ShNF-Y), composed of 9NF-YA, 18NF-YB, and 24NF-YCgenes, were identified in sugarcane (Saccharumspp.). Chromosomal distribution analysis ofShNF-Ysin aSaccharumhybrid located theNF-Ygenes on all 10 chromosomes. Multiple sequence alignment (MSA) of ShNF-Y proteins revealed conservation of core functional domains. Sixteen orthologous gene pairs were identified between sugarcane and sorghum. Phylogenetic analysis of NF-Y subunits of sugarcane, sorghum, andArabidopsisshowed that ShNF-YA subunits were equidistant while ShNF-YB and ShNF-YC subunits clustered distinctly, forming closely related and divergent groups. Expression profiling under drought treatment showed that NF-Y gene members were involved in drought tolerance in aSaccharumhybrid and its drought-tolerant wild relative,Erianthus arundinaceus.ShNF-YA5andShNF-YB2genes had significantly higher expression in the root and leaf tissues of both plant species. Similarly,ShNF-YC9had elevated expression in the leaf and root ofE. arundinaceusand in the leaf of aSaccharumhybrid. These results provide valuable genetic resources for further sugarcane crop improvement programs.