The NAC family is one of the largest families of plant-specific transcription factors. NAC proteins play important regulatory roles in a variety of developmental and stress response processes in plants. Members of the NAC family transcription factors have been shown to be important regulators of leaf senescence in a number of plant species. Here, we report the identification of the NAC family in tobacco (Nicotiana tabacum) and the characterization of the potential roles of some of the tobacco NAC transcription factors in regulating leaf senescence. A total of 154 NAC genes (NtNACs) were identified and clustered together with the Arabidopsis NAC family into fifteen groups (a-o). Transcriptome data analysis followed qRT-PCR validation and showed that the majority of the senescence-upregulated NtNAC genes fall into subgroups b and f. A number of known senescence regulators from Arabidopsis also belong to these two subgroups. Among these senescence-upregulated NtNACs, NtNAC080, a close homolog of AtNAP, is a master regulator of leaf senescence in Arabidopsis. Overexpression of NtNAC080 caused early senescence in Arabidopsis leaves and NtNAC080 mutation induced by Cas9/gRNA in tobacco led to delayed senescence.