Abstract: The NAC (No apical meristem/ATAF1, 2/cup shaped cotyledon) gene encodes an important class of transcription factor proteins in plants, and members of this family are involved in regulating plant growth and development processes such as seed development, leaf senescence, and cell division. Based on the homologous gene information of Arabidopsis, a NAC transcription factor gene (NtNAC029) was cloned from cultivated tobacco K326. The gene coding region and amino acids have conserved sequence structure and protein functional domains of the NAC family. The expression of NtNAC029 gene shows significant tissue and stage specificity. Its expression level in leaves gradually increases with leaf maturity and senescence, reaching its maximum value in the late stage of leaf senescence. However, its expression level in capsules sharply increases in the early stage of development, and then gradually decreases, indicating that this gene may be involved in regulating important physiological processes such as tobacco leaf senescence and fruit initiation and development. The expression of NtNAC029 gene is more sensitive to light, and the gene expression level is significantly reduced under light culture conditions. Darkness treatment can significantly upregulate the gene expression level. In addition, salt, abscisic acid (ABA) and methyl jasmonic acid (MeJA) could significantly induce the expression of NtNAC029 gene, indicating that NtNAC029 may play an important role in response to multiple environmental factors regulating leaf senescence.