In order to understand the regulation mechanism of NtLAR gene, we cloned the 5′terminal promoter sequence of NtLAR gene in this study using genome walking method from genomic DNA of Chinese narcissus (Narcissus tazetta var. chinensis cv. ‘Jinzhanyintai’). The sequencing result showed that the cloned fragment was 995 bp in length (GenBank number MH371155). Cisacting elements of the promoter were analyzed and predicted using plant care databases. Many cisacting elements were found, such as light response element ACE, Gbox, GATAmotif, GT1motif, hormone response element CGTCAmotif, ABRE, TGACGmotif, TGAelement, stress corresponding element and MYB binding site MBS. Two expression vectors pBI121pNtLAR∷GUS and pGreenII 0800pNtLARLuc were constructed successfully. Transient expression ofpBI121pNtLAR∷GUS in tobacco leaves showed that the cloned promoter fragment had activity. The transient expression results of pBI121pNtLAR∷GUS in Chinese narcissus showed that NtLAR promoter had different activities in different tissues of Chinese narcissus. The expression level of pNtLAR∷GUS is higher in basal plates and very low in petal and corona. When tobacco leaves were agroinfiltrated with pBI121pNtLAR∷GUS mixed with R2R3MYB genes NtMYB2 and NtMYB5 respectively, GUS staining and qRTPCR showed that the activity of NtLAR promoter could not be repressed by NtMYB2 or NtMYB5. Dural luciferase assay was also conducted in N. benthamiana leaves with pGreenII 0800pNtLARLuc mixed with NtMYB2 or NtMYB5. The results of dual luciferase assay were consistent with GUS staining and qRTPCR results.