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Brassinazole-resistant (BZR) family genes encode plant-specific transcription factors (TFs), play essential roles in the regulation of plant growth and development, and have multiple stress-resistance functions. <i>Nicotiana benthamiana</i> is a model plant widely used in basic research. However, members of the BZR family in <i>N. benthamiana</i> have not been identified, and little is known about their function in abiotic stress. In this study, a total of 14 BZR members were identified in the <i>N. benthamiana</i> genome, which could be divided into four groups according to a phylogenetic tree. <i>NbBZRs</i> have similar exon-intron structures and conserved motifs, and may be regulated by cis-acting elements such as STRE, TCA, and ARE, etc. Organ-specific expression analysis showed that <i>NbBZR</i> members have different and diverse expression patterns in different tissues, and most of the members are expressed in roots, stems, and leaves. The analysis of the expression patterns in response to different abiotic stresses showed that all the tested <i>NbBZR</i> members showed a significant down-regulation after drought treatment. Many <i>NbBZR</i> genes also responded in various ways to cold, heat and salt stress treatments. The results imply that NbBZRs have multiple functions related to stress resistance.