| Summary: | Zinc finger homeodomain (ZF-HD) transcription factors play significant roles in plant growth and responses to environmental stresses. In this study, 32 <i>ZF-HD</i> genes identified in the tobacco (<i>Nicotiana tabacum</i> L.) genome were divided into six groups according to phylogenetic analysis with <i>Arabidopsis</i> and tomato ZF-HD members. An examination of gene structures and conserved motifs revealed the relatively conserved exon/intron structures and motif organization within each subgroup. In addition, various stress-related elements are found in the promoter region of these genes. The expression profiling analysis revealed that <i>NtZF-HD</i> genes expressed in different tissues and could be induced by several abiotic stresses. Notably, <i>NtZF-HD21</i> was highly expressed in response to the drought treatments. Subcellular localization analysis and a virus-induced gene silencing (VIGS) experiment were performed to investigate the potential functions of <i>NtZF-HD21</i>. The subcellular localization indicated that NtZF-HD21 is a nuclear protein. Furthermore, gene silencing of the <i>NtZF-HD21 </i>gene reduced the drought resistance of tobacco. These findings provide insights for further biological functional analyses of the<i> NtZF-HD</i> genes in tobacco.
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