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Grape (Vitis vinifera) is among the world’s most important fruit crops and is a commonly used woody plant for genomics and post-genomics research. NAC transcription factors play central roles in plant growth and development, floral organ morphogenesis, and responses to biological stress. It is therefore important to identify key transcription factors from grape and clarify their mechanisms of action to generate genetic resources for grape molecular improvement. Our research group previously cloned a NAC transcription factor from V. vinifera ‘Yatomi Rosa’ [drought and leaf roll gene 1 (DRL1)] and demonstrated that it caused dwarfing of tobacco (Nicotiana benthamiana) plants when overexpressed. In the present work, we demonstrate that overexpression of DRL1 in transgenic tobacco delays flowering time and markedly reduces pollen viability. Furthermore, crosses between male DRL1 transgenic tobacco and female wild-type tobacco exhibit substantially lower fruit set, fruit and seed weights, fruit and seed shape indices, and seed germination rates than selfed wild-type plants or crosses with a transgenic female parent. DLR1 overexpression strongly influences flowering time and reproduction in transgenic tobacco, primarily through its effects on pollen development. These results provide a foundation for further functional characterization of DLR1 in grape.