Abstract: Tobacco transient expression is a cutting-edge bioengineering technology that rapidly introduces target genes into tobacco plants through Agrobacterium-mediated methods, enabling to obtain the target recombinant proteins in a short timeframe. Compared to traditional microbial fermentation and animal cell expression systems, the tobacco transient expression system offers several the advantages, including short production cycle, high yield, cost-effectiveness and enhanced safety. This paper reviews the historical development of tobacco transient expression technology, explores its diverse applications in agriculture, industry, and biomedicine, and highlights the supportive role of plant factories and smart agriculture in the industrialization of this technology. Additionally, it addresses the key challenges currently hindering the industrialization of tobacco transient expression technology and proposes targeted solutions and strategies, with the goal of providing valuable insights for advancing its industrial application.