Advancement in nanotechnology has improved ways for large-scale production and characterization of nanoparticles of physiologically important metals. The current study explores the impact of Zinc Oxide Nanoparticles (ZnO-NP) and Chitosan-Zinc oxide nano-bioformulation (CH-ZnO) in tissue culture raised callus of Nicotiana benthamiana. Results indicated augmented biomass in CH-ZnO treated callus, while a reduced biomass was observed in ZnO-NP treated callus, at all the concentrations tested. Higher chlorophyll and carotenoid content were recorded in callus treated with 800 ppm CH-ZnO as compared to ZnO-NP treated callus. A higher accumulation of proline was observed in CH-ZnO treated callus when compared to ZnO-NP treatment, which was significantly higher at 50, 200 and 400 ppm CH-ZnO treatment. A maximum reduction in malondialdehyde (MDA) content was recorded at 800 ppm, for both the nano-formulations tested. Likewise, a significant reduction in the H₂O₂ levels was observed in all the treatments, while the callus treated with 400 ppm ZnO-NP and 800 ppm CH-ZnO recorded the highest reduction. Phenylalanine Ammonia-Lyase (PAL), activity increased significantly in callus treated with 400 ppm concentration for both ZnO-NP and CH-ZnO with respect to control. An increased level of tannin and nicotine were recorded in callus supplemented with 50, 200 and 400 ppm CH-ZnO. Notably, a significant decline of 94 and 52% in tannin content and 25 and 50% in nicotine content was recorded in the callus treated with 800 ppm CH-ZnO and ZnO-NP, respectively. The findings of this study suggest that an optimized dosage of these nano-bioformulations could be utilized to regulate the nicotine content and stress tolerance level.

纳米技术的进步改进了生理学重要金属纳米颗粒的大规模生产和表征方法。目前的研究探讨了氧化锌纳米颗粒（ZnO-NP）和壳聚糖-氧化锌纳米生物制剂（CH-ZnO）对本氏烟草组织培养愈伤组织的影响。结果表明，在所有测试浓度下，CH-ZnO 处理的愈伤组织中的生物量增加，而 ZnO-NP 处理的愈伤组织中的生物量减少。与 ZnO-NP 处理的愈伤组织相比，用 800 ppm CH-ZnO 处理的愈伤组织记录了更高的叶绿素和类胡萝卜素含量。与 ZnO-NP 处理相比，在 CH-ZnO 处理的愈伤组织中观察到更高的脯氨酸积累，在 50、200 和 400 ppm CH-ZnO 处理时显着更高。对于所测试的两种纳米制剂，丙二醛 (MDA) 含量的最大降低量为 800 ppm。同样，在所有处理中都观察到H ₂ O _{2水平显着降低，而用400 ppm ZnO-NP 和800 ppm CH-ZnO 处理的愈伤组织记录了最高的降低。}与对照相比，用 400 ppm 浓度的 ZnO-NP 和 CH-ZnO 处理的愈伤组织中苯丙氨酸氨裂解酶 (PAL) 的活性显着增加。补充了 50、200 和 400 ppm CH-ZnO 的愈伤组织中单宁和尼古丁的含量有所增加。值得注意的是，用 800 ppm CH-ZnO 和 ZnO-NP 处理的愈伤组织中单宁含量显着下降了 94% 和 52%，尼古丁含量显着下降了 25% 和 50%。这项研究的结果表明，这些纳米生物制剂的优化剂量可用于调节尼古丁含量和应激耐受水平。