Abstract: In order to investigate any promoting effect and associated physiology by nano-carbon sol, culture experiments were carried out under solid and liquid culture conditions. The effects of nano-carbon sol application on the expression of related genes including water channel protein gene and cell division gene, biomass, nutrient accumulation and suspension cell growth characteristics of tobacco BY-2 cells were studied. The results showed that the addition of nano-carbon sol enhanced the growth of tobacco callus and increased the accumulation of N, P and K nutrients. In all the treatments, 50 mg/L concentration of nano-carbon sol had the largest increase rates of nutrient accumulation in callus, namely 41.3%, 27.6% and 47.0%, respectively. At the same time, nano-carbon sol significantly increased the expression levels of aquaporin gene NtPIP1 and cyclin gene CycB at early culture stage, which played an important role for promoting callus growth. In suspension culture experiments, nano-carbon sol presented a good biocompatibility at the early and middle culture stages without significantly affecting the density, packed cell volume and mortality of the suspension cells. At the late culture stage, nano-carbon sol had an inhibitory effect at the concentration of 10 mg/L, which decreased the cell density and cell viability, which is likely to be associated with the aging of cultured cells. In conclusion, the effects of nano-carbon sol on the growth of tobacco cells were influenced by the culture conditions and application concentration.