In order to investigate the response of soil physical and chemical properties, tobacco growth and development, and tobacco leaf quality to different carbon source biochar, this study took tobacco Yunyan 87 as the research object, with conventional fertilization as the control (CK), and set up conventional fertilization+pyrolysis biochar (T1), conventional fertilization+plant-based biochar (T2), conventional fertilization+animal based biochar (T3), and conventional fertilization+mixed carbon source biochar (T4), respectively. Explore the effects of different carbon sources of biochar on the growth, development, and quality of tobacco in southern Anhui. The results showed that T2 treatment significantly increased the available phosphorus and potassium content in the soil by 21.14% and 15.14%, respectively. The optimal sugar alkali ratio and potassium chloride ratio for the coordination of chemical composition in tobacco leaves under T3 treatment were 13.22 and 3.03, respectively. However, based on the contribution rate of biochar to the yield and output value of tobacco, T4 treatment showed the best effect, increasing the output value per hectare by 23.60%. In production practice, the growth and development of tobacco and the quality of tobacco leaves can be regulated by selecting and adding suitable carbon source biochar, providing scientific basis for achieving high-quality and high-yield tobacco.