Abstract: This objective of this research was to find out the role of arbuscular mycorrhizal fungi (AMF) inoculation on seedling growth of and nutrient uptake by tobacco grown on the soil with P deficiency and low S, as well as explore the application possibility of PG to agricultural soil. A pot simulation experiment was conducted to study the seedling growth of and phosphorus (P), sulphur (S), arsenic (As) uptake by tobacco ‘NC297’ under different phosphogypsum (PG) addition levels (0, 40 g/kg) and mycorrhizal inoculation status [none mycorrhizal(NM), Glomus mosseae (GM)]. The result showed that whether inoculation or not, the ‘NC297’ shoot and root biomass were increased by PG addition (PG40) significantly, and P concentrations in and uptake by shoot and root as well as P, S, As specific absorption rate (SAR) of ‘NC297’ were also increased by PG40 significantly, thus making the As concentrations in and uptake by, uptake ratio of P to As (P/As ratio) in roots, and S concentrations in and uptake by shoots increased. With the same phosphgypsum addition levels, inoculation GM fungi significantly increased P SAR and P/As ratio in ‘NC297’ plants and shoot P and S concentrations and uptake, but significantly decreased ‘NC297’ shoot and root As concentrations and uptake under PG0 treatments. Therefore the combination treatment of PG40 and GM inoculation was more effective for promoting plant growth through increasing P, S uptake and resisting As stress to tobacco plants.