Using floating seedling cultivation technology under artificial climate chamber conditions, the mycorrhizal effects of 10 arbuscular mycorrhizal fungi(AMF) inoculation treatments on the agronomic traits, root morphology, pigment content, root vitality, and N, P, K nutrient content of tobacco seedlings were comprehensively analyzed, and the relevant characteristics of mycorrhizal infection were explored. The results showed there were significant differences in the infection characteristics and mycorrhizal effects among different AMF isolates. Acaulospora kentinensis(AM1), Rhizophagus irregularis(AM3), and Rhizophagus intraradices(AM8) significantly increased the root vitality of tobacco seedlings; Acaulospora kentinensis(AM1), Funneliformis mosseae(AM2), Septoglomus viscosum(AM5), and Claroideoglomus lamellosum(AM6) effectively promoted tobacco seedling growth and optimized the root to shoot ratio; Glomus versiforme(AM9), Claroideoglomus etunicatum(AM4), and Acaulospora longula(AM7) markedly increased biomass accumulation; Rhizophagus intraradices(AM8), Glomus versiform(AM9), and Acaulospora excavata(AM10) have potential in promoting nutrient absorption and stress resistance of tobacco seedlings. Cluster analysis categorized the 10 AMF isolates into four effect types: “growth promoting type”, “growth regulating type”, “nutrient accumulation promoting type” and “stress enhancing type”, which were represented by Funneliformis mosseae(AM2), Claroideoglomus lamellosum(AM6), Rhizophagus intraradices(AM8) and Glomus versiforme(AM9), respectively.