The research aimed to provide a theoretical basis for the regional planning of tobacco planting and the high-quality sustainable development of tobacco in China，and to explore the spatial variation characteristics of soil nutrients and microbial community structure at different altitudes.Tobacco planting soil samples at altitudes of 500 m(T1)，1000 m(T2)and 1500 m(T3)were collected in Hanzhong，Shaanxi province.Soil nutrients were determined，and high-throughput sequencing technology was used to determine the microbial community of the soil samples.The results showed that with the increase of altitude，the soil pH increased first and then decreased，and the soil pH increased from 6.15 to 6.77，and then decreased to 5.74；Soil organic matter，alkali hydrolyzable nitrogen and available potassium increased from 14.60 g/kg to 18.52 g/kg，alkali hydrolyzable nitrogen from 150.35 mg/kg to 167.40 mg/kg，and available potassium from 134.56 mg/kg to 197.18 mg/kg；The available phosphorus in soil decreased first and then increased，from 33.72 mg/kg to 11.07 mg/kg，and then increased to 38.90 mg/kg；Soil chloride ion showed a decreasing trend，from 15.45 mg/kg to 7.28 mg/kg. The results of microbial sequencing showed that with the increase of altitude，the soil bacterial richness index first decreased and then increased，the ace index decreased from 3955.95 to 3495.43，then increased to 4107.47，the Chao index decreased from 3918.50 to 3422.21，and then increased to 4054.87. The soil bacterial diversity showed a downward trend，the Shannon index decreased from 6.48 to 6.26，and the Simpson index increased from 0.065 to 0.072；The Shannon index decreased from 6.48 to 6.26，and the Simpson index decreased slightly from 0.065 to 0.061，and then increased to 0.072；With the increase of altitude，soil fungal richness showed a trend of first increasing and then decreasing，in which ace index increased from 261.75 to 263.95，and then decreased to 247.64，Chao index decreased from 268.02 to 249.08，and soil fungal diversity showed a trend of first increasing and then decreasing，in which Shannon index increased from 3.04 to 3.24，and then decreased to 2.70，Simpson index decreased from 0.10 to 0.08，and then increased to 0.13. In conclusion，the microbial community is relatively balanced in the altitude area (500 m)，and the nutrient content is relatively low. The nutrient content is relatively high in the middle and high altitude area (1000-1500 m)，but the microbial community is relatively unbalanced. Therefore，in production，it is suggested to improve the soil nutrients and microbial community structure and improve the soil health index through rational application of organic fertilizer，carbon based fertilizer and chemical fertilizer.