Abstract: In order to determine the microbial community composition in acidic farmland soil and explore the comprehensive treatment of acidified soil, acidic soil samples collected from Lichuan City, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province, were selected as research objects, and the 16S rRNA gene V3-V4 variable region of total bacteria and culturable bacteria in each soil sample were sequenced using Illumina sequencing. Using this methodology, the effects of soil physicochemical properties on the community structure of total bacteria in soils were investigated. The results demonstrated that 4008 total bacteria and 97 OTUs culturable bacteria from Chinese cabbage, tobacco, yam, strawberry, maize and lettuce at 97% similar level were found. At phylum level, 12 bacterial groups whose relative abundance were more than 1%, whilst Proteobacteria, Acidobacteria, and Chloroflexi were the dominant phyla. There were only 3 culturable phyla bacteria whose relative abundance were more than 1%, namely Firmicutes, Proteobacteria and Bacteroidetes. At OTU level, soil total bacteria could be divided into two groups across pH gradient:pH < 5.5 (maize, yam and lettuce fields) and pH > 5.5(tobacco, cabbage and strawberry fields). RDA analysis exhibited that the relative abundance of Acidobacteria was positively related to pH, while the abundance of Firmicutes or Proteobacteria was positively correlated to w(TN), w(TP), w(TC). The phylogenetic tree showed that the top 30 OTU bacteria had higher homology with the 16S rRNA gene sequence of the bacteria found in Asia. These results showed that Illumina MiSeq sequencing can comprehensively analyze the diversity of total bacteria and culturable bacteria in acidic soils, and that the domestication process greatly reduces the species of bacteria, at the same time, the similarity of bacterial community structure may decrease with the increase of geographic distance.