The constitution of the bacterial community in Zhenjiang Hengshun vinegar acetic fermentation was analyzed by culture-independent approach. Total DNA was extracted and amplified using PCR reaction to construct 16S rDNA library, which includes 96 positive clones. Through sequencing of those clones, the phylogenetic tree is illustrated by representative sequences. Based on phylogenetic analyzing, 16 OUTs was obtained, and the composition of microorganisms in Hengshun vinegar fementation was concluded as below: five OUTs belong to genus Lactobacillus, two are genus Acetobacter, one genus Gluconacetobacter, two genus Staphylococcus, two genus Enterobacter, two genus Pseudomonas, one genus Flavobacterium, and one genus Sinorhizobium, respectively.