Production of Luzhou-flavor liquor is based on pits where ample microbial communities are living, and interactions among all sorts of microbes especially Bacilli in it make fermented sapor of Luzhou-flavor liquor very unique. In this study, 23 aerobic Bacilli were isolated from 300-year pit mud of cellar affiliate to Luzhou laojiao, Luzhou, Sichuan Province. Microscopic and colonial morphology analysis showed that the difference between these aerobic Bacilli were not significant. Through PCR amplification of 16S rDNA gene and NCBI blast, a phylogenetic tree was preliminarily constructed. The results showed that the strains of aerobic Bacilli were mainly Bacillus thuringiensis, B. cereus and B. anthracis. The genetic relationship of these strains was inconformity. As a whole, the phylogenetic relationship was close and the genetic polymorphism was low between these Bacilli isolated from 300300-year pit mud. Aged pit mud was always in the surroundings of anaerobic or facultative anaerobic, studying on the survival condition and genetic relationship of the aerobic Bacilli in aged pit mud was helpful to get a insight in the change of surrounding environment during the liquor fermentation, and improve the quality of liquor. The result provides basic data for research on the change of surrounding environment during the liquor fermentation, and improving the quality of liquor.%浓香型白酒的生产是以窖池为基础，窖池窖泥中栖息着庞大的微生物群系，微生物间相互作用形成浓香型白酒的独特风味，尤其以芽胞杆菌为主。本研究从泸州老窖酒厂300年窖池窖泥中分离出23株好氧芽胞杆菌属(Bacilli)，经光学显微镜和菌落形态观察，23株好氧芽胞杆菌形态差异不显著。通过16S rDNA基因扩增，NCBI基因比对，构建系统发育树，结果表明，23株好氧芽胞杆菌主要为苏云金芽胞杆菌(Bacillus thuringiensis, Bt)、蜡样芽胞杆菌(B. cereus, Bc)和炭疽芽胞杆菌(B. anthrax)，菌种间亲缘关系存在差异；窖泥中分离出的23株好氧芽胞杆菌亲缘关系近，遗传多样性较低。本研结果为了解白酒发酵过程中老窖泥周围环境的变化、提高白酒品质提供基础资料。
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