应用PCR-DGGE技术分析不同性状窖泥的细菌群落结构

基于聚合酶链式反应-变性梯度凝胶（PCR-DGGE）技术分析了不同性状窖泥的细菌群落DGGE指纹图谱，结果表明，通过DGGE指纹图谱能够区分新窖泥、窖壁泥和窖底泥，其中以窖壁泥退化前后图谱变化最明显。PCR-DGGE图谱优势条带测序结果显示，窖泥中细菌微生物包括拟杆菌门、放线菌门、厚壁菌门、变形菌门4类，其中新窖泥微生物种类最丰富，涵盖以上门类，特有优势微生物为Enterobacterale科、双歧杆菌属（Bifidobacterium）、假单胞菌属（Pseudomonas）及里氏杆菌属（Riemerella）。紫单孢菌科（Porphyromonadaceae）仅存在于窖底泥中，Tumebacillus属微生物只存在于窑壁泥中。通过窖泥的DGGE图谱对比分析，能够对窖泥类型做出初步判定，并能及时发现窖泥退化现象，为工厂窖泥培养、检测和窖泥优化提供了理论支持。

Application of PCR-DGGE technique in the analysis of bacterial community structure in pit mud with different characters

The DGGE fingerprint of bacterial community of pit mud with different characters was explored by PCR-DGGE. The results showed that DGGE fingerprint would distinguish the new pit mud, wall pit mud and bottom pit mud, and the change of spectra of wall pit mud before and after degradation was the most significant. The PCR-DGGE dominant bands were sequenced and the results revealed that dominant bacteria in pit muds belonged to Bacteroidetes, Actinobacteria, Firmicutes and Proteobacteria. The new pit mud covered the above categories and the special dominant bacteria were Enterobacterale, Bifidobacterium, Pseudomonas, Riemerella, while Porphyromonadaceae only existed in the bottom pit mud. Tumebacillus was only in the wall pit mud. Through the comparative analysis of DGGE chromatogram of pit mud, the initial determination of pit mud characters would be realized and pit mud degradation were timely discovered, which could provide a theoretical basis for culture, detection and optimization of pit muds in the factory.