亚麻温水脱胶液中细菌菌群结构分析

为了探讨实验室条件下模拟的工厂亚麻温水脱胶液中细菌菌群结构,采用纯培养技术和PCR-DGGE技术(Denaturing gradient gel electrophoresis)对细菌菌群结构进行了研究.用纯培养方法分离获得9类菌落,其中假单胞菌属(Pseudomonas)在有氧培养条件下总是处于优势.梭菌属(Clostridium)在厌氧培养过程中总是处于优势.微球菌属(Micrococcus)和葡萄球菌属(Staphylococcus)只有亚麻脱胶初期才有发现.PCR-DGGE指纹图谱显示,亚麻温水脱胶过程中条带数量较少且没有明显种群群落结构演替过程.通过对不同时期沤麻液中16S rDNAV3片段PCR产物d、e两个DGGE条带进行分子克隆、序列测定和Blas份析,发现e条带包含的16S rDNAV3片段除e 35外均属于假单胞菌属.d条带包含着较多不同的16S rDNAV3片段,其中有传统方法没有分离到的泛菌属(Pantoea)细菌、一些NCBI未收录的序列及一些非可培养微生物序列.纯培养技术和PCR-DGGE技术的共同使用,可以更全面准确地提供细菌多样性方面的信息.图4参15

Analysis of Bacterial Community in Water Retting of Flax

The bacterial community structure in water retting systems of Bei'an flax in Heilongjiang, China was studied by using denaturing gradient gel electrophoresis (DGGE) and culture-dependent method, respectively. The results obtained from the culture-independent method showed that Pseudomonas and Clostridium were found the dominant genera using aerobic and anaerobic cultivation in water retting, respectively. Micrococcus and Staphylococcus were only observed within initial 16 h. The profile of DGGE fingerprints in water retting of different periods revealed that the bacterial groups were relatively fewer. As a result, two major bands of 16S rDNA genes fragments from DGGE profiles were further eluted from gel, prior to be reamplified and sequenced. The sequences of these fragments were compared with the database of GenBank (NCBI). 9 genera were identified from the water retting with the culture-dependent method. Compared with the database of GenBank, the results displayed that 11 sequences of band d shared 92%～99% homology of the known sequences (Pantoea, Bacillus, unidentified bacterium and new bacteria), while the 12 sequences of band e shared 96%～100% homology of the known sequences (Pseudomonas). This suggests that a combination of molecular and culture-dependent methods can be used to analyze and monitor the community structure of water retting effectively, and will give us more information of microorganism community structure. Fig 4, Ref 15