温水沤麻与酶法沤麻液细菌群落结构分析

Analysis of Bacterial Community Structure in Water Retting and Enzymatic Retting Liquid

为了研究温水沤麻与酶法沤麻液中细菌群落结构,采用DGGE技术(Denaturing gradient gel electrophoresis)对沤麻液的细菌菌群结构进行了分析。采用DGGE分析细菌多样性时,当凝胶浓度为8%,变性剂梯度范围为25%~65%,恒温60℃、150 V电压下电泳7 h左右时,DGGE分离的效果较好。PCR-DGGE指纹图谱显示,亚麻温水脱胶及酶法脱胶过程中条带数量不同。通过对温水沤麻与酶法沤麻样品不同时期沤麻液中16S rDNA V3片段PCR产物的DGGE条带进行分子克隆、序列测定和Blast分析及建立系统发育树。从序列结果中我们发现,γ-变形菌亚门中的假单胞菌为温水沤麻与酶法沤麻过程中的共同优势菌属。芽孢杆菌属的出现伴随着整个温水沤麻和酶法沤麻过程。此外,温水沤麻与酶法沤麻样品群落结构多样性也表现了一定的差异性。

In order to investigate the bacterial community diversity of warm water retting liquid and enzymatic retting liquid, both of the retting liquid samples were studied by using Denaturing Gradient Gel Electrophoresis (DGGE). During using DGGE method to analyze the bacterial diversity, DGGE bands were separated obviously by the gel concentration of 8%, range of 25% - 65% denaturant gradient, temperature of 60?C and voltage of 150 V for electrophoresis time of about 7 h. Through analysis, the selected bands of DGGE profiles were cloned and sequenced. The obtained sequence results by Blast analysis were used to construct the phylogenetic tree. We found Pseudomonas of Gammaproteobacteria were the dominant bacteria both in warm water retting and enzymatic retting liquid samples. Bacillus of Firmicutes occurred along with the whole process of the two retting samples. However, in addition to share the dominant bacteria, the two samples also represented the differences in bacterial community structure.