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

作　　者：侯立波 刘晓兰[2] Libo Hou;Xiaolan Liu(Graduate School of Environment and Information Sciences, Yokohama National University, Yokohama, Japan;Heilongjiang Key Lab of Agricultural Product Processing, College of Food and Biology Engineering, Qiqihar University, Qiqihar)

机构地区：[1]横浜国立大学环境情报学府,横浜日本 [2]齐齐哈尔大学食品与生物工程学院,齐齐哈尔

出　　处：《生物过程》2012年第3期116-122,共7页Bioprocess

基　　金：教育部重点项目:亚麻酶法脱胶中关键酶对纤维化学组成和结构的作用机制(208038)。

摘　　要：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.

关 键 词：温水沤麻 酶法沤麻 细菌群落结构 变性梯度凝胶电泳(DGGE) 

分 类 号：S15[农业科学—土壤学]