海洋生境刀鲚菌群PCR-DGGE指纹图谱构建及分析  

Construction and analysis of PCR-DGGE gene fingerprint of bacterial community in Coilia nasus from sea

作　　者：聂志娟[1] 徐钢春[1] 程起群[2] 杜富宽[1] 张勇[3] 顾若波[1] 

机构地区：[1]中国水产科学研究院淡水渔业研究中心农业部淡水渔业和种质资源利用重点实验室,江苏无锡214081 [2]中国水产科学研究院东海水产研究所农业部海洋与河口渔业资源及生态重点开放实验室,上海200090 [3]南京农业大学渔业学院,江苏无锡214081

出　　处：《海洋科学》2014年第10期63-69,共7页Marine Sciences

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2013JBFT04);农业部东海与远洋渔业资源开发利用重点实验室开放课题;国家科技支撑计划项目(2012BAD26B05);公益性行业科研专项(201203065)

摘　　要：为了分析海洋生境刀鲚（Coilia nasus）体内及生长环境菌群结构,作者采用免培养16S rDNA 的PCR-DGGE 指纹图谱技术,对刀鲚鳃、肠道壁、肠道内容物及水体环境中的菌群结构进行了对比分析。结果表明：变性梯度为35%-55%、浓度为8%的聚丙烯酰胺凝胶,150V 60℃下电泳10 h, DGGE带谱的分离效果较好；刀鲚鳃、肠道壁、肠道内容物及水体样品指纹图谱上分别显示出24、19、14和29条信号强度不同的条带；相同样品重复组细菌结构相似度在80%以上,差异不显著；不同样品之间,刀鲚鳃与海水聚为一支,具有较高的相似度为52%,肠壁与肠内容物相似度为41%。样品菌群主要以未培养菌为主,主要包括变形菌、放线菌和厚壁菌。本文首次成功构建海洋生境刀鲚菌群16S rDNA 的PCR-DGGE 指纹图谱。In order to investigate the bacterial community structure of Coilia nasus both in vivo and in environment, the bacterial community in gill, intestinal wall, intestinal contents and the water environment of C. nasus was compared and analyzed using a culture-free 16S rDNA PCR-DGGE fingerprinting technology. The results showed that bacterial PCR amplifications could be better separated on DGGE using a denaturing gradient ranging from 35%to 55% at 50 V at 60℃ for 10h. The samples from water environment, gill, intestinal wall and intestinal contents showed 29, 24, 19, and 14 bands, respectively with different signal intensities, which might reflect the existence of rich bacteria species. The clustering analysis showed that no significant differences between repeated samples. The similarity was more than 80%. The sample from water environment and gill of C. nasus joined together to form a branch, with a similarity of 52%and the similarity between those from the intestinal wall and intestinal contents of C. nasus was 41%. The attached bacterial flora forming four samples were dominated by uncultured bacteria, including proteobacteria, actinobacteria, and firmicutes. Here, for the first time, we constructed the PCR-DGGE gene fingerprint for bacterial community in C. nasus from sea.

关 键 词：PCR-DGGE 菌群 刀鲚(Coilia nasus) PCA 

分 类 号：S917.1[农业科学—水产科学]