基于16S rRNA技术的长江口微生物分子生物学鉴定与分析

A study of the microorganism using bio-molecular technology from the Yangtze Estuary based on the technology of 16S rRNA

基于2007年12月与2008年6月长江口上海附近海域采集的海水样品,使用27F和1492R两种通用引物对海洋微生物16S rRNA基因片段进行直接扩增,克隆并测序,构建细菌16S rRNA文库,通过NCBI数据库的基因比对,区分微生物种群,并使用Mega 4.0软件利用16S rRNA序列建立微生物的分子发育树。通过直接提取DNA进行扩增的方法,共检测出17个属的53种不同微生物,微生物种群结构随温度变化明显,优势种随温度变化有所不同,在不同季节水温下,微生物优势种优势明显;冬季以不动杆菌属为优势种群,分子发育树体现的遗传差异性较小;夏季微生物种群结构较复杂,其中以希瓦氏菌属及假单胞菌属为优势种群,分子发育树体现的遗传差异性较大;微生物分布存在明显的季节变化。

This paper is mainly based on microorganism samples of seawater which were gathered around the Yangtze Estuary from Oct.2007 to June 2008.The marine microorganism species were identified by using amplifications of 16S rRNA gene with primer 27F and 1492R,cloning and sequencing techniques.Through the database of NCBI,using the software of Mega 4.0,molecular phylogenetic tree was established with the sequences of 16S rRNA.In this paper,microorganism samples of seawater were respectively gathered in winter and summer at the same destination.Seventeen genera including fifty-three strains were achieved from the analysis of the four samples.Microbial community structure changes with temperature obviously,and dominant species vary with temperature.In winter,the Acinetobacter was the dominant species,and the molecular phylogenetic tree reflects a few genetic differences;In summer,microbial community structure was more complex,in which the Shewanella and Pseudomonas were the dominant species,and the molecular phylogenetic tree reflected greater genetic differences;In the waters of different temperature,microbial species shows different advantages obviously.Molecular phylogenetic tree reflects the genetic differences with obvious seasonal variation distribution.