Culture independent analysis of bacterial community diversity associated with coastal sanddune plant

소개

사구식물분리 미생물연구, PCR, DGGE, cloning.

목차

1. Introduction

2. Materials and Methods
2.1 Clone analysis
2.1.1 Sampling of rhizosphere soil and plants root soil
2.1.2 DNA extraction
2.1.3 PCR amplification and cloning of 16S rDNA
2.1.4. ARDRA analysis
2.1.5 Sequencing and phylogenetic analysis
2.2 DGGE analysis
2.2.1 Sampling of rhizosphere soil and plants root soil
2.2.2 DNA extraction
2.2.3 16S V3 region PCR
2.2.4 DGGE
2.2.5 Reamplification and Sequencing 16S rDNA
2.2.6 Phylogenetic analysis

3. Results
3.1 Clone analysis
3.1.1 Construction of 16S rDNA Clone Library
3.1.2 Whole community ARDRA patterns of the different soils and season.
3.1.3 Identification of ARDRA types
3.2 DGGE analysis
3.2.1 DGGE fingerprints and Phylogenetic analysis of 16S rDNA sequences.
3.2.2 Specific DGGE patterns of Lysobacter sp. relatives from the sand dune soil samples.
3.3 Comparison of ARDRA analysis and DGGE analysis

4. Discussion
영문 Abstract

5. References
국문 초록

본문내용

Abstract


Increasing dependence on molecular methods to analyze community structures have made molecular biological techniques including DNA extraction and purification from environmental samples very important part of ecological studies.
In this study, the bacterial community structure of sand dune rhizosphere soil and plant root was investigated by use of the rRNA based approach. Denaturing gradient gel electrophoresis (DGGE) with eubacterial primer (GC341F) and universal primer (518R) and cloning analysis of community 16S ribosomal DNAs amplified with universal bacterial primer sets were used to describe the geographical and seasonal succession of the bacterial community structure along the Tae-an area, Chungnam in 2003 and 2004. Root and rhizosphere soil samples were collected from 4 location and 9 plant species. The DGGE analyses revealed that all of the rhizosphere and root samples possessed an apparently common band, which was identified as that of Lysobacter sp. by sequencing. The genus Lysobacter is a member of the family Xanthomonadaceae, class Gammaproteobacteria.
DGGE analysis was carried out to compare diversity among various sets of samples. The analysis of bands from roots and rhizosphere of 9 plant species indicated that the bacterial composition was taxonomically diverse, and could be grouped in to 5 bacterial phyla. Members of Gammaproteobacteria were predominant.

참고자료

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