DGGE fingerprinting of culturable soil bacterial communities complements culture-independent analyses

被引:70
作者
Edenborn, S. L. [1 ]
Sexstone, A. J. [1 ]
机构
[1] W Virginia Univ, Div Plant & Soil Sci, Morgantown, WV 26506 USA
关键词
culturable bacteria; DGGE; plate wash; bacterial community structure; culture-dependent; culture-independent;
D O I
10.1016/j.soilbio.2007.01.007
中图分类号
S15 [土壤学];
学科分类号
0903 ; 090301 ;
摘要
Culture-dependent DGGE (CD DGGE) fingerprinting of the 16S rRNA gene was used to characterize mixed bacterial communities recovered on agar plates. Using R2A Agar as a growth medium, CD DGGE analysis resulted in clear banding patterns of sufficient complexity (16-32 major bands) and reproducibility to investigate differences in bacterial communities in a silt loam soil. Replicate CD DGGE profiles from plates inoculated with less-dilute samples (10(-3)) had a higher band count and were more similar (72-77%) than profiles from more-dilute samples (51-61%). Different culture media and incubation conditions resulted in distinct community fingerprints and increased the cumulative number of unique bands detected. When CD DGGE fingerprints were compared to profiles constructed from 16S rRNA genes obtained from culture-independent clone libraries (CB DGGE profiles) 34% of the bands were unique to the culture-dependent profiles, 32% were unique to the culture-independent profiles and 34% were found in both communities. These data demonstrate that culture-independent DGGE profiles are supplemented by the distinct bands detected in culture-dependent profiles. CD DGGE can be a useful technique to follow the dynamics of distinct culturable fractions of the soil bacterial community. (c) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1570 / 1579
页数:10
相关论文
共 47 条
[41]   Bulk and rhizosphere soil bacterial communities studied by denaturing gradient gel electrophoresis: Plant-dependent enrichment and seasonal shifts revealed [J].
Smalla, K ;
Wieland, G ;
Buchner, A ;
Zock, A ;
Parzy, J ;
Kaiser, S ;
Roskot, N ;
Heuer, H ;
Berg, G .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (10) :4742-4751
[42]   Diversity and seasonal fluctuations of the dominant members of the bacterial soil community in a wheat field as determined by cultivation and molecular methods [J].
Smit, E ;
Leeflang, P ;
Gommans, S ;
van den Broek, J ;
van Mil, S ;
Wernars, K .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (05) :2284-2291
[43]   New strategies for cultivation and detection of previously uncultured microbes [J].
Stevenson, BS ;
Eichorst, SA ;
Wertz, JT ;
Schmidt, TM ;
Breznak, JA .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (08) :4748-4755
[44]   Comparative analysis of bacterial diversity in freshwater sediment of a shallow eutrophic lake by molecular and improved cultivation-based techniques [J].
Tamaki, H ;
Sekiguchi, Y ;
Hanada, S ;
Nakamura, K ;
Nomura, N ;
Matsumura, M ;
Kamagata, Y .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (04) :2162-2169
[45]   Distribution of sulfate-reducing bacteria in a stratified fjord (Mariager fjord, Denmark) as evaluated by most-probable-number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments [J].
Teske, A ;
Wawer, C ;
Muyzer, G ;
Ramsing, NB .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1996, 62 (04) :1405-1415
[46]  
Wright E.L., 1982, SOIL SURVEY MARION M
[47]  
Zuberer D.A., 1994, METHODS SOIL ANAL 2, V5, P119, DOI DOI 10.1016/J.CHEMOSPHERE.2004.05.025