DNA extraction method affects microbial community profiles from soils and sediment

被引:132
作者
Carrigg, Cora
Rice, Olivia
Kavanagh, Siobhan
Collins, Gavin
O'Flaherty, Vincent
机构
[1] Natl Univ Ireland, Dept Microbiol, Microbial Ecol Lab, Galway, Ireland
[2] Natl Univ Ireland, Environm Change Inst, Galway, Ireland
[3] Natl Univ Ireland Univ Coll Cork, Dept Microbiol, Biomerit Res Ctr, Cork, Ireland
[4] Natl Univ Ireland, DNA Lab, Natl Diagnost Ctr, Galway, Ireland
[5] Max Planck Inst Marine Microbiol, Nutrient Grp, D-28359 Bremen, Germany
关键词
soil; sediment; DNA extraction; DGGE; lysis efficiency; microbial diversity;
D O I
10.1007/s00253-007-1219-y
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
To evaluate whether different deoxyribonucleic acid (DNA) extraction procedures can affect estimates of bacterial community composition, based on the 16S ribosomal ribonucleic acid gene denaturing gradient gel electrophoresis (DGGE) profiles, we compared four in situ lysis procedures using three soils and one marine sediment. Analysis of DGGE profiles, generated by polymerase chain reaction of purified DNA extracts, demonstrated that the choice of DNA extraction method significantly influenced the bacterial community profiles generated. This was reflected both in the number of bands or ribotypes detected from each sample and in subsequent principle coordinate analysis and unweighted-pair group method using arithmetic average analyses. The methods also differed significantly in their robustness, i.e. reproducibility across multiple analyses. Two methods, both based on bead beating, were demonstrated to be suitable for comparative studies of a range of soil and sediment types.
引用
收藏
页码:955 / 964
页数:10
相关论文
共 61 条
[31]  
Miller DN, 1999, APPL ENVIRON MICROB, V65, P4715
[32]  
MORE MI, 1994, APPL ENVIRON MICROB, V60, P1572
[33]   PROFILING OF COMPLEX MICROBIAL-POPULATIONS BY DENATURING GRADIENT GEL-ELECTROPHORESIS ANALYSIS OF POLYMERASE CHAIN REACTION-AMPLIFIED GENES-CODING FOR 16S RIBOSOMAL-RNA [J].
MUYZER, G ;
DEWAAL, EC ;
UITTERLINDEN, AG .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1993, 59 (03) :695-700
[34]   Microbial diversity and soil functions [J].
Nannipieri, P ;
Ascher, J ;
Ceccherini, MT ;
Landi, L ;
Pietramellara, G ;
Renella, G .
EUROPEAN JOURNAL OF SOIL SCIENCE, 2003, 54 (04) :655-670
[35]  
NIELSEN MN, 2002, 338 NAT ENV RES I
[36]   Extraction and purification of DNA in rhizosphere soil samples for PCR-DGGE analysis of bacterial consortia [J].
Niemi, RM ;
Heiskanen, I ;
Wallenius, K ;
Lindström, K .
JOURNAL OF MICROBIOLOGICAL METHODS, 2001, 45 (03) :155-165
[37]   Spatial distribution of bacterial communities and their relationships with the micro-architecture of soil [J].
Nunan, N ;
Wu, KJ ;
Young, IM ;
Crawford, JW ;
Ritz, K .
FEMS MICROBIOLOGY ECOLOGY, 2003, 44 (02) :203-215
[38]   Epistemic and ethical duties in clinical decision-making [J].
O'Donohue, W ;
Henderson, D .
BEHAVIOUR CHANGE, 1999, 16 (01) :10-19
[39]   Soil molecular microbial ecology at age 20: methodological challenges for the future [J].
Ogram, A .
SOIL BIOLOGY & BIOCHEMISTRY, 2000, 32 (11-12) :1499-1504
[40]  
OLSEN GJ, 1986, ANNU REV MICROBIOL, V40, P337, DOI 10.1146/annurev.micro.40.1.337