Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation:: Distribution of the genes in BTEX degrading isolates and in subsurface soils of a BTEX contaminated industrial site

被引:108
作者
Hendrickx, B
Junca, H
Vosahlova, J
Lindner, A
Rüegg, I
Bucheli-Witschel, M
Faber, F
Egli, T
Mau, M
Schlömann, M
Brennerova, M
Brenner, V
Pieper, DH
Top, EM
Dejonghe, W
Bastiaens, L
Springael, D
机构
[1] Catholic Univ Louvain, Lab Soil & Water Management, B-3001 Heverlee, Belgium
[2] Flemish Inst Technol Res, Vito, B-2400 Mol, Belgium
[3] Univ Ghent, Lab Microbial Ecol & Technol, B-9000 Ghent, Belgium
[4] GBF, Biodegradat Grp, D-38124 Braunschweig, Germany
[5] Acad Sci Czech Republ, Inst Microbiol, Dept Cell & Mol Microbiol, Prague 14200 4, Czech Republic
[6] TU Bergakad Freiberg, Interdisziplinares Okol Zentrum, D-09599 Freiberg, Germany
[7] EAWAG, Dept Microbiol, CH-8600 Dubendorf, Switzerland
[8] Univ Groningen, Dept Microbiol, Ctr Ecol Evolutionary Studies, NL-9750 AA Haren, Netherlands
关键词
PCR detection; aerobic BTEX biodegradation; catabolic gene distribution; BTEX degrading isolates; BTEX contaminated site;
D O I
10.1016/j.mimet.2005.04.018
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Eight new primer sets were designed for PCR detection of (i) mono-oxygenase and dioxygenase gene sequences involved in initial attack of bacterial aerobic BTEX degradation and of (ii) catechol 2,3-dioxygenase gene sequences responsible for metacleavage of the aromatic ring. The new primer sets allowed detection of the corresponding genotypes in soil with a detection limit Of 10(3)-10(4) or 10(5)-10(6) gene copies g(-1) soil, assuming one copy of the gene per cell. The primer sets were used in PCR to assess the distribution of the catabolic genes in BTEX degrading bacterial strains and DNA extracts isolated from soils sampled from different locations and depths (vadose, capillary fringe and saturated zone) within a BTEX contaminated site. In both soil DNA and the isolates, tmoA-, xylM- and xylE1-like genes were the most frequently recovered BTEX catabolic genes. xylM and xylE1 were only recovered from material from the contaminated samples while tmoA was detected in material from both the contaminated and non-contaminated samples. The isolates, mainly obtained from the contaminated locations, belonged to the Actinobacteria or Proteobacteria (mainly Pseudomonas). The ability to degrade benzene was the most common BTEX degradation phenotype among them and its distribution was largely congruent with the distribution of the tmoA-like genotype. The presence of tmoA and xylM genes in phylogenetically distant strains indicated the occurrence of horizontal transfer of BTEX catabolic genes in the aquifer. Overall, these results show spatial variation in the composition of the BTEX degradation genes and hence in the type of BTEX degradation activity and pathway, at the examined site. They indicate that bacteria carrying specific pathways and primarily carrying tmoA/xylM/xylE1 genotypes, are being selected upon BTEX contamination. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:250 / 265
页数:16
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