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Presence, diversity and enumeration of functional genes (bssA and bamA) relating to toluene degradation across a range of redox conditions and inoculum sources

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Abstract

The study investigates two functional genes for toluene degradation across three redox conditions (nitrate and sulfate amended and methanogenic). The genes targeted include benzylsuccinate synthase α-subunit (bssA) and a gene recently identified as being a strong indicator of anaerobic aromatic degradation, called 6-oxocylcohex-1-ene-1-carbonyl-CoA hydrolase (bamA). In all, sixteen different anaerobic toluene degrading microcosms were investigated using several primers sets targeting bssA and one primer set targeting bamA. One bssA primer set (7772f/8546r) was the most successful in producing a strong amplicon (eight from sixteen) with the other bssA primers sets producing strong amplicons in six or less samples. In contrast, the bamA primer set (bam-sp9 and bam-asp1) produced a strong amplicon in DNA extracted from all except one microcosm. Partial bssA and bamA sequences were obtained for a number of samples and compared to those available in GenBank. The partial bssA sequences (from nitrate amended and methanogenic microcosms) were most similar to Thauera sp. DNT-1, Thauera aromatica, Aromatoleum aromaticum EbN1 and bssA clones from a study involving sulfate reducing toluene degradation. The bamA sequences obtained could be placed into five previously defined clades (bamA-clade 1, Georgfuchsia/Azoarcus, Magnetospirillum/Thauera Syntrophus and Geobacter clades), with the placement generally depending on redox conditions. Gene numbers were also correlated with toluene degradation and the final gene number for both genes differed considerably between the range of redox conditions. The work is the first in depth investigation of bamA diversity over a range of redox conditions and inoculum sources.

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Acknowledgments

Funding for this work was provided by a grant awarded to A. Cupples from the National Science Foundation (Grant 0853249). The authors thank Paul Fallgren (Western Research Institute), Kelvin Wong, Catherine Garnham, and Zhenbo Yue (Michigan State University) for supplying the contaminated soil sample (CSS), digester sludge (DSS), activated sludge (ASM) and anaerobic granular sludge (GSN) respectively.

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  1. Department of Civil and Environmental Engineering, A135 Research Engineering Complex, Michigan State University, East Lansing, MI, 48824, USA

    Weimin Sun, Xiaxuo Sun & Alison M. Cupples

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  1. Weimin Sun
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  2. Xiaxuo Sun
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  3. Alison M. Cupples
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Correspondence to Alison M. Cupples.

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Sun, W., Sun, X. & Cupples, A.M. Presence, diversity and enumeration of functional genes (bssA and bamA) relating to toluene degradation across a range of redox conditions and inoculum sources. Biodegradation 25, 189–203 (2014). https://doi.org/10.1007/s10532-013-9651-4

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