Abstract
The aim of this work was to investigate the susceptibility of the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) to biodegradation in a range of soils and to identify RDX-degrading organisms using stable isotope probing (SIP). RDX degradation was examined in ten soils, primarily from agricultural areas. RDX biodegradation was observed in six samples and only when the microcosms were not aerated. For one soil, 15N- and 13C-based DNA SIP was used to identify the microorganisms responsible for RDX degradation. Two RDX concentrations were examined (10 and 20 mg/L), however, only the higher concentration resulted in a significant SIP signal. In these ultracentrifugation fractions, one terminal restriction fragment length polymorphism (TRFLP) fragment (260 bp) showed a reliable trend of label uptake. This fragment was of higher relative abundance in the heavier fractions from labeled samples compared with the heavier fractions from the unlabeled control samples, indicating that the organism producing this fragment was responsible for label uptake (hence RDX degradation). Partial 16S rRNA gene sequencing indicated the organisms represented by fragment 260 bp belonged to either Sphingobacteria (phylum Bacteroidetes) or the phylum Acidobacteria. To date, these organisms have not previously been directly linked to RDX degradation. The 16S rRNA sequences were compared with the NCBI database and, in all cases, were most similar to uncultured bacteria. The results suggest SIP is a viable method for discovering novel, previously uncultured, RDX degraders.
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This project was supported by the Strategic Environmental Research and Development Program. The following people are acknowledged for their work on this project. Thanks to Dan Williams for sample collection and Yanglang Pan for analytical assistance. The authors thank Stuart Strand and Peter Andeer for supplying R. rhodochrous 11Y genomic DNA for the xplA studies.
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Jayamani, I., Manzella, M.P. & Cupples, A.M. RDX Degradation Potential in Soils Previously Unexposed to RDX and the Identification of RDX-Degrading Species in One Agricultural Soil Using Stable Isotope Probing. Water Air Soil Pollut 224, 1745 (2013). https://doi.org/10.1007/s11270-013-1745-4
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DOI: https://doi.org/10.1007/s11270-013-1745-4