In the present study, microbial toluene degradation in controlled constructed wetland model systems, planted fixed-bed reactors (PFRs), was queried with DNA-based methods in combination with stable isotope fractionation analysis and characterization of toluene-degrading microbial isolates. Two PFR replicates were operated with toluene as the sole external carbon and electron source for 2 years. The bulk redox conditions in these systems were hypoxic to anoxic. The autochthonous bacterial communities, as analyzed by Illumina sequencing of 16S rRNA gene amplicons, were mainly comprised of the families Xanthomonadaceae, Comamonadaceae, and Burkholderiaceae, plus Rhodospirillaceae in one of the PFR replicates. DNA microarray analyses of the catabolic potentials for aromatic compound degradation suggested the presence of the ring monooxygenation pathway in both systems, as well as the anaerobic toluene pathway in the PFR replicate with a high abundance of Rhodospirillaceae. The presence of catabolic genes encoding the ring monooxygenation pathway was verified by quantitative PCR analysis, utilizing the obtained toluene-degrading isolates as references. Stable isotope fractionation analysis showed low-level of carbon fractionation and only minimal hydrogen fractionation in both PFRs, which matches the fractionation signatures of monooxygenation and dioxygenation. In combination with the results of the DNA-based analyses, this suggests that toluene degradation occurs predominantly via ring monooxygenation in the PFRs. Biology-based remediation technologies (bioremediation) for the treatment of groundwater and soils polluted with organic compounds have been receiving high interest due to their low cost, high efficiency, and relative operational simplicity (1). Rhizoremediation is one such effective bioremediation approach, where the transformation of contaminants to innocuous products may be enhanced by plant-microbe interactions occurring in the rhizosphere (2-4). The plants provide the rhizospheric microbial community with root exudates such as carbohydrates, amino acids/amines, and organic acids (5) and thus promote the microbes' establishment and proliferation. Rhizoremediation is particularly effective in constructed wetlands. These treatment systems have been applied for the removal of even high loads of contaminants present in inflow waters (6-8). In addition to the input of labile organic carbon, the helophytes used in these systems have the capacity of channeling significant amounts of oxygen from the atmosphere through a specific tissues, the aerenchyma, into their roots, and thereby foster aerobic microbial activities in the rhizosphere (9).In order to enhance the performance of constructed wetlands through engineering optimizations of the systems, it is deemed necessary to understand the functionality of the relevant microbial community present in the rhizosphere, and some efforts have been pursued in this direction (7, 10). However, due to the presence of steep chemical gradients and variable environmental ...
Previously, Planted Fixed-Bed Reactors (PFRs) have been used to investigate microbial toluene removal in the rhizosphere of constructed wetlands. Aerobic toluene degradation was predominant in these model systems although bulk redox conditions were hypoxic to anoxic. However, culture-independent approaches indicated also that microbes capable of anaerobic toluene degradation were abundant. Therefore, we aimed at isolating anaerobic-toluene degraders from one of these PFRs. From the obtained colonies which consisted of spirilli-shaped bacteria, a strain designated 15–1 was selected for further investigations. Analysis of its 16S rRNA gene revealed greatest similarity (99%) with toluene-degrading Magnetospirillum sp. TS-6. Isolate 15–1 grew with up to 0.5 mM of toluene under nitrate-reducing conditions. Cells reacted to higher concentrations of toluene by an increase in the degree of saturation of their membrane fatty acids. Strain 15–1 contained key genes for the anaerobic degradation of toluene via benzylsuccinate and subsequently the benzoyl-CoA pathway, namely bssA, encoding for the alpha subunit of benzylsuccinate synthase, bcrC for subunit C of benzoyl-CoA reductase and bamA for 6-oxocyclohex-1-ene-1-carbonyl-CoA hydrolase. Finally, most members of a clone library of bssA generated from the PFR had highest similarity to bssA from strain 15–1. Our study provides insights about the physiological capacities of a strain of Magnetospirillum isolated from a planted system where active rhizoremediation of toluene is taking place.
Pink disease is a major problem in the pineapple canning industry. Affected fruit acquire a brownish pigment after pasteurization and can contaminate non-affected fruit before they are released to the consumer. In the last few years, Pantoea citrea has been described as the causative agent of pink disease. In this study, over 300 bacterial isolates from pineapple plants, growing in Mexican commercial fields, and from soil close to plant roots were recovered. Over 250 isolates showed a very high similarity in their phenotypic and genotypic traits with Tatumella ptyseos, a close relative of Pantoea. These isolates exhibited typical pathogenicity reactions in pineapple juice tests, pineapple slices and fruit. On this basis, molecular identification procedures for the Tatumella isolates associated with pink disease were implemented. In affected fruit populations around 10 6 CFU ⁄ g of fresh tissue were recovered. This is first time that T. ptyseos is demonstrated as a causal agent of pink disease.
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