RESUMEN
To cope with heterogeneous subsurface environments mycelial microorganisms have developed a unique ramified growth form. By extending hyphae, they can obtain nutrients from remote places and transport them even through air gaps and in small pore spaces, repectively. To date, studies have been focusing on the role that networks play in the distribution of nutrients. Here, we investigated the role of mycelia for the translocation of nonessential substances, using polycyclic aromatic hydrocarbons (PAHs) as model compounds. We show that the hyphae of the mycelial soil oomycete Pythium ultimum function as active translocation vectors for a wide range of PAHs. Visualization by two-photon excitation microscopy (TPEM) demonstrated the uptake and accumulation of phenanthrene (PHE) in lipid vesicles and its active transport by cytoplasmic streaming of the hyphae ('hyphal pipelines'). In mycelial networks, contaminants were translocated over larger distances than by diffusion. Given their transport capacity and ubiquity, hyphae may substantially distribute remote hydrophobic contaminants in soil, thereby improving their bioavailability to bacterial degradation. Hyphal contaminant dispersal may provide an untapped potential for future bioremediation approaches.
Asunto(s)
Micelio/metabolismo , Hidrocarburos Policíclicos Aromáticos/metabolismo , Pythium/metabolismo , Biodegradación Ambiental , Transporte Biológico Activo , Microscopía de Fluorescencia por Excitación Multifotónica , Fenantrenos/metabolismo , Factores de TiempoRESUMEN
Mycelia of fungi and soil oomycetes have recently been found to act as effective paths boosting bacterial mobility and bioaccessibility of contaminants in vadose environments. In this study, we demonstrate that mycelia can be used for targeted separation and isolation of contaminant-degrading bacteria from soil. In a 'proof of concept' study we developed a novel approach to isolate bacteria from contaminated soil using mycelia of the soil oomycete Pythium ultimum as translocation networks for bacteria and the polycyclic aromatic hydrocarbon naphthalene (NAPH) as selective carbon source. NAPH-degrading bacterial isolates were affiliated with the genera Xanthomonas, Rhodococcus and Pseudomonas. Except for Rhodococcus the NAPH-degrading isolates exhibited significant motility as observed in standard swarming and swimming motility assays. All steps of the isolation procedures were followed by cultivation-independent terminal 16S rRNA gene terminal fragment length polymorphism (T-RFLP) analysis. Interestingly, a high similarity (63%) between both the cultivable NAPH-degrading migrant and the cultivable parent soil bacterial community profiles was observed. This suggests that mycelial networks generally confer mobility to native, contaminant-degrading soil bacteria. Targeted, mycelia-based dispersal hence may have high potential for the isolation of bacteria with biotechnologically useful properties.
Asunto(s)
Bacterias/aislamiento & purificación , Bacterias/metabolismo , Técnicas Microbiológicas/métodos , Micelio/química , Naftalenos/metabolismo , Microbiología del Suelo , Contaminantes del Suelo/metabolismo , Bacterias/clasificación , Bacterias/genética , Biodegradación AmbientalRESUMEN
Contaminant biodegradation in soil is frequently limited by hindered physical access of bacteria to the contaminants. In the frame of the development of novel bioremediation approaches based on ecological principles, we tested the hypothesis that fungal networks facilitate the movement of bacteria by providing continuous liquid films in which gradients of chemoattractants can form and chemotactic swimming can take place. Unlike bacteria, filamentous fungi spread with ease in water-unsaturated soil. In a simple laboratory model of a water-unsaturated environment, we studied the movement of polycyclic aromatic hydrocarbon-degrading Pseudomonas putida PpG7 (NAH7) along a mycelium of Pythium ultimum. Some undirected dispersal was observed in the absence of a chemoattractant or when the non-chemotactic derivative strain P. putida G7.C1 (pHG100) was used. The bacterial movement became fourfold more effective and clearly directed when the chemotactic wild type was used and salicylate was present as a chemoattractant. No dispersal of bacteria was found in the absence of the fungus. These findings point at a role of mycelia for the translocation of chemicals and microorganisms. The results suggest that fungi improve the accessibility of contaminants in water-unsaturated environments.