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Exploiting the fungal highway: development of a novel tool for the in situ isolation of bacteria migrating along fungal mycelium

2015, Simon, Anaele, Bindschedler, Saskia, Job, Daniel, Wick, Lukas Y., Filippidou, Sevasti, Kooli, Wafa M., Verrecchia, Eric P., Junier, Pilar

Fungi and bacteria form various associations that are central to numerous environmental processes. In the so-called fungal highway, bacteria disperse along fungal mycelium. We developed a novel tool for the in situ isolation of bacteria moving along fungal hyphae as well as for the recovery of fungi potentially involved in dispersal, both of which are attracted towards a target culture medium. We present the validation and the results of the first in situ test. Couples of fungi and bacteria were isolated from soil. Amongst the enriched organisms, we identified several species of fast-growing fungi (Fusarium sp. and Chaetomium sp.), as well as various potentially associated bacterial groups, including Variovorax soli, Olivibacter soli, Acinetobacter calcoaceticus, and several species of the genera Stenotrophomonas, Achromobacter and Ochrobactrum. Migration of bacteria along fungal hyphae across a discontinuous medium was confirmed in most of the cases. Although the majority of the bacteria for which migration was confirmed were also positive for flagellar motility, not all motile bacteria dispersed using their potential fungal partner. In addition, the importance of hydrophobicity of the fungal mycelial surface was confirmed. Future applications of the columns include targeting different types of microorganisms and their interactions, either by enrichment or by state of the art molecular biological methods.

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Mitchell, E. A. D., Belbahri, L., Job, D., Pawlowski, J, Lara, E. Exploring the Terra incognita of unknown eukaryotic diversity in Soils - A major challenge we now have the tools to tackle!

2011-10-10, Job, Daniel

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Guggiari M., Bloque R., Aragno M., Verrecchia E.P., Job D. Role and dynamics of calcium oxalate production by selected Fungi

2010-6-15, Job, Daniel

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Fungal implication in secondary calcium carbonate accumulation in soils and caves

2009, Bindschedler, Saskia, Milliere, Laure, Cailleau, Guillaume, Job, Daniel, Verrecchia, Eric P.

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Isolation of oxalotrophic bacteria able to disperse on fungal mycelium

2013, Bravo, Daniel, Cailleau, Guillaume, Bindschedler, Saskia, Simon, Anaele, Job, Daniel, Verrecchia, Eric, Junier, Pilar

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Edith Joseph, Anaëlle Simon, Silvia Prati, Marie Wörle, Daniel Job, Rocco Mazzeo. Development of an analytical procedure for evaluation of the protective behavior of innovative fungal patinas on archaeological and artistic metal artefacts

2011-6-15, Job, Daniel

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Khammar N. Gaëtan M. Ferro K. Job D, Verrecchia E. and Aragno M. 2009. Use of the frc gene as a molecular marker to characterize oxalate-oxidizing bacterial abundance and diversity structure in soil

2009-6-15, Job, Daniel

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Guggiari M., Bloque R., Aragno M., Verrecchia E., Job D., Junier P.Experimental calcium oxalate crystals production and metabolization by selected wood rot fungi

2011-10-21, Job, Daniel

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Saskia Bindschedler, L. Millière, G. Cailleau, D. Job and E. P. Verrecchia. Calcitic nanofibres in soils and caves: a putative fungal contribution to carbonatogenesis

2010-6-15, Job, Daniel

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Use of the frc gene as a molecular marker to characterize oxalate-oxidizing bacterial abundance and diversity structure in soil

2009, Khammar, Nadia, Martin, Gaëtan, Ferro, Katia, Job, Daniel, Aragno, Michel, Verrecchia, Eric

Oxalate catabolism, which can have both medical and environmental implications, is performed by phylogenetically diverse bacteria. The formyl-CoA-transferase gene was chosen as a molecular marker of the oxalotrophic function. Degenerated primers were deduced from an alignment of frc gene sequences available in databases. The specificity of primers was tested on a variety of frc-containing and frc-lacking bacteria. The frc-primers were then used to develop PCR-DGGE and real-time SybrGreen PCR assays in soils containing various amounts of oxalate. Some PCR products from pure cultures and from soil samples were cloned and sequenced. Data were used to generate a phylogenetic tree showing that environmental PCR products belonged to the target physiological group. The extent of diversity visualised on DGGE pattern was higher for soil samples containing carbonate resulting from oxalate catabolism. Moreover, the amount of frc gene copies in the investigated soils was detected in the range of 1.64x10(7) to 1.75x10(8)/g of dry soil under oxalogenic tree (representing 0.5 to 1.2% of total 16S rRNA gene copies), whereas the number of frc gene copies in the reference soil was 6.4x10(6) (or 0.2% of 16S rRNA gene copies). This indicates that oxalotrophic bacteria are numerous and widespread in soils and that a relationship exists between the presence of the oxalogenic trees Milicia excelsa and Afzelia africana and the relative abundance of oxalotrophic guilds in the total bacterial communities. This is obviously related to the accomplishment of the oxalate-carbonate pathway, which explains the alkalinization and calcium carbonate accumulation occurring below these trees in an otherwise acidic soil. The molecular tools developed in this study will allow in-depth understanding of the functional implication of these bacteria on carbonate accumulation as a way of atmospheric CO2 sequestration. (c) 2008 Elsevier B.V. All rights reserved.