Up-dating the Cholodny method using PET films to sample microbial communities in soil
The aim of this work was to investigate the use of PET (polyethylene terephtalate) films as a modern development of Cholodny’s glass slides, to enable microscopy and molecular-based analysis of soil communities where spatial detail at the scale of microbial habitatsis essential to understand microb...
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| Datum: | 2011 |
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| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | English |
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Інститут молекулярної біології і генетики НАН України
2011
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| Schriftenreihe: | Вiopolymers and Cell |
| Schlagworte: | |
| Online Zugang: | http://dspace.nbuv.gov.ua/handle/123456789/156399 |
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| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Zitieren: | Up-dating the Cholodny method using PET films to sample microbial communities in soil / O.V. Moshynets, A. Koza, P.Dello Sterpaio, V.A. Kordium, A.J. Spiers // Вiopolymers and Cell. — 2011. — Т. 27, № 3. — С. 199-205. — Бібліогр.: 29 назв. — англ. |
Institution
Digital Library of Periodicals of National Academy of Sciences of Ukraine| Zusammenfassung: | The aim of this work was to investigate the use of PET (polyethylene terephtalate) films as a modern development of Cholodny’s glass slides, to enable microscopy and molecular-based analysis of soil communities where
spatial detail at the scale of microbial habitatsis essential to understand microbial associations and interactions
in this complex environment. Methods. Classical microbiological methods; attachment assay; surface tension
measurements; moleculartechniques: DNA extraction, PCR; confocal laserscanning microscopy (CLSM); micro-focus X-ray computed tomography (µCT). Results. We first show, using the model soil and rhizosphere bacteria Pseudomonas fluorescens SBW25 and P. putida KT2440, that bacteria are able to attach and detach from
PET films, and that pre-conditioning with a filtered soil suspension improved the levels of attachment. Bacteria
attached to the films were viable and could develop substantial biofilms. PET films buried in soil were rapidly
colonised by microorganisms which could be investigated by CLSM and recovered onto agar plates. Secondly,
we demonstrate that µCT can be used to non-destructively visualise soil aggregate contact points and pore spaces across the surface of PET films buried in soil. Conclusions. PET films are a successful development of Cholodny’s glass slides and can be used to sample soil communities in which bacterial adherence, growth, biofilm
and community development can be investigated. The use of these films with µCT imaging in soil will enable a
better understanding of soil micro-habitats and the spatially-explicit nature of microbial interactions in this
complex environment.
Keywords: Pseudomonas, soil, buried slide, PET film. |
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