scholarly journals Potential microbial contamination during sampling of permafrost soil assessed by tracers

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Toke Bang-Andreasen ◽  
Morten Schostag ◽  
Anders Priemé ◽  
Bo Elberling ◽  
Carsten S. Jacobsen

Abstract Drilling and handling of permanently frozen soil cores without microbial contamination is of concern because contamination e.g. from the active layer above may lead to incorrect interpretation of results in experiments investigating potential and actual microbial activity in these low microbial biomass environments. Here, we present an example of how microbial contamination from active layer soil affected analysis of the potentially active microbial community in permafrost soil. We also present the development and use of two tracers: (1) fluorescent plastic microspheres and (2) Pseudomonas putida genetically tagged with Green Fluorescent Protein production to mimic potential microbial contamination of two permafrost cores. A protocol with special emphasis on avoiding microbial contamination was developed and employed to examine how far microbial contamination can penetrate into permafrost cores. The quantity of tracer elements decreased with depth into the permafrost cores, but the tracers were detected as far as 17 mm from the surface of the cores. The results emphasize that caution should be taken to avoid microbial contamination of permafrost cores and that the application of tracers represents a useful tool to assess penetration of potential microbial contamination into permafrost cores.

2012 ◽  
Vol 77 (6) ◽  
pp. 603-608 ◽  
Author(s):  
T. V. Komarova ◽  
E. V. Sheval ◽  
D. V. Pozdyshev ◽  
V. S. Kolesnikova ◽  
Yu. L. Dorokhov

2011 ◽  
pp. 391-406
Author(s):  
Susan Schlegel ◽  
Mirjam Klepsch ◽  
Dimitra Gialama ◽  
David Wickström ◽  
David Drew ◽  
...  

2010 ◽  
Vol 77 (4) ◽  
pp. 1263-1267 ◽  
Author(s):  
Charissa de Bekker ◽  
G. Jerre van Veluw ◽  
Arman Vinck ◽  
L. Ad Wiebenga ◽  
Han A. B. Wösten

ABSTRACTThe fungusAspergillus nigerforms (sub)millimeter microcolonies within a liquid shaken culture. Here, we show that such microcolonies are heterogeneous with respect to size and gene expression. Microcolonies of strains expressing green fluorescent protein (GFP) from the promoter of the glucoamlyase geneglaAor the ferulic acid esterase genefaeAwere sorted on the basis of diameter and fluorescence using the Complex Object Parametric Analyzer and Sorter (COPAS) technology. Statistical analysis revealed that the liquid shaken culture consisted of two populations of microcolonies that differ by 90 μm in diameter. The population of small microcolonies of strains expressing GFP from theglaAorfaeApromoter comprised 39% and 25% of the culture, respectively. Two populations of microcolonies could also be distinguished when the expression of GFP in these strains was analyzed. The population expressing a low level of GFP consisted of 68% and 44% of the culture, respectively. We also show that mRNA accumulation is heterogeneous within microcolonies ofA. niger. Central and peripheral parts of the mycelium were isolated with laser microdissection and pressure catapulting (LMPC), and RNA from these samples was used for quantitative PCR analysis. This analysis showed that the RNA content per hypha was about 45 times higher at the periphery than in the center of the microcolony. Our data imply that the protein production ofA. nigercan be improved in industrial fermentations by reducing the heterogeneity within the culture.


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