Pseudomonas frederiksbergensis

About: Pseudomonas frederiksbergenis is a gram-negative bacteria that was originally identified in Frederiksberg, Copenhagen, Denmark at a coal gasification site. This bacteria is rod shaped, and colonies are known to be a pale yellow. Pseudomonas is known to be a species that is important for soil, food and water decomposition.

Date Collected: 2/8/2017

Methods of isolation and characterization: 

  • P. frederiksbergenis was isolated from a specific soil sample at the Environmental Education Center (EEC) located in Farmville, VA.
  • The soil sample was taken directly from Buffalo Creek, a small creek that flows into the Appomattox river.
  • Once we collected our sample we plated it onto three agar plates: direct sample, 1/10 and 1/100 dilution.
  • We observed growth at 24hrs and 48hrs and selected a specific colony to analyze that was not overlapping other colonies.
  • We ran DNA isolation and purification through Nano Drop to ensure we had a pure sample to then proceed to PCR amplification of 16s rRNA.
  • After isolating 16s rRNA of our colony we ran gel electrophoresis against MSp1 to ensure we isolated 16s rRNA to send off for sequencing.
  • Sadly we were unable to draw any conclusions from our gel due to clarity issues, never the less we went ahead and sent off our sample off to be sequenced.
  • Once sequenced, we than took our sequence and ran it through BLAST analysis which gave us a 99% match
  • Using our data analysis from BLAST as well as the color, form, and size of our colony we identified our sample to be P. frederiksbergenis.

Results:

 

Figure 1. Chromatogram of our Buffalo Creek soil sample, which we identified as P. frederiksbergenis.

 

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Figure 2. Buffalo Creek soil sample and location in which we isolated P. frederiksbergenis.

 

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Figure 3. Our colony of P. frederiksbergenis under a microscope. This photo highlights the pale yellow color that we used to further support our classification.

 

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Figure 4.   The 1/100 dilution plate of the Buffalo Creek soil shows the specific colony we isolated P. frederiksbergenis.

 

References:

Pseudomonas frederiksbergensis sp. nov., isolated from soil at a coal gasification site.
S. M. Andersen, K. Johnsen, J. Sørensen, P. Nielsen, C. S. Jacobsen
Int J Syst Evol Microbiol. 2000 Nov; 50(Pt 6): 1957–1964. doi: 10.1099/00207713-50-6-1957