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Hindi Special Issue Article
Ameliorating Risk: Culturable and Metagenomic Monitoring of the 14 Year Decline of a Genetically Engineered Microorganism at a Bioremediation Field Site
Alice C. Layton1*, Abby E. Smartt1, Oya Tekeli2, Archana Chauhan1, Steven Ripp1, Daniel E. Williams1, Whitney Burton1, Scott Moser1, Jana Phillips2, Anthony V. Palumbo2 and Gary S. Sayler1,21The University of Tennessee Center for Environmental Biotechnology, 676 Dabney Hall, Knoxville, Tennessee 37996, USA
2Oak Ridge National Laboratory, Biosciences Division, Oak Ridge, Tennessee, 37831, USA
- *Corresponding Author:
- Dr. Alice C. Layton
The University of Tennessee
Center for Environmental Biotechnology
676 Dabney Hall, Knoxville, Tennessee 37996, USA
Tel: 865-974-8072
Fax: 865-974-8086
E-mail: alayton@utk.edu
Received February 28, 2012; Accepted April 20, 2012; Published April 22, 2012
Citation: Layton AC, Smartt AE, Chauhan A, Ripp S, Williams DE, et al. (2012) Ameliorating Risk: Culturable and Metagenomic Monitoring of the 14 Year Decline of a Genetically Engineered Microorganism at a Bioremediation Field Site. J Bioremed Biodegrad S1:009 doi: 10.4172/2155-6199.S1-009
Copyright: © 2012 Layton AC, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
In 1996, the first EPA sanctioned release of a recombinant microbe (Pseudomonas fluorescens HK44) into the subsurface soil environment was initiated in a replicated semi-contained array of soil lysimeters. With an aim to access the survivability/environmental fate of HK44, soil sampling was performed 14 years post release. Although after extensive sampling culturable HK44 cells were not found, qPCR and metagenomic analyses indicated that genetic signatures of HK44 cells still persisted in the soils, with genes diagnostic for the bioluminescent transposon carried by strain HK44 (luxA and tetA) being found at low concentrations (< 5000 copies/g). Additionally, metagenome analysis of lysimeter 2 using amplicon pyrosequencing showed that Burkholderia was more abundant in the sample extracted before storage at 4°C than after storage at 4°C (79% and 5.6% Burkholderia sequences, respectively).
