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Vol. 72, No: 5, 2006; Pages: 3111–3118

Horizontal Gene Transfer of PIB-Type ATPases among Bacteria Isolated from Radionuclide- and Metal-Contaminated Subsurface Soils

Robert J. Martinez,1 Yanling Wang,2 Melanie A. Raimondo,1 Jonna M. Coombs,3
Tamar Barkay,4 and Patricia A. Sobecky1*

School of Biology, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, GA 30332-0230.


Aerobic heterotrophs were isolated from subsurface soil samples obtained from the U.S. Department of Energy’s (DOE) Field Research Center (FRC) located at Oak Ridge, Tenn. The FRC represents a unique, extreme environment consisting of highly acidic soils with cooccurring heavy metals, radionuclides, and high nitrate concentrations. Four hundred isolates obtained from contaminated soil were assayed for heavy metal resistance, and a smaller subset was assayed for tolerance to uranium. The vast majority of the isolates were gram-positive bacteria and belonged to the high-G+C- and low-G+C-content genera Arthrobacter and Bacillus, respectively. Genomic DNA from a randomly chosen subset of 50 Pb-resistant (Pbr) isolates was amplified with PCR primers specific for PIB-type ATPases (i.e., pbrA/cadA/zntA). A total of 10 pbrA/cadA/zntA loci exhibited evidence of acquisition by horizontal gene transfer. A remarkable dissemination of the horizontally acquired PIB-type ATPases was supported by unusual DNA base compositions and phylogenetic incongruence. Numerous Pbr PIB-type ATPase-positive FRC isolates belonging to the genus Arthrobacter tolerated toxic concentrations of soluble U(VI) (UO22+) at pH 4. These unrelated, yet synergistic, physiological traits observed in Arthrobacter isolates residing in the contaminated FRC subsurface may contribute to the survival of the organisms in such an extreme environment. This study is, to the best of our knowledge, the first study to report broad horizontal transfer of PIB-type ATPases in contaminated subsurface soils and is among the first studies to report uranium tolerance of aerobic heterotrophs obtained from the acidic subsurface at the DOE FRC.

Keywords:Aerobic heterotrophs;Field Research Center;Arthrobacter;Bacillus;Genomic DNA;pbrA/cadA/zntA;Arthrobacter tolerated toxic;Desulfovibrio sp;Geobacter sp;Shewanella sp;radionuclide.

Corresponding author: Tel (404) 894-5819; Fax (404) 385-4440.

E-mail: patricia.sobecky@biology.gatech.edu


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