Reverse-transcriptase quantitative PCR method to detect uptake of hydrogen produced from cyanobacteria by Alcaligenes hy
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S H O R T CO M MU N I C A T I O N
Reverse-transcriptase quantitative PCR method to detect uptake of hydrogen produced from cyanobacteria by Alcaligenes hydrogenophilus, an aerobic hydrogen-oxidising bacterium Sarah Schroeder · Anthony Ranchou-Peyruse · Magali Ranchou-Peyruse · Jim C. Spain
Received: 4 October 2010 / Revised: 6 February 2011 / Accepted: 29 April 2011 / Published online: 27 May 2011 © Springer-Verlag 2011
Abstract Hydrogen-oxidising bacteria play a key ecological role in a variety of habitats including the rhizosphere and hot springs. To investigate the possibly of interspecies hydrogen exchange between cyanobacteria and hydrogenoxidising bacteria, we developed a sensitive and reliable reverse-transcriptase qPCR assay for up-regulation of the hupS gene in the knallgas bacterium Alcaligenes hydrogenophilus DSM2625. The assay detected up-regulation of the gene at initial hydrogen concentrations as low as 0.12 M. Expression of hupS also increased in the presence of hydrogen-producing cyanobacteria, both when Ah DSM2625 was directly added to a hydrogen-producing culture of the cyanobacteria, and when cultures were physically separated in a vessel that allowed gas exchange. Additional reWnements and development of the sensitive assay will lead to a better understanding of hydrogen exchange in aerobic ecosystems and development of reporter strains to detect hydrogen-producing organisms. Keywords Hydrogen exchange · Reverse-transcriptase real time PCR · Cyanobacteria · Hydrogen-oxidising
Communicated by William Metcalf. S. Schroeder · A. Ranchou-Peyruse · M. Ranchou-Peyruse · J. C. Spain (&) School of Civil and Environmental Engineering, Georgia Institute of Technology, Atlanta, GA, USA e-mail: [email protected] A. Ranchou-Peyruse · M. Ranchou-Peyruse Equipe Environnement et Microbiologie -UMR IPREM 5254, Université de Pau et des Pays de l’Adour, Bâtiment IBEAS, BP1155, 64013 Pau Cedex, France
Introduction Hydrogen-oxidising bacteria are a phylogenetically diverse group of aerobic, chemolithoautotrophic bacteria, able to derive energy from the oxidation of molecular hydrogen and carbon from CO2. Although they are unable to consume atmospheric H2 due to their elevated concentration thresholds (Conrad et al. 1983), hydrogen-oxidising bacteria are found in a wide range of diverse habitats such as oceans and hot springs, sediment, activated sludge and often in soil surrounding legume nodules (Aragno and Schlegel 1999; Constant et al. 2008; Zhang et al. 2009). The apparent dominance of H2-metabolizing organisms in certain springs in Yellowstone National Park suggests that H2 is the main source of energy for primary production in the ecosystem (Spear et al. 2005; Meyer-Dombard et al. 2005). Hydrogen concentrations in the hot springs ranged up to 300 nM, which is consistent with those reported for other oxygen-poor environments such as lake sediment (36 nM), rice paddies (28 nM) and sewage sludge (203 nM) (Zinder 1993). Cyanobacteria dominating hot spring mats (Boyd et al. 2009; Ley et al. 2006) are known t
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