Background: Natural populations of the teleost fish Fundulus heteroclitus tolerate a broad range of environmental conditions including temperature, salinity, hypoxia and chemical pollutants. Strikingly, populations of Fundulus inhabit and have adapted to highly polluted Superfund sites that are contaminated with persistent toxic chemicals. These natural populations provide a foundation to discover critical gene pathways that have evolved in a complex natural environment in response to environmental stressors.
To study the mechanisms soil bacteria use to cope with elevated concentrations of heavy metals in the environment, a mutagenesis with the lacZ-based reporter gene transposon Tn5B20 was performed. Random gene fusions in the genome of the common soil bacterium Pseudomonas fluorescens strain ATCC 13525 were used to create a bank of 5,000 P. fluorescens mutants. This mutant bank was screened for differential gene expression in the presence of the toxic metal cadmium. Fourteen mutants were identified that responded with increased or reduced gene expression to the presence of cadmium. The mutants were characterized with respect to their metal-dependent gene expression and their metal tolerance. Half the identified mutants reacted with differential gene expression specifically to the metal cadmium, whereas some of the other mutants also responded to elevated concentrations of copper and zinc ions. One of the mutants, strain C8, also showed increased gene expression in the presence of the solvent ethanol, but otherwise no overlap between cadmium-induced gene expression and general stress response was detected. Molecular analysis of the corresponding genetic loci was performed using arbitrary polymerase chain reaction (PCR), DNA sequencing and comparison of the deduced protein products with sequences deposited in genetic databases. Some of the genetic loci targeted by the transposon did not show any similarities to any known genes; thus, they may represent 'novel' loci. The hypothesis that genes that are differentially expressed in the presence of heavy metals play a role in metal tolerance was verified for one of the mutants. This mutant, strain C11, was hypersensitive to cadmium and zinc ions. In mutant C11, the transposon had inserted into a genetic region displaying similarity to genes encoding the sensor/regulator protein pairs of two-component systems that regulate gene expression in metal-resistant bacteria, including czcRS of Ralstonia eutropha, czrRS of Pseudomonas aeruginosa and copRS of Pseudomonas syringae. Although the P. fluorescens strain used in this study had not been isolated from a metal-rich environment, it nevertheless contained at least one genetic region enabling it to cope with elevated concentrations of heavy metals.
Background: Mesophotic corals (light-dependent corals in the deepest half of the photic zone at depths of 30 -150 m) provide a unique opportunity to study the limits of the interactions between corals and endosymbiotic dinoflagellates in the genus Symbiodinium. We sampled Leptoseris spp. in Hawaii via manned submersibles across a depth range of 67 -100 m. Both the host and Symbiodinium communities were genotyped, using a non-coding region of the mitochondrial ND5 intron (NAD5) and the nuclear ribosomal internal transcribed spacer region 2 (ITS2), respectively.
The Kalamazoo River Superfund site in Michigan is contaminated with polychlorinated biphenyls (PCBs), which were heavily discharged into the river from several paper companies as part of the deinking process in the 1950s through 1970s. We characterized biomarkers of chronic PCB exposure in a resident fish population using real-time reverse transcriptase-polymerase chain reaction to examine mRNA expression levels of multiple genes in carp (Cyprinus carpio) liver from PCB contaminated and reference sites in the Kalamazoo River. We also measured these same genes in juvenile carp exposed to dietary PCBs for 4 months. Kalamazoo River carp had significantly increased levels of cytochrome P450 1A (CYP1A) mRNA as did carp fed PCBs in the laboratory. No significant mRNA upregulation occurred in the specific oxidative stress genes (gamma-glutamylcysteine synthetase and magnesium superoxide dismutase) and metabolic genes (phosphoenolpyruvate carboxykinase and nucleolin) examined. These data are consistent with the idea that carp from the Kalamazoo River Superfund Site are responding to PCB exposure via upregulation of CYP1A independent of activation of the oxidative stress response genes normally thought to be co-regulated with CYP1A.
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