ABSTRACT: Enrichment of Marinobacter sp. and Halophilic Homoacetogens at the Biocathode of Microbial Electrosynthesis System Inoculated With Red Sea Brine Pool
Project description:Halophilic microorganisms have long been known to survive within the brine inclusions of salt crystals, as evidenced by their pigmentation. However, the molecular mechanisms allowing this survival has remained an open question for decades. While protocols for the surface sterilization of halite (NaCl) have enabled isolation of cells and DNA from within halite brine inclusions, “-omics” based approaches have faced two main technical challenges: (1) removal of all contaminating organic biomolecules (including proteins) from halite surfaces, and (2) performing selective biomolecule extractions directly from cells contained within halite brine inclusions with sufficient speed to avoid modifications in gene expression during extraction. In this study, we present methods to resolve these two technical challenges. In addition, we apply these methods to perform the first examination of the early acclimation of a model haloarchaeon (Halobacterium salinarum NRC-1) to halite brine inclusions. Examinations of the proteome of Halobacterium cells two months post-evaporation revealed a high degree of similarity with stationary phase liquid cultures, but with a sharp down-regulation of ribosomal proteins. Low quantities of RNA from halite brine inclusions corroborate the hypothesis of low transcriptional and translational activities. While proteins for central metabolism were part of the shared proteome between liquid cultures and halite brine inclusions, proteins involved in cell mobility (archaellum, gas vesicles) were either absent or less abundant in halite samples. Proteins unique to cells within brine inclusions included transporters, suggesting modified interactions between cells and the surrounding brine inclusions microenvironment. The methods and hypotheses presented here enable future studies of the survival of halophiles in both culture model and natural halite systems.
Project description:The goal of this study was to identify the key functions of the six main symbionts that are hosted in gills of the marine bivalve, Idas modiolaeformis, which lives at deep-sea hydrocarbon seeps and wood falls in the Eastern Atlantic Ocean and the Mediterranean Sea. These symbionts include the main autotrophic methane- and sulfur-oxidizing lineages (Methyloprofundus, Thioglobus, Thiodubillierella), as well as a Methylophagaceae methylotrophic autotroph, a flavobacterial degrader of complex polysaccharides Urechidicola and a Nitrincolaceae heterotroph that specializes in degradation of nitrogen-rich compounds such as peptides and nucleosides. Four I. modiolaeformis individuals were preserved in RNAlater following retrieval from a brine pool habitat in the Eastern Mediterranean at 1,150 m water depth (32° 13.4' N 34° 10.7' E), using a remotely-operated vehicle. RNAlater was discarded after 24 hours, and the specimens were kept at -80°C until DNA/RNA/protein co-extraction using the AllPrep DNA/RNA/Protein Mini Kit (Cat. No. 80004, Qiagen).
Project description:Bathymodiolus childressi is a species of deep-sea mussels found predominantly in the Gulf of Mexico. It colonizes cold seeps such as brine pool and oil seeps. The success of these animals in such environment is thought to be due to the symbiotic association of the mussel host with several species of bacteria. The aim of this study is to understand the role of the different partners involved in the symbiotic system using various “-omics” approaches. In addition to protein identification we used the mass spectrometry data generated and submitted with this project to derive the stable carbon isotope ratios for the different members of the symbiosis using the direct Protein-SIF method. The respective isotope pattern file and SIF computation files are included with this submission.