Great Salt Lake halophilic microorganisms as models for astrobiology: evidence for desiccation tolerance and ultraviolet irradiation resistance

Great Salt Lake (GSL) is home to halophiles, salt-tolerant Bacteria and Archaea, which live at 2-5M NaCl. In addition to salt tolerance, GSL halophiles exhibit resistance to both ultraviolet (UV) irradiation and desiccation. First, to understand desiccation resistance, we sought to determine the diversity of GSL halophiles capable of surviving desiccation in either recently formed GSL halite crystals or GSL Artemia (brine shrimp) cysts. From these desiccated environments, surviving microorganisms were cultured and isolated, and genomic DNA was extracted from the individual species for identification by 16S rRNA gene homology. From the surface-sterilized cysts we also extracted DNA of the whole microbial population for non-cultivation techniques. We amplified the archaeal or bacterial 16S rRNA gene from all genomic DNA, cloned the cyst population amplicons, and sequenced. These sequences were compared to gene databases for determination of closest matched species. Interestingly, the isolates from the crystal dissolution are distinct from those previously isolated from GSL brine. The cyst population results reveal species not found in crystals or brine, and may indicate microorganisms that live as endosymbionts of this hypersaline arthropod. Second, we explored UV resistance in a GSL haloarchaea species, "H. salsolis." This strain resists UV irradiation an order of magnitude better than control species, all of which have intact repair systems. To test the hypothesis that halophiles have a photoprotection system, which prevents DNA damage from occurring, we designed an immunoassay to detect thymine dimers following UV irradiation. "H. salsolis" showed remarkable resistance to dimer formation. Evidence for both UV and desiccation resistance in these salt-tolerant GSL halophiles makes them well-suited as models for Astrobiological studies in pursuit of questions about life beyond earth.

[1]  B. Austin,et al.  Microbiology of laboratory-hatched brine shrimp (Artemia) , 1982 .

[2]  W. Moore,et al.  Taxonomic Note : A Place for DNA-DNA Reassociation and 16 s rRNA Sequence Analysis in the Present Species Definition in , 2022 .

[3]  J. Clegg Desiccation Tolerance in Encysted Embryos of the Animal Extremophile, Artemia1 , 2005, Integrative and comparative biology.

[4]  P. DasSarma,et al.  Microbial Diversity of Great Salt Lake , 2005 .

[5]  L. Marcello,et al.  Repair of UV damage in Halobacterium salinarum. , 2003, Biochemical Society transactions.

[6]  Aaas News,et al.  Book Reviews , 1893, Buffalo Medical and Surgical Journal.

[7]  P. Liang,et al.  The synthesis of a small heat shock/alpha-crystallin protein in Artemia and its relationship to stress tolerance during development. , 1999, Developmental biology.

[8]  D. Gevers,et al.  Phylogeny and Molecular Identification of Vibrios on the Basis of Multilocus Sequence Analysis , 2005, Applied and Environmental Microbiology.

[9]  Jean Swings,et al.  Phenotypic diversity amongst Vibrio isolates from marine aquaculture systems , 2003 .

[10]  S. McCready A dot blot immunoassay for UV photoproducts. , 1999, Methods in molecular biology.

[11]  T. Onstott,et al.  Isolation of Halobacterium salinarum retrieved directly from halite brine inclusions. , 2003, Environmental microbiology.

[12]  D. W. Powers,et al.  Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal , 2000, Nature.

[13]  Erko Stackebrandt,et al.  Taxonomic Note: A Place for DNA-DNA Reassociation and 16S rRNA Sequence Analysis in the Present Species Definition in Bacteriology , 1994 .

[15]  E. Roedder The fluids in salt , 1984 .

[16]  S. Lory,et al.  Complete genome sequence of Pseudomonas aeruginosa PAO 1 , an opportunistic pathogen , 2000 .

[17]  M. Maurel,et al.  The search for traces of life: the protective effect of salt on biological macromolecules , 2002, Extremophiles.

[18]  Min Pan,et al.  Low-pass sequencing for microbial comparative genomics , 2004, BMC Genomics.