Ancient Microorganism Communities in Fluid Inclusions in Halite and Gypsum

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Intellectual merit: The proposed research explores the fundamental problem of long-term survival of microorganism communities and preservation of biomaterials in fluid inclusions in halite and gypsum. The goal of the proposed research is to determine the distribution, survival, and diversity of microorganism communities and biomaterials that have been in the subsurface for periods of thousands to hundreds of millions of years. The research plan will follow the successful interdisciplinary approach recently used for the study of halite from the subsurface of Death Valley, CA, but extended to older halite deposits, 105to 108 Ma in age, and for the first time to gypsum. New emphasis will be placed on: (1) Molecularbiological techniques involving amplification of fragments of DNA by the polymerase chain reaction (PCR), followed by cloning and sequencing, which will characterize the phylogenetic diversity of microorganisms in fluid inclusions in saline minerals. (2) Raman spectroscopy, which has the potential to characterize the nature of biomolecules in fluid inclusions, in situ. All haliteand gypsum to be examined contains preserved primary textures and fluid inclusions originally formed during crystallization in ancient brine bodies. Complementary cultivation experiments will attempt to isolate halophilic and halotolerant Archaea and Bacteria from fluid inclusionsin halite and gypsum. Environmental SEM of filtrates from dissolved crystals will help confirmidentification of microorganisms and biomaterials made by in situ microscopy and Raman spectroscopy. A second goal of the proposed research will be to attempt to replicate experiments from this project and from published reports claiming amplification of DNA and cultivation of Archaea and Bacteria from ancient halite. Replication of results, an important criterion needed for verifying the authenticity of cultured ancient microorganisms and DNA, will be conducted at a laboratory at the University of Otago, New Zealand, using high throughput next generation DNA sequencing.Broader Impacts: The proposed research will promote interdisciplinary training and the excitement of scientific discovery at the undergraduate and graduate level in geoscience, anthropology, and biology at Binghamton, Virginia Tech and Otago. Both PIs will incorporate their research into their undergraduate courses, and are committed to K-12 education.Lowenstein?s group will collaborate with the Kopernik Observatory & Science Center ?Link Summer Science Explorations Program? for students in grades 1-12, where research results and hands-on activities related to long-term survival of microbes and preservation of DNA in salt crystals and the implications for life on Mars will be presented. A website will be developed describing the details of the current project, including a searchable database of phylogenetic sequences from organisms sequenced in this project.