Ancient Microorganism Communities in Fluid Inclusions in Halite and Gypsum

学术价值：拟议的研究探索了微生物群落的长期生存和盐岩和石膏中流体包裹体中生物材料的保存这一基本问题。这项拟议研究的目标是确定地下微生物群落和生物材料的分布、生存和多样性，这些微生物群落和生物材料在地下已经存在了数千到数亿年。该研究计划将遵循最近用于研究加州死亡谷地下岩盐的成功的跨学科方法，但扩展到年龄为105-108 Ma的较老的岩盐矿床，并首次扩展到石膏。新的重点将放在：(1)分子生物学技术，包括通过聚合酶链式反应(PCR)扩增DNA片段，然后克隆和测序，这将表征盐类矿物流体包裹体中微生物的系统发育多样性。(2)原位拉曼光谱，它有可能表征流体包裹体中生物分子的性质。所有待检验的盐岩和石膏都含有保存完好的原生结构和流体包裹体，这些包裹体最初是在古卤水体内结晶过程中形成的。补充性培养实验将试图从岩盐和石膏中的流体包裹体中分离出嗜盐和耐盐的古生菌和细菌。溶解晶体滤液的环境扫描电子显微镜将有助于确认通过原位显微镜和拉曼光谱制作的微生物和生物材料的鉴定。这项拟议研究的第二个目标将是试图复制该项目和发表的报告中的实验，这些报告声称扩增了DNA并从古代岩盐中培养出古生菌和细菌。结果的复制是验证培养的古代微生物和DNA真实性所需的一个重要标准，将在新西兰奥塔哥大学的一个实验室进行，使用高通量的下一代DNA测序。广泛的影响：拟议的研究将促进宾厄姆顿、弗吉尼亚理工大学和奥塔哥的地学、人类学和生物学本科生和研究生水平的跨学科培训和科学发现的兴奋。这两位PI都将把他们的研究纳入他们的本科课程，并致力于K-12教育。洛文斯坦？S团队将与科珀尼克天文台和科学中心？链接夏季科学探索计划？合作。对于1-12年级的学生，将介绍与微生物的长期生存和盐晶体中DNA的保存以及对火星生命的影响有关的研究结果和实践活动。将开发一个网站，描述当前项目的细节，包括一个可搜索的数据库，其中包括该项目中测序的生物体的系统发育序列。

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.