Ancient Microorganism Communities in Fluid Inclusions in Halite and Gypsum

智力优点：该研究探讨了微生物群落长期生存和生物材料在岩盐和石膏流体包裹体中保存的基本问题。拟议研究的目标是确定在地下存在了数千至数亿年的微生物群落和生物材料的分布、生存和多样性。 该研究计划将遵循最近成功的跨学科方法，用于研究加利福尼亚州死亡谷地下的岩盐，但扩展到年龄为105至108 Ma的较老的岩盐矿床，并首次研究石膏。新的重点将放在：（1）分子生物学技术，包括通过聚合酶链反应（PCR）扩增DNA片段，随后进行克隆和测序，这将表征盐水矿物流体包裹体中微生物的系统发育多样性。(2)拉曼光谱，它有潜力表征流体包裹体中生物分子的性质，在原位。所有待检查的岩盐和石膏都含有保存完好的原生结构和古卤水体结晶过程中形成的流体包裹体。补充培养实验将试图从岩盐和石膏中的流体包裹体中分离嗜盐和耐盐的微生物和细菌。溶解晶体中微生物的环境扫描电镜将有助于确认原位显微镜和拉曼光谱所做的微生物和生物材料的鉴定。拟议研究的第二个目标将是试图复制该项目的实验，以及声称扩增DNA和培养古岩盐细菌的已发表报告。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。拟议的研究将促进跨学科培训和科学发现的兴奋在本科和研究生水平的地球科学，人类学，宾汉姆顿、弗吉尼亚理工大学和奥塔哥大学的生物学教授。这两个PI将把他们的研究纳入他们的本科课程，并致力于K-12教育。的小组将与科珀尼克天文台科学中心合作？链接夏季科学探索计划？为1-12年级的学生举办的研讨会，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成果和实践活动。将建立一个网站，介绍目前项目的细节，包括一个可检索的数据库，提供本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.