Biomaterials in Ancient Salt Formations: Examination of Primary Crystals for Biological Materials

智力优势：这一活动的智力价值涉及继续寻找困在具有重要地质年代的原生岩盐晶体中的古代生物材料。这些活的生物体、DNA和/或病毒代表了盐所代表的每个时代的进化状态。它们也是研究微生物物种长期生存和微生物在空间中从一个行星到另一个行星的可能运动问题的重要材料。最近的研究表明，嗜盐微生物可以从各种年龄的盐晶体中的盐水包裹体中分离出来（Norton和Grant 1988，Vreeland等人2000），细菌和盐古菌DNA片段可以从古岩盐中的流体包裹体中回收（Fish等人2002，Park等人2008 PNAS提交）。这些报告只是上个世纪一系列公告中的最新消息（Namyslowski，1919; Dombrowski，1961; Tasch，1963;雷泽和Tasch，1960）。一项研究（Vreeland等人，2000年）因采用极端灭菌技术避免现代微生物污染而受到广泛关注。Vreeland等人（2000年）报道的2.5亿年前的Virgibacillus（菌株2-9-3）对地球上的长期生存和在火星等行星上寻找生命具有深远的意义。该项目将继续推进这些迷人的发现。这项研究将在两个不同的二叠纪盐层中进行，试图复制最初NSF资助的发现。此外，该项目还将从更古老的地层（泥盆纪3.2亿年;志留纪4.12亿年）中收集更多样本，试图记录原始岩盐矿物更长期生存的可靠证据。由于这些地点保存完好，还将从整个蒸发周期中获得垂直样本，以检查古湖泊的盐化学随时间变化可能发生的种群差异。此外，该项目还将在每个矿井的不同水平样本中检查这些相同的地层，以便获得关于生物材料在每个地层中的空间分布的数据。这些不同地层中的微生物种群将使用培养物依赖性和独立测定法进行研究。因此，该项目将提供关于能够在盐中生存的微生物类型的数据，同时提供第一次系统的基于DNA的地层内微生物多样性调查，以及盐地层最初沉淀期间可能存在的微生物多样性。该项目还包括首次检查这些原生盐卤水中是否存在被困在古代卤水中的细菌病毒，这些卤水形成了晶体内的流体包裹体。更广泛的影响：该项目有许多广泛的影响领域，包括支持和培训几个研究实习生，支持培训几个K-12教师在PI和堪萨斯地下盐博物馆之间的联合教育努力，并为目前的NSF少数族裔提供地球生物学培训，博士研究员，他真诚地希望参与快速发展的地质微生物学领域，对以下方面特别感兴趣：对古代生物材料的研究。此外，这项研究的重点是一个公众非常感兴趣的领域，因为它试图打破目前已知的最古老的活生物体的记录，同时试图复制以前的工作，分离出目前的记录，二叠纪年龄的微生物（约100岁）。2.5亿年）。

Intellectual Merit: The intellectual merit of this activity relates to continuing the search for ancient biomaterials trapped in primary halite crystals of significant geological age. These live organisms, DNA and/or viruses represent the state of evolution during each of the ages represented by the salts. They are also significant study materials for issues of long-term survival of microbial species and the possible movement of microorganisms from planet to planet across space. Recent research has shown that halophilic microorganisms can be isolated from brine inclusions in salt crystals of variousages (Norton and Grant 1988, Vreeland et al. 2000) and that fragments of bacterial and haloarcheal DNA can be recovered from fluid inclusions in ancient halite (Fish et al. 2002, Park et al. 2008 PNAS submitted). These reports are only the latest in a series of announcements made over the last century (Namyslowski 1919, Dombrowski 1961, Tasch 1963, Reiser and Tasch 1960). One study (Vreeland et al. 2000) has received significant publicity because of the extreme sterilization techniques used to avoid contamination by modern microorganisms. The 250-million-year-old Virgibacillus (strain 2-9-3) reported by Vreeland et al. (2000) has profound implications for long-term survival on Earth and for finding life on planets such as Mars. This project will continue to advance these fascinating discoveries. The research will be conducted in two different Permian-aged salt formations in an attempt to duplicate the original NSF funded discovery. Further, the project will gather additional samples from even older formations (Devonian 320 million years of age; Silurian 412 MYA) in an attempt to document sound evidence for even longer term survival in primary halite minerals. Due to the excellent preservation of the sites, vertical samples will also be obtained from an entire evaporation cycle in order to examine population differences that might have occurred as the saline chemistry of the ancient lakes changed over time. In addition, the project will examine these same layers in different horizontal samples of each of the mines in order to develop data regarding the spatial distribution of biological materials in each of the formations. The microbial populations in these various formations will be studied using both culture dependent and independent assays. Consequently, this project will provide data on the types of microbes that are able to survive within the salts while concurrently providing the first systematic DNA-based survey of microbial diversity within the formations and that may have been present during the original precipitation of the salt formations. The project also includes the first examination of these primary salt brines for the presence of bacterial viruses trapped within the ancient brines that formed the fluid inclusions within the crystals. Broader Impact: The project has many broad areas of impact including the support and training of several research interns, supporting training for several K-12 teachers in a joint educational effort between the PI and the Kansas Underground salt museum, and providing training in geobiology to a current NSF minority Post-Doctoral fellow who has expressed a sincere desire to become involved in the rapidly developing field of geological microbiology with a specific interest in the study of ancient biomaterials. In addition, the research focuses on an area of great public interest as it seeks to break the current record for the oldest live organism known while at the same time attempting to duplicate previous work that isolated the current record, a Permian aged microbe (ca. 250 MYA).