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

智力价值：这项活动的智力价值与继续寻找困在地质年代显著的原生岩盐晶体中的古代生物材料有关。这些活的有机体、DNA和/或病毒代表了盐所代表的每个年龄段的进化状态。它们也是研究微生物物种长期生存和微生物可能在太空中从一个星球移动到另一个星球的重要研究材料。最近的研究表明，可以从各种盐晶中的卤水包裹体中分离出嗜盐微生物(Norton和Grant 1988，Vreeland等人)。2000年)，可以从古岩盐中的流体包裹体中恢复细菌和卤虫DNA片段(Fish等人。2002年，Park等人。提交了2008年国家行动计划)。这些报告只是上个世纪(纳米斯罗斯基1919年、东布朗斯基1961年、塔希1963年、赖瑟和塔希1960年)公布的一系列报告中的最新一份。一项研究(Vreeland等人2000年)因为使用极端的灭菌技术来避免现代微生物的污染而得到了广泛的宣传。Vreeland等人报道了2.5亿年前的Virgibacillus2-9-3菌株。(2000)对地球上的长期生存和在火星等行星上发现生命有着深远的影响。该项目将继续推进这些令人着迷的发现。这项研究将在两个不同的二叠纪年代的盐岩地层中进行，试图复制美国国家科学基金会最初的发现。此外，该项目将从更古老的地层(泥盆纪3.2亿年；志留纪412年)收集更多样本，试图证明原始岩盐矿物中存在更长时间的可靠证据。由于遗址保存良好，还将从整个蒸发循环中获取垂直样本，以检查随着时间的推移古代湖泊的盐分化学变化可能出现的种群差异。此外，该项目将在每个矿场的不同水平样本中检查这些相同的层，以便开发关于每个矿层中生物材料的空间分布的数据。这些不同地层中的微生物种群将使用依赖培养和独立分析两种方法进行研究。因此，该项目将提供关于能够在盐层内生存的微生物类型的数据，同时提供首次基于DNA的对地层内微生物多样性的系统调查，这些微生物可能存在于盐层的原始沉淀过程中。该项目还包括对这些初级盐卤水进行第一次检查，以确定在形成晶体内流体包裹体的古代盐水中是否存在细菌病毒。更广泛的影响：该项目有许多广泛的影响领域，包括支持和培训几名研究实习生，支持在PI和堪萨斯地下盐博物馆之间的联合教育努力中对几名K-12教师进行培训，以及为目前NSF的一名少数民族博士后提供地球生物学方面的培训，他真诚地希望参与快速发展的地质微生物学领域，并对古代生物材料研究特别感兴趣。此外，这项研究集中在一个极具公众兴趣的领域，因为它试图打破已知的最古老的活生物体的当前纪录，同时试图复制分离出当前记录的先前工作，一种二叠纪的古老微生物(约250MYA)。

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).