Microbial Observatories: The Galapagos Microbial Observatory: Analysis of Novel Archaeal and Bacterial Diversity Associated with Iguanas Across Environmental Gradients

由微生物观测站计划和NSF国际科学与工程办公室资助的一项赠款已授予伊利诺伊大学的R.Mackie和I.Cann博士，以及普林斯顿大学的M.Wikelski，使用经典培养和现代分子方法相结合的方法，鉴定和表征独特食草性鬣蜥肠道中新的古菌和细菌的多样性和动态。加拉帕戈斯群岛有两个大型食草性蜥蜴的特有属，即海洋蜥蜴(Amblyriunchus)和陆地鬣蜥(Conolophus)。这些岛屿位于厄瓜多尔以西1000公里处的火山热点地区，从未与任何大陆大陆接触过。加拉帕戈斯群岛于1984年被宣布为联合国教科文组织生物圈保护区。他们将利用一系列经典的基于培养的技术来丰富、分离和表征未培养和未描述的厌氧细菌和古生菌。为了评估分子多样性和系统发育，他们将利用一种独立于培养的方法，基于直接从鬣蜥肠道样本中检索基因组DNA。DNA指纹图谱技术(PCR-DGGE)以及从混合微生物群落DNA中编码16S rDNA和大基因组片段以使每个克隆片段包含系统发育信息端的克隆文库(SuperPhylo-BAC)将用于这一目的。样本将在凉干季节(9月)和热湿季节(2月)收集，用于为期两年的“多样性证明”奖。长期目标是将鬣蜥的系统发育和功能微生物多样性与这个进化生物学自然实验室内存在的环境和生态梯度联系起来。这些结果有望揭示细菌和古菌通过依赖培养和独立的方法获得的新的生化和遗传能力。该奖项将为未来筛选新基因和潜在有用的产品提供分子和培养数据库。该项目将通过培训美国和厄瓜多尔的本科生和研究生，促进与厄瓜多尔旧金山大学和查尔斯·达尔文研究站的合作，并为加拉帕戈斯世界遗产的长期保护做出重大贡献。

A grant funded by the Microbial Observatories program and the NSF Office of International Science and Engineering has been awarded to Drs R. Mackie, and I. Cann of the University of Illinois, and M. Wikelski of Princeton University to identify and characterize novel archaeal and bacterial diversity and dynamics in the intestinal tract of unique herbivorous iguanas using a combination of classical cultivation and modern molecular methods. The Galapagos archipelago harbors two endemic genera of large herbivorous lizards, the marine (Amblyrhynchus) and land (Conolophus) iguanas. These islands are situated over a volcanic hotspot 1000 km west of Ecuador and have never been in contact with any continental landmass. The Galapagos Islands were declared a UNESCO Biosphere Reserve in 1984. They will utilize an array of classical cultivation based techniques for enrichment, isolation and characterization of uncultured and undescribed anaerobic bacteria and archaea. To assess molecular diversity and phylogeny, they will utilize a culture independent approach based on retrieval of genomic DNA directly from the iguana intestinal samples. DNA fingerprinting technique (PCR-DGGE) as well as clone libraries encoding16S rDNA and large genomic fragments from mixed microbial community DNA such that each cloned fragment contains a phylogenetically-informative end (SuperPhylo-BAC) libraries will be used for this purpose. Samples will be collected in the cool-dry season (September) and the hot-wet season (February) for this two-year "proof of diversity " award.The long-term objective is to link phylogenetic and functional microbial diversity in iguanas with the environmental and ecological gradients that exist within this natural laboratory of evolutionary biology. The results are expected to reveal novel biochemical and genetic capacity of the bacteria and archaea accessed by the culture-dependent and independent approaches. This award will provide a molecular and culture database for future screening of novel genes and potentially useful products. The project will foster collaboration with the Universidad San Francisco de Quito, Ecuador and Charles Darwin Research Station through training of US and Ecuadorian undergraduate and graduate students, and contribute significantly to the long-term conservation of the Galapagos world heritage site.