The Development of Microbial Associations in Major Reef Building Corals of the Pacific Ocean

珊瑚-微生物关联研究的基础是了解这种关系何时以及如何建立，以及它们的特异性。最近，研究人员首次提供了珊瑚（Pocillopora meandrina）的早期生活史阶段与特定细菌群（Alphaproteobacteria玫瑰杆菌分支的Jannaschia谱系）之间特定关联的证据。他们将通过研究来自北太平洋夏威夷和南太平洋法属波利尼西亚的Moorea的主要造礁珊瑚中微生物关联的开始来扩展这项工作。了解与这些珊瑚物种相关的微生物群落的发生、特异性和功能，对于理解和预测珊瑚对变化环境的响应是必要的。本建议的主要目标是：1。使用独立培养技术来识别和量化与北太平洋夏威夷几种主要造礁珊瑚相关的微生物，这些微生物代表了各种繁殖策略（孵卵和广播产卵）和虫黄藻共生传播的不同模式（垂直与水平），贯穿每种繁殖周期、早期发育阶段和定居后阶段。收集和分析在南太平洋法属波利尼西亚Moorea地区发现的相同（或类似）珊瑚物种的类似样本，以评估目标1中记录的关联是否局限于夏威夷，还是广泛分布于整个太平洋，并可能代表珊瑚发育的共同特征。利用荧光原位杂交技术枚举在夏威夷收集的玫瑰杆菌分支的Jannaschia谱系的整个发育周期。指数将扩大这一目标，包括其他珊瑚物种、目标细菌和/或地理位置，因为它们确定了更多的联系。通过应用新的培养技术，分离出在不依赖于培养调查的meandrina相关微生物群落（例如Jannaschia sp.）中普遍存在的微生物，以便开发用于调查珊瑚-微生物相互作用的模型系统。由于人为活动造成的环境压力不断增加，珊瑚礁正在减少，现在有相当多的证据表明，它们正受到海面温度上升和海洋酸化的威胁。与珊瑚相关的微生物被认为在维持珊瑚宿主的健康和弹性方面发挥着各种潜在的重要作用，主要由分子生物学领域的发展推动的方法学的进步正在促进对这种联系的深入了解。许多珊瑚微生物研究都集中在微生物对疾病和白化的作用上，而且几乎完全集中在成年珊瑚群落上。这项研究将提供独特的信息，了解微生物在整个发育周期中与健康珊瑚相互作用的方式，这些关系的特殊性，它们是如何启动的，以及它们在自然界中的分布和频率。(1)本提案将支持培养一名博士后学者，研究分子微生物生态学技术、新型栽培方法和珊瑚生物学。(2)研究人员将开发一个网站，其中包含有关珊瑚-微生物相互作用的信息，其内容旨在作为K-12学生的教育资源，并作为对珊瑚-微生物关联感兴趣的其他科学家的资源。这项研究产生的数据，包括出版物、报告、微生物菌株、基因序列、显微照片和系统发育分析，将通过网站公开。(3)博士后和项目负责人将设计并每年提供为期5天的ARB序列分析程序使用强化培训讲习班，包括荧光原位显微镜在海洋细菌和珊瑚样品中的应用。(4)该提案将支持培养博士后学者指导下的本科生，并将利用现有的项目和关系，专门为夏威夷大学代表性不足的学生提供这些机会，如夏威夷原住民和太平洋岛民。

Fundamental to the study of coral-microbial associations is an understanding of when and how the relationships are established, and their specificity. Recently, the investigators provided the first evidence of a specific association between an early life history stage of a coral (Pocillopora meandrina) and a particular group of bacteria (Jannaschia lineage of the Roseobacter clade of Alphaproteobacteria). They will be expanding this work by examining the onset of microbial associations in key reef building corals from Hawaii in the North Pacific Ocean and Moorea, French Polynesia, in the South Pacific Ocean. Understanding the onset, specificity and function of the microbial community associated with these coral species is necessary to understand and predict the coral holobiont response to a changing environment.The main objectives of this proposal are to: 1. Use cultivation independent techniques to identify and quantify microorganisms associated with several major reef building corals of Hawaii in the North Pacific Ocean that represent a variety of reproductive strategies (brooding and broadcast spawning) and differing modes of zooxanthellae symbiont transmission (vertical vs. horizontal), throughout the reproductive cycle, early developmental stages, and post-settlement stages of each.2. Collect and analyze similar samples from the same (or similar) species of coral found in the South Pacific Ocean in Moorea, French Polynesia, in order to assess whether the associations documented in objective 1 are localized to Hawaii, or broadly distributed across the Pacific and likely to represent common features of coral development.3. Use fluorescence in situ hybridization to enumerate cells of the Jannaschia lineage of the Roseobacter clade throughout the development cycle of P. meandrina collected in Hawaii. The PIs will expand this objective to include other coral species, target bacteria, and/or geographic location as they identify additional associations.4. Isolate microorganisms prevalent in cultivation-independent surveys of P. meandrina-associated microbial communities (e.g. Jannaschia sp.) by the application of novel culturing techniques, in order to develop model systems for the investigation of coral-microbe interactions.Intellectual merit Coral reefs are in decline as a result of increasing environmental stress due to anthropogenic activity, and there is now considerable evidence indicating that they are under threat from the effects of rising sea surface temperature and ocean acidification. Microorganisms associated with corals are thought to play a variety of potentially important roles in maintaining the health and resiliency of the coral host, and advances in methodology primarily driven by developments in the field of molecular biology are facilitating growing insight into this association. Much of coral-microbial research is focused on the contribution of microorganisms to disease and bleaching, and is focused almost exclusively on adult coral colonies. This study will provide unique information on the manner in which microorganisms interact with healthy corals throughout their developmental cycle, the specificity of these relationships, how they are initiated, and their distribution and frequency in nature.Broader impacts (1) This proposal will support the training of one postdoctoral scholar in advancing molecular microbial ecology techniques, novel cultivation-based methodology, and coral biology. (2) The investigators will develop a website that will contain information on coral-microbial interactions, with content designed to serve as an educational resource for K-12 students, and to serve as a resource for other scientists interested in coral-microbial associations. Data generated from this study, including publications, presentations, microbial strains, gene sequences, photomicrographs and phylogenetic analyses, will be made publicly available via the website. (3) The postdoc and PI will design and annually offer a five-day intensive training workshop on the use of ARB sequence analysis program, including the application of fluorescence in situ microscopy to marine bacteria and coral samples. (4) The proposal will support the training of undergraduate students under the mentorship of the postdoctoral scholar, and will take advantage of existing programs and relationships to specifically offer these opportunities to underrepresented students at the University of Hawaii such as Native Hawaiians and Pacific Islanders.