Collaborative Research: Iron and phosphorus balanced limitation of nitrogen fixation in the oligotrophic ocean

海洋蓝细菌能够利用或“固定”大气中的氮气，因此提供了支持开放海洋食物链的大部分必需营养氮。海洋学家通常认为，这些固氮蓝藻的生长在任何特定的时间和地点都受到铁或磷供应的限制。然而，初步的实验表明，这些固氮菌在铁和磷同时缺乏的情况下生长得最好。在该项目中，研究人员将使用专门针对铁和磷限制的细胞指标来确定这种类型的固氮蓝细菌的“平衡限制”在控制热带大西洋海洋食物链生产力方面的重要性。两名研究生将在南加州大学（USC）和伍兹霍尔海洋研究所接受培训，还有一名博士后研究员将在南加州大学接受培训。教育推广工作将在洛杉矶市中心的一所高中进行，该高中的学生中超过98%是西班牙裔和非洲裔美国人，南加州大学全球环境微生物学课程的本科生人数不足。此外，本科生的两个研究经验将被监督夏季研究项目，以帮助他们了解科学职业选择。研究人员将调查这种独特的平衡铁/磷限制表型的生物学和地球化学后果，使用实验室和野外工作方法。在该项目的第一年，固氮蓝藻将在铁和/或磷限制下培养，随后应用蛋白质组学和转录组学来鉴定作为铁/磷平衡限制的潜在诊断生物标志物的基因。初步工作已经在一种蓝细菌中确定了一种有希望的候选生物标志物，这是一种EzrA蛋白结构域，似乎与平衡限制下特别观察到的细胞大小减少有关，研究人员已经确定了许多其他类似生物标志物的潜在候选者。在第二年，这些新的共同限制生物标志物和其他以前验证的铁限制（IsiB）和磷限制（SphX）将被用来调查研究巡航期间的平衡限制横断相对高铁，低磷北大西洋沃茨，相对高磷，低铁南大西洋。这一实地考察的组成部分将调查固氮蓝藻种群在这个自然的铁/磷梯度的遗传，蛋白质组学和生理指标的平衡限制，以及测试他们的反应，铁和磷的操作在船上孵化实验。第三年将致力于样品分析，并出版物探讨海洋固氮生物对铁和磷同时限制的反应。

Marine cyanobacteria are able to use or "fix" atmospheric nitrogen gas, and so supply much of the essential nutrient nitrogen that supports open ocean food chains. Oceanographers have usually thought that the growth of these nitrogen-fixing cyanobacteria is limited at any particular time and place by the supply of either iron, or of phosphorus. Preliminary experiments have shown, though, that these nitrogen fixers instead grow best when both iron and phosphorus are scarce at the same time. In this project, the researchers will use cellular indicators that are specific for iron and phosphorus limitation to determine how important this type of "balanced limitation" of nitrogen-fixing cyanobacteria is in controlling the productivity of ocean food chains in the tropical Atlantic Ocean. Two graduate students will be trained at the University of Southern California (USC) and Woods Hole Oceanographic Institution, as well as a postdoctoral researcher at USC. Educational outreach efforts will take place at a Los Angeles inner city high school with a student body that is over 98% Hispanic and African-American, and with underrepresented undergraduates in the USC Global Environmental Microbiology course. In addition, two Research Experiences for Undergraduates students will be supervised for summer research projects to help them learn about science career options. The researchers will investigate the biological and biogeochemical consequences of this unique balanced iron/phosphorus-limited phenotype, using both laboratory and fieldwork approaches. During the first year of this project, the nitrogen-fixing cyanobacteria will be cultured under iron and/or phosphorus limitation, followed by application of proteomics and transcriptomics to identify genes that are potential diagnostic biomarkers for iron/phosphorus balanced limitation. Preliminary work has already identified one promising candidate biomarker in one cyanobacterium, an EzrA protein domain that appears to be associated with the cell size decreases seen specifically under balanced limitation, and the researchers have identified numerous other potential candidates for similar biomarkers. During the second year, these new co-limitation biomarkers and others previously validated for iron limitation (IsiB) and phosphorus limitation (SphX) will be used to investigate balanced limitation during a research cruise transecting from relatively high-iron, low-phosphorus North Atlantic waters, to the relatively high-phosphorus, low-iron South Atlantic. This fieldwork component will survey nitrogen fixing cyanobacteria populations across this natural iron/phosphorus gradient for genetic, proteomic, and physiological indicators of balanced limitation, as well as testing their responses to iron and phosphorus manipulations in shipboard incubation experiments. The third year will be devoted to sample analysis, and publications exploring the responses of oceanic nitrogen fixers to simultaneous limitation by both iron and phosphorus.