EN-GEN: Transcriptional and Proteomic Analyses of Multiple Environmental Stressors in Marine Diatoms (TP-AMES)

硅藻是一种数量丰富且分布广泛的浮游植物功能群，在海洋中负责大量的初级生产。因此，它们对全球碳循环产生了深远的影响。认识到这一意义，硅藻假海藻是第一个真核海洋浮游植物模型选择全基因组测序。这种模式硅藻的基因组突出了假单胞藻代谢的意想不到的方面（例如，来自植物和动物祖先的途径马赛克），并强调尽管它们很重要，但我们对硅藻生理学和这些生物如何在海洋中发挥作用的理解存在根本差距。在许多海洋系统中，微量营养素（如铁）和大量营养素（如磷）的供应被认为是硅藻生物量的主要驱动因素。事实上，模型预测，在大西洋和太平洋的大片地区，硅藻受到这些营养物质的限制。本研究项目将使用转录和蛋白质组学分析来研究海洋硅藻对铁和磷限制反应的潜在分子机制。由于多种因素可能限制环境中的生长，我们还建议通过李比希定律的分子测试来解剖营养物质共同限制对海洋硅藻的影响。这项基因组支持的工作将首先与假单胞菌一起进行，使用转录组分析和先进的质谱分析进行蛋白质组分析，并将假单胞菌基因组作为基因发现和注释的框架。然后，这些研究人员将应用这些方法来了解营养限制如何影响其他生态上重要的、未测序的硅藻物种。该项目将解决我们对硅藻生理生态学理解上的关键空白，并将四位早期职业研究者和他们在海洋功能基因组学方面的综合专业知识汇集在一起，共同研究这一重要但尚未充分研究的海洋学概念。研究人员还将为其他对浮游植物基因组学和硅藻生物学感兴趣的人提供一个获取基因表达和蛋白质表达数据的公共数据库，并通过一个每天有2.5万用户的互动教育网站（www.whyville.net）促进儿童（8-14岁）的科学学习。通过这一努力，研究人员将有机会向数百万儿童教授浮游植物生物学和海洋学的关键概念，以及科学探究的过程。该项目还将促进研究生教育和NSF ADVANCE资助的研究人员的指导，以扩大女性参与科学并为研究生提供指导支持。

Diatoms are an abundant and widespread functional group of phytoplankton, responsible for significant amounts of primary production in the ocean. As such, they exert a profound influence on the global cycling of carbon. In recognition of this significance, the diatom Thalassiosira pseudonana was the first eukaryotic marine phytoplankton model selected for whole genome sequencing. The genome of this model diatom highlighted unexpected aspects of T. pseudonana metabolism (e.g. a mosaic of pathways derived from plant and animal ancestors), and emphasized that despite their importance, there are fundamental gaps in our understanding of diatom physiology and how these organisms function in the sea. The supply of micronutrients (e.g. iron) and macronutrients (e.g. phosphorus) are considered to be primary drivers of diatom biomass in many marine systems. In fact, models predict there are large regions of both the Atlantic and the Pacific where diatoms are limited by these nutrients. This research project will use transcriptional and proteomic profiling to examine the underlying molecular mechanisms involved in the response of marine diatoms to iron and phosphorus limitation. Since multiple factors can be limiting for growth in the environment, we are also proposing experiments to dissect the impact of nutrient co-limitation on marine diatoms with a molecular test of Liebig's Law. This genome-enabled work will first be pursued with T. pseudonana using transcriptome profiling and advanced mass spectrometry for the proteome profiling, with the T. pseudonana genome as a frame-work for gene discovery and annotation. These investigators will then apply these methods to understand how nutrient limitation impacts other ecologically important, unsequenced diatom species. This project will address key gaps in our understanding of diatom physiological ecology, and bring together four early-career investigators and their combined expertise in marine functional genomics towards this important yet understudied oceanographic concept of co-limitation. The investigators will also provide a public database with access to the gene expression and protein expression data for others interested in phytoplankton genomics and the biology of diatoms, and promote science learning in children (ages 8-14) using an interactive educational website (www.whyville.net) that has 25,000 users daily. Through this effort the investigators will have the opportunity to teach key concepts in phytoplankton biology and oceanography, and the process of scientific inquiry, to millions of children. This project will also promote graduate education and mentoring by NSF ADVANCE funded investigators that serves to broaden participation of women in science and provides mentoring support for graduate students.