Collaborative Research: From phylogeny to biomolecules: a cross-scale approach to understand the making of a unique carbon-concentrating mechanism in hornworts

合作研究：从系统发育到生物分子：一种跨尺度的方法来了解金鱼藻独特的碳浓缩机制的形成

• 批准号: 2213841
• 负责人: Fay-Wei Li
• 金额: $ 50.37万
• 依托单位: Boyce Thompson Institute Plant Research
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2213841&HistoricalAwards=false
• 关键词: Collaborative; Research; phylogeny; biomolecules; cross

Nature’s carbon fixing enzyme, Rubisco, is notoriously inefficient - often limiting the growth rate of photosynthetic organisms, including crop species. Some organisms have found a way around this problem by packaging Rubisco into compartments, called pyrenoids, which concentrate substrate CO2 around Rubisco active sites. Hornworts are the only land plants that have a pyrenoid. This project seeks to identify the hornwort pyrenoid components, characterize the hornwort carbon-fixing enzyme (i.e. Rubisco) housed within the pyrenoids, and determine how the various pyrenoid components interact with each other. The results of this study will provide evolutionary, biochemical, and mechanistic insights into the hornwort pyrenoid, and make a first step towards gathering the requisite understanding to transplant hornwort pyrenoids into crop plants to boost carbon fixation. This project will also train diverse undergraduates in a range of computational and experimental techniques, including plant tissue culture and transformation and bioinformatics. In addition, a three-part video series will be produced to provide a fun and informative introduction to hornwort diversity, evolution and pyrenoids, and distributed on YouTube and other online platforms. The importance of hornwort and Rubisco research will be further promoted through established podcast series.Photosynthesis is the source of energy for almost every life on earth, and is often limited by the carbon fixing enzyme, Rubisco. Some organisms have evolved ingenious ways to circumvent these limitations, such as pyrenoids, which are a kind of membrane-less organelle that liquid-liquid phase separates Rubisco from the stroma. Pyrenoids actively concentrate substrate CO2 around Rubisco active sites, thereby greatly enhancing the efficiency of carbon fixation. Hornworts are the only land plants with a pyrenoid, and these have translational potential for improving C3 crop efficiency. The fact that hornwort pyrenoids have been gained and lost over the course of evolution offers a unique phylogenetic replication for comparative studies. Putative pyrenoid components will be identified using comparative genomics, RNA-seq, and proteomics, while leveraging repeated pyrenoid-present/absent transitions. The subcellular localization of putative pyrenoid components and Rubisco subunits will be validated using in vivo localization approaches, and Rubisco populations structurally characterized. A constructionist biology approach will identify and characterize interactions between Rubisco and recombinantly-produced putative pyrenoid components in vitro, and will be used to assess the potential for Rubisco phase separation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

众所周知，大自然的碳固定酶Rubisco效率低下 - 通常限制了包括作物物种在内的光合生物的生长速度。一些生物体通过将Rubisco包装到隔室（称为pyrenoids）中找到了解决这个问题的方法，该隔室浓缩了Rubisco活性位点浓度底物CO2。霍恩沃尔特是唯一具有类酚的陆地植物。该项目旨在鉴定霍恩沃特的荷马类成分，表征了霍恩沃特碳固定酶（即rubisco）所容纳的荷兰酶，并确定各种类型类分量如何相互作用。这项研究的结果将为霍恩沃特荷马菌提供进化，生化和机械洞察力，并迈出第一步，朝着收集必要的理解以移植霍恩沃特的荷马植物植物中，以增强碳固定。该项目还将在一系列计算和实验技术中训练多样化的本科生，包括植物组织培养，转化以及生物信息学。此外，还将制作一个由三部分组成的视频系列，以提供有关Hornwort多样性，进化论和类型类动物的有趣且内容丰富的介绍，并在YouTube和其他在线平台上分发。 Hornwort和Rubisco Research的重要性将通过既定的播客系列进一步促进。斑点合成是地球上几乎所有生命的能量来源，通常受到碳固定酶Rubisco的限制。一些生物已经进化出巧妙的方法来规避这些局限性，例如类拟南芥，这是一种无膜细胞器，液 - 液相可将Rubisco与基质分开。类似力型在Rubisco活跃部位周围积极浓缩底物CO2，从而大大提高了碳固定的效率。霍恩沃尔特（Hornworts）是唯一具有类酚的陆地植物，这些植物具有提高C3作物效率的潜力。在进化过程中，霍恩沃特荷马菌已经获得并丢失的事实为比较研究提供了独特的系统发育复制。假定的类拟南芥成分将使用比较基因组学，RNA-SEQ和蛋白质组学鉴定，同时利用重复的pyrenoid-trecresent/不存在过渡。推定的类拟南芥成分和Rubisco亚基的亚细胞定位将使用体内定位方法进行验证，而Rubisco种群在结构上表征了。建筑主义生物学方法将在体外识别和表征鲁比斯科与重组产生的推定的类拟南芥组件之间的相互作用，并将用于评估Rubisco阶段分离的潜力。这项奖项反映了NSF的法定任务，并被认为是通过该基金会的知识分子功能和广泛影响的评估来评估CRETERIA的评估。