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.

自然界的碳固定酶Rubisco是出了名的低效--经常限制包括农作物在内的光合作用有机体的生长速度。一些生物已经找到了一种绕过这个问题的方法，将Rubisco包装成称为类蛋白的隔间，这些隔间将底物二氧化碳集中在Rubisco活性部位周围。金鱼草是唯一一种具有类蛋白核的陆地植物。该项目致力于鉴定类蛋白核成分，鉴定类蛋白核中的角藻固碳酶(即Rubisco)，并确定不同的类蛋白核组分如何相互作用。这项研究的结果将为从进化、生化和机理上深入了解类胡萝卜素提供依据，并向收集必要的了解迈出第一步，以便将类胡萝卜素移植到农作物中以促进固碳。该项目还将在一系列计算和实验技术方面培训不同的本科生，包括植物组织培养和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，以有趣和翔实的方式介绍金鱼藻的多样性、进化和类蛋白核，并在YouTube和其他在线平台上分发。霍恩沃特和Rubisco研究的重要性将通过建立的播客系列得到进一步推广。光合作用是地球上几乎所有生命的能量来源，而且经常受到碳固定酶Rubisco的限制。一些生物体已经进化出巧妙的方法来绕过这些限制，例如蛋白核，这是一种无膜细胞器，液-液相将Rubisco与基质分开。蛋白核在Rubisco活性中心附近活跃地浓缩底物二氧化碳，从而极大地提高了固碳效率。金鱼藻是唯一一种具有类蛋白核的陆地植物，这些植物具有提高C3作物效率的翻译潜力。在进化过程中，金鱼草蛋白核类化合物的获得和丢失为比较研究提供了独特的系统发育复制。将使用比较基因组学、RNA-seq和蛋白质组学，同时利用重复的有/无类蛋白核转换来鉴定假定的类蛋白核成分。假想的蛋白核成分和Rubisco亚基的亚细胞定位将使用体内定位方法进行验证，并对Rubisco种群进行结构表征。一种建构主义生物学方法将在体外识别和表征Rubisco和重组产生的推定的蛋白核成分之间的相互作用，并将用于评估Rubisco相分离的可能性。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。