RESEARCH-PGR: Discovering new metabolic constraints and regulatory nodes in oilseeds engineered for enhanced fatty acid synthesis and seed oil

RESEARCH-PGR：发现油籽中新的代谢限制和调节节点，旨在增强脂肪酸合成和种子油

• 批准号: 1829365
• 负责人: Jay Thelen
• 金额: $ 343.76万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-11-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1829365&HistoricalAwards=false
• 关键词: RESEARCH; PGR; Discovering; new; metabolic

Plant oils are a renewable source of food, fuels, and chemicals; world output of plant oils must double by 2030 to meet projected demand. A major source of plant oils comes from seeds. While research has yielded improvements in seed oil quantity and quality, there are tradeoffs for other seed attributes such as protein content and yield. For example, plants that are bred or engineered to produce high seed oil often have decreased protein content or make fewer seeds. The molecular mechanism driving such tradeoffs remains unclear and creates a barrier for making improved crops with specific amounts of oil and protein content. This project seeks to unravel the genetic and molecular mechanisms related to the tradeoffs between oil, protein content, seed size, and seed set in order to enhance seed oil content of crops. By using an integrated approach, this project will investigate biochemical steps used by seeds to prioritize protein or oil production. Using this knowledge, this project will generate plants with improved seed oil production. In addition, this project promotes STEM education using the science of seed oils to train tomorrow's scientists as well as develops a science literacy program using hands-on research experience for students who are training to become future STEM teachers. The scientific and educational outcomes of this project will have broad impacts for both research and society through improved oil production in crop plants.Quantitative, comparative profiling of high-oil soybean, Camelina, and Arabidopsis will reveal the coordinated metabolic response to de-regulated de novo FAS. These findings will reveal the next metabolic constraint for engineering seed oil content, discover new genes involved in acyl lipid metabolism, and provide the plant lipid community with characterized, high-oil germplasm for gene stacking and comparative genomics. This project will integrate comparative transcriptomics, proteomics, and translatomics during three stages of seed development of FAS-engineered, high-oil plants to develop correlative and kinetic models to make metabolic predictions, and discover coregulated gene networks. Flux analyses and clustering models will be used to link high-oil germplasm to causal effects within combined central and lipid metabolic networks and used to make hypotheses for next-generation crop improvements. Aspects of in vivo FAS rates, the uncoupling of FAS from TAG synthesis that can lead to futile cycles of FAS and breakdown; and acyl fluxes through the lipid metabolic network will be investigated to elucidate new bottlenecks for TAG accumulation as a result of de-regulating FAS. Web resources (fatplants.net) developed as a part of this project will serve to educate the community and provide an interactive tool to integrate metabolism and gene regulation Integrated tools are necessary to inspire tomorrow's scientists and engineers and enable multidisciplinary training of all ages. This project will also develop a hands-on science literacy program for future educators of science and technology (Sci-FEST) and high school science teaching modules on plant metabolism and biotechnology will be created by the program participants.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.

植物油是一种可再生的食品、燃料和化学品来源；到2030年，世界植物油产量必须翻一番，才能满足预计的需求。植物油的一个主要来源是种子。虽然研究已经在种子油的数量和质量方面取得了改善，但在蛋白质含量和产量等其他种子属性方面也存在权衡。例如，经过培育或改造以产生高籽油的植物通常蛋白质含量较低或种子较少。驱动这种权衡的分子机制尚不清楚，并为培育特定数量的脂肪和蛋白质含量的改良作物创造了障碍。该项目旨在揭示与油、蛋白质含量、种子大小和结实率之间的权衡相关的遗传和分子机制，以提高作物的种子含油量。通过使用综合方法，该项目将研究种子用于优先生产蛋白质或油脂的生化步骤。利用这一知识，该项目将培育出种子油产量更高的植物。此外，该项目利用种子油科学促进STEM教育，以培训未来的科学家，并利用实践研究经验为正在培训成为未来STEM教师的学生开发科学素养计划。该项目的科学和教育成果将通过提高作物的含油量对研究和社会产生广泛的影响。高油大豆、山茶和拟南芥的定量比较图谱将揭示对去调控的从头Fas的协调代谢反应。这些发现将揭示下一个控制种子含油量的代谢限制因素，发现与脂酰化代谢有关的新基因，并为基因堆积和比较基因组学提供具有代表性的高油种质。该项目将在Fas工程高油植物种子发育的三个阶段整合比较转录组学、蛋白质组学和翻译组学，以开发相关的动力学模型来进行代谢预测，并发现共调控基因网络。通量分析和聚类模型将用于将高油种质与中央和脂肪代谢网络相结合的因果效应联系起来，并用于为下一代作物改良做出假设。在体内的Fas速率，Fas与Tag合成的解偶联，可能导致Fas和破坏的无用循环；以及通过脂代谢网络的酰基通量将被研究，以阐明由于去调节Fas而导致TAG积累的新瓶颈。作为该项目的一部分开发的网络资源(FATPLANS.net)将用于教育社区，并提供一种整合新陈代谢和基因调控的互动工具。综合工具对于激励未来的科学家和工程师是必要的，并使所有年龄段的多学科培训成为可能。该项目还将为未来的科学和技术教育工作者开发一个实践科学素养计划(Sci-fest)，该计划的参与者将创建有关植物新陈代谢和生物技术的高中科学教学模块。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。

项目成果

The BADC and BCCP subunits of chloroplast acetyl-CoA carboxylase sense the pH changes of the light–dark cycle

叶绿体乙酰辅酶 A 羧化酶的 BADC 和 BCCP 亚基感知光暗循环的 pH 变化

• DOI: 10.1074/jbc.ra120.012877
• 发表时间: 2020
• 期刊: Journal of Biological Chemistry
• 影响因子: 4.8
• 作者: Ye, Yajin;Fulcher, Yan G.;Sliman, David J.;Day, Mizani T.;Schroeder, Mark J.;Koppisetti, Rama K.;Bates, Philip D.;Thelen, Jay J.;Van Doren, Steven R.
• 通讯作者: Van Doren, Steven R.

A General Method for Quantification and Discovery of Acyl Groups Attached to Acyl Carrier Proteins in Fatty Acid Metabolism Using LC-MS/MS

使用 LC-MS/MS 定量和发现脂肪酸代谢中酰基载体蛋白上附着的酰基的通用方法

• DOI: 10.1105/tpc.19.00954
• 发表时间: 2020
• 期刊: The Plant Cell
• 作者: Nam, Jeong-Won;Jenkins, Lauren M.;Li, Jia;Evans, Bradley S.;Jaworski, Jan G.;Allen, Doug K.
• 通讯作者: Allen, Doug K.

Normal phase HPLC method for combined separation of both polar and neutral lipid classes with application to lipid metabolic flux

• DOI: 10.1016/j.jchromb.2020.122099
• 发表时间: 2020-05-15
• 期刊: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
• 影响因子: 3
• 作者: Kotapati, Hari Kiran;Bates, Philip D.
• 通讯作者: Bates, Philip D.

Quantification of Acyl-Acyl Carrier Proteins for Fatty Acid Synthesis Using LC-MS/MS

• DOI: 10.1007/978-1-0716-1362-7_13
• 发表时间: 2021-01-01
• 期刊: PLANT LIPIDS
• 作者: Jenkins, Lauren M.;Nam, Jeong-Won;Allen, Doug K.
• 通讯作者: Allen, Doug K.

An efficient LC-MS method for isomer separation and detection of sugars, phosphorylated sugars, and organic acids

用于糖、磷酸化糖和有机酸异构体分离和检测的高效 LC-MS 方法

• DOI: 10.1093/jxb/erac062
• 发表时间: 2021
• 期刊: Journal of Experimental Botany
• 影响因子: 6.9
• 作者: Koley, Somnath;Chu, Kevin L;Gill, Saba S;Allen, Doug K
• 通讯作者: Allen, Doug K