Collaborative Research: EDGE FGT: Transformation and Genomic Resources to Advance Diverse, Emerging Model Angiosperms

合作研究：EDGE FGT：促进多样化、新兴模型被子植物的转化和基因组资源

• 批准号: 2128195
• 负责人: Elena Kramer
• 金额: $ 31.47万
• 依托单位: Harvard University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2128195&HistoricalAwards=false
• 关键词: Collaborative; Research; EDGE; FGT; Transformation

The flowering plants, also known as angiosperms, encompass the most successful living group of land plants, and include all of our major domesticated crop plants. For these reasons, understanding the evolution, development, genetics and physiology of diverse angiosperms is a major focus for plant biologists. However, successful pursuit of such research programs requires the existence of a specific set of investigative tools, particularly a well-annotated genome sequence and methods that allow for the stable insertion of experimental DNA, a process known as transformation. This project will use cutting-edge approaches to both optimize effective transformation protocols and improve the genomic resources available for a set of promising experimental angiosperm systems that represent poorly understood lineages of the flowering plant tree of life. The generation of these tools in the selected model systems has considerable potential to improve the ability to conduct comparative studies of plant biology. Moreover, dissemination of protocols developed will create a template that can be applied by other researchers to even more angiosperm models, thereby having an even broader positive impact on the field. With respect to outreach and training, activities include creating a YouTube-based video explaining how modern plant transformation works, supporting diverse training opportunities at all academic career levels from undergraduate students through postdoctoral fellows, and developing undergraduate course resources. Recently, major breakthroughs in several species have shown that overexpressing genes that regulate morphogenesis, specifically Baby Boom/Wuschel2 (Bbm/Wus2) and Growth Regulating Factor 4/GRF Interacting Factor 1 (GRF4-GIF1), results in significantly enhanced plant transformation and regeneration. This project strives to adapt these methods on a set of diverse angiosperms, targeted because they occupy the phylogenetic gap between well-studied crop and model systems. The primary target taxa are Aquilegia coerulea (columbine, Ranunculaceae); Spirodela polyrhiza line 7498 (duckweed, Araceae); and Asparagus officinalis (asparagus, Asparagaceae). If time allows, efforts will also extend to Nymphaea thermarum (Rwandan waterlily, Nymphaeaceae) and Aristolochia fimbriata (dutchman’s pipe, Aristolochiaceae) to further circumscribe the phylogenetic backbone of the angiosperms. As a complement to the development of these key tools, the project will use next-generation sequencing to improve genomic resources for the three primary model taxa, particularly in regard to the annotation of small RNA producing loci. Education on and distribution of the developed methodologies and resources are a key component of the project. Gene constructs will be deposited in nonprofit global plasmid repositories and protocol videos will be published to enable users unfamiliar with transformation to successfully repeat the process. All sequencing data will be made publicly available as soon as completed with no embargo. Additionally, funds will support 1) the training of undergraduate researchers, leveraging already successful resources such as the NSF-supported Research Experiences for Undergraduates (REU) summer programs and 2) the creation of public science communication videos to improve public understanding of transgenic approaches.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.

开花植物，也称为被子植物，包括最成功的陆地植物群，包括我们所有主要的驯化作物植物，因此，了解不同被子植物的进化、发育、遗传学和生理学是研究的重点。然而，成功开展此类研究项目需要一套特定的研究工具，特别是注释良好的基因组序列和允许实验 DNA 稳定插入的方法，这一过程称为转化。使用尖端方法来优化有效的转化方案并改善一组有前途的实验被子植物系统可用的基因组资源，这些系统代表了人们对开花植物生命之树的谱系知之甚少。这些工具在选定的模型系统中的产生具有相当大的意义。此外，所开发的协议的传播将创建一个模板，可供其他研究人员应用于更多的被子植物模型，从而对该领域产生更广泛的积极影响。外展和培训，活动包括创建一个基于 YouTube 的视频，解释现代植物转化的工作原理，支持从本科生到博士后的所有学术职业级别的多样化培训机会，并开发本科课程资源。最近，几个物种的重大突破表明，过度表达调节形态发生的基因。 ，特别是 Baby Boom/Wuschel2 (Bbm/Wus2) 和生长调节因子 4/GRF 相互作用因子 1 (GRF4-GIF1)，可显着增强植物转化和再生。该项目致力于适应这些。针对一组不同被子植物的方法，因为它们占据了经过充分研究的作物和模型系统之间的系统发育差距，主要目标分类群是耧斗菜（耧斗菜，毛茛科）；紫萍属 7498（浮萍，天南星科）； （芦笋，天门冬科）如果时间允许，努力还将延伸到睡莲属。 thermarum（卢旺达睡莲，睡莲科）和马兜铃（荷兰管，马兜铃科）进一步界定被子植物的系统发育骨架作为对这些关键工具开发的补充，该项目将使用下一代测序来改善基因组资源。三个主要模型分类单元，特别是在小 RNA 产生位点的注释方面，以及已开发方法的教育和分布。资源是该项目的关键组成部分，基因构建体将存放在非营利性全球质粒存储库中，并且将发布协议视频，以使不熟悉转化的用户能够成功地重复该过程，所有测序数据将在完成后立即公开。没有禁运，资金将支持 1) 培训本科生研究人员，利用已经成功的资源，例如 NSF 支持的本科生研究经验 (REU) 暑期项目，以及 2) 制作公共科学传播视频，以提高公众对科学传播的理解该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。