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

开花植物，也被称为被子植物，包括最成功的陆地植物群体，并包括我们所有主要的驯化农作物。由于这些原因，了解不同被子植物的进化、发育、遗传学和生理学是植物生物学家关注的主要问题。然而，成功地进行这类研究项目需要存在一套特定的调查工具，特别是注释良好的基因组序列和方法，允许稳定地插入实验DNA，这一过程被称为转化。该项目将使用尖端方法来优化有效的转化方案，并改善可用于一系列有前途的被子植物系统的基因组资源，这些系统代表了鲜为人知的开花植物生命树的谱系。在选定的模型系统中生成这些工具具有相当大的潜力，可以提高进行植物生物学比较研究的能力。此外，开发的方案的传播将创建一个模板，可以被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。在外联和培训方面，活动包括制作一个基于YouTube的视频，解释现代植物转化是如何工作的，支持从本科生到博士后研究员的各种学术职业水平的培训机会，以及开发本科课程资源。最近在几个物种中的重大突破表明，过表达调控形态发生的基因，特别是Baby Boom/Wuschel2(BBM/Wus2)和GRF4/GRF相互作用因子1(GRF4-GIF1)，可以显著促进植物的转化和再生。该项目努力使这些方法适用于一组不同的被子植物，目标是因为它们占据了经过充分研究的作物和模式系统之间的系统发育差距。主要的目标分类群是蓝花草(毛茅科)、多根螺旋藻品系7498(天南星科浮萍)和芦笋(芦笋、芦笋科)。如果时间允许，还将努力扩大到睡莲(卢旺达睡莲，睡莲科)和马兜铃(荷兰管，马兜铃科)，以进一步界定被子植物的系统发育主干。作为开发这些关键工具的补充，该项目将使用下一代测序来改进三个主要模式分类群的基因组资源，特别是在小RNA生产基因座的注释方面。关于制定的方法和资源的教育和分发是该项目的一个关键组成部分。基因构建物将被存放在非营利性全球质粒库中，并将发布协议视频，使不熟悉转化的用户能够成功重复这一过程。所有测序数据将在没有禁运的情况下，在完成后尽快公开。此外，资金将支持1)本科生研究人员的培训，利用已经成功的资源，如NSF支持的本科生研究经验(REU)暑期项目和2)创建公共科学交流视频，以提高公众对转基因方法的理解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.