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

基本信息

批准号：
2128196
负责人：
Alex Harkess
金额：
$ 23.69万
依托单位：
Auburn 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 2023-10-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2128196&HistoricalAwards=false
关键词：
Collaborative Research EDGE FGT Transformation

项目摘要

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，涵盖了最成功的陆地植物群，并包括我们所有主要的驯化作物植物。由于这些原因，了解潜水员被子植物的进化，发展，遗传学和生理学是植物生物学家的主要重点。但是，成功地追求此类研究计划需要存在一组特定的研究工具，尤其是允许稳定插入实验DNA的良好基因组序列和方法，这是一种称为转化的过程。该项目将使用尖端的方法来优化有效的转化方案，并改善用于一组承诺的实验被子植物系统的基因组资源，这些系统代表了开花植物生命树的谱系不足的谱系。所选模型系统中这些工具的生成已经考虑了提高进行植物生物学比较研究的能力的潜力。此外，开发的协议的传播将创建一个模板，其他研究人员可以将其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。关于宣传和培训，活动包括创建基于YouTube的视频，解释了现代工厂转型的工作方式，并在本科生通过博士后研究员提供了所有学术职业级别的潜水员培训机会，并开发了本科课程资源。最近，几个物种的主要突破表明，过表达调节形态发生的基因，特别是婴儿繁荣/wuschel2（BBM/WUS2）和调节生长因子4/GRF相互作用因子1（GRF4-GIF1）的基因，导致显着增强的植物转化和再生。该项目努力将这些方法适应一组潜水员被子植物，其目标是因为它们占据了良好的作物和模型系统之间的系统发育差距。主要的目标分类单元是Aquilegia Coerulea（哥伦拜科，ranunculaceae）； Spirodela Polyrhiza系列7498（Araceae Duckweed）；和Asparagus officinalis（Asparagus，Asparagaceae）。如果时间允许，努力也将延伸到若虫热（rwandan Waterlily，Nymphaeaceae）和Aristolochia fimbriata（荷兰人的管道，Aristolochiaceae），以进一步限制该血管植物的系统发育骨干。作为这些关键工具的完整开发，该项目将使用下一代测序来改善三个主要模型类群的基因组资源，尤其是在小型RNA生产场所的注释方面。关于开发方法和资源的教育和分配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，并将发布协议视频，以使不熟悉转型的用户成功重复该过程。所有测序数据将在没有禁运的情况下立即公开提供。此外，资金将支持1）对本科研究人员的培训，利用已经成功的资源（例如NSF支持的本科生（REU）夏季计划和2）创建公共科学沟通视频来提高公众对转基因方法的理解，这反映了NSF的法定任务和审查的良好依据，这是通过评估良好的支持。