IOS EDGE: Generating Transgenic Cuscuta as a Tool for Studying Plant Interactions

IOS EDGE：生成转基因菟丝子作为研究植物相互作用的工具

• 批准号: 1645027
• 负责人: James Westwood
• 金额: $ 56万
• 依托单位: Virginia Polytechnic Institute and State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1645027&HistoricalAwards=false
• 关键词: IOS; EDGE; Generating; Transgenic; Cuscuta

Cuscuta (commonly known as dodder) is a parasitic plant with an unusual growth habit. As a parasite it lives in association with at least one other plant, from which it derives all its water and nutritional needs, while providing nothing to its host in return. Cuscuta obtains its food from other plants, so it has no need for roots, leaves or photosynthesis, and appears as a network of yellow vining stems that wind around host plants. Cuscuta provides a fresh perspective for understanding plant science because its evolution to parasitism has resulted in exaggerated features that push the boundaries of plants capabilities. For example, Cuscuta seedlings can identify host locations and grow toward them, demonstrating an ability to detect and respond to other plants in their environment. Also, the ability of Cuscuta to feed off other plants creates a system in which to explore plant interactions at the cell level and to shape concepts of plant disease. Finally, the Cuscuta habit of exchanging RNA and proteins with hosts challenges our understanding of genetic regulation of individual organisms. This EDGE project aims to expand the use of Cuscuta as a research model system through development of plant transformation methods to genetically test gene function, and to provide techniques and tips on growing Cuscuta to other scientists. Growing the Cuscuta research community will promote progress in basic plant science with potential impacts in disease resistance, agricultural ecology and weed control.The major barrier preventing the use of Cuscuta as a model plant system is the lack of a protocol for transformation and regeneration of the plant that would enable studies of gene function. The primary objective of this project is to develop transgenic Cuscuta. A few previous studies have established conditions for either Cuscuta callus transformation or regeneration from callus, but never both with the same tissue. This project focuses on systematically optimizing Cuscuta culture conditions, transformation, and regeneration to produce stably transformed parasitic plants. This will include evaluation of several Cuscuta species for amenability to transformation, along with testing Agrobacterium strains for efficiency of Cuscuta transformation. Additional approaches that aim for rapid transformation of Cuscuta without a tissue culture step (i.e., floral dip) will be tested as well. All protocols and resources will be made accessible to the broader plant community through a project website and through publications. Parties interested in obtaining germplasm used or generated in this project are encouraged to first contact USDA-APHIS regarding how to obtain permission to work with these species.

菟丝子（俗称菟丝子）是一种寄生植物，具有特殊的生长习性。作为一种寄生虫，它与至少一种其他植物生活在一起，从那里获得所有的水和营养需求，而不向宿主提供任何回报。菟丝子从其他植物中获取食物，因此它不需要根，叶或光合作用，并表现为围绕宿主植物缠绕的黄色藤蔓茎的网络。菟丝子为理解植物科学提供了一个新的视角，因为它进化为寄生性，导致了夸大的特征，推动了植物能力的界限。例如，菟丝子幼苗可以识别寄主位置并向它们生长，表现出检测和响应环境中其他植物的能力。此外，菟丝子以其他植物为食的能力创造了一个系统，在该系统中，探索植物在细胞水平上的相互作用，并形成植物疾病的概念。最后，菟丝子与宿主交换RNA和蛋白质的习惯挑战了我们对个体生物遗传调控的理解。该EDGE项目旨在通过开发植物转化方法来遗传测试基因功能，并为其他科学家提供种植菟丝子的技术和技巧，以扩大菟丝子作为研究模型系统的使用。菟丝子研究群体的发展将促进基础植物科学的进步，并对抗病性、农业生态学和杂草控制产生潜在影响。阻碍菟丝子作为模式植物系统使用的主要障碍是缺乏一种能够进行基因功能研究的植物转化和再生方案。本项目的主要目标是培育转基因菟丝子。之前的一些研究已经建立了菟丝子愈伤组织转化或愈伤组织再生的条件，但从未同时使用相同的组织。本项目的重点是系统地优化菟丝子的培养条件、转化和再生，以生产稳定转化的寄生植物。这将包括评价几种菟丝子属物种对转化的适应性，沿着测试土壤杆菌菌株对菟丝子属转化的效率。旨在快速转化菟丝子而无需组织培养步骤的其它方法（即，花浸）也将被测试。所有协议和资源将通过项目网站和出版物向更广泛的工厂社区提供。鼓励有兴趣获得本项目中使用或产生的种质的各方首先联系USDA-APHIS，了解如何获得使用这些物种的许可。

项目成果

An artificial host system enables the obligate parasite Cuscuta campestris to grow and reproduce in vitro

• DOI: 10.1093/plphys/kiac106
• 发表时间: 2022-03-16
• 期刊: PLANT PHYSIOLOGY
• 影响因子: 7.4
• 作者: Bernal-Galeano, Vivian;Beard, Keely;Westwood, James H.
• 通讯作者: Westwood, James H.

MicroRNAs from the parasitic plant Cuscuta campestris target host messenger RNAs

• DOI: 10.1038/nature25027
• 发表时间: 2018-01-04
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Shahid, Saima;Kim, Gunjune;Axtell, Michael J.
• 通讯作者: Axtell, Michael J.