NSF Postdoctoral Fellowship in Biology: Plant Cell Networking

本行动资助2023财年美国国家科学基金会植物基因组生物学博士后研究奖学金。该奖学金支持奖学金获得者在主办实验室的研究和培训计划，该奖学金获得者还提出了扩大生物学参与的计划。Imani Madison博士的研究和培训计划的标题是“植物细胞网络”。该奖学金的主办机构是北卡罗莱纳州立大学，赞助科学家是Rosangela Sozzani博士。植物细胞之间通过各种信号分子和途径相互沟通，协调生长，建立细胞身份，传递胁迫反应。植物的细胞间通讯是由胞间连丝或细胞壁上的门控通道介导的。控制胞间连丝的形成、门控和运输是调节细胞间通讯的关键。总体而言，本项目将研究胞间连丝如何在细胞壁形成并调节细胞间分子运动。在整株植物中，由于胞间连丝体积小且难以建立突变体研究系统，对其进行研究具有挑战性。该项目将通过使用创新的3D生物打印技术来创建源自大豆的植物单细胞系统，以研究细胞壁的形成，从而解决这一挑战。该项目还将研究基因组编辑或复合治疗如何控制细胞间通讯。由于间连丝存在于所有植物物种中，它们是在各种非生物或生物胁迫下影响细胞通讯的普遍靶点。为了成功完成这些目标，该研究员将继续接受共聚焦显微镜技术的培训，并获得单细胞基因组编辑以及数据和项目管理方面的专业知识。这项研究还将为社区外展和历史上被排斥的科学家的参与提供机会。为了促进这些机会，奖学金获得者将在赞助机构接受指导和领导培训以及领导讲习班，向更广泛的社区介绍这项工作的重要性。为了建立研究系统，大豆细胞将通过细胞壁消化分离出来，然后进行生物打印。然后用电子显微镜观察生物打印细胞以检测胞间连丝。为了补充这一技术，荧光探针和基因组编辑将被用于荧光标记胞间连丝，以检测胞间连丝在细胞分裂和重组细胞壁时的生物发生和定位动态。为了量化细胞间通讯的动态，荧光移动探针将应用于生物打印细胞，并测量其通量率，以估计大分子和小分子的运动。最后，为了操纵细胞间通讯，将进行高通量化合物筛选，以量化通讯可以增强、抑制或减少的程度，以及这些化合物是否具有永久性影响。总的来说，这将提供一个全面的了解基本的细胞通信和如何广泛地控制它。展望未来，这将有助于在诸如抑制细胞间病原体传播和改善依赖细胞通信的植物发育和环境响应方面改善作物生长。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This action funds an NSF Plant Genome Postdoctoral Research Fellowship in Biology for FY 2023. The fellowship supports a research and training plan in a host laboratory for the Fellow who also presents a plan to broaden participation in biology. The title of the research and training plan for this fellowship to Dr. Imani Madison is “Plant Cell Networking” The host institution for the fellowship is North Carolina State University and the sponsoring scientist is Dr. Rosangela Sozzani. Plant cells communicate with each other to coordinate growth, establish cell identity, and transmit stress responses through various signaling molecules and pathways. Plant intercellular communication is mediated by plasmodesmata, or gated channels in cell walls. Harnessing control over plasmodesmata formation, gating, and trafficking is key to regulating intercellular communication. Overall, this project will investigate how plasmodesmata form in cell walls and regulate intercellular molecular movement. In whole plants, it is challenging to study plasmodesmata due to their small size and the difficulty of creating mutant study systems. This project will address this challenge through the use of innovative 3D bioprinting technology to create plant single-cell systems, derived from soybean, to study cell walls as they form. This project will also investigate how either genome editing or compound treatments can control intercellular communication. As plasmodesmata are present across all plant species, they present a universal target for influencing cell communication under various abiotic or biotic stressors. To successfully complete these goals, the Fellow will continue her training in confocal microscopy techniques and gain expertise in single-cell genome editing as well as data and project management. This research will also provide opportunities for community outreach and participation of historically excluded scientists. To facilitate these opportunities, the Fellow will take mentorship and leadership training as well as lead workshops at the sponsor institution to introduce the importance of this work to the broader community.To create the study system, soybean cells will be isolated by cell wall digestion and then bioprinted. Bioprinted cells will then be visualized using electron microscopy to detect plasmodesmata. To complement this technique, fluorescent probes and genome editing will be used to fluorescently label plasmodesmata to detect the dynamics of plasmodesmata biogenesis and localization in cells as they divide and reform cell walls. To quantify the dynamics of intercellular communication, fluorescent mobile probes will be applied to bioprinted cells and their flux rate will be measured to estimate the movement of both large and small molecules. Finally, to manipulate intercellular communication, a high throughput compound screen will be performed to quantify to what extent communication can be enhanced, inhibited, or decreased and whether these compounds have permanent effects. Overall, this will provide a comprehensive understanding of the fundamentals of cell communication and how extensively it can be controlled. Going forward, this will be useful in efforts to improve crop growth in areas such as inhibiting intercellular pathogen spread and improving aspects of plant development and environmental responses that are reliant on cell communication.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.