EAGER: Multiscale multiplex spatial-omics: Illuminating molecular pathways and architecture in plant cells and tissues

该项目将开发先进的显微镜技术，用于同时观察植物细胞内的许多不同分子，从而有助于了解植物如何生长，繁殖以及与致病微生物相互作用。这项对开发多重显微镜的投资将为更大的科学界提供新的植物聚焦和其他非生物医学应用。该项目将用于培训各级科学家，直至主要研究人员。明天的科学家将通过积极参与唐纳德·丹福思植物科学中心（DDPSC）NSF本科生研究经验（REU）计划，教授高级成像短期课程，招募高中生进行成像项目，并参与高中生区域实习计划，称为学生和教师作为研究科学家（STARS）。 此外，非常成功的DDPSC教育和推广计划将为教师举办研讨会，并提供互动指导，将我们的科学家与附近的学校，包括哈里斯斯托大学，历史上的黑人学院和大学。与植物生物学生物分析资源合作，这项工作将在线共享植物数据，并通过NSF资助的植物细胞图谱倡议积极交流和传播新开发的多重技术，这是一种社区资源，旨在建立一个综合的高分辨率植物细胞分子图谱。这项工作的成果将通过同行评审的出版物、科学会议、实践/在线研讨会、培训视频和可搜索的在线数据库传播。生物学的一个当代挑战是了解大量不同分子如何分布在细胞和组织中。为了解决这个问题，空间组学技术正在改变我们理解复杂生物体的方式。一种形式，“多重”显微镜，已被广泛应用于生物医学应用，以了解人类疾病，但几乎没有在植物中使用。 该项目将开发一个变革性的多功能管道，用于多尺度植物组织阵列，使用多重显微镜，探测核酸，蛋白质和多糖，用于同一样品的相关光，超分辨率和电子显微镜，同时保留细胞超微结构。多重技术将在选择的植物系统上验证，以创建以下的高维多重数据集：1）授粉期间的细胞/细胞壁修饰，2）小RNA生物发生期间花药中的蛋白质、mRNA和sRNA定位，和3）主机-该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准。

This project will develop advanced microscopy techniques for visualizing many different molecules at the same time within plant cells, thus contributing to understanding of how plants grow, reproduce and interact with disease causing microbes. This investment in developing multiplex microscopy will provide novel plant focused and other non-biomedical applications for the greater scientific community. This project will be leveraged for training scientists at all ranks up to principal investigators. Tomorrow’s scientists will be trained through active participation in the Donald Danforth Plant Science Center (DDPSC) NSF-Research Experiences for Undergraduates (REU) Program, teaching an advanced imaging short course, recruitment of high school students for imaging projects and participation in a regional internship program for high school students called Students and Teachers As Research Scientists (STARS). Additionally, the highly successful DDPSC Education and Outreach program will host workshops for teachers and provide interactive mentoring that connects our scientists with nearby schools including Harris Stowe University, a Historically Black College and University. In collaboration with The Bio-Analytic Resource for Plant Biology, this effort will share plant data online, and actively communicate and disseminate newly developed multiplex techniques through the NSF funded Plant Cell Atlas initiative, a community resource with the goal to build an integrated high-resolution molecular map of plant cells. The results from this effort will be disseminated in peer-reviewed publications, at scientific meetings, in hands-on/online workshops, training videos and a searchable online database.One contemporary challenge in biology is to understand how large numbers of diverse molecules are distributed within cells and tissues. To address this, spatial-omics techniques are transforming the way we understand complex organisms. One form, “multiplex” microscopy, has been applied largely for biomedical applications to understand human disease, but is virtually unused in plants. This project will develop a transformative and versatile pipeline for multiscale plant tissue arrays using multiplex microscopy, probing nucleic acids, proteins and polysaccharides for correlated light, super-resolution and electron microscopy on the same sample, while preserving cell ultrastructure. The multiplex technique will be validated on select plant systems to create high-dimensional multiplex datasets of 1) cellular/cell wall modifications during abscission, 2) protein, mRNA and sRNA localization in anthers during small RNA biogenesis, and 3) host-pathogen interactions at infection sites.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.