RESEARCH-PGR: Single-cell Transcriptomic Analyses of Shoot Meristem Ontogeny and Function

RESEARCH-PGR：芽分生组织个体发育和功能的单细胞转录组分析

• 批准号: 2016021
• 负责人: Michael Scanlon
• 金额: $ 187.29万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2016021&HistoricalAwards=false
• 关键词: RESEARCH; PGR; Single; cell; Transcriptomic

The vast majority of cells, tissues and organs that comprise the adult maize plant are formed long after embryonic stages, due to the activity and proliferation of plant-specific stem cells called shoot meristems. Shoot meristems generate organ initial cells, which undergo specific patterns of gene expression during growth and development to give rise to the complex pattern of cells and tissues found in the mature plant body. In this way, shoot meristems arise during embryonic stages and generate all the vegetative structures of the maize shoot. New advances in genomic technology now enable analyses of gene expression within individual cells obtained at all stages of maize development, which promises to generate novel insight into the genetic and genomic mechanisms of shoot development in this agronomically important crop plant. Embryos make shoot meristems and shoot meristems make organs. This study will examine the specific patterns of gene expression in individual cells throughout shoot development, from early embryonic stages before, during and after shoot meristem initiation, and during formation of vegetative organs such as leaves, stems, and branches. These studies will advance our understanding of the cell-specific mechanisms underlying pattern formation in the maize vegetative shoot, and of the structure and function of shoot meristems. This project will generate original data for public release, while providing a framework for scientific training and teaching of graduate students, undergraduates who are under-represented in science, and people incarcerated in upstate New York State prisons.Pattern formation in the maize shoot begins early in embryogenesis and occurs across multiple scales, from individual cells and tissues, to all the organs within the plant. Recent advances now enable analyses of maize pattern formation at single-cell resolution. The maize shoot apical meristem (SAM) is a stem cell reservoir responsible for the development of all shoot-derived organs. This study will generate transcriptomic data from individual cells in the developing maize shoot, and will decipher the effects of cell position, cell signaling, rare cell types, and stochasticity on single-cell gene expression. Single-cell transcriptomic data will be obtained from key ontogenetic stages ranging from the establishment of embryonic axes, to formation of the SAM and the first lateral organs of the shoot, to the development of the seedling SAM and foliar leaf primordia, and during morphogenesis of the lateral branch meristems that will ultimately give rise to ears. Likewise, this project will investigate the single-cell transcriptomic networks within specific, functional domains of the vegetative SAM, and in wild type versus mutant shoot apices defective in leaf outgrowth to investigate the transcriptomic networks underlying maize shoot pattern formation throughout shoot ontogeny, at single-cell resolution. These studies will provide scientific training and release of original data, and will host minority undergraduate and high school students for summer research internships. Lastly, the Scanlon lab will continue teaching at the Elmira Correctional Facility, as part of the Cornell Prison Education Program.This award was co-funded by the Plant Genome Research Program and the Plant, Fungal and Microbial Developmental Mechanisms Program in the Division of Integrative Organismal Systems.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.

构成成年玉米植物的绝大多数细胞、组织和器官在胚胎阶段之后很久形成，这是由于被称为茎分生组织的植物特异性干细胞的活性和增殖。茎分生组织产生器官原始细胞，其在生长和发育期间经历特定的基因表达模式，以产生成熟植物体中发现的复杂的细胞和组织模式。以这种方式，芽分生组织在胚胎阶段出现并产生玉米芽的所有营养结构。基因组技术的新进展，现在能够分析在玉米发育的所有阶段获得的单个细胞内的基因表达，这有望产生新的见解，在这个农艺学上重要的作物植物的芽发育的遗传和基因组机制。胚形成芽分生组织，芽分生组织形成器官。本研究将研究在整个芽发育过程中，从早期胚胎阶段之前，期间和之后，芽分生组织的启动，并在营养器官，如叶，茎，和分支的形成过程中，在单个细胞中的基因表达的特定模式。这些研究将促进我们对玉米营养芽中图案形成的细胞特异性机制以及芽分生组织的结构和功能的理解。该项目将产生原始数据供公众发布，同时为研究生、在科学领域代表性不足的本科生和被监禁在北部纽约州监狱的人提供科学培训和教学框架。玉米芽中的模式形成始于胚胎发生的早期，并发生在多个尺度上，从单个细胞和组织到植物内的所有器官。最近的进展，现在能够在单细胞分辨率的玉米图案形成的分析。 玉米茎尖分生组织（SAM）是一个干细胞库，负责所有茎源器官的发育。这项研究将从发育中的玉米芽中的单个细胞中产生转录组数据，并将破译细胞位置，细胞信号传导，稀有细胞类型和随机性对单细胞基因表达的影响。单细胞转录组学数据将获得从关键的个体发育阶段，从胚轴的建立，形成的SAM和第一个横向器官的拍摄，幼苗SAM和叶原基的发展，并在形态建成的横向分支分生组织，最终会引起耳朵。同样，本项目将调查单细胞转录组网络内的特定功能域的营养SAM，并在野生型与突变体芽尖缺陷的叶生长调查转录组网络的玉米芽模式形成整个拍摄个体发育，在单细胞分辨率。这些研究将提供科学培训和发布原始数据，并将接待少数民族本科生和高中生进行暑期研究实习。最后，作为康奈尔监狱教育计划的一部分，斯坎伦实验室将继续在埃尔迈拉教养所教学。该奖项由植物基因组研究计划和植物，真菌和微生物发育机制计划在综合有机体系统的部门。这个奖项反映了NSF的法定使命，并已被认为是值得支持的，通过评估使用基金会的智力价值和更广泛的影响审查标准。

项目成果

A Wox3-patterning module organizes planar growth in grass leaves and ligules.

• DOI: 10.1038/s41477-023-01405-0
• 发表时间: 2023-05
• 期刊: NATURE PLANTS
• 影响因子: 18
• 作者: Satterlee, James W.;Evans, Lukas J.;Conlon, Brianne R.;Conklin, Phillip;Martinez-Gomez, Jesus;Yen, Jeffery R.;Wu, Hao;Sylvester, Anne W.;Specht, Chelsea D.;Cheng, Jie;Johnston, Robyn;Coen, Enrico;Scanlon, Michael J.
• 通讯作者: Scanlon, Michael J.