Regulatory Hierarchies and Roles of Non-Coding RNAs in Maize Anthers

玉米花药中非编码 RNA 的调控层次和作用

• 批准号: 1649424
• 负责人: Blake Meyers
• 金额: $ 412.88万
• 依托单位: Donald Danforth Plant Science Center
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1649424&HistoricalAwards=false
• 关键词: Regulatory; Hierarchies; Roles; Non; Coding

PI: Blake C. Meyers (University of Delaware)CoPIs: Virginia Walbot (Stanford University) and Greg Abrams (Texas Advanced Computing Center/University of Texas - Austin) Information generated from this project will provide new insights into the regulation of developmental events in the anther, the male reproductive organ, of maize and related grasses - crops that feed much of the world. New insights and resources of broad utility should ultimately contribute to controlling plant fertility in agriculture, particularly in the cereal crops. The project builds and improves on the varied and extensive outreach and education components of the PIs and their contributions via scientific leadership to continue a strong, positive impact on local and scientific communities, and on the next generation of scientists. Implementation of the "Virtual Anther" will make the project data widely accessible while helping students and the public understand how anthers develop. An undergraduate-focused project to sequence and annotate a basal grass genome will provide invaluable data for the project and to all those employing phylogenomics analysis in flowering plants, while training the next generation of plant biologists. All project outcomes will be accessible to the research community through a project website and long-term repositories such as GEO and MaizeGDB. All biological resources such as antibodies, clones and vectors will be available upon request. Seed will be deposited and distributed long-term through the Maize Genetics Cooperative. Software for the "Virtual Anther" will be made open-source and available via the Texas Advanced Computing Center website (https://www.tacc.utexas.edu/tacc-software/).Male sterility is a common phenotype in natural species, functioning as one mechanism to promote outcrossing. Furthermore, control of pollen production by plant breeders is a fundamental technology that continues to underpin hybrid seed production in many crops. In the agricultural context, there is also a growing concern over climate disruption: both heat and cold cause male sterility in crop plants. This study seeks to more broadly understand the coordination of anther development and underlying cellular processes, including chromatin remodeling, as plant cells switch from mitosis to meiosis. The specific goals are to: (1) organize hierarchies of gene expression in individual anther cell types and (2) elucidate functions of small RNAs, particularly two unusual classes of "phased" secondary small RNAs (phasiRNAs) that are highly enriched in grass anthers. These phasiRNAs are processed from an extensive set of coordinately-expressed long non-coding RNA precursors into secondary small RNAs of 21 or 24 nucleotides. Using precisely staged maize anthers and laser microdissected cell types, the project will generate copious transcriptomic and proteomic data, organize transcriptional hierarchies of transcription factors, generate extensive data on small RNAs in a spatiotemporal context, and analyze the function and biogenesis of phasiRNAs. Genetic analysis using existing and engineered mutations will be used to test for essential roles of key transcription and phasiRNA biogenesis factors in anther development. Genetic and "omics" experiments will test new hypotheses about phasiRNA functions in anther cells and in meiosis. Most of the analysis will be conducted in maize because of its exquisite developmental regularity, the large size and ease of anther dissection, and existing molecular and genetic knowledge about maize anthers. Key insights will be evaluated in rice to distinguish general and species-specific points, with additional comparisons to other grasses. To generate new hypotheses, all data will be integrated into a "Virtual Anther" visualization system.

首席研究员：Blake C. Meyers（特拉华大学） 首席研究员：Virginia Walbot（斯坦福大学）和 Greg Abrams（德克萨斯州高级计算中心/德克萨斯大学奥斯汀分校） 该项目产生的信息将为玉米和相关草类（为世界大部分地区提供粮食的作物）的花药（雄性生殖器官）的发育事件的调控提供新的见解。 具有广泛用途的新见解和资源最终应有助于控制农业中的植物肥力，特别是谷类作物。该项目建立并改进了 PI 的各种广泛的外展和教育部分以及他们通过科学领导力做出的贡献，以继续对当地和科学界以及下一代科学家产生强大、积极的影响。 “虚拟花药”的实施将使项目数据可以广泛访问，同时帮助学生和公众了解花药如何发育。一个以本科生为重点的对基础草基因组进行测序和注释的项目将为该项目以及所有在开花植物中使用系统发育组学分析的人员提供宝贵的数据，同时培训下一代植物生物学家。研究界可以通过项目网站和长期存储库（例如 GEO 和 MaizeGDB）访问所有项目成果。所有生物资源，如抗体、克隆和载体将根据要求提供。种子将通过玉米遗传合作社长期存放和分发。 “虚拟花药”的软件将开源并可通过德克萨斯高级计算中心网站 (https://www.tacc.utexas.edu/tacc-software/) 获得。雄性不育是自然物种中的一种常见表型，是促进异型杂交的一种机制。此外，植物育种者控制花粉生产是一项基本技术，继续支撑许多作物的杂交种子生产。在农业方面，人们越来越担心气候破坏：炎热和寒冷都会导致农作物雄性不育。这项研究旨在更广泛地了解植物细胞从有丝分裂转变为减数分裂时花药发育和潜在细胞过程（包括染色质重塑）的协调。 具体目标是：(1) 组织个体花药细胞类型中基因表达的层次结构；(2) 阐明小 RNA 的功能，特别是在草花药中高度富集的两类不寻常的“定相”次级小 RNA (phasiRNA)。这些 phasiRNA 由大量协调表达的长非编码 RNA 前体加工成 21 或 24 个核苷酸的次级小 RNA。使用精确分级的玉米花药和激光显微切割的细胞类型，该项目将产生大量的转录组和蛋白质组数据，组织转录因子的转录层次结构，在时空背景下产生有关小RNA的大量数据，并分析phasiRNA的功能和生物发生。使用现有和工程突变的遗传分析将用于测试关键转录和 phasiRNA 生物发生因子在花药发育中的重要作用。遗传和“组学”实验将测试有关 phasiRNA 在花药细胞和减数分裂中功能的新假设。大多数分析将在玉米中进行，因为其精致的发育规律、花药体积大且易于解剖，以及关于玉米花药的现有分子和遗传知识。将在水稻中评估关键见解，以区分一般点和物种特异性点，并与其他禾本科植物进行额外比较。为了产生新的假设，所有数据将被集成到“虚拟花药”可视化系统中。