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.

合作伙伴：弗吉尼亚·沃尔伯特（斯坦福大学）和格雷格·艾布拉姆斯（德克萨斯高级计算中心/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牧草的花药（雄性生殖器官）发育事件的调控提供新的见解，这些作物为世界上大部分地区提供食物。具有广泛用途的新见解和资源最终将有助于控制农业中的植物肥力，特别是谷类作物。该项目建立和改进了pi的各种广泛的外展和教育组成部分，以及他们通过科学领导的贡献，继续对当地和科学界以及下一代科学家产生强大的积极影响。“虚拟花药”的实施将使项目数据广泛可访问，同时帮助学生和公众了解花药是如何发展的。一个以本科生为重点的基生草基因组测序和注释项目将为该项目和所有在开花植物中使用系统基因组学分析的人员提供宝贵的数据，同时培养下一代植物生物学家。所有项目成果将通过项目网站和GEO和MaizeGDB等长期存储库向研究社区开放。所有的生物资源，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”的软件将开放源代码，并可通过德克萨斯高级计算中心网站（https://www.tacc.utexas.edu/tacc-software/）.Male)获得。不育是自然物种的一种常见表型，是促进异交的一种机制。此外，植物育种家控制花粉产量是一项基础技术，它继续支撑着许多作物的杂交种子生产。在农业方面，人们对气候破坏的担忧也越来越大：炎热和寒冷都会导致农作物的雄性不育。本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，特别是两种不寻常的“阶段性”次级小rna (phasiRNAs)，它们在草花药中高度富集。这些相RNA从大量协调表达的长链非编码RNA前体加工成21或24个核苷酸的次级小RNA。利用精确分期的玉米花药和激光显微解剖的细胞类型，该项目将生成丰富的转录组学和蛋白质组学数据，组织转录因子的转录层次，在时空背景下生成小rna的广泛数据，并分析相rna的功能和生物发生。利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。遗传学和“组学”实验将检验关于phasiRNA在花药细胞和减数分裂中的功能的新假设。由于玉米的发育规律较好，花药体积大，易于解剖，以及已有的关于玉米花药的分子和遗传知识，因此大部分分析将在玉米中进行。将评估水稻的关键见解，以区分一般和特定物种的点，并与其他禾本科植物进行额外的比较。为了产生新的假设，所有数据将被整合到“虚拟花药”可视化系统中。