Inactivation of microRNAs in Crop Plants Using Short Tandem Target Mimic (STTM) Technology

PI：Guiliang Tang（Michigan Technological University，霍顿）CoPI：Xuemei Chen and Wenbo Ma（University of加州，滨江），Harold N. Trick（堪萨斯州立大学）和Hairong Wei（密歇根理工大学，霍顿）主要合作者：Kan Wang（爱荷华州州立大学）小RNA，包括microRNA（miRNAs）和小干扰RNA（siRNAs），在植物的生长和发育、表观遗传学、基因组完整性、抵御病原体感染以及对环境变化的反应中发挥着核心作用。miRNA通过在转录后水平负调控基因表达，在控制植物发育、生产力以及对生物和非生物胁迫的响应中特别重要。已经从几十种植物中鉴定出数百至数千种miRNAs。然而，它们在植物发育和对各种病原体和其他胁迫的反应中的作用在很大程度上是未知的。 该项目将最新的尖端基因组学技术整合到作物中的miRNAs研究中。 各级教育的学生将参加各种活动，包括教学，培训，学习研讨会和技术推广。该项目将为博士后、研究生和本科生提供小串联靶模拟（STTM）技术、水稻转化和miRNA功能研究方面的研究培训。 代表人数不足的少数民族和妇女以及来自五大湖上半岛地区的学生将成为招聘对象。该项目还将组织和举办一个STTM建设培训讲习班。 该项目提供的知识将通过在适当的社区场所举行的非正式会谈向广大公众传播。这项工作将通过回答重要的问题来解决关于miRNA进化的关键机制假设：为什么植物需要这么多的miRNA？它们是必不可少的吗？它们在不同植物谱系中的功能保守性和多样性是什么？该项目将使用最近开发的STTM技术在选定的农业重要作物中开发miRNA“敲低”群体作为试点项目。这些突变群体将进一步用于研究6个高度保守的植物miRNAs家族的功能保守性和多样性，以及miRNAs在植物-微生物相互作用中的调控作用。该项目将提供一个资源数据库、应用程序和服务，用于网上材料的传输和分发。 所有序列数据将保存在Gene Expression Omnibus（GEO）。 STTM构建体，水稻，玉米，拟南芥和大豆的STTM种子用于植物科学和作物改良的基础生物学将根据要求提供。

PI: Guiliang Tang (Michigan Technological University, Houghton) CoPIs: Xuemei Chen and Wenbo Ma (University of California, Riverside), Harold N. Trick (Kansas State University), and Hairong Wei (Michigan Technological University, Houghton) Key Collaborator: Kan Wang (Iowa State University) Small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs), play central roles in growth and development, epigenetics, genome integrity, defense against pathogen infection, and responses to environmental changes in plants. miRNAs are especially important in controlling plant development, productivity, and responses to biotic and abiotic stresses by negatively regulating gene expression at the post-transcriptional level. Hundreds to thousands of miRNAs have been identified from dozens of plant species. However, their roles in plant development and response against various pathogens and other stresses are largely unknown. This project will integrate the most recent cutting-edge genomics technology into the study of miRNAs in crop plants. Students at all levels of education will be engaged in a variety of activities including teaching, training, learning workshops, and technology-extension. The project will provide research training in small tandem target mimic (STTM) technology, rice transformation, and functional studies of miRNAs for postdoctoral associates, graduate and undergraduate students. Underrepresented minorities and women, as well as students from the Upper Peninsula region of Great Lakes, will be targeted and recruited. The project will also organize and hold a training workshop in STTM construction. The knowledge provided by this project will be disseminated to the public at large through informal talks at appropriate community venues. This work will address key mechanistic hypotheses regarding miRNA evolution by answering important questions: Why do plants need so many miRNAs? Are they essential? What are their functional conservation and diversification in different plant lineages? This project will develop miRNA "knockdown" populations in selected agriculturally important crops as a pilot project using the recently developed STTM technology. These mutant populations will be further used to investigate the functional conservation and diversification of six highly conserved families of plant miRNAs, and to study the regulatory role of miRNAs in plant-microbe interactions. The project will provide a resource database, application and service for web-based materials transfer and distribution. All sequence data will be deposited at Gene Expression Omnibus (GEO). STTM constructs, STTM seeds of rice, maize, Arabidopsis and soybean for use in basic biology in plant science and crop improvement will be available upon request.