Functional Analyses of Genes Involved in Meristem Organization and Leaf Initiation

参与分生组织和叶子起始的基因的功能分析

• 批准号: 0638770
• 负责人: Michael Scanlon
• 金额: $ 181.17万
• 依托单位: Cornell Univ - State: AWDS MADE PRIOR MAY 2010
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0638770&HistoricalAwards=false
• 关键词: Functional; Analyses; Genes; Involved; Meristem

All above ground organs of higher plants are ultimately derived from specialized organogenic structures called shoot apical meristems (SAMs). The SAM exhibits distinctive structural organization, marked by tissue zonation and cell layering. The structure of plant SAMs is correlated with their function, such that new leaves are initiated from the peripheral zone of the SAM and the central zone replenishes new meristematic cells that are lost during organogenesis. Establishment and maintenance of these structurally and functionally distinct zones of the SAM is achieved by differential expression patterns and interactions of thousands of plant genes. Breakthrough technologies have emerged that enable the identification and analysis of genes required for meristem function in maize. Laser dissection microscopy (LCM) is a powerful technique that permits the isolation of RNA from specific cell types within fixed plant tissues immobilized on slides. RNA collected from 1,000-10,000 cells is sufficient for use in microarray analyses of gene expression, which permit the simultaneous examination of expression profiles of 15,000 to 30,000 genes. The relatively large size of the maize vegetative meristem, approximately 250 meristematic cells are recruited into the incipient maize leaf, renders this plant especially tractable for this experimental system. The laser-capture microdissection/microarray technique will be used to capture cells from specific domains of the maize meristem and newly formed leaf primordia for use in comparative analyses of global gene expression. The differential expression patterns of candidate genes will be verified by more traditional analyses (RT-PCR, RNA gel blot hybridization and in situ hybridization) of transcript accumulation in maize tissues. These experiments will microdissect gene expression patterns in meristems and leaf primordia, and will provide novel insight into mechanisms of plant development. This research project will generate the following set of deliverables:1. LCM will be optimized and improved for use on maize SAMs. Information will be available on a project web site at ISU and in publications.2. Microarray profiles of global gene expression in the SAM will be deposited in public web sites, maizeGDB. 3. An EST collection from meristem-enriched cDNA library will be developed beginning in the second year of the project, depending on need. If developed, all EST sequences will be deposited in Genbank and in maizeGDB.4. Sequence generated by students mining for maize equivalents to already known genes from other plants will be sent directly to Genbank and maizeGDB.Outreach and training will be enhanced in three ways:1. Undergraduate students from Truman State University will engage in cutting edge genomics research. 2. PI Scanlon will deliver guest lectures at local high schools with predominantly minority enrollment. 3. The University of Georgia and the CSHL Dolan DNA Learning Center (DNALC) will train teachers from underrepresented minority high schools and university settings in plant genomics and bioinformatics.

最终，所有高于较高植物的地面器官都源自称为芽顶分生组织（SAM）的专门有机生成结构。 SAM表现出独特的结构组织，其标志是组织分区和细胞分层。 植物SAM的结构与它们的功能相关，因此从SAM的外围带开始了新的叶子，并且中央区域补充了在器官发生过程中丢失的新分生物细胞。 这些结构和功能上不同区域的建立和维护是通过数千种植物基因的差异表达模式和相互作用来实现的。 突破性技术已经出现，可以识别和分析玉米分生组织功能所需的基因。 激光解剖显微镜（LCM）是一种强大的技术，可以从固定在载玻片上的固定植物组织中的特定细胞类型中分离RNA。 从1,000-10,000个细胞收集的RNA足以用于基因表达的微阵列分析，该分析允许同时检查15,000至30,000个基因的表达谱。 玉米植物分生组织的相对较大，将大约250个分生组织细胞募集到初期的玉米叶中，这使该植物在该实验系统中尤其可处理。 激光捕获微分解/微阵列技术将用于从玉米分生组织的特定结构域中捕获细胞和新形成的叶子原始素，用于用于全局基因表达的比较分析。 候选基因的差异表达模式将通过对玉米组织中转录本积累的更多传统分析（RT-PCR，RNA凝胶印迹杂交和原位杂交）进行验证。 这些实验将在分生组织和叶子原始基因上进行显微解剖的基因表达模式，并将为植物发育机制提供新的见解。 该研究项目将生成以下一组可交付成果：1。 LCM将被优化并改进，以用于玉米SAM。信息将在ISU和Publications的项目网站上找到2。 SAM中全球基因表达的微阵列概况将存放在玉米公共网站上。 3。根据需求，将从项目的第二年开始开发富含分生组织的cDNA库的EST藏品。如果开发，所有EST序列将沉积在GenBank和MaizeGDB.4中。 由学生在玉米等效物中开采与其他植物的基因产生的顺序将直接发送给GenBank，并以三种方式增强玉米和培训。 杜鲁门州立大学的本科生将从事尖端基因组学研究。 2。PiScanlon将在当地高中的嘉宾讲座中，主要是少数派。 3。佐治亚大学和CSHL Dolan DNA学习中心（DNALC）将培训来自代表性不足的少数民族中学和大学植物基因组学和生物信息学的教师。