CAA: Differential Gene Expression Related to Calcium Oxalate Formation in Grape

CAA：与葡萄中草酸钙形成相关的差异基因表达

• 批准号: 9707817
• 负责人: Mary Webb
• 金额: $ 6.12万
• 依托单位: Purdue Research Foundation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 1999-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9707817&HistoricalAwards=false
• 关键词: CAA; Differential; Gene; Expression; Related

In most higher plants, certain cells become specialized to accumulate calcium and precipitate it as crystalline calcium oxalate. These cells appear to function in bulk regulation of calcium by sequestering excess calcium, thereby reducing levels of free ionic calcium within the plant. Plant cells that sequester calcium oxalate exhibit distinct structural and biochemical differences as they differentiate for this role. Dr. Webb's research goals focus on understanding what stimulates plant cells to develop for this specialized function, and how the cell controls the crystallization process. The primary objective of her current research is the application of molecular approaches to identify genes that are differentially expressed at early stages in the development of crystal-forming cells. Molecular studies will be initiated with two tissues in grape, endosperm and root cortex, which have contrasting patterns of crystal morphology and development. Differentially expressed genes will be identified using mRNA differential display to compare distinct populations of mRNA from each of these tissues. Messages from endosperm will be compared at selected developmental stages, and those from roots after manipulating calcium nutrition to alter crystal development. After differential expression of selected cDNAs is confirmed, they will be sequenced and compared to known sequences in computer databases. Full-length cDNA clones will be isolated and characterized. Clones that have potential roles in calcium transport, oxalate metabolism, or vacuole function will be of particular interest. To identify genes specifically expressed in crystal idioblasts, the cellular localization of expression will also be determined for selected cDNAs using in situ hybridization. These studies will form the basis for future research to determine how expression of particular genes and gene products are important in calcium oxalate crystallization and the differentiation of crystal-forming cells. Correlated studies will examine the structural development of crystal-forming cells in grape roots during their differentiation. A variety of approaches will be explored using scanning confocal microscopy to observe unique features of these cells. A primary objective in this work will be to examine the development of the membrane systems that compartmentalize calcium oxalate crystals within the vacuole. All of the research to be conducted under this Career Advancement Award emphasizes activities and techniques that are new to Dr. Webb and will significantly enhance her research and advisory capabilities.

在大多数高等植物中，某些细胞专门积累钙并将其沉淀为结晶草酸钙。 这些细胞似乎通过螯合过量的钙来大量调节钙，从而降低植物内游离钙离子的水平。 螯合草酸钙的植物细胞表现出明显的结构和生化差异，因为它们分化为这种作用。 韦伯博士的研究目标集中在了解是什么刺激植物细胞发展这种专门的功能，以及细胞如何控制结晶过程。 她目前研究的主要目标是应用分子方法来鉴定在晶体形成细胞发育早期阶段差异表达的基因。 分子研究将开始与两个组织在葡萄，胚乳和根皮层，这有不同的晶体形态和发展模式。 将使用mRNA差异显示来鉴定差异表达的基因，以比较来自这些组织中的每一种的不同mRNA群体。 胚乳的信息将在选定的发育阶段进行比较，那些从根后操纵钙营养，以改变晶体的发展。 在确认所选cDNA的差异表达后，将对它们进行测序并与计算机数据库中的已知序列进行比较。 全长cDNA克隆将被分离和表征。 在钙转运、草酸盐代谢或液泡功能中具有潜在作用的克隆将特别感兴趣。 为了鉴定在晶体异细胞中特异性表达的基因，还将使用原位杂交确定所选cDNA的表达的细胞定位。 这些研究将为未来的研究奠定基础，以确定特定基因和基因产物的表达如何在草酸钙结晶和晶体形成细胞的分化中发挥重要作用。 相关的研究将检查晶体形成细胞在葡萄根的分化过程中的结构发展。 各种方法将探索使用扫描共聚焦显微镜观察这些细胞的独特功能。 这项工作的一个主要目标将是检查的膜系统，划分草酸钙晶体内的液泡的发展。 在这个职业发展奖下进行的所有研究都强调对韦伯博士来说是新的活动和技术，并将大大提高她的研究和咨询能力。