Microtome and knife maker for structure and function analysis of Arabidopsis and canola seed coats

用于拟南芥和油菜籽皮结构和功能分析的切片机和刀具制造机

• 批准号: 390268-2010
• 负责人: Belmonte, Mark
• 金额: $ 2.33万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2009
• 资助国家: 加拿大
• 起止时间: 2009-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=443148
• 关键词: Microtome; knife; maker; structure; function

My research interest focuses on the developmental and molecular genetic events associated with seed coat development in oilseeds. One of the underlying mechanisms of development is through the localized expression of genes encoding transcription factors (TFs) at specific times and places. Identification of TF genes that are essential for growth and development of oilseed crops such as canola (Brassica napus) by conventional genetic approaches has been limiting. Understanding the transcriptional mechanisms that control seed development in the closely related model species Arabidopsis thaliana will very likely provide the insight necessary to solve this issue. Growth of oilseeds like Arabidopsis and canola relies on the coordinated development of the embryo, endosperm and seed coat. Unlike the embryo and endosperm, the seed coat has received little attention when it comes to understanding the structural and genetic mechanisms that underlie its function. For example, the function of the proximal seed coat, which comprises the chalazal proliferating tissue and chalazal seed coat is unknown. However, histological observations suggest a transport function due to transfer cell morphology. Moreover, rapid differentiation of this tissue makes it an ideal candidate to study cell differentiation. Based on these results I will carry out extensive histological studies to observe the effects of genetic perturbations on proximal seed coat development. The requested semi-motorized rotary microtome and glass knife maker will be used for (1) generating paraffin-embedded thick sections for laser capture microdissection and RNA profiling of the chalazal proliferating tissue in Arabidopsis and (2) anatomical observations of the proximal seed coat following genetic perturbations using thin plastic-embedded sections. Since this instrument can produce ribbons using both paraffin and plastic embedded materials, it provides the advantage to perform organized high-throughput reconstruction of seed parts. No microtome with these features is currently available in our department. Moreover, this instrument will be used to train highly qualified personnel in the art of histology which contribute greatly to thesis and publication quality figures.

我的研究兴趣主要集中在油料种子种皮发育相关的发育和分子遗传事件。发育的潜在机制之一是通过编码转录因子（tf）的基因在特定时间和地点的局部表达。对油籽作物如油菜（Brassica napus）的生长发育至关重要的TF基因的鉴定，通过传统的遗传方法是有限的。了解在密切相关的模式物种拟南芥中控制种子发育的转录机制将很可能为解决这一问题提供必要的见解。拟南芥、油菜等油籽的生长依赖于胚、胚乳和种皮的协调发育。与胚胎和胚乳不同，种皮在结构和遗传机制方面的研究很少受到关注。例如，包括合点增殖组织和合点种皮的近端种皮的功能是未知的。然而，组织学观察表明，由于转移细胞形态的运输功能。此外，该组织的快速分化使其成为研究细胞分化的理想候选者。基于这些结果，我将进行广泛的组织学研究，观察遗传扰动对近端种皮发育的影响。所要求的半电动旋转切片机和玻璃刀制造商将用于(1)生成石蜡包埋的厚切片，用于激光捕获显微解剖和拟南芥合点间增殖组织的RNA分析；(2)使用薄塑料包埋切片对遗传扰动后近端种皮进行解剖观察。由于该仪器可以使用石蜡和塑料嵌入材料生产条带，因此它提供了对种子部分进行有组织的高通量重建的优势。目前我们部门还没有具备这些功能的切片机。此外，该仪器将用于培养组织学艺术方面的高素质人才，这些人才对论文和出版物质量数字做出了巨大贡献。