High performance liquid chromatography system for compound analysis in tree embryogenesis

用于树木胚胎发生中化合物分析的高效液相色谱系统

• 批准号: 345275-2007
• 负责人: VonAderkas, Patrick
• 金额: $ 3万
• 依托单位: University of Victoria
• 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: 2006
• 资助国家: 加拿大
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=325581
• 关键词: performance; liquid; chromatography; system; compound

A key instrument for the separation of biologically important molecules is a high performance liquid chromatography system (HPLC). In our laboratory, we regularly use an HPLC to separate small molecular weight compounds, such as hormones, and large compounds such as polypeptides and proteins. This system will be used in the various research projects that are used to train graduate students and post-doctoral fellows in our lab. The HPLC will be applied to purification and identification of defense compounds produced by ovules during pollination. Ovules secrete liquids that intercept pollen, then transport the pollen into the ovule where it germinates. Proteins found within these secretions have been shown by us to constitute an innate defense, killing bacteria and fungi that indiscriminately blow in with the pollen. Ovules are also subject to insect attack. In particular, we have studied the effects of a seed chalcid - Megastigmus spermotrophus - which redirects Douglas-fir seed development by altering the hormone metabolism of developing seed. Much of this work was possible because of superior separation and fractionation by HPLC of hormones and their metabolites. We also have worked extensively on the profiles of hormones and their metabolites during seed development and in vitro, that is, during somatic embryogenesis, which is a method of artificial seed production. The diode array detector will be essential for obtaining information on the suite of metabolites produced during by these types of seed development. Lastly, we have a project in which we are characterizing hormone metabolomic profiles during cone induction. It is possible to manipulate conifer seed cone yield by physical treatments such as root pruning, and by chemical application of plant growth regulating substances such as gibberellic acid. HPLC is critical for separation and analysis of these compounds. These projects currently provide training for two post-docs and two graduate students, whose work will be compromised if we do not get a new instrument, as our current instrument is 21 years old.

高效液相色谱系统（HPLC）是分离生物重要分子的关键工具。在我们的实验室中，我们经常使用高效液相色谱分离小分子量化合物，如激素，和大分子量化合物，如多肽和蛋白质。本系统将用于本实验室培养研究生和博士后的各种研究项目。高效液相色谱法将应用于花粉授粉过程中胚珠产生的防御化合物的纯化和鉴定。胚珠分泌液体拦截花粉，然后将花粉输送到胚珠中发芽。在这些分泌物中发现的蛋白质已经被我们证明构成了一种天生的防御，杀死不加区分地与花粉一起吹进来的细菌和真菌。胚珠也容易受到昆虫的攻击。特别地，我们研究了一种名为Megastigmus spermotrophus的种子酸对道格拉斯冷杉种子发育的影响，它通过改变种子发育过程中的激素代谢来重定向种子发育。由于高效液相色谱法对激素及其代谢物进行了有效的分离和分选，这项工作得以实现。我们还广泛研究了种子发育和体外（即体细胞胚胎发生，这是一种人工种子生产方法）过程中激素及其代谢物的概况。二极管阵列检测器对于获得这些类型种子发育过程中产生的一系列代谢物的信息至关重要。最后，我们有一个项目，我们是表征激素代谢组谱在锥体诱导。通过物理处理（如根修剪）和化学施用植物生长调节物质（如赤霉素酸）来控制针叶树种子球果产量是可能的。高效液相色谱法对这些化合物的分离和分析至关重要。这些项目目前培养了两名博士后和两名研究生，如果我们没有新的仪器，他们的工作将受到影响，因为我们现有的仪器已经使用了21年。