High performance liquid chromatography system for compound analysis in tree embryogenesis

用于分离生物学上重要分子的关键仪器是高效液相色谱系统（HPLC）。在我们的实验室中，我们经常使用HPLC来分离小分子量化合物，如激素，以及大分子化合物，如多肽和蛋白质。该系统将用于本实验室培养研究生和博士后研究员的各种研究项目中。高效液相色谱法可用于纯化和鉴定胚珠授粉过程中产生的防御物质。胚珠分泌液体拦截花粉，然后将花粉运送到胚珠中萌发。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先天防御，杀死不分青红皂白地与花粉一起吹进来的细菌和真菌。胚珠也会受到昆虫的攻击。特别是，我们已经研究了种子小孢子虫- Megastigmus spermotrophus的影响，它通过改变发育种子的激素代谢来重定向花旗松种子的发育。由于高效液相色谱法对激素及其代谢物的上级分离和分级作用，这项工作大部分是可能的。我们还广泛研究了种子发育过程中和体外（即体细胞胚胎发生过程中，这是一种人工种子生产方法）激素及其代谢产物的概况。二极管阵列检测器将是必不可少的，以获得信息的套件中产生的代谢产物在这些类型的种子发展。最后，我们有一个项目，其中我们正在表征激素代谢谱在锥诱导。可以通过物理处理如根修剪和通过化学施用植物生长调节物质如赤霉酸来操纵针叶树种球产量。HPLC对于这些化合物的分离和分析至关重要。这些项目目前为两名博士后和两名研究生提供培训，如果我们没有新的仪器，他们的工作将受到影响，因为我们目前的仪器已经使用了21年。

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.