High performance liquid chromatography system for compound analysis in tree embryogenesis

分离生物重要分子的一个关键仪器是高效液相色谱系统(HPLC)。在我们的实验室中，我们经常使用高效液相色谱法来分离小分子化合物，如激素，以及大分子化合物，如多肽和蛋白质。该系统将用于我们实验室用于培养研究生和博士后研究员的各种研究项目。高效液相色谱法将应用于胚珠在授粉过程中产生的防御物质的纯化和鉴定。胚珠分泌液体，拦截花粉，然后将花粉输送到胚珠中，在那里萌发。我们已经证明，在这些分泌物中发现的蛋白质构成了一种先天防御，杀死了不分青红皂白地随花粉吹进来的细菌和真菌。胚珠也容易受到昆虫的攻击。特别是，我们研究了一种种子查尔剂-巨藻精子营养细胞-通过改变正在发育的种子的激素代谢来重定向道格拉斯-冷杉种子的发育。由于荷尔蒙及其代谢物的高效分离和分级，这项工作中的大部分都是可能的。我们还对种子发育和体外培养过程中激素及其代谢产物的分布进行了广泛的研究，即在体细胞胚胎发生期间，体细胞胚胎是一种人工种子生产方法。二极管阵列探测器对于获得有关这些类型种子发育过程中产生的代谢物的信息至关重要。最后，我们有一个项目，在这个项目中，我们正在描述锥体诱导过程中的激素代谢谱。通过根部修剪等物理处理和赤霉酸等植物生长调节物质的化学应用，可以控制针叶树的球果产量。高效液相色谱法是分离和分析这些化合物的关键。这些项目目前为两名博士后和两名研究生提供培训，如果我们没有新的乐器，他们的工作将受到影响，因为我们目前的乐器已有21年的历史。

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.