Arabidopsis 2010: Phenylpropanoid Pathway Networks: An Integrated Approach to Establishing Protein/Enzyme Function in Arabidopsis and their Associated Networks

本拟南芥2010项目的目标是确定248种拟南芥酶和蛋白质的生理功能，这些酶和蛋白质被认为参与了各种苯丙酸-醋酸酯代谢网络。有两个主要目标：确定与苯丙烷偶联/聚合相关的网络（例如导致木质素，木脂素，亚木质素，孢粉蛋白等），包括这些酶/蛋白质的功能。第二个目标是精确识别存在于拟南芥中参与苯丙氨酸转化为单脂醇的不同网络。在这两个目标中，功能将在体外得到证明，我们还将通过展示与所涉及的代谢途径网络片段的时间和空间相关性来确定这是真正的生理功能。因此，这项工作将定义拟南芥在其整个生命周期中各种苯丙酸自由基-自由基偶联和相关代谢过程的组织。对科学界的好处将包括通过一个与拟南芥网站相连的网站快速传播结果（在发表之前），以及根据需要提供研究材料（基因、结构、重组蛋白质、转基因和突变植物）。另一个重要的好处将是获得涉及大分子组装的迄今为止困难的系统（例如偶联/聚合）的新知识，以及将获得的新见解。对于选择的最终不参与这些途径的酶/基因，认为代谢物分析将提供功能的线索，然后也将进行检查。此外，除了木质素化作用，本研究还将揭示体内其他高度调控的自由基-自由基酚偶联系统，包括：种皮及其代谢产物的构建；生成（加强的）毛状体基质；在形成羽化组织和加强花茎方面；在生物合成孢粉素（花粉粒中非常稳定的成分）；强化角质组织；通过羟基肉桂酸（酚类）偶联交联细胞壁碳水化合物，产生过多的防御化合物，可能还有其他有待发现的用途。除了同行评议的期刊，我们的研究成果、信息和材料将提供给公共数据库，定期（每月）更新，通过在一个专门的网站（http://ibc.wsu.edu/lewislab/nsf/index.html）上发布数据/信息，该网站将链接到与该项目相关的拟南芥网络网站。这些信息将包括：我们实验室构建的重组蛋白表达载体；基因鉴定与基因功能分析；动力学数据结果；基因表达谱；代谢物和木质素分析；原位杂交数据；光学显微镜记录。

The goal of this Arabidopsis 2010 Project is to establish the physiological function of 248 Arabidopsis enzymes and proteins presumed to be involved in various networks of phenylpropanoid-acetate metabolism. There are two main objectives: identifying networks associated with phenylpropanoid coupling/polymerization (e.g. leading to lignins, lignans, suberins, sporopollenins, etc.), including how these enzymes/proteins function. The second objective is to precisely identify the different networks that exist in Arabidopsis that are involved in the conversion of phenylalanine through to the monolignols. In both objectives, functions will be demonstrated in vitro, and we will also establish that this is the true physiological function by demonstrating temporal and spatial correlation with the segments of the metabolic pathway networks involved.This work will thus define the organization of the various phenylpropanoid radical-radical coupling and related metabolic processes in Arabidopsis through its entire life cycle. The benefits to the scientific community will include rapid dissemination of results (prior to publication) through a website linked to the Arabidopsis sites, and the provision of research materials (genes, constructs, recombinant proteins, transgenic and mutant plants) as needed. Another important benefit will be the new knowledge gained on these hitherto difficult systems (e.g. coupling/polymerization) involving macromolecular assemblies, and the new insights that will be gained. For enzymes/genes chosen that are ultimately not involved in these pathways, it is considered that metabolite profiling will provide a clue as to function, and this will then be examined also.Further, in addition to lignification, this study will shed important light on other highly regulated radical-radical phenolic coupling systems in vivo including: the construction of seed coats and metabolites therein; in generating the (strengthened) matrix of trichomes; in forming suberized tissue and in strengthening flower stalks; in biosynthesizing sporopollenin (a remarkably stable component of pollen grains); in reinforcing cutinized tissue; in cross-linking cell wall carbohydrates through hydroxycinnamic acid (phenolic) coupling, in producing a plethora of defense compounds and presumably for other purposes awaiting discovery. In addition to peerreviewed journals, our research findings, information and materials generated by this research will be made available to public databases, updated periodically on a (monthly) basis, by posting data/information on a dedicated website (http://ibc.wsu.edu/lewislab/nsf/index.html) that will be linked up to the Arabidopsis network sites relevant to the project. This information will include: recombinant protein expression vectors constructed by our laboratory; gene identification and gene function analysis; kinetic data results; gene expression profiles; metabolite and lignin analysis; in situ hybridization data; and light microscopy documentation.