Trafficking and Sequestration of Anthocyanins

知识价值与有关植物新陈代谢的广泛知识相比，人们对植物如何将有毒或高活性的化学物质从其合成地点运输到最终储存的地方知之甚少。植物化合物(植物化学物质)的适当运输和储存往往是旨在对植物新陈代谢进行理论工程的努力的一个瓶颈。因此，了解植物化学物质转运的细胞和分子机制具有极其重要的意义。拟南芥中花青素色素的形成为研究植物细胞内植物化学物质的运输和隔离提供了一个出色的系统。这项研究的假设是，花青素首先积累在内质网内，由这个细胞器产生的小泡将这些化学物质运输到液泡，在液泡中吸收后，定位于亚液泡结构。本项目将首先研究内质网中花青素生物合成酶和可能的花青素转运蛋白的定位。然后，将从蛋白质和作为其中一部分的花青素的角度来表征积累花青素的亚空泡状结构。最后，结合细胞标记和各种遗传工具，将确定花青素从内质网移动到液泡的途径。总之，这些研究将为植物在细胞内移动植物化学物质的机制提供基本的见解。广泛影响这是一项跨学科的建议，独特地整合了细胞生物学和生物化学，以解决植物如何管理植物化学物质亚细胞运输的基本生物学问题。本文提出的这些研究可能会产生深远的影响，为植物代谢工程的新细胞结构的形成和影响策略提供洞察力。这可能会产生重大的社会效益，例如，有利于生物燃料作物的发展和提高植物产品的营养价值。除了两个PI都积极参与的标准课堂教学和实验室人员培训之外，该项目还以一种独特的方式在化学和生物之间的前沿界面上整合了对学生和博士后的研究和教育培训。通过举办研讨会和会议，如戈登·凯南研究生研究研讨会和戈登植物代谢工程会议，私人投资机构在很大程度上将教育和研究联系起来，这些活动将在本项目期间继续开展。

Intellectual MeritIn contrast to the extensive knowledge available on plant metabolism, very little is known on how plants transport often toxic or highly reactive chemicals from their site of synthesis to where they are ultimately stored. Proper trafficking and storage of plant compounds (phytochemicals) is often a bottleneck in efforts aimed at the rationale engineering of plant metabolism. Thus, understanding the cellular and molecular mechanisms involved in phytochemical trafficking is of the utmost significance. The formation of anthocyanin pigments in Arabidopsis provides an outstanding system to investigate how phytochemicals are transported and sequestered inside plant cells. This research is geared by the hypothesis that anthocyanins first accumulate inside the endoplasmic reticulum, and that vesicles derived from this organelle transport these chemicals to the vacuole, where upon uptake, localize in sub-vacuolar structures.This project will first investigate the localization of anthocyanin biosynthetic enzymes and putative anthocyanin transporters on the endoplasmic reticulum. Then, the sub-vacuolar structures that accumulate anthocyanins will be characterized from the perspective of the proteins and anthocyanins that are part of them. Finally, using a combination of cellular markers and various genetic tools, the pathways by which anthocyanins move from the endoplasmic reticulum to the vacuole will be determined. Together, these studies will provide fundamental insights on the mechanisms used by plants to move phytochemicals within cells.Broader ImpactsThis is an interdisciplinary proposal that uniquely integrates cell biology and biochemistry to address the fundamental biological problem of how plants manage the sub-cellular trafficking of phytochemicals. The implications derived from the studies proposed here are likely to have far-reaching consequences, providing insights into the formation of new cellular structures and impacting strategies for plant metabolic engineering. This could result in significant societal benefits, for example, benefiting the development of biofuel crops and improving the nutritional value of plant products. Beyond the standard classroom teaching and lab personnel training in which both PIs are actively involved, this project integrates research and educational training for students and postdocs in a singular way at the cutting-edge interface between chemistry and biology. By organizing workshops and conferences, such as the Gordon Kenan Graduate Research Seminars and the Gordon Conference in Plant Metabolic Engineering, the PIs have significantly bridged education and research, activities that will be continued during the duration of this project.