EAGER: Advancing auxin transport research with patch clamp electrophysiology

EAGER：利用膜片钳电生理学推进生长素转运研究

• 批准号: 1360751
• 负责人: Edgar Spalding
• 金额: $ 18.92万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1360751&HistoricalAwards=false
• 关键词: EAGER; Advancing; auxin; transport; research

Auxin is a hormone in plants that governs many basic physiological processes including cell growth. A special transport process moves auxin from cell to cell through plant organs. Understanding how this happens in detail, at the molecular and physical levels, has been a long-standing goal in the field of plant biology. If the auxin transport process were better understood, it could be possible to manipulate it to improve the growth of crops and other plants on which society depends. The project will use a biophysical technique called patch-clamp electrophysiology to determine how proteins belonging to the B subfamily of ATP-Binding-Cassette Transporters, contribute to the auxin transport process. Previous research has shown such proteins are required for auxin transport, but exactly what makes them necessary is not known, because it is not known if they actually transport auxin by themselves, assist other proteins, or in some other way are necessary for the process. By expressing such ABCB transporter proteins in animal cells, which do not possess auxin transport capability themselves, and then using the patch clamp technique to measure the flow of ions (electrically charged atoms) across the membranes of the animal cells, with and without auxin being present, the investigators will learn how ABCB transporters function, and this will lead to new, testable ideas about how auxin moves from cell to cell. The patch-clamp technique has not been used to study auxin transport before, so this project represents a new combination of biophysics and plant genetics. One expected result is a direct recording of the electric current caused by the charged auxin molecule as it passes through the ABCB transporter, or a definitive demonstration of its absence. A postdoctoral student will be trained to work at the interdisciplinary interface of biophysics and plant molecular biology during this project.

生长素是植物中的一种激素，控制许多基本的生理过程，包括细胞生长。生长素通过一种特殊的运输过程在植物器官中从一个细胞转移到另一个细胞。在分子和物理水平上详细了解这是如何发生的，一直是植物生物学领域的长期目标。如果能更好地理解生长素的运输过程，就有可能操纵它来改善作物和社会所依赖的其他植物的生长。该项目将使用称为膜片钳电生理学的生物物理技术来确定属于ATP结合盒转运蛋白B亚家族的蛋白质如何促进生长素的转运过程。以前的研究表明，这些蛋白质是生长素运输所必需的，但确切地说，是什么使它们成为必要的是不知道的，因为它是不知道，如果他们实际上运输生长素本身，协助其他蛋白质，或以其他方式是必要的过程。通过在动物细胞中表达这种ABCB转运蛋白，这些细胞本身不具有生长素转运能力，然后使用膜片钳技术测量离子（带电原子）穿过动物细胞膜的流动，有和没有生长素存在，研究人员将了解ABCB转运蛋白的功能，这将导致生长素如何在细胞间移动的新的，可测试的想法。膜片钳技术还没有被用于研究生长素的运输之前，所以这个项目代表了生物物理学和植物遗传学的一个新的结合。一个预期的结果是直接记录带电生长素分子通过ABCB转运蛋白时产生的电流，或者明确证明它的缺失。在本项目期间，将培养一名博士后学生从事生物物理学和植物分子生物学的跨学科工作。