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使用流通系统测量植物根和假根急性暴露于化合物期间的电反应。

Measuring Electrical Responses during Acute Exposure of Roots and Rhizoids of Plants to Compounds Using a Flow-Through System.

作者信息

Cooper Robin Lewis, Thomas Matthew A, Vascassenno Rachael M, Brock Kaitlyn E, McLetchie David Nicholas

机构信息

Department of Biology, University of Kentucky, Lexington, KY 40506-0225, USA.

Department of Biology, Eastern Kentucky University, Richmond, KY 40475-3102, USA.

出版信息

Methods Protoc. 2022 Jul 18;5(4):62. doi: 10.3390/mps5040062.

Abstract

Monitoring electrical signals in plants allows the examination of their acute and chronic physiological changes and responses to stimuli. Understanding how plant roots/rhizoids respond to chemical cues in their environment will provide insight into how these structures acquire resources. Chronic exposure to L-glutamate alters root growth and is known to alter Ca flux inside roots. The ionic flux can be detected by electrical changes. A rapid and relatively easy approach is presented to screen the electrical sensitivity of roots/rhizoids to compounds such as amino acids and known agonists/antagonists to receptors and ion channels. The approach uses a background-flow system of basal salt or water; then, the administered compounds are added to the roots/rhizoids while monitoring their electrical responses. As a proof of concept, the response to flow-through of glutamate (1 mM) was targeted at the root/rhizoids of three plants (, and ). Both and produced rapid depolarization upon exposure to glutamate, while did not show an electrical response. In some experiments, simultaneous recordings with impedance measures for acute changes and glass electrodes for chronic electrical potential changes were used. The effect of potassium chloride (300 mM) as a depolarizing stimulus produced responses in both and The protocol presented can be used to screen various compounds in a relatively rapid manner for responsiveness by the roots/rhizoids of plants.

摘要

监测植物中的电信号有助于检测其急性和慢性生理变化以及对刺激的反应。了解植物根/假根如何对其环境中的化学信号作出反应,将有助于深入了解这些结构如何获取资源。长期暴露于L-谷氨酸会改变根的生长,并且已知会改变根内的钙通量。离子通量可以通过电变化来检测。本文介绍了一种快速且相对简便的方法,用于筛选根/假根对氨基酸等化合物以及已知的受体和离子通道激动剂/拮抗剂的电敏感性。该方法使用基础盐或水的背景流动系统;然后,在监测根/假根电反应的同时,将施用的化合物添加到根/假根中。作为概念验证,针对三种植物(、和)的根/假根,研究了其对1 mM谷氨酸流经的反应。和在暴露于谷氨酸后均产生快速去极化,而未表现出电反应。在一些实验中,同时使用阻抗测量来记录急性变化,使用玻璃电极来记录慢性电位变化。300 mM氯化钾作为去极化刺激物,对和均产生了反应。本文介绍的方案可用于以相对快速的方式筛选各种化合物,以检测植物根/假根的反应性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a6/9351672/b5a6d5893486/mps-05-00062-g001.jpg

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