Chen Zhong-Hua, Wu Dezhi, Eisenach Cornelia, Hills Adrian, Zhang Guoping, Blatt Michael R
School of Natural Sciences, University of Western Sydney, Sydney, NSW, Australia.
Methods Mol Biol. 2012;913:19-33. doi: 10.1007/978-1-61779-986-0_2.
Voltage clamp is one of the key techniques for the dissection, identification, and monitoring of ion transporters in plant cells. Voltage clamp-based research work on salinity stress in plants enables the characterization of many plant ATP-dependent pumps, ion channels, and ion-coupled carriers through heterologous expression in Xenopus laevis oocytes and in vivo measurements in salt-tolerant and salt-sensitive giant green algae such as Chara and many plant species. We have modified and developed a reliable set of procedures for voltage clamp analysis in intact guard cells and root epidermal cells from Arabidopsis thaliana with potentially broad applications in the salinity response of plants. These procedures greatly extend the duration of measurements and scope for analysis of the predominant K(+) and anion channels.
电压钳是剖析、鉴定和监测植物细胞中离子转运体的关键技术之一。基于电压钳的植物盐胁迫研究工作,能够通过在非洲爪蟾卵母细胞中的异源表达以及在耐盐和盐敏感的大型绿藻(如轮藻)和许多植物物种中的体内测量,来表征许多植物ATP依赖性泵、离子通道和离子偶联载体。我们已经修改并开发了一套可靠的程序,用于对拟南芥完整保卫细胞和根表皮细胞进行电压钳分析,这些程序在植物盐胁迫响应方面具有潜在的广泛应用。这些程序极大地延长了测量持续时间,并扩大了对主要钾离子和阴离子通道的分析范围。