Li Bin, Takahashi Daisuke, Kawamura Yukio, Uemura Matsuo
Cryobiofrontier Research Center, Faculty of Agriculture, Iwate University, Morioka, Japan.
College of Plant Science, Jilin University, Changchun, China.
Methods Mol Biol. 2018;1696:185-194. doi: 10.1007/978-1-4939-7411-5_12.
Arabidopsis thaliana suspension-cultured cells (T87 line) are important model system for studies of responses to biotic and abiotic stresses at the cellular level in vitro since the cells have certain advantages compared with the whole plant system. However, the physiological and morphological characteristics of the cells are influenced by the progress of the growth phase of cells, which may result in different stress tolerance. To obtain comprehensive proteome profiles of the plasma membrane of Arabidopsis thaliana T87 suspension-cultured cells at the lag, log, or stationary growth phase, a shotgun proteomics method using nano-LC-MS/MS is used. The results obtained indicate that proteome profiles of the plasma membrane with the progress of the growth phase of cells dynamically changed, which may be associated with the physiological and morphological characteristics of the plasma membrane of the suspension-cultured cells. The proteomics results are further applied to explain different responsive patterns in the plasma membrane to cold acclimation and ABA treatment, which lead to understanding of different freezing tolerance associated with the growth phase of the cells.
拟南芥悬浮培养细胞(T87系)是体外细胞水平上研究对生物和非生物胁迫响应的重要模型系统,因为与整个植物系统相比,这些细胞具有一定优势。然而,细胞的生理和形态特征受细胞生长阶段进程的影响,这可能导致不同的胁迫耐受性。为了获得处于滞后、对数或稳定生长阶段的拟南芥T87悬浮培养细胞质膜的全面蛋白质组图谱,采用了基于纳升液相色谱-串联质谱的鸟枪法蛋白质组学方法。所得结果表明,随着细胞生长阶段的推进,质膜的蛋白质组图谱动态变化,这可能与悬浮培养细胞质膜的生理和形态特征有关。蛋白质组学结果进一步用于解释质膜对冷驯化和脱落酸处理的不同响应模式,从而有助于理解与细胞生长阶段相关的不同抗冻性。