Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-okubo, Sakura-ku, Saitama, Japan.
Department of Biochemistry, University of Cambridge, Cambridge, UK.
J Exp Bot. 2020 Sep 19;71(18):5414-5424. doi: 10.1093/jxb/eraa236.
Arabinogalactan-proteins (AGPs) are a family of plant extracellular proteoglycans implicated in many physiological events. AGP is decorated with type II arabinogalactans (AGs) consisting of a β-1,3-galactan backbone and β-1,6-galactan side chains, to which other sugars are attached. Based on the fact that a type II AG-specific inhibitor, β-Yariv reagent, perturbs growth and development, it has been proposed that type II AGs participate in the regulation of cell shape and tissue organization. However, the mechanisms by which type II AGs participate have not yet been established. Here, we describe a novel system that causes specific degradation of type II AGs in Arabidopsis, by which a gene encoding a fungal exo-β-1,3-galactanase that specifically hydrolyzes β-1,3-galactan backbones of type II AGs is expressed under the control of a dexamethasone-inducible promoter. Dexamethasone treatment increased the galactanase activity, leading to a decrease in Yariv reagent-reactive AGPs in transgenic Arabidopsis. We detected the typical oligosaccharides released from type II AGs by Il3GAL in the soluble fraction, demonstrating that Il3GAL acted on type II AG in the transgenic plants. Additionally, this resulted in severe tissue disorganization in the hypocotyl and cotyledons, suggesting that the degradation of type II AGs affected the regulation of cell shape.
阿拉伯半乳聚糖蛋白(AGP)是一类植物细胞外糖蛋白,参与许多生理过程。AGP 上连接有 II 型阿拉伯半乳聚糖(AG),由β-1,3-半乳糖主链和β-1,6-半乳糖侧链组成,其上还连接有其他糖分子。由于 II 型 AG 特异性抑制剂β-Yariv 试剂会干扰生长和发育,因此有人提出 II 型 AG 参与了细胞形态和组织形成的调控。然而,II 型 AG 参与的机制尚未确定。在这里,我们描述了一种新的系统,该系统可在拟南芥中特异性降解 II 型 AG,其方法是在皮质酮诱导型启动子的控制下表达一种编码真菌外切-β-1,3-半乳糖苷酶的基因,该酶特异性水解 II 型 AG 的β-1,3-半乳糖主链。皮质酮处理增加了半乳糖苷酶活性,导致转基因拟南芥中 Yariv 试剂反应性 AGP 的减少。我们在可溶性部分检测到 Il3GAL 从 II 型 AG 释放的典型低聚糖,证明 Il3GAL 在转基因植物中作用于 II 型 AG。此外,这导致下胚轴和子叶的组织严重紊乱,表明 II 型 AG 的降解影响了细胞形态的调控。