Nurani Alif Meem, Kondo Yuki, Fukuda Hiroo
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Methods Mol Biol. 2018;1830:149-159. doi: 10.1007/978-1-4939-8657-6_10.
Vascular system is vital for the transport of water and nutrients as well as for providing mechanical support in many land plants. Plant transcription factors play a central role in regulating vascular development downstream of hormones and peptide signaling pathways. Particularly, cell culture systems have contributed to isolating such key transcription factors for xylem differentiation. However, there had been no efficient systems that can mimic phloem differentiation in the model plant Arabidopsis, preventing the identification of phloem-related transcription factors. We have recently established Vascular cell Induction culture System Using Arabidopsis Leaves (VISUAL), which concomitantly generates both xylem and phloem cells in the cotyledon of Arabidopsis. This system can be used to take a closer look at the bi-directional differentiation mechanism of (pro)cambial cells into xylem and phloem cells. Here, we report the methods of microscopic, genetic, and molecular analysis using VISUAL, which can help in decrypting the transcriptional networks that regulate vascular cell differentiation.
在许多陆生植物中，维管系统对于水分和养分的运输以及提供机械支撑至关重要。植物转录因子在激素和肽信号通路下游调节维管发育过程中发挥着核心作用。特别是，细胞培养系统有助于分离出用于木质部分化的此类关键转录因子。然而，一直没有能够模拟模式植物拟南芥韧皮部分化的有效系统，这阻碍了韧皮部相关转录因子的鉴定。我们最近建立了利用拟南芥叶片的维管细胞诱导培养系统（VISUAL），该系统能在拟南芥子叶中同时生成木质部和韧皮部细胞。该系统可用于更深入地研究（原）形成层细胞向木质部和韧皮部细胞的双向分化机制。在此，我们报告使用VISUAL进行显微、遗传和分子分析的方法，这些方法有助于解密调节维管细胞分化的转录网络。
