Oda Yoshihisa
Center for Frontier Research, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka, 411-8540, Japan.
Department of Genetics, SOKENDAI (Graduate University for Advanced Studies), 1111 Yata, Mishima, Shizuoka, 411-8540, Japan.
Methods Mol Biol. 2017;1544:67-73. doi: 10.1007/978-1-4939-6722-3_6.
In vitro xylem differentiation is a powerful technique that can be used to elucidate the process of xylem development that occurs deep inside plant tissues in nature. The experimental procedure described here is designed to induce metaxylem vessel differentiation at exceptionally high frequency and synchronicity using genetically engineered Arabidopsis cell suspensions. By triggering a transcriptional switch, over 80 % of the cells synchronously differentiate into xylem cells within 32 h of treatment with estradiol. Exogenous marker genes can be transiently introduced into the cells by coculturing them with transformed Agrobacterium before inducing xylem differentiation. This system is fast, easy to handle, and highly compatible with molecular and cell biology techniques used to explore xylem cell differentiation.
体外木质部分化是一种强大的技术，可用于阐明自然界中发生在植物组织深处的木质部发育过程。这里描述的实验程序旨在使用基因工程拟南芥细胞悬浮液，以极高的频率和同步性诱导后生木质部导管分化。通过触发转录开关，超过80%的细胞在用雌二醇处理32小时内同步分化为木质部细胞。在诱导木质部分化之前，通过将细胞与转化的农杆菌共培养，可以将外源标记基因短暂引入细胞。该系统快速、易于操作，并且与用于探索木质部细胞分化的分子和细胞生物学技术高度兼容。
