Kärkönen Anna, Santanen Arja, Iwamoto Kuninori, Fukuda Hiroo
Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland.
Methods Mol Biol. 2011;715:1-20. doi: 10.1007/978-1-61779-008-9_1.
Plant tissue cultures are an efficient system to study cell wall biosynthesis in living cells in vivo. Tissue cultures also provide cells and culture medium where enzymes and cell wall polymers can easily be separated for further studies. Tissue cultures with tracheary element differentiation or extracellular lignin formation have provided useful information related to several aspects of xylem and lignin formation. In this chapter, methods for nutrient medium preparation, callus culture initiation, and its maintenance, as well as those for protoplast isolation and viability observation, are described. As a case study, we describe the establishment of a xylogenic culture of Zinnia elegans mesophyll cells.
植物组织培养是研究活体细胞内细胞壁生物合成的高效系统。组织培养还提供了细胞和培养基，在其中酶和细胞壁聚合物能够轻易分离以便进一步研究。具有管状分子分化或细胞外木质素形成的组织培养为木质部和木质素形成的多个方面提供了有用信息。在本章中，将描述营养培养基制备、愈伤组织培养起始及其维持的方法，以及原生质体分离和活力观察的方法。作为一个案例研究，我们描述了百日草叶肉细胞木质化培养的建立。
