Plant Tissue Cultures

Abstract

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.

Formation of plant tracheary elements in vitro – a review

“…Cell viability can be assessed following staining with Evans blue, 4′,6-diamidino-2-phenylindole (DAPI), or fluorescein diacetate (FDA) staining (Groover et al 1997). Evans blue is an indicator for non-viable cells while DAPI or FDA staining can be performed to differentiate between still living, differentiating TEs and dead TEs (Kärkönen et al (2011).…”

“…The system for generating Z. elegans single mesophyll cell cultures is simple and well-defined: the first appearing pair of leaves of Zinnia elegans plantlets is mixed with mannitol and the mixture ground gently in a mortar and pestle or using a blender. Steps for mechanical isolation and culture of mesophyll cells are described in detail by Kärkönen et al (2011). An adaptation of the process of TE differentiation in the Z. elegans xylogenic culture is presented in Figure 2.…”