Establishment of a new Zinnia experimental system including separation of the distinct cell population with flow cytometry and its culture

Abstract

An in vitro culture system of Zinnia mesophyll cells has best used for studies on xylogenesis. Our previous studies with Zinnia cell culture demonstrated the presence of different cell types in the culture and their cell-cell communication. As the first step to investigate such cell-cell communication, we intended to establish a method to separate different cell populations and to culture a specific one. Zinnia mesophyll cells were stained with fluorescein diacetate, and then the fluorescence-positive and negative-cells were separated successfully using flow cytometric cell sorting. The fluorescence-positive cells differentiated into tracheary elements in the presence of an antibiotic, nalidixic acid. These procedures will be useful not only for the analysis of cell-cell interactions in Zinnia culture, but also for separating cells with intact cell walls in other plant species.