Analysis of the Distribution of Symplasmic Tracers During Zygotic and Somatic Embryogenesis

Plasmodesmata (PD) are membraneous channels that span cell walls of adjacent cells to establish the symplasm. These connections are unique to plants and enable the cell-to-cell exchange of information via the symplasm. However, not every plant cell is connected to its neighbor. Absence of PD and lack of communication (symplasmic isolation) are important regulators of cell differentiation. To determine cell-to-cell symplasmic connectivity, the distribution of fluorescent tracers can be analyzed. Here, we describe in detail the entire procedure for conducting such analysis using fluorescence and confocal microscopy to study molecular fluxes in fluorescence recovery after photobleaching (FRAP) experiments. Studies using fluorochromes and fluorescent-labeled dextrans successfully inform the degree of symplasmic connectivity between cells in zygotic and somatic embryos. Small molecules, such as water and ions, travel through PD but also transcription factors and different types of RNA. Studies of symplasmic communication are important to determine the spatio-temporal correlation between cell differentiation and the exchange of information between cells. This information is necessary to determine the role of symplasmic communication during embryogenesis, which is a very important stage in plant development and morphogenesis.