Tracking Intercellular Movement of Fluorescent Proteins in Bryophytes

An important approach to investigate intercellular connectivity via plasmodesmata is to visualize and track the movement of fluorescent proteins between cells. The intercellular connectivity is largely controlled by the size exclusion limit of the pores. Over the past few decades, the technique to observe and analyze intercellular movement of a fluorescent protein has been developed mainly in angiosperms such as Arabidopsis thaliana. We recently applied the corresponding system to track the intercellular movement of the fluorescent protein Dendra2 in the moss Physcomitrium (Physcomitrella) patens. The protonemal tissues are particularly suited for observation of the intercellular movement due to the simple organization. Here, we describe a protocol suitable for the analysis of Dendra2 movement between cells in P. patens.