Abstract
The actin microfilaments (MFs) have essential roles in plant cell division and morphogenesis. However, since MFs are very sensitive to fixation, conventional actin staining involving fixation has possibility to give artificial rearrangement of MFs. To observe MFs in vivo without fixation, we have made reporter construct by fusion of GFP and an actin binding domain of AtFim1. The transgenic cell lines were established and selected. The most suitable one was designated as BY-GF11 (BY-2 cells stably expressing GFP-Fimbrin line 11). During cell cycle progression, the GFP fluorescence was in agreement with well-known MF structures, such as cortical filaments and phragmoplast. On the other hand, along the cytoplasmic strands, MFs localized at the periphery of the strands rather than in the center of strands. These MF structures that may have been collapsed by fixation seem to associate to membrane structures.
