e martë, shkurt 08, 2005

Cell-to-cell movement of GFP during embryogenesis and early seedling

There is increasing evidence for intercellular trafficking of macromolecules through plasmodesmata (PD) during plant development. Here we study the ability of PD to traffic proteins during embryogenesis and early seedling development in Arabidopsis. Transgenic lines that induce GFP expression only in meristems, MSG (meristem-specific GFP), were used to monitor GFP movement. Cell-to-cell movement of different-sized GFP reporters reveals that embryos and young seedlings traffic proteins at least 54 kDa in size. Although 27-kDa soluble GFP (1xsGFP) freely moves between cells throughout the entire embryo during all stages analyzed, 2xsGFP movement becomes more restricted as development proceeds. After germination, cells near the apical meristem in seedlings show a higher size exclusion limit (SEL), whereas the SEL becomes more restricted as surrounding tissues develop identities. Although 1xsGFP moves throughout leaf primordia, as the leaf develops only the basal part of leaf petioles, main vascular tissues, and leaf veins (not blades) allow 1xsGFP movement. Although previous studies showed that embryos allow movement of small symplastic tracers (0.5 kDa), the present data demonstrate that the embryo constitutes a single symplast that allows transport of macromolecules as well. Even 2xsGFP moves from its site of expression at the apical meristem in embryos and seedlings, yet the extent of movement is more limited than 1xsGFP. Thus, PD have distinct SELs in different subregions of the embryo and seedling. These studies support the general concept that PD in younger tissues are more dilated and less restrictive than PD in older (nonvascular) tissues.