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

  1. Insoon Kim,
  2. Euna Cho,
  3. Katrina Crawford,
  4. Frederick D. Hempel*, and
  5. Patricia C. Zambryski
  1. Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720
  1. Contributed by Patricia C. Zambryski, December 9, 2004

Abstract

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 (1×sGFP) freely moves between cells throughout the entire embryo during all stages analyzed, 2×sGFP 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 1×sGFP moves throughout leaf primordia, as the leaf develops only the basal part of leaf petioles, main vascular tissues, and leaf veins (not blades) allow 1×sGFP 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 2×sGFP moves from its site of expression at the apical meristem in embryos and seedlings, yet the extent of movement is more limited than 1×sGFP. 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.

Footnotes

  • To whom correspondence should be addressed. E-mail: zambrysk{at}nature.berkeley.edu.

  • * Present address: Mendel Biotechnology, Inc., 21375 Cabot Boulevard, Hayward, CA 94545.

  • Author contributions: I.K., E.C., K.C., and F.D.H. performed research; and I.K. and P.C.Z. wrote the paper.

  • Abbreviations: PD, plasmodesmata; ER, endoplasmic reticulum; SEL, size exclusion limit; MSG, meristem-specific GFP; sGFP, soluble GFP; SAM, shoot apical meristem; RAM, root apical meristem; UAS, upstream activation sequence; dpg, days postgermination; F-dextran, FITC-conjugated dextran.

  • See Commentary on page 1817.

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