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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Evidence for Unidirectional Flow through Plasmodesmata¹

Institute of Molecular Plant Sciences, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom

The leaf trichome of tobacco (Nicotiana tabacum) represents a unique secretory structure in which the basal trichome cell is connected to the epidermis by numerous plasmodesmata (PD). Small fluorescent probes microinjected into the basal trichome cell moved apically into distal trichome cells but not into the subtending epidermal cell. In marked contrast, the same probes moved apically into trichome cells when injected into the epidermal cell. Noninvasive methods of dye loading, including ester loading into the apical secretory cell by trichome "capping" and by infiltration of caged fluorescein, produced the same result. In transgenic tobacco plants constitutively expressing photoactivatable green fluorescent protein (PAGFP), activation of PAGFP above the epidermal/trichome (e/t) boundary resulted in movement of protein apically into the distal trichome cells but not across the e/t boundary, while PAGFP activated in the epidermal cell moved apically across the e/t boundary. Experiments with apoplastic tracers also revealed the presence of a distinct apoplastic barrier to solute movement at the e/t interface. These data point to unidirectional transport of solutes through PD. PAGFP activated in individual cells equidistant between the basal cell and the apical cell moved bidirectionally from these cells, suggesting that mass flow was not the driving force for unidirectional transport. We found that unidirectional transport across the e/t boundary was not affected by virus infection or by addition of the actin inhibitor latrunculin but could be dissipated completely by addition of sodium azide. Collectively, our data suggest that active, unidirectional transport of molecules may occur through PD located at unique interfaces in the plant.

² These authors contributed equally to the article.

³ Present address: Risø National Laboratory for Sustainable Energy, Technical University of Denmark, DK–4000 Roskilde, Denmark.

⁴ Present address: Department of Disease and Stress Biology, John Innes Centre, Norwich NR4 7UH, United Kingdom.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Karl Oparka (karl.oparka{at}ed.ac.uk).

www.plantphysiol.org/cgi/doi/10.1104/pp.109.137083

^* Corresponding author; e-mail karl.oparka{at}ed.ac.uk.

Received February 18, 2009; accepted March 1, 2009; published March 6, 2009.