A trafficking pathway for anthocyanins overlaps with the endoplasmic reticulum-to-vacuole protein-sorting route in Arabidopsis and contributes to the formation of vacuolar inclusions.
Plants produce a very large number of specialized compounds that must be transported from their site of synthesis to the sites of storage or disposal. Anthocyanin accumulation has provided a powerful system to elucidate the molecular and cellular mechanisms associated with the intracellular trafficking of phytochemicals. Benefiting from the unique fluorescent properties of anthocyanins, we show here that in Arabidopsis (Arabidopsis thaliana), one route for anthocyanin transport to the vacuole involves vesicle-like structures shared with components of the secretory pathway. By colocalizing the red fluorescence of the anthocyanins with green fluorescent protein markers of the endomembrane system in Arabidopsis seedlings, we show that anthocyanins are also sequestered to the endoplasmic reticulum and to endoplasmic reticulum-derived vesicle-like structures targeted directly to the protein storage vacuole in a Golgi-independent manner. Moreover, our results indicate that vacuolar accumulation of anthocyanins does not depend solely on glutathione S-transferase activity or ATP-dependent transport mechanisms. Indeed, we observed a dramatic increase of anthocyanin-filled subvacuolar structures, without a significant effect on total anthocyanin levels, when we inhibited glutathione S-transferase activity, or the ATP-dependent transporters with vanadate, a general ATPase inhibitor. Taken together, these results provide evidence for an alternative novel mechanism of vesicular transport and vacuolar sequestration of anthocyanins in Arabidopsis.
Poustka
Frantisek
F
Department of Plant Cellular and Molecular Biology and Plant Biotechnology Center, Ohio State University, Columbus, Ohio 43210, USA.
Irani
Niloufer G
NG
Feller
Antje
A
Lu
Yuhua
Y
Pourcel
Lucille
L
Frame
Kenneth
K
Grotewold
Erich
E
D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors; D000872:Anthocyanins / Q000378:metabolism*; D017360:Arabidopsis / Q000187:drug effects / Q000378:metabolism*; D020126:Brefeldin A / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005453:Fluorescence; D005982:Glutathione Transferase / Q000378:metabolism; D021382:Protein Sorting Signals; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D036226:Seedlings / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology; D021601:trans-Golgi Network / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D000872:Anthocyanins; D021382:Protein Sorting Signals; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; D014638:Vanadates; D005982:Glutathione Transferase