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Evidence for a putative flavonoid translocator similar to mammalian bilitranslocase in grape berries (Vitis vinifera L.) during ripening

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Abstract

During maturation, Vitis vinifera berries accumulate a large amount of several anthocyanins in the epidermal tissue, whereas their precursors and intermediates are ubiquitously synthesized within the fruit. Up to date, several mechanisms of flavonoid transport at subcellular level have been hypothesized, but it is not possible to identify a general model applicable in every plant tissue and organ. Recently, a putative anthocyanin carrier, homologue to mammalian bilitranslocase (BTL) (TC 2.A.65.1.1), was found in Dianthus caryophyllus petal microsomes. In the present paper, an immunohistochemical and immunochemical analysis, using an antibody raised against a BTL epitope, evidences the expression and function of such a transporter in V. vinifera berries (cv. Merlot). Specific localisations of the putative carrier within berry tissues together with expression changes during different developmental stages are shown. Water stress induces an increase in protein expression in both skin and pulp samples. A bromosulfalein (BSP) uptake activity, inhibitable by the BTL antibody, is detected in berry mesocarp microsomes, with K m = 2.39 μM BSP and V max = 0.29 μmol BSP min−1 mg−1 protein. This BSP uptake is also competitively inhibited by quercetin (K i = 4 μM). A putative role for this carrier is discussed in relation to the membrane transport of secondary metabolites.

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Abbreviations

ABC proteins:

ATP-binding cassette proteins

BSA:

Bovine serum albumin

BTL:

Bilitranslocase

BSP:

Bromosulfalein

DTE:

Dithioerythritol

EDTA:

Ethylenediaminetetraacetic acid

FCCP:

Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone

FITC:

Fluorescein isothiocyanate

GST:

Glutathione S-transferases

GS-X:

Glutathione S-conjugate pumps

MRPs:

Multidrug resistance-associated proteins

Pas:

Proanthocyanidins

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Acknowledgments

This research was supported by “Tenuta Villanova”, Vineyard and Winery at Farra d’Isonzo (GO), Friuli-Venezia Giulia, Italy, and by the Regione Autonoma Friuli-Venezia Giulia (L.R. 26/2005 art. 17).

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Authors and Affiliations

  1. Department of Biology and Plant Protection, University of Udine, via Cotonificio 108, 33100, Udine, Italy

    E. Braidot, E. Petrussa, A. Bertolini, C. Peresson, P. Ermacora, N. Loi, F. Macrì & A. Vianello

  2. Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Via L. Giorgieri, 1, 34127, Trieste, Italy

    M. Terdoslavich & S. Passamonti

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  1. E. Braidot
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  2. E. Petrussa
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  3. A. Bertolini
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  4. C. Peresson
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  5. P. Ermacora
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  6. N. Loi
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  9. F. Macrì
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  10. A. Vianello
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Correspondence to A. Vianello.

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Braidot, E., Petrussa, E., Bertolini, A. et al. Evidence for a putative flavonoid translocator similar to mammalian bilitranslocase in grape berries (Vitis vinifera L.) during ripening. Planta 228, 203–213 (2008). https://doi.org/10.1007/s00425-008-0730-4

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