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JXB Advance Access originally published online on July 12, 2009
Journal of Experimental Botany 2009 60(13):3861-3871; doi:10.1093/jxb/erp225
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

Identification and localization of the bilitranslocase homologue in white grape berries (Vitis vinifera L.) during ripening

Alberto Bertolini1, Carlo Peresson1, Elisa Petrussa1, Enrico Braidot1, Sabina Passamonti2, Francesco Macrì1 and Angelo Vianello1,*

1Dipartimento di Biologia e Protezione delle Piante, Sezione di Biologia Vegetale, Università di Udine, via delle Scienze 91, I-33100 Udine, Italy
2Dipartimento di Scienze della Vita, Università di Trieste, via L. Giorgieri, 1, I-34127 Trieste, Italy

* To whom correspondence should be addressed: E-mail: angelo.vianello{at}uniud.it

A homologue of the mammalian bilirubin transporter bilitranslocase (BTL) (TCDB 2.A.65.1.1), able to perform an apparent secondary active transport of flavonoids, has previously been found in carnation petals and red grape berries. In the present work, a BTL homologue was also shown in white berries from Vitis vinifera L. cv. Tocai/Friulano, using anti-sequence antibodies specific for rat liver BTL. This transporter, similarly to what found in red grape, was localized in the first layers of the epidermal tissue and in the vascular bundle cells of the mesocarp. In addition, a strong immunochemical reaction was detected in the placental tissue and particularly in peripheral integuments of the seed. The protein was expressed during the last maturation stages in both skin and pulp tissues and exhibited an apparent molecular mass of c. 31 kDa. Furthermore, the transport activity of such a carrier, measured as bromosulphophthalein (BSP) uptake, was detected in berry pulp microsomes, where it was inhibited by specific anti-BTL antibodies. The BTL homologue activity exhibited higher values, for both Km and Vmax, than those found in the red cultivar. Moreover, two non-pigmented flavonoids, such as quercetin (a flavonol) and eriodictyol (a flavanone), inhibited the uptake of BSP in an uncompetitive manner. Such results strengthen the hypothesis that this BTL homologue acts as a carrier involved also in the membrane transport of colourless flavonoids and demonstrate the presence of such a carrier in different organs and tissues.

Key words: Bilitranslocase, flavonoid transport, immunohistochemistry, seed, Vitis vinifera L., white berry cultivar

Received 8 May 2009; Revised 23 June 2009 Accepted 24 June 2009


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