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An H+ P-ATPase on the tonoplast determines vacuolar pH and flower colour

Nature Cell Biology volume 10, pages 1456–1462 (2008)Cite this article

Abstract

The regulation of pH in cellular compartments is crucial for intracellular trafficking of vesicles and proteins and the transport of small molecules, including hormones. In endomembrane compartments, pH is regulated by vacuolar H+-ATPase1 (V-ATPase), which, in plants, act together with H+-pyrophosphatases2 (PPase), whereas distinct P-type H+-ATPases in the cell membrane control the pH in the cytoplasm and energize the plasma membrane3. Flower colour mutants have proved useful in identifying genes controlling the pH of vacuoles where anthocyanin pigments accumulate4,5. Here we show that PH5 of petunia encodes a P3A-ATPase proton pump that, unlike other P-type H+-ATPases, resides in the vacuolar membrane. Mutation of PH5 reduces vacuolar acidification in petals, resulting in a blue flower colour and abolishes the accumulation of proanthocyanindins (condensed tannins) in seeds. Expression of PH5 is directly activated by transcription regulators of the anthocyanin pathway, in conjunction with PH3 and PH4. Thus, flower coloration, a key-factor in plant reproduction, involves the coordinated activation of pigment synthesis and a specific pathway for vacuolar acidification.

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Figure 1: Characterization and isolation of PH5.
Figure 2: PH5 is a P3AH+-ATPase.
Figure 3: Expression pattern and genetic control of PH5.
Figure 4: Immunogold localization of P3A-ATPases in petal cells.
Figure 5: Intracellular localization of PH5–GFP.

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Acknowledgements

We are grateful to. Inhwan Hwang (Pohang, Korea) and Katrin Czempinski (Golm, Germany) for AHA2–GFP and KCO1–GFP constructs, and to Dorus Gadella (University of Amsterdam, The Netherlands) for the use of confocal microscopy facilities. We thank Pieter Hoogeveen, Martina Meesters and Daisy Kloos for plant care. This work was supported by a grant from The Netherlands Organization of Scientific Research (NWO-ALW).

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Author notes

  1. Walter Verweij and Cornelis Spelt: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Molecular Cell Biology, Graduate School of Experimental Plant Sciences, Vrije Universiteit, de Boelelaan 1087, 1081HV Amsterdam, The Netherlands.,

    Walter Verweij, Cornelis Spelt, Ronald Koes & Francesca Quattrocchio

  2. Laboratorio di Botanica - Di.S.Te.B.A., Università di Lecce, Campus ECOTEKNE, 73100 Lecce, Italy.,

    Gian-Pietro Di Sansebastiano

  3. Centre for Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 316, 1098SM Amsterdam, The Netherlands.,

    Joop Vermeer

  4. Department of Applied Biology, Universita' di Perugia, Borgo XX Giugno 74, 06121 Perugia, Italy.,

    Lara Reale & Francesco Ferranti

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  1. Walter Verweij
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Contributions

W.V. and C.S. contributed equally to the experimental work; G.P.S. and J.V. contributed to the GFP localization studies; L.R and F.F. ; performed the immunogold experiments and electron microscopy; F.Q. and R.K. ; conceived the project, performed the genetic analysis and wrote the manuscript.

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Correspondence to Francesca Quattrocchio.

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Verweij, W., Spelt, C., Di Sansebastiano, GP. et al. An H+ P-ATPase on the tonoplast determines vacuolar pH and flower colour. Nat Cell Biol 10, 1456–1462 (2008). https://doi.org/10.1038/ncb1805

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