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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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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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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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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DOI: https://doi.org/10.1038/ncb1805
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