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Self-incompatibility in Papaver targets soluble inorganic pyrophosphatases in pollen

Nature volume 444, pages 490–493 (2006)Cite this article

Abstract

In higher plants, sexual reproduction involves interactions between pollen and pistil. A key mechanism to prevent inbreeding is self-incompatibility through rejection of incompatible (‘self’) pollen1. In Papaver rhoeas, S proteins encoded by the stigma interact with incompatible pollen, triggering a Ca2+-dependent signalling network2,3,4,5 resulting in pollen tube inhibition and programmed cell death6. The cytosolic phosphoprotein p26.1, which has been identified in incompatible pollen, shows rapid, self-incompatibility-induced Ca2+-dependent hyperphosphorylation in vivo3. Here we show that p26.1 comprises two proteins, Pr-p26.1a and Pr-p26.1b, which are soluble inorganic pyrophosphatases (sPPases). These proteins have classic Mg2+-dependent sPPase activity, which is inhibited by Ca2+, and unexpectedly can be phosphorylated in vitro. We show that phosphorylation inhibits sPPase activity, establishing a previously unknown mechanism for regulating eukaryotic sPPases. Reduced sPPase activity is predicted to result in the inhibition of many biosynthetic pathways, suggesting that there may be additional mechanisms of self-incompatibility-mediated pollen tube inhibition. We provide evidence that sPPases are required for growth and that self-incompatibility results in an increase in inorganic pyrophosphate, implying a functional role for Pr-p26.1.

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Figure 1: Expression analysis of Pr-p26.1a and Pr-p26.1b sPPases.
Figure 2: Pr-p26.1a and Pr-p26.1b show sPPase activity and are phosphorylated, and phosphorylation affects sPPase activity.
Figure 3: Functional evidence for Pr-p26.1 and sPPase involvement in modulating pollen tube growth.

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Acknowledgements

We thank G. Wullems for the ntp303 pollen promoter; G. Jones for help and advice on statistical analysis; and E. Sanchez-Moran for help with imaging. Work in the laboratories of F.C.H.F. and V.E.F-T. is funded by the UK Biotechnology and Biological Sciences Research Council. Author Information Sequences have been deposited in the EMBL Nucleotide Sequence Database (http://www.ebi.ac.uk/embl/) under accession codes AM162550 and AM162551. The authors declare no competing financial interests.

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Author notes

  1. Jason J. Rudd

    Present address: Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK

  2. Michael J. Wheeler

    Present address: Institute of Cancer Studies, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

  3. Barend H. J. de Graaf, Jason J. Rudd and Michael J. Wheeler: These authors contributed equally to this work.

Authors and Affiliations

  1. School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Barend H. J. de Graaf, Jason J. Rudd, Michael J. Wheeler, Ruth M. Perry, Elizabeth M. Bell, Kim Osman, F. Christopher H. Franklin & Vernonica E. Franklin-Tong

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  1. Barend H. J. de Graaf
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  8. Vernonica E. Franklin-Tong
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Corresponding author

Correspondence to Vernonica E. Franklin-Tong.

Supplementary information

Supplementary Figure Legends

This file contains text to accompany Supplementary Figures 1–4. (DOC 32 kb)

Supplementary Figure 1

Model for sPPase phosphorylation, effect on activity and biological consequences. (PDF 63 kb)

Supplementary Figure 2

Deduced amino acid sequences of the Pr-p26.1a/b cDNAs. (PDF 138 kb)

Supplementary Figure 3

sPPase activities in different tissues from Papaver rhoeas (PDF 66 kb)

Supplementary Figure 4

Antisense oligonucleotides show that Pr-p26.1a/b play a key role in pollen tube growth (PDF 144 kb)

Supplementary Methods

This file contains additional details on the methods used in this study. (DOC 51 kb)

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de Graaf, B., Rudd, J., Wheeler, M. et al. Self-incompatibility in Papaver targets soluble inorganic pyrophosphatases in pollen. Nature 444, 490–493 (2006). https://doi.org/10.1038/nature05311

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