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The phosphoproteome of rice leaves responds to water and nitrogen supply

Sara Hamzelou, Vanessa J. Melino, Darren C. Plett, Karthik Shantharam Kamath, Arkadiusz Nawrocki, Martin R. Larsen, Brian J. Atwell, View ORCID ProfilePaul A. Haynes
doi: https://doi.org/10.1101/2021.03.15.435047
Sara Hamzelou
1Department of Molecular Sciences, Macquarie University, North Ryde, NSW 2109, Australia
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Vanessa J. Melino
2King Abdullah University for Science and Technology, 2955-6990, The Kingdom of Saudi Arabia
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Darren C. Plett
3The Plant Accelerator, Australian Plant Phenomics Facility, The University of Adelaide, PMB 1, Glen Osmond, SA, 5064, Australia
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Karthik Shantharam Kamath
1Department of Molecular Sciences, Macquarie University, North Ryde, NSW 2109, Australia
4Australian Proteome Analysis Facility, Macquarie University, North Ryde, NSW 2109, Australia
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Arkadiusz Nawrocki
5Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK 5230 Odense M, Denmark
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Martin R. Larsen
5Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK 5230 Odense M, Denmark
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Brian J. Atwell
6Department of Biological Sciences, Macquarie University, North Ryde, NSW 2109, Australia
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Paul A. Haynes
1Department of Molecular Sciences, Macquarie University, North Ryde, NSW 2109, Australia
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  • ORCID record for Paul A. Haynes
  • For correspondence: paul.haynes@mq.edu.au
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Abstract

The scarcity of freshwater is an increasing concern in flood-irrigated rice, whilst excessive use of nitrogen fertilizers is both costly and contributes to environmental pollution. To co-ordinate growth adaptation under prolonged exposure to limited water or excess nitrogen supply, plants have processes for signalling and regulation of metabolic processes. There is limited information on the involvement of one of the most important post-translational modifications (PTMs), protein phosphorylation, on plant adaptation to long-term changes in resource supply. Oryza sativa cv. Nipponbare was grown under two regimes of nitrogen from the time of germination to final harvest. Twenty-five days after germination, water was withheld from half the pots in each nitrogen treatment and low water supply continued for an additional 26 days, while the remaining pots were well watered. Leaves from all four groups of plants were harvested after 51 days in order to test whether phosphorylation of leaf proteins responded to prior abiotic events. The dominant impact of these resources is exerted in leaves, where PTMs have been predicted to occur. Proteins were extracted and phosphopeptides were analysed by nanoLC-MS/MS analysis, coupled with label-free quantitation. Water and nitrogen regimes triggered extensive changes in phosphorylation of proteins involved in membrane transport, such as the aquaporin OsPIP2-6, a water channel protein. Our study reveals phosphorylation of several peptides belonging to proteins involved in RNA-processing and carbohydrate metabolism, suggesting that phosphorylation events regulate the signalling cascades that are required to optimize plant response to resource supply.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Reorganised figures for better readability, and corrected details in several references.

  • Abbreviations

    ABA
    Abscisic acid
    ACN
    Acetonitrile
    Ar
    Assimilation rate
    DAP
    Days after planting
    FC
    Field capacity
    GO
    Gene ontology
    LFQ
    Label-free quantitation
    N
    Nitrogen
    nLC–MS/MS
    Nanoflow liquid chromatography-tandem mass spectrometry
    PIP
    Plasma membrane Intrinsic Protein
    PTM
    Post-translational modification
    R:S
    root-to-shoot ratio
    Tr
    Transpiration rate
    TFA
    Trifluoroacetic acid
    WD
    Water deficit
    WW
    Well-watered
  • Copyright 
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    Posted March 18, 2021.
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    The phosphoproteome of rice leaves responds to water and nitrogen supply
    Sara Hamzelou, Vanessa J. Melino, Darren C. Plett, Karthik Shantharam Kamath, Arkadiusz Nawrocki, Martin R. Larsen, Brian J. Atwell, Paul A. Haynes
    bioRxiv 2021.03.15.435047; doi: https://doi.org/10.1101/2021.03.15.435047
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    The phosphoproteome of rice leaves responds to water and nitrogen supply
    Sara Hamzelou, Vanessa J. Melino, Darren C. Plett, Karthik Shantharam Kamath, Arkadiusz Nawrocki, Martin R. Larsen, Brian J. Atwell, Paul A. Haynes
    bioRxiv 2021.03.15.435047; doi: https://doi.org/10.1101/2021.03.15.435047

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