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Rapid turnover of polyphosphoinositides in carnation flower petals

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Abstract

Carnation (Dianthus caryophyllus L. cv. White Sim) petal discs were radiolabelled with [32P]orthophosphate and the lipids were extracted and analysed by thin-layer chromatography and autoradiography. Phospholipids were identified by co-migration with standards using thin-layer chromatography with different solvent systems. Results showed that [32P]orthophosphate was rapidly incorporated into the minor lipids phosphatidic acid (PtdOH), phosphatidylinositol monophosphate (PtdInsP) and phosphatidylinositol bisphosphate (PtdInsP2), and relatively slowly into the structural lipids phosphatidylcholine, -ethanolamine, -glycerol and -inositol. Pulse-chase experiments revealed that the label was rapidly lost from PtdOH, PtdInsP and PtdInsP2 while the structural lipids remained radiolabelled. The amount of PtdInsP and PtdInsP2 was found to constitute 0.45% and 0.013%, respectively, of the total phospholipids, on a molar basis. Together these results show that the turnover of the chemically low-abundant polyphosphoinositides is relatively high compared with the major structural phospholipids. Phosphatidylinositol monophosphate was further characterized by showing that it incorporates myo[3H]inositol and that its major fatty-acid constituents are palmitic acid and linoleic acid. Furthermore, we present evidence for the presence of both phosphatidylinositol 3-phosphate and phosphatidylinositol 4-phosphate isomers. The significance of these results is discussed with respect to plant phosphoinositide signal transduction.

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Abbreviations

AU:

arbitrary units

DAG:

diacylglycerol

GroPIns(3)P:

glycerophosphoinositol 3-phosphate

GroPIns(4)P:

glycerophosphoinositol 4-phosphate

PtdCho:

phosphatidylcholine

PtdEtn:

phosphatidylethanolamine

PtdGro:

phosphatidylglycerol

PtdIns:

phosphatidylinositol

PtdInsP:

phosphatidylinositol monophosphate

PtdInsP2 :

phosphatidylinositol bisphosphate

PtdOH:

phosphatidic acid

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Author notes

  1. Teun Munnik

    Present address: Department of Molecular Cell Biology, University of Amsterdam, Kruislaan 318, NL-1098, SM Amsterdam, The Netherlands

Authors and Affiliations

  1. Agrotechnological Research Institute (ATO-DLO), P.O. Box 17, 6700, AA Wageningen, The Netherlands

    Teun Munnik & Truus de Vrije

  2. Department of Molecular Cell Biology, University of Amsterdam, Kruislaan 318, 1098, SM Amsterdam, The Netherlands

    Alan Musgrave

Authors
  1. Teun Munnik
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  2. Alan Musgrave
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  3. Truus de Vrije
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Additional information

We thank Theo de Rijk and Gern Huijberts (ATO-DLO) for the GC-MS analysis of fatty acids, Eddy Smid (ATO-DLO) for yeast cultures, Hans Klerk (University of Amsterdam) for his discussions concerning isotopic labelling, and Jeanine Brederoo (University of Amsterdam) and Ernst Woltering (ATO-DLO) for critical reading of this manuscript. We are indebted to Bernard Payrastre (INSERM Unité 326, Toulouse, France) for analyzing the PtdInsP isomers by HPLC.

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Munnik, T., Musgrave, A. & de Vrije, T. Rapid turnover of polyphosphoinositides in carnation flower petals. Planta 193, 89–98 (1994). https://doi.org/10.1007/BF00191611

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