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JXB Advance Access published online on April 18, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri163
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received December 10, 2004
Accepted March 16, 2005

RESEARCH PAPER

Phosphoinositides and phosphatidic acid regulate pollen tube growth and reorientation through modulation of [Ca2+]c and membrane secretion

David Monteiro 1 *, Qunlu Liu 1 *, Saskia Lisboa 2, G. E. F. Scherer 3, Hartmut Quader 2, and Rui Malhó 1*

1 Universidade de Lisboa, Faculdade de Ciências de Lisboa, ICAT, 1749-016 Lisboa, Portugal
2 Institute of General Botany, University of Hamburg, Ohnhorst-Straße 18, D-22609 Hamburg, Germany
3 University of Hannover, Institut für Zierpflanzenanbau, Baumschule u. Pflanzenzüchtung, Herrenhäuser Str. 2, D-30419 Hannover, Germany

* To whom correspondence should be addressed.
Rui Malhó, E-mail: r.malho{at}fc.ul.pt


  Abstract

The maintenance of a calcium gradient and vesicle secretion in the apex of pollen tubes is essential for growth. It is shown here that phosphatidylinositol-4,5-bisphosphate (PIP2) and D-myo-inositol-1,4,5-trisphosphate (IP3), together with phosphatidic acid (PA), play a vital role in the regulation of these processes. Changes in the intracellular concentration of both PIP2 and IP3 (induced by photolysis of caged-probes), modified growth and caused reorientation of the growth axis. However, measurements of cytosolic free calcium ([Ca2+]c) and apical secretion revealed significant differences between the photorelease of PIP2 or IP3. When released in the first 50 µm of the pollen tube, PIP2 led to transient growth perturbation, [Ca2+]c increases, and inhibition of apical secretion. By contrast, a concentration of IP3 which caused a [Ca2+]c transient of similar magnitude, stimulated apical secretion and caused severe growth perturbation. Furthermore, the [Ca2+]c transient induced by IP3 was spatially different causing a pronounced elevation in the sub-apical region. These observations suggest different targets for the two phosphoinositides. One of the targets is suggested to be PA, a product of PIP2 hydrolysis via phospholipase C (PLC) or phospholipase D (PLD) activity. It was found that antagonists of PA accumulation (e.g. butan-1-ol) and inhibitors of PLC and PLD reversibly halted polarity. Reduction of PA levels caused the dissipation of the [Ca2+]c gradient and inhibited apical plasma membrane recycling. It was also found to cause abolition of the apical zonation. These data suggest that phosphoinositides and phospholipids regulate tip growth through a multiple pathway system involving regulation of [Ca2+]c levels, endo/exocytosis, and vesicular trafficking.

Keywords: Ins(1,4,5)P3; phosphatidic acid; phospholipases; PIP2; secretion.

*These authors have contributed equally to this work.


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