This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 20/3/614    most recent tpc.106.047423v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Certal, A. C. Articles by Feijó, J. A. PubMed PubMed Citation Articles by Certal, A. C. Articles by Feijó, J. A. Agricola Articles by Certal, A. C. Articles by Feijó, J. A.

Exclusion of a Proton ATPase from the Apical Membrane Is Associated with Cell Polarity and Tip Growth in Nicotiana tabacum Pollen Tubes^[W]

^a Instituto Gulbenkian de Ciência, Centro de Biologia do Desenvolvimento, 2780-156 Oeiras, Portugal
^b Departamento Biologia Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1700 Lisboa, Portugal
^c Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, Massachusetts, 01003
^d Centre de Medicina Regenerativa de Barcelona, E-08003 Barcelona, Spain

¹ Address correspondence to jfeijo{at}fc.ul.pt.

Polarized growth in pollen tubes results from exocytosis at the tip and is associated with conspicuous polarization of Ca²⁺, H⁺, K⁺, and Cl^– -fluxes. Here, we show that cell polarity in Nicotiana tabacum pollen is associated with the exclusion of a novel pollen-specific H⁺-ATPase, Nt AHA, from the growing apex. Nt AHA colocalizes with extracellular H⁺ effluxes, which revert to influxes where Nt AHA is absent. Fluorescence recovery after photobleaching analysis showed that Nt AHA moves toward the apex of growing pollen tubes, suggesting that the major mechanism of insertion is not through apical exocytosis. Nt AHA mRNA is also excluded from the tip, suggesting a mechanism of polarization acting at the level of translation. Localized applications of the cation ionophore gramicidin A had no effect where Nt AHA was present but acidified the cytosol and induced reorientation of the pollen tube where Nt AHA was absent. Transgenic pollen overexpressing Nt AHA-GFP developed abnormal callose plugs accompanied by abnormal H⁺ flux profiles. Furthermore, there is no net flux of H⁺ in defined patches of membrane where callose plugs are to be formed. Taken together, our results suggest that proton dynamics may underlie basic mechanisms of polarity and spatial regulation in growing pollen tubes.