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THE PLANT CELL, Vol 7, Issue 8 1173-1184, Copyright © 1995 by American Society of Plant Biologists
Calcium Channel Activity during Pollen Tube Growth and Reorientation
R. Malho, N. D. Read, A. J. Trewavas and M. S. Pais
Departamento de Biologia Vegetal, Faculdade de Ciencias de Lisboa, R. Ernesto de Vasconcelos, Bloco C2, 1700 Lisbon, Portugal
We have shown previously that the inhibition of pollen tube growth and its
subsequent reorientation in Agapanthus umbellatus are preceded by an
increase in cytosolic free calcium ([Ca2+]c), suggesting a role for Ca2+ in
signaling these processes. In this study, a novel procedure was used to
measure Ca2+ channel activity in living pollen tubes subjected to various
growth reorienting treatments (electrical fields and ionophoretic
microinjection). The method involves adding extracellular Mn2+ to quench
the fluorescence of intracellular Indo-1 at its ca2+-insensitive wavelength
(isosbestic point). The spatial and temporal kinetics of Ca2+ channel
activity correlated well with measurements of [Ca2+]c dynamics obtained by
fluorescence ratio imaging of Indo-1. Tip-focused gradients in Ca2+ channel
activity and [Ca2+]c were observed and quantified in growing pollen tubes
and in swollen pollen tubes before reoriented growth. In nongrowing pollen
tubes, Ca2+ channel activity was very low and [Ca2+]c gradients were
absent. Measurements of membrane potential indicated that the growth
reorienting treatments induced a depolarization of the plasma membrane,
suggesting that voltage-gated Ca2+ channels might be activated.
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