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Plant Cell, Vol. 10, 1267-1276, August 1998, Copyright © 1998, American Society of Plant Physiologists

Ca2+, Annexins, and GTP Modulate Exocytosis from Maize Root Cap Protoplasts

Andrew D. Carrolla, Christelle Moyena, Pim Van Kesterenb, Fiona Tookeb, Nicholas H. Batteyb, and Colin Brownleea
a Marine Biological Association, Citadel Hill, Plymouth PL1 2PB, United Kingdom
b Plant Science Laboratories, University of Reading, Whiteknights, Reading RG6 6AS, United Kingdom

Correspondence to: Nicholas H. Battey, n.h.battey{at}reading.ac.uk (E-mail), 44-118-9750630 (fax).

Protoplasts isolated from root cap cells of maize were shown to secrete fucose-rich polysaccharides and were used in a patch–clamp study to monitor changes in whole-cell capacitance. Ca2+ was required for exocytosis, which was measured as an increase in cell capacitance during intracellular dialysis with Ca2+ buffers via the patch pipette. Exocytosis was stimulated significantly by small increases above normal resting [Ca2+]. In the absence of Ca2+, protoplasts decreased in size. In situ hybridization showed significant expression of the maize annexin p35 in root cap cells, differ-entiating vascular tissue, and elongating cells. Dialysis of protoplasts with maize annexins stimulated exocytosis at physiological [Ca2+], and this could be blocked by dialysis with antibodies specific to maize annexins. Dialysis with milli-molar concentrations of GTP strongly inhibited exocytosis, causing protoplasts to decrease in size. GTP{gamma}S and GDPßS both caused only a slight inhibition of exocytosis at physiological Ca2+. Protoplasts were shown to internalize plasma membrane actively. The results are discussed in relation to the regulation of exocytosis in what is usually considered to be a constitutively secreting system; they provide direct evidence for a role of annexins in exocytosis in plant cells.




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