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AOBPreview originally published online on April 15, 2004
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Annals of Botany 93: 741-753, 2004
© 2004 Annals of Botany Company

Calcium Channels are Involved in Calcium Oxalate Crystal Formation in Specialized Cells of Pistia stratiotes L.

GAYLE M. VOLK{dagger},1, LENORA J. GOSS1 and VINCENT R. FRANCESCHI*,1

1 School of Biological Sciences, Washington State University, Pullman, WA 99164-4236, USA

* For correspondence. E-mail vfrances{at}mail.wsu.edu
{dagger} Present address: United States Department of Agriculture, Agricultural Research Service, National Center for Genetic Resources Preservation, 1111 S. Mason St, Fort Collins, CO 80521, USA

Received: 20 December 2003; Returned for revision: 22 January 2004; Accepted: 12 February 2004; Published electronically: 15 April 2004

• Background and Aims Pistia stratiotes produces large amounts of calcium (Ca) oxalate crystals in specialized cells called crystal idioblasts. The potential involvement of Ca2+ channels in Ca oxalate crystal formation by crystal idioblasts was investigated.

• Methods Anatomical, ultrastructural and physiological analyses were used on plants, fresh or fixed tissues, or protoplasts. Ca2+ uptake by protoplasts was measured with 45Ca2+, and the effect of Ca2+ channel blockers studied in intact plants. Labelled Ca2+ channel blockers and a channel protein antibody were used to determine if Ca2+ channels were associated with crystal idioblasts.

• Key Results 45Ca2+ uptake was more than two orders of magnitude greater for crystal idioblast protoplasts than mesophyll protoplasts, and idioblast number increased when medium Ca was increased. Plants grown on media containing 1–50 µM of the Ca2+ channel blockers, isradipine, nifedipine or fluspirilene, showed almost complete inhibition of crystal formation. When fresh tissue sections were treated with the fluorescent dihydropyridine-type Ca2+ channel blocker, DM-Bodipy-DHP, crystal idioblasts were intensely labelled compared with surrounding mesophyll, and the label appeared to be associated with the plasma membrane and the endoplasmic reticulum, which is shown to be abundant in idioblasts. An antibody to a mammalian Ca2+ channel {alpha}1 subunit recognized a single band in a microsomal protein fraction but not soluble protein fraction on western blots, and it selectively and heavily labelled developing crystal idioblasts in tissue sections.

• Conclusions The results demonstrate that Ca oxalate crystal idioblasts are enriched, relative to mesophyll cells, in dihydropyridine-type Ca2+ channels and that the activity of these channels is important to transport and accumulation of Ca2+ required for crystal formation.

Key words: Calcium, calcium channel, calcium oxalate, crystals, dihydropyridine, Pistia stratiotes.


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