Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on June 24, 2005; 10.1104/pp.105.062703

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Received March 11, 2005
Returned for revision April 17, 2005
Accepted April 19, 2005

A Gibberellin-Regulated Calcineurin B in Rice Localizes to the Tonoplast and Is Implicated in Vacuole Function

Yong-sic Hwang , Paul C. Bethke , Yong Hwa Cheong , Hur-Song Chang , Tong Zhu , and Russell L. Jones *

Department of Plant and Microbial Biology, University of California, Berkeley, California 94720-3102
Torrey Mesa Research Institute, Syngenta Research and Technology, San Diego, California 92121

* Corresponding author; email: rjones{at}nature.berkeley.edu.

Many developmental and environmental signals are transduced through changes in intracellular calcium concentrations, yet only a few calcium-binding proteins have been identified in plants. Calcineurin B-like (CBL) proteins are calcium-binding proteins that are thought to function as plant signal transduction elements. RNA profiling using a rice (Oryza sativa cv Nipponbare) oligonucleotide microarray was used to monitor gene expression in de-embryonated rice grains. This analysis showed that a putative rice CBL gene responded to gibberellic acid, but not abscisic acid, treatment. The CBL gene family in rice contains at least 10 genes and these have extensive similarity to the CBLs of Arabidopsis (Arabidopsis thaliana). In yeast (Saccharomyces cerevisiae) two-hybrid assays, rice CBLs interact with the kinase partners of Arabidopsis CBLs. Only one rice CBL gene, OsCBL2, is up-regulated by GA in the aleurone layer. A homolog with 91% sequence identity to OsCBL2 was cloned from barley (Hordeum vulgare cv Himalaya), and designated HvCBL2. We examined the localization and function of OsCBL2 and HvCBL2 in rice and barley aleurone because changes in cytosolic calcium have been implicated in the response of the aleurone cell to GA. Green fluorescent protein translational fusions of OsCBL2 and OsCBL3 were localized to the tonoplast of aleurone cell protein storage vacuoles and OsCBL4-green fluorescent protein was localized to the plasma membrane. Data from experiments using antisense expression of OsCBL2 and HvCBL2 are consistent with a role for OsCBL2 in promoting vacuolation of barley aleurone cells following treatment with GA.




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