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CELL BIOLOGY AND SIGNAL TRANSDUCTION

A Gibberellin-Regulated Calcineurin B in Rice Localizes to the Tonoplast and Is Implicated in Vacuole Function¹

Department of Plant and Microbial Biology, University of California, Berkeley, California 94720–3102 (Y.-s.H., P.C.B., Y.H.C., R.L.J.); and Torrey Mesa Research Institute, Syngenta Research and Technology, San Diego, California 92121 (H.-S.C., T.Z.)

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.

² Present address: Diversa Corporation, 4955 Directors Place, San Diego, CA 92121.

³ Present address: Syngenta Biotechnology, Inc., 3054 Cornwallis Road, Research Triangle Park, NC 27709.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.062703.

^* Corresponding author; e-mail rjones{at}nature.berkeley.edu; fax 510–642–4995.

Received March 11, 2005; returned for revision April 17, 2005; accepted April 19, 2005.

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