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Plant Physiol, July 2001, Vol. 126, pp. 1092-1104

The Ca2+ Status of the Endoplasmic Reticulum Is Altered by Induction of Calreticulin Expression in Transgenic Plants1

Staffan Persson,2 Sarah E. Wyatt,2 John Love,2 William F. Thompson, Dominique Robertson, and Wendy F. Boss*

North Carolina State University, Department of Botany, Raleigh, North Carolina 27695-7612

To investigate the endoplasmic reticulum (ER) Ca2+ stores in plant cells, we generated tobacco (Nicotiana tabacum; NT1) suspension cells and Arabidopsis plants with altered levels of calreticulin (CRT), an ER-localized Ca2+-binding protein. NT1 cells and Arabidopsis plants were transformed with a maize (Zea mays) CRT gene in both sense and antisense orientations under the control of an Arabidopsis heat shock promoter. ER-enriched membrane fractions from NT1 cells were used to examine how altered expression of CRT affects Ca2+ uptake and release. We found that a 2.5-fold increase in CRT led to a 2-fold increase in ATP-dependent 45Ca2+ accumulation in the ER-enriched fraction compared with heat-shocked wild-type controls. Furthermore, after treatment with the Ca2+ ionophore ionomycin, ER microsomes from NT1 cells overproducing CRT showed a 2-fold increase in the amount of 45Ca2+ released, and a 2- to 3-fold increase in the amount of 45Ca2+ retained compared with wild type. These data indicate that altering the production of CRT affects the ER Ca2+ pool. In addition, CRT transgenic Arabidopsis plants were used to determine if altered CRT levels had any physiological effects. We found that the level of CRT in heat shock-induced CRT transgenic plants correlated positively with the retention of chlorophyll when the plants were transferred from Ca2+-containing medium to Ca2+-depleted medium. Together these data are consistent with the hypothesis that increasing CRT in the ER increases the ER Ca2+ stores and thereby enhances the survival of plants grown in low Ca2+ medium.

1 This work was supported in part by the North Carolina State University-National Aeronautics and Space Administration Specialized Center of Research and Training (grant no. NAGW-4984) and in part by funding from the North Carolina Agricultural Research Service (to W.F.T., D.R., and W.F.B.).

2 These authors contributed equally to the paper.

* Corresponding author; email wendy_boss{at}ncsu.edu; fax 919-515-3436.

© 2001 American Society of Plant Physiologists


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