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Plant Physiol, November 1999, Vol. 121, pp. 705-714

Distinct Calcium Signaling Pathways Regulate Calmodulin Gene Expression in Tobacco1

Arnold H. van der Luit,* Claudio Olivari, Ann Haley, Marc R. Knight, and Anthony J. Trewavas

Institute for Molecular Cell Biology, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands (A.H.v.d.L.); Institute of Cell and Molecular Biology, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JH, United Kingdom (A.H., A.J.T.); Dipartimento di Biologia, Sezione Biochimica e Fisiologia delle Piante, University of Milan, Via Celoria 26, 20133 Milano, Italy (C.O.); and Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, United Kingdom (M.R.K.)

Cold shock and wind stimuli initiate Ca2+ transients in transgenic tobacco (Nicotiana plumbaginifolia) seedlings (named MAQ 2.4) containing cytoplasmic aequorin. To investigate whether these stimuli initiate Ca2+ pathways that are spatially distinct, stress-induced nuclear and cytoplasmic Ca2+ transients and the expression of a stress-induced calmodulin gene were compared. Tobacco seedlings were transformed with a construct that encodes a fusion protein between nucleoplasmin (a major oocyte nuclear protein) and aequorin. Immunocytochemical evidence indicated targeting of the fusion protein to the nucleus in these plants, which were named MAQ 7.11. Comparison between MAQ 7.11 and MAQ 2.4 seedlings confirmed that wind stimuli and cold shock invoke separate Ca2+ signaling pathways. Partial cDNAs encoding two tobacco calmodulin genes, NpCaM-1 and NpCaM-2, were identified and shown to have distinct nucleotide sequences that encode identical polypeptides. Expression of NpCaM-1, but not NpCaM-2, responded to wind and cold shock stimulation. Comparison of the Ca2+ dynamics with NpCaM-1 expression after stimulation suggested that wind-induced NpCaM-1 expression is regulated by a Ca2+ signaling pathway operational predominantly in the nucleus. In contrast, expression of NpCaM-1 in response to cold shock is regulated by a pathway operational predominantly in the cytoplasm.

1 This work was funded by the Research Training Grant Body of the European Commission and the Biotechnology and Biological Sciences Research Council.

* Corresponding author; e-mail luit{at}bio.uva.nl; fax 31-20-5257934.

© 1999 American Society of Plant Physiologists


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