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Analysis of Promoter Activity for the Gene Encoding Pyruvate Orthophosphate Dikinase in Stably Transformed C4 Flaveria Species1

Elke Rosche2, Julie Chitty, Peter Westhoff, and William C. Taylor*

Cooperative Research Centre for Plant Science, G.P.O. Box 475, Canberra 2601, Australia (E.R., W.C.T.); Commonwealth Scientific and Industrial Research Organisation Plant Industry, G.P.O. Box 1600, Canberra 2601, Australia (J.C., W.C.T.); and Institut für Entwicklungs und Molekularbiologie der Pflanzen, Heinrich-Heine Universität Düsseldorf, D-40225 Düsseldorf, Germany (P.W.)

The C4 enzyme pyruvate orthophosphate dikinase is encoded by a single gene, Pdk, in the C4 plant Flaveria trinervia. This gene also encodes enzyme isoforms located in the chloroplast and in the cytosol that do not have a function in C4 photosynthesis. Our goal is to identify cis-acting DNA sequences that regulate the expression of the gene that is active in the C4 cycle. We fused 1.5 kb of a 5' flanking region from the Pdk gene, including the entire 5' untranslated region, to the uidA reporter gene and stably transformed the closely related C4 species Flaveria bidentis. beta -Glucuronidase (GUS) activity was detected at high levels in leaf mesophyll cells. GUS activity was detected at lower levels in bundle-sheath cells and stems and at very low levels in roots. This lower-level GUS expression was similar to the distribution of mRNA encoding the nonphotosynthetic form of the enzyme. We conclude that cis-acting DNA sequences controlling the expression of the C4 form in mesophyll cells and the chloroplast form in other cells and organs are co-located within the same 5' region of the Pdk gene.

1   The isolation of the Pdk promoter was done in Düsseldorf, Germany, and was supported by a grant from the Deutsche Forschungsgemeinschaft via Sonderforschungsbereicht 189. E.R. was supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft.
2   Present address: Department of Biological Sciences, University of Newcastle, Newcastle NSW 2308, Australia.
*   Corresponding author; e-mail bt{at}pi.csiro.au; fax 61-2-6246-5000.

Plant Physiol. (1998) 117: 821-829
Copyright Clearance Center:   0032-0889/98/117/0821/09
© 1998 American Society of Plant Physiologists




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Copyright © 1998 by the American Society of Plant Biologists