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PLANT PHYSIOLOGY , Vol 114, Issue 3 881-886, Copyright © 1997 by American Society of Plant Biologists
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GENE REGULATION AND MOLECULAR GENETICS |
Antisense Expression and Overexpression of Biotin Carboxylase in Tobacco Leaves
D. Shintani, K. Roesler, B. Shorrosh, L. Savage and J. Ohlrogge
Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan 48824
The plastid acetyl-coenzyme A carboxylase (ACCase) catalyzes the first
committed step of fatty acid synthesis and in most plants is present as a
heteromeric complex of at least four different protein subunits: the biotin
carboxylase (BC), the biotin carboxyl carrier protein, and the [alpha] and
[beta] subunits of the carboxyltransferase. To gain insight into the
subunit organization of this heteromeric enzyme complex and to further
evaluate the role of ACCase in regulating fatty acid synthesis, BC
expression was altered in transgenic plants. Tobacco (Nicotiana tabacum)
was transformed with antisense-expression and overexpression tobacco BC
constructs, which resulted in the generation of plants with BC levels
ranging from 20 to 500% of wild-type levels. Tobacco plants containing
elevated or moderate decreases in leaf BC were phenotypically
indistinguishable from wild-type plants. However, plants with less than 25%
of wild-type BC levels showed severely retarded growth when grown under
low-light conditions and a 26% lower leaf fatty acid content than wild-type
plants. A comparison of leaf BC and biotin carboxyl carrier protein levels
in plants with elevated and decreased BC expression revealed that these two
subunits of the plastid ACCase are not maintained in a strict
stoichiometric ratio.
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