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First published online December 13, 2002; 10.1105/tpc.005975

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The Plant Cell, Vol. 15, 151-163, January 2003, Copyright © 2003,
American Society of Plant Biologists

Overexpression of a Novel Class of Gibberellin 2-Oxidases Decreases Gibberellin Levels and Creates Dwarf Plants

Fritz M. Schomburg1,a, Colleen M. Bizzell1,a, Dong Ju Leeb, Jan A. D. Zeevaart2,b,c and Richard M. Amasino2,a

a Department of Biochemistry, University of Wisconsin–Madison, Madison, Wisconsin 53706
b Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48824-1312
c Department of Plant Biology, Michigan State University, East Lansing, Michigan 48824-1312

2 To whom correspondence should be addressed. E-mail amasino{at}biochem.wisc.edu; fax 608-262-3453

Degradation of active C19-gibberellins (GAs) by dioxygenases through 2{beta}-hydroxylation yields inactive GA products. We identified two genes in Arabidopsis (AtGA2ox7 and AtGA2ox8), using an activation-tagging mutant screen, that encode 2{beta}-hydroxylases. GA levels in both activation-tagged lines were reduced significantly, and the lines displayed dwarf phenotypes typical of mutants with a GA deficiency. Increased expression of either AtGA2ox7 or AtGA2ox8 also caused a dwarf phenotype in tobacco, indicating that the substrates for these enzymes are conserved. AtGA2ox7 and AtGA2ox8 are more similar to each other than to other proteins encoded in the Arabidopsis genome, indicating that they may constitute a separate class of GA-modifying enzymes. Indeed, enzymatic assays demonstrated that AtGA2ox7 and AtGA2ox8 both perform the same GA modification: 2{beta}-hydroxylation of C20-GAs but not of C19-GAs. Lines containing increased expression of AtGA2ox8 exhibited a GA dose–response curve for stem elongation similar to that of the biosynthetic mutant ga1-11. Double loss-of-function Atga2ox7 Atga2ox8 mutants had twofold to fourfold higher levels of active GAs and displayed phenotypes associated with excess GAs, such as early bolting in short days, resistance to the GA biosynthesis inhibitor ancymidol, and decreased mRNA levels of AtGA20ox1, a gene in the GA biosynthetic pathway.




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