First published online May 21, 2004; 10.1104/pp.104.039677
Plant Physiology 135:978-988 (2004)
© 2004 American Society of Plant Biologists
DEVELOPMENT AND HORMONE ACTION
A Family of Auxin-Conjugate Hydrolases That Contributes to Free Indole-3-Acetic Acid Levels during Arabidopsis Germination1
Rebekah A. Rampey,
Sherry LeClere2,
Mariusz Kowalczyk,
Karin Ljung,
Göran Sandberg and
Bonnie Bartel*
Department of Biochemistry and Cell Biology, Rice University, Houston, Texas 77005 (R.A.R., S.L., B.B.); and Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE–901 83 Umea, Sweden (M.K., K.L., G.S.)
Auxins are hormones important for numerous processes throughout plant growth and development. Plants use several mechanisms to regulate levels of the auxin indole-3-acetic acid (IAA), including the formation and hydrolysis of amide-linked conjugates that act as storage or inactivation forms of the hormone. Certain members of an Arabidopsis amidohydrolase family hydrolyze these conjugates to free IAA in vitro. We examined amidohydrolase gene expression using northern and promoter- -glucuronidase analyses and found overlapping but distinct patterns of expression. To examine the in vivo importance of auxin-conjugate hydrolysis, we generated a triple hydrolase mutant, ilr1 iar3 ill2, which is deficient in three of these hydrolases. We compared root and hypocotyl growth of the single, double, and triple hydrolase mutants on IAA-Ala, IAA-Leu, and IAA-Phe. The hydrolase mutant phenotypic profiles on different conjugates reveal the in vivo activities and relative importance of ILR1, IAR3, and ILL2 in IAA-conjugate hydrolysis. In addition to defective responses to exogenous conjugates, ilr1 iar3 ill2 roots are slightly less responsive to exogenous IAA. The triple mutant also has a shorter hypocotyl and fewer lateral roots than wild type on unsupplemented medium. As suggested by the mutant phenotypes, ilr1 iar3 ill2 imbibed seeds and seedlings have lower IAA levels than wild type and accumulate IAA-Ala and IAA-Leu, conjugates that are substrates of the absent hydrolases. These results indicate that amidohydrolases contribute free IAA to the auxin pool during germination in Arabidopsis.
1 This work was supported by the National Institutes of Health (grant no. R29–GM54749 to B.B.), by the Robert A. Welch Foundation (grant no. C–1309 to B.B.), and by the Swedish Research Council and the Swedish Foundation for Strategic Research (grant to G.S.). Partial support was provided by an NIH Training Grant (grant no. T32–GM08362 to S.L.).
2 Present address: Stoller Enterprises, 4001 W. Sam Houston Parkway N., Suite 100, Houston, TX 77043.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.039677.
* Corresponding author; e-mail bartel{at}rice.edu; fax 713–348–5154.
Received February 1, 2004;
returned for revision March 26, 2004;
accepted March 30, 2004.
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