Abstract
Turnover rate is an important aspect of the regulation of plant processes by plant growth substances. To study turnover of indole-3-acetic acid (IAA), two [alpha]-methyltryptophan-resistant lines (MTR1 and MTR2) of Lemna gibba were generated by nitrosomethyl urea treatment of an inbred line derived from L. gibba G-3. In this report we describe: (a) the development of a selection system using this near isogenic line of L. gibba; (b) techniques for chemical mutation of the lines and selection for [alpha]-methyltryptophan resistance; and (c) the partial characterization of the selected lines. MTR lines contained 3-fold higher levels of anthranilate synthase activity. The enzyme in the MTR lines required higher levels of tryptophan for feedback inhibition. MTR lines also contained 8-fold higher levels of tryptophan, 3-fold higher levels of free IAA, and similar levels of total IAA compared to the inbred line. Turnover rates in the inbred and selected lines were calculated, using the first-order rate equation, based on the decrease over time in isotopic enrichment of I3C6-IAA introduced into L. gibba during a 1-h pulse period. Isotope enrichment in IAA was determined by using gas chromatography-mass spectrometry. Both MTR lines had an approximately 10-fold higher rate of IAA turnover than the parent inbred line.
Full Text
The Full Text of this article is available as a PDF (847.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baldi B. G., Maher B. R., Slovin J. P., Cohen J. D. Stable Isotope Labeling, in Vivo, of d- and l-Tryptophan Pools in Lemna gibba and the Low Incorporation of Label into Indole-3-Acetic Acid. Plant Physiol. 1991 Apr;95(4):1203–1208. doi: 10.1104/pp.95.4.1203. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bialek K., Cohen J. D. Free and conjugated indole-3-acetic Acid in developing bean seeds. Plant Physiol. 1989 Oct;91(2):775–779. doi: 10.1104/pp.91.2.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen K. H., Miller A. N., Patterson G. W., Cohen J. D. A Rapid and Simple Procedure for Purification of Indole-3-Acetic Acid Prior to GC-SIM-MS Analysis. Plant Physiol. 1988 Mar;86(3):822–825. doi: 10.1104/pp.86.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cleland C. F., Briggs W. R. Flowering Responses of the Long-day Plant Lemna gibba G3. Plant Physiol. 1967 Nov;42(11):1553–1561. doi: 10.1104/pp.42.11.1553. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Epstein E., Cohen J. D., Bandurski R. S. Concentration and Metabolic Turnover of Indoles in Germinating Kernels of Zea mays L. Plant Physiol. 1980 Mar;65(3):415–421. doi: 10.1104/pp.65.3.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kreps J. A., Town C. D. Isolation and Characterization of a Mutant of Arabidopsis thaliana Resistant to alpha-Methyltryptophan. Plant Physiol. 1992 May;99(1):269–275. doi: 10.1104/pp.99.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Last R. L., Fink G. R. Tryptophan-Requiring Mutants of the Plant Arabidopsis thaliana. Science. 1988 Apr 15;240(4850):305–310. doi: 10.1126/science.240.4850.305. [DOI] [PubMed] [Google Scholar]
- MOYED H. S. False feedback inhibition: inhibition of tryptophan biosynthesis by 5-methyltryptophan. J Biol Chem. 1960 Apr;235:1098–1102. [PubMed] [Google Scholar]
- Michalczuk L., Ribnicky D. M., Cooke T. J., Cohen J. D. Regulation of indole-3-acetic Acid biosynthetic pathways in carrot cell cultures. Plant Physiol. 1992 Nov;100(3):1346–1353. doi: 10.1104/pp.100.3.1346. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niyogi K. K., Fink G. R. Two anthranilate synthase genes in Arabidopsis: defense-related regulation of the tryptophan pathway. Plant Cell. 1992 Jun;4(6):721–733. doi: 10.1105/tpc.4.6.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Normanly J., Cohen J. D., Fink G. R. Arabidopsis thaliana auxotrophs reveal a tryptophan-independent biosynthetic pathway for indole-3-acetic acid. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10355–10359. doi: 10.1073/pnas.90.21.10355. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ranch J. P., Rick S., Brotherton J. E., Widholm J. M. Expression of 5-Methyltryptophan Resistance in Plants Regenerated from Resistant Cell Lines of Datura innoxia. Plant Physiol. 1983 Jan;71(1):136–140. doi: 10.1104/pp.71.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slovin J. P., Cohen J. D. Levels of Indole-3-Acetic Acid in Lemna gibba G-3 and in a Large Lemna Mutant Regenerated from Tissue Culture. Plant Physiol. 1988 Feb;86(2):522–526. doi: 10.1104/pp.86.2.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Widholm J. M. Anthranilate synthetase from 5-methyltryptophan-susceptible and -resistant cultured Daucus carota cells. Biochim Biophys Acta. 1972 Aug 18;279(1):48–57. doi: 10.1016/0304-4165(72)90240-1. [DOI] [PubMed] [Google Scholar]
- Widholm J. M. Cultured Nicotiana tabacum cells with an altered anthranilate synthetase which is less sensitive to feedback inhibition. Biochim Biophys Acta. 1972 Jan 28;261(1):52–58. doi: 10.1016/0304-4165(72)90312-1. [DOI] [PubMed] [Google Scholar]
- Widholm J. M. Tryptophan biosynthesis in Nicotiana tabacum and Daucus carota cell cultures: site of action of inhibitory tryptophan analogs. Biochim Biophys Acta. 1972 Jan 28;261(1):44–51. doi: 10.1016/0304-4165(72)90311-x. [DOI] [PubMed] [Google Scholar]