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Quantitative analysis of indole-3-acetic acid metabolites in Arabidopsis.
Plant Physiol. 2001 Dec; 127(4):1845-53.PP

Abstract

A general gas chromatography/mass spectrometry (MS)-based screen was performed to identify catabolites and conjugates of indole-3-acetic acid (IAA) during vegetative growth of Arabidopsis. This experiment revealed the existence of two new conjugates: N-(indole-3-acetyl)-alfa-alanine (IA-Ala) and N-(indole-3-acetyl)-alfa-leucine (IA-Leu). A method for quantitative analysis of IAA metabolites in plant extracts by liquid chromatography-electrospray tandem MS has been developed. The accuracy and precision of the new method are better than 10% for standards close to the detection limit, and are between 6% and 16% for the entire protocol applied to plant extracts. The low detection limits, 0.02 to 0.1 pmol for the different metabolites, made it possible to use as little as 50 to 100 mg of tissue for quantitative analysis. The analysis was performed on different tissues of an Arabidopsis plant at two stages of development, using heavy labeled internal standards of the catabolite 2-oxoindole-3-acetic acid as well as IAA conjugated to amino acids: aspartate, glutamate, Ala, and Leu. Expanding leaves and roots that generally contain high amounts of the free hormone also contained the highest levels of IA-aspartate, IA-glutamate, and 2-oxoindole-3-acetic acid, supporting their role as irreversible catabolic products. The levels of IA-Leu and IA-Ala did not follow the general distribution of IAA. Interestingly, the level of IA-Leu was highest in roots and IA-Ala in the aerial tissues.

Authors+Show Affiliations

Umeå Plant Science Center, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11743128

Citation

Kowalczyk, M, and G Sandberg. "Quantitative Analysis of Indole-3-acetic Acid Metabolites in Arabidopsis." Plant Physiology, vol. 127, no. 4, 2001, pp. 1845-53.
Kowalczyk M, Sandberg G. Quantitative analysis of indole-3-acetic acid metabolites in Arabidopsis. Plant Physiol. 2001;127(4):1845-53.
Kowalczyk, M., & Sandberg, G. (2001). Quantitative analysis of indole-3-acetic acid metabolites in Arabidopsis. Plant Physiology, 127(4), 1845-53.
Kowalczyk M, Sandberg G. Quantitative Analysis of Indole-3-acetic Acid Metabolites in Arabidopsis. Plant Physiol. 2001;127(4):1845-53. PubMed PMID: 11743128.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitative analysis of indole-3-acetic acid metabolites in Arabidopsis. AU - Kowalczyk,M, AU - Sandberg,G, PY - 2001/12/18/pubmed PY - 2002/4/17/medline PY - 2001/12/18/entrez SP - 1845 EP - 53 JF - Plant physiology JO - Plant Physiol VL - 127 IS - 4 N2 - A general gas chromatography/mass spectrometry (MS)-based screen was performed to identify catabolites and conjugates of indole-3-acetic acid (IAA) during vegetative growth of Arabidopsis. This experiment revealed the existence of two new conjugates: N-(indole-3-acetyl)-alfa-alanine (IA-Ala) and N-(indole-3-acetyl)-alfa-leucine (IA-Leu). A method for quantitative analysis of IAA metabolites in plant extracts by liquid chromatography-electrospray tandem MS has been developed. The accuracy and precision of the new method are better than 10% for standards close to the detection limit, and are between 6% and 16% for the entire protocol applied to plant extracts. The low detection limits, 0.02 to 0.1 pmol for the different metabolites, made it possible to use as little as 50 to 100 mg of tissue for quantitative analysis. The analysis was performed on different tissues of an Arabidopsis plant at two stages of development, using heavy labeled internal standards of the catabolite 2-oxoindole-3-acetic acid as well as IAA conjugated to amino acids: aspartate, glutamate, Ala, and Leu. Expanding leaves and roots that generally contain high amounts of the free hormone also contained the highest levels of IA-aspartate, IA-glutamate, and 2-oxoindole-3-acetic acid, supporting their role as irreversible catabolic products. The levels of IA-Leu and IA-Ala did not follow the general distribution of IAA. Interestingly, the level of IA-Leu was highest in roots and IA-Ala in the aerial tissues. SN - 0032-0889 UR - https://www.unboundmedicine.com/medline/citation/11743128/Quantitative_analysis_of_indole_3_acetic_acid_metabolites_in_Arabidopsis_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/11743128/ DB - PRIME DP - Unbound Medicine ER -