A microtechnique for the analysis of free and conjugated indole-3-acetic acid in milligram amounts of plant tissue using a benchtop gas chromatograph-mass spectrometer |
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Authors: | David M Ribnicky Todd J Cooke Jerry D Cohen |
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Affiliation: | (1) Horticultural Crops Quality Laboratory, United States Department of Agriculture, Agricultural Research Service, Beltsville, MD 20705-2350, USA, US;(2) Department of Plant Biology, University of Maryland, College Park, MD 20742, USA, US |
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Abstract: | A microtechnique was developed for the quantification of indole-3-acetic acid (IAA) in plant samples of one milligram fresh
weight or less. The method permitted quantification of both free and conjugated IAA using a benchtop gas chromatograph-mass
spectrometer. New methods for sample purification with high recovery at microscale levels, together with simple changes that
result in enhanced sensitivity of the instrumentation, allowed for a significant reduction in the amount of plant material
required for analysis. Single oat (Avena sativa L.) coleoptile tips could be studied with this method and were found to contain free and total IAA levels of 137 and 399
pg · mg−1 fresh weight, respectively. A single 5-d-old Arabidopsis thaliana (L.) Heynh. seedling was shown to contain 61 pg · mg−1 fresh weight free IAA and 7850 pg · mg−1 fresh weight of total IAA following basic hydrolysis. This microtechnique provides a way to accurately measure IAA levels
in very small structures and individual seedlings, thus making it a valuable research tool for elucidating the role and distribution
of auxin in relation to growth and development.
Received: 1 May 1994 / Accepted: 25 June 1997 |
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Keywords: | :Arabidopsis (auxin analysis) Auxin analysis Avena (auxin analysis) Mass spectrometry Stable isotopes |
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