Indole-3-acetic Acid Levels of Plant Tissue as Determined by a New High Performance Liquid Chromatographic Method

Sweetser, Philip B.; Swartzfager, D. G.

doi:10.1104/pp.61.2.254

Cited by 66 publications

(29 citation statements)

References 14 publications

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“…Leaves can also synthesize cytokinins (Singh et al 1992). Auxin is synthesized in young leaves, apical buds of stems, and immature seeds (Sweetser and Swartzfager 1978). Auxin is transported to the other parts of plants, and acts to suppress leaf senescence and abscission (Nooden et al 1979).…”

Section: Discussionmentioning

confidence: 99%

Interaction of Scion and Stock on Leaf Senescence of Soybean Plants Grafted at Mid-Stem during Ripening

Ookawa

Tomita

Hirasawa

2005

Plant Production Science

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Section: Discussionmentioning

confidence: 99%

Interaction of Scion and Stock on Leaf Senescence of Soybean Plants Grafted at Mid-Stem during Ripening

Ookawa

Tomita

Hirasawa

2005

Plant Production Science

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“…Sweetser and Swartzfager (16) and Guinn (9) used microparticulate SAX columns for IAA and ABA, respectively. Both used 10-,gm particles.…”

Section: Methodsmentioning

confidence: 99%

Purification and Measurement of Abscisic Acid and Indoleacetic Acid by High Performance Liquid Chromatography

Guinn¹,

Brummett²,

Beier³

1986

Plant Physiol.

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ABSTRACIA procedure was selected for the simultaneous extraction and purification of abscisic acid (ABA) and indoleacetic acid (IAA). Unnecessary steps were eliminated and an accumulation of aqueous phase was avoided. The superior performance of diethyl ether (compared to ethyl acetate) for bulk purification and the superior resolution provided by 250 millimeter columns packed with 5-micrometer spherical particles of strong anion exchanger and octadecylsilane (C18) greatly facilitated the purification of samples. A fixed-wavelength (254 nanometer) ultraviolet detector and a fluorescence detector connected in series on a high performance liquid chromatograph permitted nondestructive monitoring and measurement of ABA and IAA. Derivatization was not necessary for chromatography or for detection. Isocratic elution with simple mobile phases gave sharp peaks. A few simple precautions minimized losses. Recoveries through the entire procedure averaged about 75% for ABA and about 50% for IAA. Purifled ABA and IAA fractions were usually free of interfering contaminants. Identities were confirmed by gas chromatography-mass spectrometry.Poor recovery is a frequent problem with IAA determination because IAA is easily oxidized and is subject to photodecomposition (11,14). ABA is also somewhat labile, although not as labile as IAA. Because of variable losses during extraction and purification, the use of internal standards is essential for accurate quantification of ABA and IAA (4). trans-ABA has been used as an internal standard for the estimation of ABA (9, 19), but the acceptability of its use depends upon its absence from the tissue being analyzed and upon lack of interconversion of ABA and trans-ABA during the procedure. The use of radioactive ABA as an internal standard circumvents these potential problems. Indolebutyric acid has been used as an internal standard for IAA (5,19), but its value is questionable because of differences in partitioning and chromatographic behavior and possible differences in stability. Labeled IAA is preferable as an internal standard.HPLC and GLC are frequently used in the final stages of purification and for quantification of ABA and IAA. HPLC has advantages over GLC in quantification because UV and fluorescence detectors, used with HPLC, are nondestructive, and the entire sample can readily be collected for measurement of radioactive internal standards. Also, it is not necessary to form volatile derivatives (e.g. by methylation or silylation) for HPLC as it is for GLC. The UV detector, however, is not very specific and impurities are likely to interfere with quantification unless the sample is almost pure.The objectives of the work reported here were to select procedures that would (a) permit simultaneous extraction and purification of ABA and IAA, (b) provide purification sufficient that ABA could be quantified by UV absorbance and IAA could be quantified by natural fluorescence with the detectors connected in series on an HPLC, (c) give a high percentage recovery so that small amounts of ...

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“…Several physicochemical methods for determining IAA in the ng range have been proposed in the last decade to replace bioassay systems but, for various reasons, they are of limited use for plant extracts and are not readily applicable to physiological experiments (24).…”

Section: Scribedmentioning

confidence: 99%

“…The indolo-a-pyrone fluorescence method of determining indole-3-acetic acid (IAA) is improved by adding butylated hydroxytoluene (BHT) (24).…”

Section: Introductionmentioning

confidence: 99%

“…

ABSTRACTThe indolo-a-pyrone fluorescence method of determining indole-3-acetic acid (IAA) is improved by adding butylated hydroxytoluene (BHT) (24).The indolo-a-pyrone fluorescence method, which was first devised by Stoessl and Venis (23), has been shown to be highly sensitive and specific (17, 23) and used to determine IAA in crude acidic extracts prepared by liquid partitioning (6,10,11,23). This procedure suffers from the disadvantage that 4-Cl-IAA' and 5-OH-IAA, which occur naturally in plant tissues (8,15,25) can produce fluorescence indistinguishable from that of IAA after

…”

mentioning

confidence: 99%

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Improved Procedure for the Estimation of Nanogram Quantities of Indole-3-acetic Acid in Plant Extracts using the Indolo-α-pyrone Fluorescence Method

Iino

Yu²,

Carr³

1980

Plant Physiol.

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The indolo-a-pyrone fluorescence method of determining indole-3-acetic acid (IAA) is improved by adding butylated hydroxytoluene (BHT) (24).The indolo-a-pyrone fluorescence method, which was first devised by Stoessl and Venis (23), has been shown to be highly sensitive and specific (17, 23) and used to determine IAA in crude acidic extracts prepared by liquid partitioning (6,10,11,23). This procedure suffers from the disadvantage that 4-Cl-IAA' and 5-OH-IAA, which occur naturally in plant tissues (8,15,25) can produce fluorescence indistinguishable from that of IAA after

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Indole-3-acetic Acid Levels of Plant Tissue as Determined by a New High Performance Liquid Chromatographic Method

Cited by 66 publications

References 14 publications

Interaction of Scion and Stock on Leaf Senescence of Soybean Plants Grafted at Mid-Stem during Ripening

Interaction of Scion and Stock on Leaf Senescence of Soybean Plants Grafted at Mid-Stem during Ripening

Purification and Measurement of Abscisic Acid and Indoleacetic Acid by High Performance Liquid Chromatography

Improved Procedure for the Estimation of Nanogram Quantities of Indole-3-acetic Acid in Plant Extracts using the Indolo-α-pyrone Fluorescence Method

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