Abstract
A specific solid-phase enzyme immunoassay for the detection of as little as 3–4 pg of indole-3-acetic acid (IAA) is described. The assay involves minimal procedural efforts and requires only standard laboratory equipment. Up to 50 samples in triplicate, processed simultaneously, can be assayed and evaluated in 2.5 h. As little as 1 mg oat coleoptile tissue is sufficient for a quantitative IAA analysis and little or no extract purification is necessary. Using this assay, levels of IAA have been determined in coleoptiles of maize and oat. The distribution of IAA within single coleoptiles was quantitated and the production of IAA during the regeneration of the physiological tip in Avena coleoptiles was investigated. The changes in levels of IAA and other major phytohormones were quantitated during the growth of oat coleoptiles.
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Abbreviations
- ABA:
-
abscisic acid
- BHT:
-
butylated hydroxytoluene
- BSA:
-
bovine serum albumin
- IAA:
-
indole-3-acetic acid
- TBS:
-
Trishydroxymethylaminomethane buffered saline
References
Anker, L. (1973) The auxin production of the physiological tip of the Avena coleoptile and the repression of tip regeneration by indoleacetic acid (not by naphthylacetic acid and 2,4-dichlorophenoxyacetic acid). Acta Bot. Neerl. 22, 221–227
Bandurski, R.S. (1979) Homeostatic concentrations of indole-3-acetic acid. In: Plant growth substances, pp. 37–49, Skoog, F., ed. Springer-Verlag, Berlin Heidelberg New York
Bandurski, R.S., Schulze, A. (1974) Concentrations of indole-3-acetic acid and its esters in Avena and Zea. Plant Physiol. 54, 257–262
Bandurski, R.S., Schulze, A. (1977) Concentration of indole-3-acetic acid and its derivatives in plants. Plant Physiol. 60, 211–213
Crozier, A. Loferski, K., Zaerr, J., Morris, O. (1980) Analysis of picogram quantities of indole-3-acetic acid by high performance liquid chromatography-fluorescence procedures. Planta 150, 366–370
Gillesie, B., Thimann, K.V. (1976) Transport and distribution of auxin during tropistic response. I. The lateral migration of auxin in geotropism. Plant Physiol. 38, 214–225
Greenwood, M.S., Shaw, S., Hillman, J.R., Ritchie, A., Wilkins, M.B. (1972) Identification of auxin from Zea coleoptile tips by mass spectrometry. Planta 108, 179–183
Hemberg, T., Tillberg, E. (1980) The influence of the extraction procedure on yield of indole-3-acetic acid in plant extracts. Physiol. Plant. 50, 176–182
Hurn, B.A.L., Chantler, S.M. (1980) Production of reagent antibodies. Meth. Enzymol. 70, 104–141
Libbert, E., Wichner, S., Duerst, E., Kunert, R., Kaiser, W., Manicki, A., Manteuffel, R., Ricke, E., Schroder, R. (1968) Auxin content and auxin synthesis in sterile and non-sterile plants, with special regard to the influence of epiphytic bacteria. In: Biochemistry and physiology of plant growth substances. pp. 213–230, Wightman, F. and Setterfield, G. ed., Runge Press, Ottawa, Canada
Marschner, I., Dobry, H., Erhardt, F., Landersdorfer, T., Popp, B., Ringel, C., Scriba, P.C. (1974) Berechnung radioimmunologischer Meßwerte mittels Spline-Funktion. Ärztl. Lab. 20, 184–191
Mer, C.L. (1968) Plant growth in relation to endogenous auxin with special reference to cereal seedlings. New Phytol. 68, 275–294
Mousdale, D.M.A., Butcher, D.N., Powell, R.G. (1978) Spectrofluorimetric methods of determining indole-3-acetic acid. In: Isolation of plant growth substances pp. 27–31, Hillman, J.R., ed. University Cambridge, Press
Pengelly, W., Meins, F., Jr., (1977) A specific radioimmunoassay for nanogram quantities of the auxin, indole-3-acetic acid. Planta 136, 173–180
Sheldrake, A.R. (1973) Do coleptile tips produceauxin? New Phytol. 72, 433–447
Thimann, K.V. (1934) Studies on the growth hormones of plants. VI. The distribution of the growth substances in plant tissues. J. Gen. Physiol. 18, 23–34
Walton, D., Dashek, W., Galson, E. (1979) A radioimmunoassay for abscisic acid. Planta 146, 139–145
Weiler, E.W. (1979) Radioimmunoassay for the determination of free and conjugated abscisic acid. Planta 144, 255–263
Weiler, E.W. (1980a) Radioimmunoassay for the differential and direct analysis of free and conjugated abscisic acid in plant extracts. Planta 148, 262–272
Weiler, E.W. (1980b) Radioimmunoassay for trans-zeatin and related cytokinins. Planta 149, 155–162
Weiler, E.W. (1981) Radioimmunoassay for pico-mol quantities of indole-3-acetic acid for use with highly stable [125I]- and [3H]IAA derivatives asradiotracers. Planta 153, 319–325
Weiler, E.W., Spanier, K. (1981) Phytohormones in the formation of crown-gall tumors. Planta 153, 326–337
Weiler, E.W., Wieczorek, U. (1981) Determination of femto-molquantities of gibberellic acid by radioimmunoassay. Planta 152, 159–167
Weiler, E.W., Ziegler, H. (1981) Determination of phytohormones in phloem exudate of tree species by radioimmunoassay. Planta 152, 168–170
Went, F.W. (1928) Wuchsstoff und Wachstum. Rec. Trav. Bot. Neerl. 25, 1–116
Went, F.W., Thimann, K.V. (1937) Phytohormones. Macmillan Co., New York
Zenk, M.H. (1970) Phytohormone and Genaktivität. Ber. Dtsch. Bot. Ges. 83, 325–344
Zimmermann, H., Siegert, Chr., Karl, R. (1976) Veränderungen des Wuchsstoffes Indolyl-3-essigsäure bei der Keimung von Avena sativa L. Z. Pflanzenphysiol. 80, 225–235
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Part 21 in the series “Use of Immunoassay in Plant Science”
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Weiler, E.W., Jourdan, P.S. & Conrad, W. Levels of indole-3-acetic acid in intact and decapitated coleoptiles as determined by a specific and highly sensitive solid-phase enzyme immunoassay. Planta 153, 561–571 (1981). https://doi.org/10.1007/BF00385542
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DOI: https://doi.org/10.1007/BF00385542