Abstract
Following application of 3H-Gibberellin A20 (GA20) to roots of G2 pea seedlings and homogenization of the roots, about 3% of the radioactivity in the tissue could be precipitated from a 30,000 × g supernatant with trichloroacetic acid (TCA) (soluble fraction) while about 5% of the radioactivity pelleted at 30,000 × g (particulate fraction). The radioactivity in the particulate fraction was soluble in sodium dodecyl sulfate (SDS), but was not dialyzable and was insoluble in ethanol. Electrophoresis of the soluble fraction gave only one band of radioactivity, while that of the particulate fraction gave multiple bands. Acid hydrolysis of the soluble fraction released radioactivity that ran coincident with acid-treated GA20 on silicic-acid column chromatography. The particulate fraction gave numerous radioactive peaks following acid hydrolysis, two of which were coincident with GA20 and GA29 (hydroxylation product of GA20) on silicic acid chromatography. Treatment of the particulate and soluble fractions with RNase, DNase, and proteases showed a significant solubilization of radioactivity only with the proteases, suggesting that the GA is bound to a proteinaceous macromolecule. Complete proteolytic hydrolyis followed by thin layer chromatography showed 65% of the radioactivity from the soluble fraction running separately from free GAs or the individual amino acids; the particulate fraction gave mainly (60%) free GAs on enzymatic hydrolysis and much smaller amounts (17%) in a position separate from that of the GAs or amino acids. Binding of 3H-GA to protease-sensitive material was obtained with biologically active 3H-GA20 and 3H-GA1.
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Bialek, K and Cohen, JD (1986) Isolation and partial characterization of the major amide linked conjugate of indole-3-acetate acid from Phaseolus vulgaris L. Plant Physiol 80: 99–104
Butcher, DN (1963) The presence of gibberellins in excised tomato roots. J Exp Bot 14:272–280
Grozier, A and Reid, DM (1971) Do roots synthesize gibberellins? Can J Bot 49:967–975
Davies, PJ, Emshwiller, E, Gianfagna, T and Proebsting, W (1982) The endogenous gibberellins of vegetative and reproductive tissue of G2 peas. Planta 154:266–272
Hill, RL and Schmidt, WR (1962) The complete hydrolysis of proteins. J Biol Chem 237:389–396
Ingram, TJ, Reid, JB and MacMillan, J (1985) Internode length in Pisum sativum L. The kinetics of growth and 3H-gibberellin A20 metabolism in genotype na le. Planta 164:429–438
Jones, RL and Phillips, IDJ (1966) Organs of GA synthesis in light-grown sunflower plants. Plant Physiol 41:1381–1386
Jones, RL (1968) Gibberellin extracts from peas. Plant 81:97–105
Katsumi M and Phinney BO (1969) Biosynthesis and metabolism of gibberellins. In: Tamura S, ed. Gibberellins, pp. 195–219. University of Tokyo Press
Keith, B and Srivastava, LM (1980) In vivo binding of GA1 in dwarf pea epicotyl. Plant Physiol 66:962–967
Konjevic, R, Grubisic, D, Markovic, R, Petrovic, J (1976) Gibberellic acid-binding proteins from pea stems. Planta 131:125–128
Laemmli, UK (1970) Cleavage of structural protein during assembly of the head of the bacteriophage T4. Nature 227:680–685
Lehnardt, J and Winzler, D (1968) Determination of neutral sugars in glycoproteins by GLC. J Chromatography 34:471–479
Musgrave, A, Day, SE, Kende, H (1969) In vivo binding of radioactive gibberellins in dwarf pea shoot. Planta 89:165–177
Nadeau, R and Rappaport, L (1974) An amphoteric conjugate of 3H-GA1. Plant Physiol 54:809–812
Powell, L and Tautvidas, KJ (1967) Chromatography of gibberellins on silica gel partition columns. Nature 213:292–293
Reid, DM and Crozier, A (1971) Effect of waterlogging on the gibberellin content and growth of tomato plants. J Exp Bot 22:39–48
Sitton, DA, Richmond, A and Vaadia, Y (1967a) On the synthesis of gibberellin in roots. Phytochem 6:1101–1105
Stoddart, JL and Lang (1968) The effect of daylength on gibberellin biosynthesis in leaves of red clover (Trifolium pratense L) In: FWightman and GSetterfield, ed. Biochemistry and Physiology of Plant Growth Regulators, pp 1371–1383. Ottawa: Runge Press
Stoddart, JL, Breidenbach, W, Nadeau, R and Rappaport, L (1974) Selective binding of 3H-Gibberellin A1 by protein fractions from dwarf pea epicotyls. Proc Natl Acad Sci USA 71:3255–3259
Stoddart, JL (1979a) Interaction of 3H-Gibberellin Al with a subcellular fraction from lettuce (Lactuca sativa L.) hypocotyls. I. Kinetics of labeling. Planta 146:353–361
Stoddart, JL (1979b) Interaction of 3H-Gibberellin Al with a subcellular fraction from lettuce (Lactuca sativa L.) hypocotyls. II. Stability and properties of the association. Planta 146:363–368
Stoddart, JL and Williams, PD (1979) Interaction of 3H-Gibberellin A1 with a subcellular fraction from lettuce (Lactuca sativa L.) hypocotyls requirement for protein synthesis. Planta 147:264–268
Verbiscar, AJ, Gragg, G, Geissman, TA and Phinney, BO (1967) Studies on the biosynthesis of gibberellins-II. The biosynthesis of gibberellins from (−)-kaurenol and the conversion of gibberellins 14C-GA4 and 14C-GA7 into 14C-GA3 by Gibberella fujikuroi. Phytochem 6:807–814
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Delorme, E.O. Incorporation of 3H-Gibberellin A20 in roots of G2 peas. Plant Growth Regul 5, 125–140 (1987). https://doi.org/10.1007/BF00024740
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DOI: https://doi.org/10.1007/BF00024740