Summary
An extract from 6000 dark-grown Phaseolus coccineus seedlings was purified by countercurrent distribution and G-10 Sephadex followed by gradient elution from a silicic acid partition column with increasing amounts of ethyl actetate in n-hexane. 25 fractions were collected and tested with the barley-aleurone, ‘Tan-ginbozu’ dwarf-rice, lettuce, cucumber, dwarf-pea, d-1, d-2, d-3 and d-5 maize, oat first-internode, and sugarcane-spindle bioassays. Major gibberellin (GA)-like activity was detected in fractions 4 (500μg GA3-equivalents) and 12–13 (270 μg GA3-equivalents) with smaller amounts in fractions 6, 8–9, 15–16, 18, 20, 23 and 25. The extracts were also applied to AMO-1618=dwarfed Ph.-coccineus seedlings. Fractions 4, 8 and 12 promoted the growth of both light- and dark-grown seedlings. GA1, GA3, GA4 and GA8 were active in the Phaseolus bioassay but GA8-glucoside was inactive.
The biological and chromatographic properties of fractions 4, 8–9 and 12–13 correspond with those of GA4, GA19 and GA1. The identity of GA4 in fraction 4 was conclusively established by combined gas chromatography-mass spectrometry (GC-MS) of the methyl ester and the trimethylsilyl ether of the methyl ester. Gasliquid-chromatography peaks corresponding to these derivatives of GA19 and GA1 were detected on QF-1 and SE-33 columns but their intensities were too weak to permit conclusive identification by GC-MS.
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References
Binks, R., MacMillan, J., Pryce, R. J.: Plant hormones. VIII. Combined gas chromatography-mass spectrometry of gibberellins A1 to A24 and their trimethylsilyl ethers. Phytochem. 8, 271–284 (1969).
Brian, P. W., Hemming, H. G., Lowe, D.: Comparative potency of nine gibberellins. Ann. Bot. 28, 369–389 (1964).
Cavell, B. D., MacMillan, J., Pryce, R. J., Sheppard, A. C.: Plant hormones-V. Thin-layer and gas-liquid chromatography of the gibberellins; direct identification of the gibberellins in a crude plant extract by gas-liquid chromatography. Phytochem. 6, 867–874 (1967).
Crozier, A., Aoki, H., Pharis, R. P.: Efficiency of countercurrent distribution, sephadex G-10 and silicic acid column chromatography in the purification and separation of gibberellin-like substances from plant tissue. J. exp. Bot. 20, 786–795 (1969).
— Audus, L. J.: Biological and chromatographic properties of two gibberellin-like compounds from etiolated Phaseolus multiflorus seedlings. Phytochem. 7, 1923–1931 (1968).
—— Distribution of gibberellin-like substances in light- and dark-grown seedlings of Phaseolus multiflorus. Planta (Berl.) 83, 207–217 (1968a).
— Kuo, C. C., Durley, R. C., Pharis, R. P.: The biological activities of 26 gibberellins in nine plant bioassays. Canad. J. Bot. 48, 867–877 (1970).
Durley, R. C.: Chemistry and biochemistry of plant gibberellins; quinones from Aspergillus melleus. Doct. dissect. Univ. of Bristol, England (1968).
Durley, R. C. MacMillan, J., Pryce, R. J.: Investigation of gibberellins and other growth substances in the seed of Phaseolus multiflorus and Phaseolus vulgaris by gas chromatography and by gas chromatography-mass spectrometry. Phytochem. (1971), in press.
jones, D. F.: Examination of the gibberellins of Zea mays and Phaseolus multiflorus using thin layer chromatography. Nature (Lond.) 202, 1309–1310 (1964).
Jones, R. L.: Aqueous extraction of gibberellins from peas. Planta (Berl.) 81, 97–105 (1968).
— Varner, J. E.: The bioassay of gibberellins. Planta (Berl.) 72, 155–161 (1967).
Kende, H., Lang, A.: Gibberellins and light inhibition of stem growth in peas. Plant Physiol. 39, 435–440 (1964).
Köhler, D., Lang, A.: Evidence for substances in higher plants interfering with response of dwarf peas to gibberellins. Plant Physiol. 38, 555–560 (1963).
MacMillan, J., Pryce, R. J.: Recent studies of endogenous plant growth substances using combined gas chromatography-mass spectrometry. In: Plant growth regulators. Soc. Chem. Ind., Monogr. No 31, p. 36–50 (1968).
Most, B. H.: A sugar spindle bioassay for gibberellins and its use in detecting diffusible gibberellins from sugarcane. In: Biochemistry and physiology of plant growth regulators (F. Wightman and G. Setterfield, eds.) p. 1619–1633. Ottawa: Runge Press.
Murakami, Y.: A new rice seedling bioassay for gibberellins, “Microdrop method” and its use for testing extracts of rice and morning glory. Bot. Mag. (Tokyo) 81, 33–43 (1968).
Musgrave, A., Kende, H.: Radioactive gibberellin A5 and its metabolism in dwarf pea. Plant Physiol. 45, 56–61 (1970).
Nitsch, J. P., Nitsch, C.: Studies of the growth of coleoptile and first internode sections. A new, sensitive, straight growth test for auxins. Plant Physiol. 31, 94–111 (1956).
Nitsch, J. P., Nitsch, C.: Activités comparées de neuf gibberellines sur trois tests biologiques. Ann. Physiol. vég. 4, 85–97 (1962).
Phinney, B. O., West, C. A.: Gibberellins and plant growth. Handbuch der Pflanzenphysiologie (W. Ruhland, ed.), vol. 14, p. 1184–1227. Berlin-Göttingen-Heidelberg: Springer 1961.
Powell, L. E., Tautvydas, K. J.: Chromatography of gibberellins on silica gel partition columns. Nature (Lond.) 213, 292–293 (1967).
Sembdner, G., Weiland, J., Aurich, O., Schreiber, K.: Isolation structure and metabolism of a gibberellin glucoside. In: Plant growth regulators. Soc. Chem. Ind. Monogr. No 31, p. 70–86 (1968).
Skene, K. G. M.: The gibberellins of developing bean seeds. J. exp. Bot. 21, 236–246 (1970).
Zeevaart, J. A. D.: Reduction in gibberellin content of Pharbitis seeds by CCC and after effects in the progeny. Plant Physiol. 51, 856–862 (1966).
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Supported by an S.R.C. Studentship
Supported by a NATO Grant.
Supported by NRC Grant A-5727.
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Crozier, A., Bowen, D.H., MacMillan, J. et al. Characterization of gibberellins from dark-grown Phaseolus coccineus seedlings by gas-liquid chromatography and combined gas chromatography-mass spectrometry. Planta 97, 142–154 (1971). https://doi.org/10.1007/BF00386762
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DOI: https://doi.org/10.1007/BF00386762