Chromatography of Gibberellins on Silica Gel Partition Columns

Powell, Loyd E.; Tautvydas, Kestutis J.

doi:10.1038/213292a0

Cited by 78 publications

(30 citation statements)

References 8 publications

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“…), and heterozygous dwarf (D8/+), 960 g f.w.]. Each acidic EtOAc fraction from greenhouse-grown plants was further purified by partition chromatography on silicic acid (11). The fractions eluting from 30o to 65% EtOAc in n-hexane were combined and passed through a Sep-Pak C18 cartridge (Waters Associates), eluted with MeOH/H20 (4:1, vol/vol, 4 ml twice).…”

Section: Methodsmentioning

confidence: 99%

The dominant non-gibberellin-responding dwarf mutant ( D8 ) of maize accumulates native gibberellins

Fujioka

Yamane

Spray

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

137

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Section: Methodsmentioning

confidence: 99%

The dominant non-gibberellin-responding dwarf mutant ( D8 ) of maize accumulates native gibberellins

Fujioka

Yamane

Spray

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

137

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“…Plants were grown in a cooled greenhouse (21.5 C day, 15.5 C night) under natural photoperiod from September to December, 1973. Plant parts sampled are given in Table I. Batches of plant tissue, as described in Table I, were extracted in cold (<0 C) 80% methanol (10 g methanol to 1 g tissue) for GAs as described previously (3). Purification of the acidic, ethyl acetate-soluble fraction on Polyclar AT (PVP), an insoluble form of poly-N-vinylpyrrolidone (11), and charcoal-Celite columns (26) was followed by chromatography on Woelm silica gel partition columns (5,20) which were gradient-eluted using a four-chamber Varigrad system, chambers 1 to 4 being 50%, 65%, 85%, and 100% ethyl acetate in hexane (w/w), respectively. The 26 fractions collected were diluted serially over a wide range (1/150 to 1/3600 depending upon the dry weight and biological activity present) and bioassayed for GA-like activity in 0.5 al of 95% ethanol at each diluiton on 10 dwarf rice cv.…”

Section: Methodsmentioning

confidence: 99%

Analysis of Native Gibberellins in the Internode, Nodes, Leaves, and Inflorescence of Developing Avena Plants

et al. 1976

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The native gibbereilins (GAs) of various organs of the Avena plant were analyzed by bioassay and gas chromatography-mass spectrometry (GC-MS) after silicic acid partition column chromatography. The major GA of the inflorescence was identified as GA3 by GC-MS, and this GA also forms the major component of the nodes, p-i internode, and roots as determined by GLC or chromatography/bioassay. The inflorescence and nodes are the major sources of native GAs, the last two leaves, internode, and roots having dgnificantly lower amounts of GA-like substances. In the internode, less polar GAs predominated at the lag stage of development, whereas by the log and plateau stages, the more polar GAs increased significantly.Since less polar GAs are early in oxidative interconversion sequences, this fnding indicates sequential conversion to more polar and probably more active GAs, during log phase growth of the p-i internode.Excised stem segments of Avena plants are known to be very sensitive in their growth response to exogenously supplied gibberellins (GAs), especially GA, and GA3 (13,15). In contrast, they show no significant growth promotion by other plant hormones such as IAA, kinetin, and ABA (14). In light of this marked specificity of oat stem segments to GAs, especially the more oxidized GA, and GA3, it is germane to ask the following questions: Where in the Avena plant are the primary sources of native GAs that may be regulating internodal elongation in situ? Are these GAs produced mainly in the internodes, or are they derived from other parts of the plant? What are the predominant forms (i.e. degree of oxidation) of GAs found in the Avena plant during rapid internode elongation?Past studies on native GAs in other cereals are relevant to the present investigation. Radley and co-workers (12, 21, 22) have characterized GA1-and GA3-like substances in barley by means of TLC, fluorimetry, and bioassay. Faull et al. (9) reached similar conclusions for barley GAs using paper, TLC, GLC, paper electrophoresis, and bioassay. Murphy and Briggs (18) identified GA3 and GA7 in germinating barley by novel means using crystallization to constant specific radioactivity with unlabeled carrier GA methyl ester after initial methylation of the endogenous GA with '4C-diazomethane. By techniques they estimated GA3 levels to be 1.5 ng/grain, a level which correlates well with bioassay estimates. Faull et al. (10) showed that 11-day-old barley seedlings synthesized an A,-like GA de novo from 14C02, and that the labeled GA was metabolized within 12 hr, implying a high rate of turnover of GAs in the seedlings. Nicholls (19) found very high levels (15 to 48 Ag/g, dry weight) of GAs in barley inflorescences.In the present study native GAs of Avena nodes, internodes (at 3 stages of elongation), roots, leaves, and inflorescences are analyzed by silicic acid partition column chromatography, being quantitated by bioassay, and in certain cases, GLC. Identification of one GA was made by GC-MS.4 These analyses have allowed the questions pose...

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“…These procedures significantly reduced the dry weight of the acidic fraction. Gibberellin-like substances in the sample were separated on a silica gel partition column (8,37) (8). The more polar acidic GAs such as GA32 and GA39 would not be "seen" in our study should they be present since they can only be extracted from the acidic, aqueous phase with butanol.…”

Section: Methodsmentioning

confidence: 99%

Endogenous Gibberellins of Pine Pollen

Kamieńska

Pharis²

1975

Plant Physiol.

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Thie endogenous gibberellins (GAs) of pollen of Pinus attenuata, P. coulteri, and P. ponderosa were bioassayed at hour 0, 3, 13, 24, 48 and 72 of germination. Dormant pollen showed relatively high GA activity throughout the elution spectrum (i.e. ranging from relatively nonpolar to highly polar). The maximum GA activity was obtained at hour 13 in more polar regions and especially in the zone corresponding to GA3 (for P. attenuata estimated as 230 micrograms of GA3/kilogram pollen). It is probable that the "nonpolar"GAs present in high quantities in dormant pollen and in early stages of germination were converted to "more polar"GAs as germination progressed. The amount of all GAs decreased after hour 13 of germination and by hour 72 no GAs could be detected. Among the species tested P. attenuata showed the highest over-all GA activity.

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Chromatography of Gibberellins on Silica Gel Partition Columns

Cited by 78 publications

References 8 publications

The dominant non-gibberellin-responding dwarf mutant ( D8 ) of maize accumulates native gibberellins

The dominant non-gibberellin-responding dwarf mutant ( D8 ) of maize accumulates native gibberellins

Analysis of Native Gibberellins in the Internode, Nodes, Leaves, and Inflorescence of Developing Avena Plants

Endogenous Gibberellins of Pine Pollen

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