Use of insoluble polyvinylpyrrolidone for purification of plant extracts and chromatography of plant hormones

Glenn, James L.; Kuo, C. C.; Durley, Richard C.; Pharis, Richard P.

doi:10.1016/s0031-9422(00)90013-x

Cited by 166 publications

(54 citation statements)

References 16 publications

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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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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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“…In our case the plants not only remained intact throughout the course of the experiment, but care was taken not to damage the seeds during application of [3H]GA20 and tests were subsequently conducted to demonstrate the viability of the seed. The labeled GA was not applied in doses which exceeded the endogenous levels of the seed (18) since we used GA20 of high specific activity, thus enabling the exogenous carrier GA20 to be applied in amounts (< 0.5,ug/pod-about 5 seeds p/ pod) much lower than the endogenous level of GA20 in the seed [6 ILg/seedI.…”

Section: Discussionmentioning

confidence: 99%

Metabolism of Tritiated Gibberellin A₂₀ in Immature Seeds of Dwarf Pea, cv. Meteor

Durley

Sassa²,

Pharis³

1979

Plant Physiol.

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Tritium-labeled gibberellin A20 (13HIGA2o) was applied via the pedicel to immature pods and seeds of dwarf peas and three harvests were made at days 5, 10, and 23 (mature) after application. Of the five metabolites of 13HIGA20, the three in highest yield were GA29, an a,fl-unsaturated ketone, and a compound (B), whose structure was only tentatively assigned.The metabolic sequence GA20 --GA29 -* compound B -* the ketone was indicated. The amount of 13HIGA29 in both seeds and pods was highest at day 5 and declined to its lowest level at maturity. The amount of the I3Hlketone in the seed increased with time to its highest level at maturity.It is suggested that compound B and the ketone represent the major pathway of catabolism of GA2,, a 2#-hydroxylated GA of low biological activity, and that the ketone is not metabolized, or only slowly metabolized, during seed maturation.The roles of gibberellins (GAs) in the growth of seeds and fruit have yet to be adequately resolved. Many GAs are present in substantially higher levels in immature seeds (4, 7) than in other parts of the plant. These high levels of GAs may be causally correlated with seed and fruit growth (1, 10) or may ensure that adequate levels are stored in the seed for use in growth processes during germination (1). Evidence for the latter role has been cast in doubt by recent work with 3H-GAs (cited in ref. 1).The major biologically active GA in immature pod and seed of pea is GA20 (7,8). In seeds of dwarf pea high levels of the less biologically active GA29 are present together with much smaller amounts of GA9, GA17, GAu, GA44, and GA51 (18).

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“…Young leaves were sampled, weighed and immediately frozen with liquid nitrogen and stored in deep freezer (for a period not exceeding 3 weeks) until extraction and determination of endogenous hormones. The methods adapted for extraction and separation of plant growth substances in leaves of gerbera treated plants and control was that described by [13] and [14] respectively.…”

Section: Endogenous Hormones (µG/gfw)mentioning

confidence: 99%

Studies on the Effect of Gamma, Laser Irradiation and Progesterone Treatments on Gerbera Leaves

Metwally¹

2015

EJB

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Abstract:The present study, we aimed to evaluate the effect of progesterone (prog.) concentrations (10, 20 and 30 ppm), gamma rays (γ) doses (100,150 and 250 rad), helium neon laser rays (He-Ne) exposure time (1.0, 5.5 and 11.5 min.) and argon laser rays (Ar) exposure time (1.0, 7.5 and 15.5 min.) and some interaction treatments between progesterone and irradiation treatments on biochemical constituents i.e. growth regulators and some enzymes activity in gerbera leaves. For this reason, at the greenhouse of the National Research Centre, Dokki, Giza, Egypt. Pot experiments were carried out two successive seasons of 2006 and 2007, and growth regulators (IAA, GA 3 , Cytok. and ABA), some enzymes activity in gerbera leaves (POD, CAT, Alk. phosph and Acid. phosph) and protein content were determined. The results showed that, low conc. of progesterone hormone, low exposure time of argon and He-Ne laser has a great effect on increasing POD activities and Cytokine content. Gamma rays at 150 rad increased ABA, POD, CAT and Alkaline and acid phosphatase, while 250 rad increased IAA and GAlike substances. The least value of alkaline phosphatase activity was recorded by progesterone treatments.

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Use of insoluble polyvinylpyrrolidone for purification of plant extracts and chromatography of plant hormones

Cited by 166 publications

References 16 publications

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

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

Metabolism of Tritiated Gibberellin A₂₀ in Immature Seeds of Dwarf Pea, cv. Meteor

Studies on the Effect of Gamma, Laser Irradiation and Progesterone Treatments on Gerbera Leaves

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Use of insoluble polyvinylpyrrolidone for purification of plant extracts and chromatography of plant hormones

Cited by 166 publications

References 16 publications

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

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

Metabolism of Tritiated Gibberellin A20 in Immature Seeds of Dwarf Pea, cv. Meteor

Studies on the Effect of Gamma, Laser Irradiation and Progesterone Treatments on Gerbera Leaves

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Metabolism of Tritiated Gibberellin A₂₀ in Immature Seeds of Dwarf Pea, cv. Meteor