Gibberellin-like Substances from the Developing Banana Fruit

The occurrence of 2 gibberellin-like substances was demonstrated in the developing banana fruit, Musa sapientum, Linn. Chemical and biological evidence led to the tentative identification of the 2 compounds as GA₇ and GA_x (previously isolated from citrus fruits). Support for such identification was obtained from thin layer chromatography, gradient elution column chromatography, spectrofluorometry, the dwarf maize test, and the cucumber hypocotyl test. Significance of the GA_x -designated compound increased since it is believed to occur in the fungus Fusarium moniliforme, Sheld. in addition to 2 different species of higher plants. It does not resemble any of the known gibberellins as far as chromatography is concerned.     

The Gibberellin-like Substances and Growth Inhibitors in Developing Strawberry Leaves

Gibberellin-like substances and inhibitors present in extractsof runner buds, and unfolding and mature leaves of strawberryhave been fractionated and compared. The runner-bud water-solubleresidue was more strongly inhibitory than that from unfoldingand mature leaves. The crude acid fraction from unfolding andmature leaves contained appreciable amounts of an inhibitor,probably abscisic acid, which was not detected in the buds.The unfolding leaves had a higher concentration of acidic gibberellin-likesubstances than either buds or mature leaves; they also containeda number of neutral gibberellin-like substances. Mature leaveshad a low concentration of gibberellin-like substances, allof which were acidic. A number of these were highly non-polartypes, not found in the buds or unfolding leaves.     

Alterations in Endogenous Levels of Gibberellin-like Substances during Germination of Phaseolus vulgaris Seeds

The endogenous gibberellin-like substances were determined in mature dry and germinating bean seeds, Phaseolus vulgaris L. cv. Alabaster. Methanol extracts were partitioned against ethyl acetate and butanol at neutral and acid pH. Each phase was individually chromatographed on a silica gel column. The gibberellin activity was measured with the Tan-ginbozu dwarf rice micro-drop bioassay. Each extract was tested in two dilutions. Extracts from dry seeds showed the highest gibberellin activity, largely attributable to ethyl acetate-soluble substances. The activity was considerably reduced in extracts from seeds imbibed for 1 day. Gibberellin-like substances soluble in butanol appeared in extracts from seeds soaked for 1 or more days.     

Changes in Levels of Naturally Occurring Gibberellin-like Substances during Germination of Seed of Lycopersicon esculentum Mill.

Naturally occurring gibberellin-like substances possessing acidic,basic, and neutral properties were detected, by paper partitionchromatography, in ethanolic extracts of tomato seed and ofetiolated seedlings after 72 and 116 hours' growth. Dwarf maizemutants of the d-1 and d-5 types, ‘Meteor’ pea seedlingsand young ‘Potentate’ tomato plants were used asbioassay material. Hydrolysis of seed and seedling proteinsby ficin in phosphate buffer, pH 6.2, after removal of ethanol-solublesubstances, liberated more and different ‘bound’gibberellin-like substances. It is suggested that protein hydrolysisduring germination is an important means of liberating thesesubstances at different stages of seedling development. Acidic substances were present in all the extracts prepared,but in general two with Rfs 0.25 and 0.55 in iso-propanol: ammonia:water : : 10:1:1 v/v were differentiated on d-2 and d-5 maizerespectively. Neutral substances in dry seed extracts chromatographedin the same solvent, had Rfs of 0.05, 0.35, and 0.95 and thesewere found only in the ethanolic (‘free’) extracts.They were active on d-1 and d-2 maize and ‘Meteor’pea. Basic gibberellin-like substances with Rfs of 0.05 and0.35 were found in ‘free’ extracts of both dry seedand etiolated seedlings after 116 hours' growth which were activeon d-2 maize only. Two others with Rfs 0.45 and 0.95 were extractedfrom seedlings after 72 hours' growth and these were activeon young ‘Potentate’ tomato plants. It is suggested that certain gibberellin-like substances, capableof reversing dwarfism in test plants, may be responsible formorphogenetic or other responses not involving stem extensionin the parent species. Changes were found in the levels of gibberellin-likesubstances but there was no evidence of changes in levels ofseed inhibitors relative to seed growth substances.     

Gibberellin-like Substances in Developing Barley Grain and Their Relation to Dry Weight Increase

Gibberellin-like activity of two varieties of barley (Hordeum vulgare L.) at different stages of grain development was determined by barley endosperm bioassay (acid phosphatase bioassay). Activity of the acidic ethyl-acetate fraction (“free” GAs) in both varieties displayed two maxima, a first maximum at the 9th day and a second one 20 to 21 days after pollination. Activity of the n-butanol fraction (“bound” GAs) first dropped to a minimum level at the 9th day, then increased to reach a maximum 32 days after pollination, and finally decreased again towards maturity. From the 9th day after pollination, a conversion of “free” GAs to “bound” GAs has probably occurred. From the 12th day after pollination, the curves of rate of dry weight increase and of “free” GAs run nearly parallel, but the latter reached its maximum about 2–4 days earlier than the former. The results indicate that gibberellins may participate in the regulation of the accumulation processes into the barley grain.     

Changes of Endogenous Gibberellin-like Substances with Sex Reversal of the Apical Inflorescence of Corn

In developing apical meristems of corn, the level of acidic, ethyl acetate-soluble gibberellin (GA)-like substances increased to a maximum of 108 micrograms GA₃-equivalents per kilogram dry weight of tissue at inflorescence initiation, and then fell rapidly. At anthesis, only a trace (0.2 microgram per kilogram) of GA-like activity remained in the apical (male) inflorescences, whereas moderate activity (32 micrograms per kilogram), mostly of a nonpolar nature, was present in lateral, female, inflorescences.     

Gibberellin-like Substances From Vegetative Tissue of a Conifer, Arizona Cypress

Vegetative shoots of Arizona cypress (Cupressus arizonica Greene) were found to contain an estimated 40 to 70 μg of gibberellin-like activity per kg. Based on elution patterns of silicic acid and celite partition columns, mobilities on thin layer chromatograms and specificity of the cucumber, d-3 dwarf maize, dwarf pea, and barley half seed bioassays it was possible to determine that the tissue contained at least 5 acidic, ethyl acetate-soluble gibberellin-like substances. The major one would appear to be GA₃. In addition, GA₉, GA₄, and/or GA₇-like compounds, and 2 unidentified gibberellin-like substances are present.     

Gibberellin-like Activity in the Developing Avocado Fruit

Gibberellin-like activity in avocado (Persea americana) fruit extracts was measured by the barley endosperm bioassay. Fruit tissues were anlyzed separately during fruit growth. The level of the activity found was very high in the endosperm and in the seed coats, but decreased in the latter during fruit growth. No measurable gibberellin-like activity was detected in the mesocarp or in the embryo. It is assumed that the seed coats are a site of production of gibberellin-like substances in the avocado fruit.     

Gibberellin-like Substances in the Transpiration Stream of Apple and Pear Trees

Gibberellin-like substances have been detected in sap exudingfrom decapitated apple and pear trees and also in the xylemsap sucked from stems of apple. The quantity of gibberellinin the sap appears sufficient to produce important effects onshoot development, and this result is discussed in relationto rootstock effects of fruit trees.     

Seed Number, Seed Size and Seed Diversity in Washington Lupin (Lupinus polyphyllus Lindl.)

Over 20 000 seeds of the Washington lupin (Lupinus polyphyllusLindl.) were examined and measured in an experiment carriedout over a 10 year period (1989–1999). Four differentgroups of Washington lupin seeds were found: dark, patterned,grey and light seeds. During the 10 year experiment, the totalaverage number of seeds per plant decreased from 2654 (1990)to 1220 (1999), there was a slight decrease in seed weight perplant and an increase in the average weight per seed. Therewas a clear seed size/number trade-off at the intraspecificlevel. The relative proportions of each seed group also changedwith patterned seeds becoming dominant (50% at the beginningand 90% at the end of the experiment), grey seeds remained constant(constituting 10% of the total seeds) and the proportion ofboth dark (33% at the beginning and 5% at the end) and light(10% at the beginning and 5% at the end of the experiment) seedsdecreased. Six hundred and sixty nine seeds were found to havea different testa ornamentation; they were distributed amongthe different groups as follows: 48% patterned seeds, 29% darkseeds, 12% grey seeds and 11% light seeds. There were no significantdifferences in the physical dimensions of the ornamented seedscompared with all other seeds. The results suggest that theWashington lupin is a mixture of different species and botanicalforms; this is discussed in relation to possible selection pressuresto produce both smaller and larger seeds. The possibility thatchanges in testa ornamentation are influenced by genes controllingthe synthesis of the seed coat pigment is discussed. Copyright2001 Annals of Botany Company Lupinus polyphyllus Lindl., lupine, plant biotechnology, plant parameters, selection pressure, seed size/number trade-off, testa ornamentation     

Gibberellin-like Substances in the Sporophores of Agaricus bisporus (Lange) Imbach

Sporophores of cultivated Agaricus bisporus (Lange) Imbach,were shown to contain a gibberellin-like substance active inthe dwarf maize (d-5), -amylase and other bioassays. Ethyl-acetateextraction followed by paper, column, and thin-layer chromatographyrevealed the presence of one major active substance. Ficin hydrolysisof dried sporophore powder, after the complete removal of freesubstances, released more gibberellin-like substances, one ofwhich appeared identical to the free compound. The free substance was predominantly in the lamellae and residualpileus tissue. The major active substance released by ficinoccurred mostly in the lamellae but also in substantial equalamounts in both stipes and pilei. No activity was found in extractsof dikaryotic vegetative mycelium on malt agar. The level ofactivity in extracts from sporophores stored at – 20 °Cfell sharply after 7 d, and then remained constant over a periodof 6 weeks. The content of gibberellin-like substances in youngand old whole sporophores showed wide variation between experiments.In most cases young 2-d tissue had higher levels than old, 11-dtissue on a fresh-weight basis. Purified sporophore extractsand authentic gibberellins had no stimulating effect on growthof sporophores or of cultured vegetative mycelium. The inhibitorsof diterpene biosynthesis, CCC, and AMO-1618 induced a smallincrease in mycelial growth rate. Ethyl-acetate extraction ofhorse-straw compost prior to inoculation with Agaricus bisporusshowed the presence of gibberellin-like activity in significantamounts.     

Gibberellin-like Substances in Immature Seeds of Guava (Psidium guajava L.)

From immature seeds of guava fruits, Psidium guajava, L. eightgibberellin-like substances designated X1, X2, X3, X4, X5, X6,X7, and X8 were isolated. Chemical and biological evidence ledto the tentative identification of X1, X2, X3, X4, X5, X6, andX7 as gibberellins A₆, A₅, A₁, A₃, A₇, A₉, and A₄ respectively.Support for such identification was obtained from paper chromatography,thin-layer chromatography, gradient elution column chromatography,and the lettuce hypocotyl test. The compound X8 has not beenidentified chemically.     

Changes in Endogenous Gibberellin-like Substances in the Vegetative Shoot and Inflorescences in Solanum sisymbrifolium Lam. in Relation to Potassium Content of the Plant

Several acidic gibberellin-like substances were detected inmethanolic extracts from the vegetative shoot and inflorescencesof S. sisymbrifolium by thin layer chromatography and two bioassays.Plants with a relatively low but not a deficient content ofpotassium were shown to contain higher levels of the GA-likesubstances in both tissues than those having a higher K content.Also qualitative changes in the spectrum of the biologicallyactive substances are affected by varying the K content of theplant. The changes in the GA-like substances are found to correlatewith the previously reported morphogenetic effects induced byvarying the K content.     

Studies on two Gibberellin-like Substances in Young Shoots of Tomato (Lycopersicon esculentum Mill.)

Two gibberellin-like substances were found in the acidic fractionof shoot extracts of the tomato (Lycopersicon esculentum Mill.,cultivar Potentate). These were resolved by paper chromotographywith iso-propanol/ammonia/water (10:1:1) as the developing solventbut not with n-butanol/1.5 N ammonia (3:1). Both substanceswere active in the dwarf maize bioassay on mutants d-1, d-2,d-3, and d-5, and appeared to be more active on d-5 than d-1.Neither was active in the Meteor Pea assay. Neutral and basicfractions were inactive. The relative amounts of these two substances varied accordingto the age of the tissues from which they were extracted andthis feature is discussed in relation to future studies on thephysiology of gibberellin-like substances in vivo.     

Auxins and Gibberellin-like Substances in Parthenocarpic and Non-parthenocarpic Syconia of Ficus carica L., cv. King

In the King cultivar of fig, the first crop is parthenocarpic, develops on previous year's growth, and a series of supernumerary ovules develops within the original ovules. The second crop, formed on current-season's growth, requires fertilization. To determine whether the 2 crops differed in types, and in patterns of concentrations of total `free' auxins and acidic gibberellins, they were extracted from weekly fruit samples. Timing of the 3 peaks of total auxins and the single peak of gibberellins was identical in the 2 crops. The first auxin peak in both occurred at the end of fruit growth period I (first rapid growth period), the second shortly before the end of period II (period of slow growth), and the rise and fall in concentrations of the third peak accompanied the rise and fall of the fruit growth rate in period III. The end of period II was marked by the single gibberellin peak. Additional peaks before the first sampling dates, of auxins in the first crop, of gibberellins in the second, were indicated by high concentrations in the first samples and subsequent rapid decline. The same 4 individual auxins appeared in both crops. Auxins I and II were highest in concentration in the first total auxin peak of both crops. In the second peak of the first crop, auxins II and III were highest, whereas in that peak of the second crop auxins II and IV were highest. Qualitative differences in gibberellins occurred in the 2 crops. In general, auxin concentrations were higher in the first than the second crop, and gibberellin concentrations higher in the second. High concentrations of gibberellins coincided with low ones of auxins, and vice versa.     

Indoleacetic Acid Levels in Phaseolus, Zea, and Pincus during Seed Germination

The content of indoleacetic acid (IAA) was determined in dry and germinating seeds of French bean (Phaseolus vulgaris), sugar maize (Zea mays), and Scots pine (Pinus silvestris). IAA was found in both the dry and the soaked seeds of the three species examined. The amount of IAA per gram fresh weight was extremely different in the three species whereas the variation between different harvests of the same species was small. Maize contained the highest level of IAA and bean the smallest. The time of imbibition was of decisive importance for the level of IAA. In all three species the content of IAA increased considerably during the initial 4 hours of swelling. The highest level of IAA was found in seeds that had swelled for 24 to 48 hours, during which period the radicles began to emerge from the seed coat. Later, during the period of rapid root growth, the content of IAA declined.     

Changes in Abscisic Acid Content in Axis and Cotyledons of Developing Phaseolus vulgaris Embryos and their Physiological Consequences

Prevost, I. and Le Page–Degivry, M. Th. 1985. Changesin absicisic acid content in axis and cotyledons of developingPhaseolus vulgaris embryos and their physiological consequences.—J.exp. Bot. 36: 1900–1905.Changes in abscisic acid (ABA)content with time were measured in embryonic axes and in cotyledonsof Phaseolus vulgaris embryos using a radio–immunoassay.During embryogenesis, a similar pattern was observed in bothtissues: ABA increased to a maximum 29 d after an thesis, followedby a decrease as the seed matured. The level of ABA in the cotyledonswas always much higher than that in the axes. In in vitro cultures,the duration of the lag phase before germination of isolatedembryonic axes increased with ABA content. The presence of cotyledonsalways lengthened the lag phase; longer lag phases were associatedwith greater concentrations of ABA in the cotyledons. Moreoverthe presence of cotyledons stimulated the growth of seedlings. Key words: ABA distribution, embryo maturation, axis and embryo germinability     

Efficiency of Countercurrent Distribution, Sephadex G-10, and Silicic Acid Partition Chromatography in the Purification and Separation of Gibberellin-like Substances from Plant Tissue

The effectiveness of countercurrent distribution, Sephadex G-10column chromatography and silicic acid partition column chromatographyin the purification of gibberellin-like substances from extractsof etiolated Phaseoltu multiflorus seedlings, and elongatingvegetative shoots of Arizona cypress (Cupressus arizonica Greene)and coastal Douglas-fir (Pseudotsuga menziesii var. menziesii)was followed by the use of the barley half-seed -amylase bioassayand dry-weight measurements. Countercurrent distribution resultedin a 10- to 30-fold reduction in the dry weight of the acidic,ethyl acetate-soluble fraction. Sephadex G-10 column chromatographyfurther lowered the dry weight by about two-thirds. Silicicacid partition column chromatography separated gibberellin-likesubstances from each other and again reduced the dry weight.Enhancement of bioassay activity was noted at each step of thepurification procedure. It is concluded that the above proceduresconstitute useful and efficient tools for the initial purificationof gibberellin-like substances from plant tissue.