Endogenous gibberellins and inhibitors in caryopses of rye

Gibberellins A₈, A₁₆, A₂₄, and abscisic acid were identified by GC-MS of derivatized extracts from immature fruits of Secale cereale. Mature caryopses contained ABA and trans-ABA in a ratio 1:1 as well as 4′-dihydrophaseic acid. During milk ripeness a neutral GA conjugate was detected. Free GA, afforded by enzymatic hydrolysis of the conjugate, was chromatographically identified as GA₁₆     

The identification of gibberellins in immature seeds of Vicia faba,and some chemotaxonomic considerations

GA₁₇, GA₁₉, GA₂₀, GA₂₉, GA₄₄ and 13-hydroxy-GA₁₂, now named GA₅₃, were identified by GC-MS in immature seeds of Vicia faba (broad bean). Also identified were a GA catabolite, two polyhydroxykauranoic acids, and abscisic, phaseic and dihydrophaseic acids. The GAs of Vicia are hydroxylated at C-13, in common with those of other legumes. However the GAs of Vicia are not hydroxylated at C-3, nor do they appear to be readily conjugated. In these respects Vicia resembles Pisum, another member of the tribe Viciae. Vicia differs from Phaseolus and Vigna, of the tribe Phaseoleae, in both these respects.Abbreviations ABA abscisic acid - DPA dihydrophaseic acid - GA_n gibberellin A_n - GC gas chromatography - GC-MS gas chromatography mass spectrometry - KA kauranoic acid - PA phaseic acid - TLC thin layer chromatography     

The isolation of abscisic acid (ABA) deficient mutants by selection of induced revertants in non-germinating gibberellin sensitive lines of Arabidopsis thaliana (L.) heynh.

Summary By selecting for germinating seeds in the progeny of mutagen-treated non-germinating gibberellin responsive dwarf mutants of the ga–1 locus in Arabidopsis thaliana, germinating lines (revertants) could be isolated. About half of the revertants were homozygous recessive for a gene (aba), which probably regulates the presence of abscisic acid (ABA). Arguments for the function of this gene were obtained from lines homozygous recessive for this locus only, obtained by selection from the F₂ progeny of revertant X wild-type crosses. These lines are characterized by a reduced seed dormancy, symptoms of withering, increased transpiration and a lowered ABA content in developing and ripe seeds and leaves.Abbreviations ABA Abscisic acid - GA₄₊₇ Mixture of gibberellin A₄ and A₇ - EMS Ethylmethanesulfonate - NG Non-germinating - G Germinating     

Hormonal Response and Seed Viabil ity of Parteresia coarctata,a Brackish Water Species

Stimulation of α-amylase activity was observed in Porteresia coarctata immature seeds (20-day-old) when de-embryonated prewashed half seeds were incubated in media containing gibberellic acid (GA₃, 10^?5M). No such activity was observed in mature seeds even when GA₃ concentration was increased up to five fold. ABA suppressed the GA₃ enhanced α-amylase synthesis up to nearly 70% in the immature seeds. Absence of this enzyme activity in mature seeds may be due to high levels of ABA. The immature aleurone showed a 23 kD polypeptide induced by ABA.     

Influence of an inhibitor of gibberellin biosynthesis, paclobutrazol, on apple seed gibberellin content

Tissue-culture-propagated own-rooted cv. Spartan apple trees (Malus domestica Borkh.) planted in 1979 were treated in 1983 and 1985 via a soil-line trunk drench with the plant growth retardant paclobutrazol [(2RS, 3RS)-1-(4-chlorophenyl)-4.4-dimethyl-2-(1,2, 4-triazol-1-yl) pentan-3-ol]. Seeds of immature fruits from untreated and treated trees were sampled in 1989 ca 75 days after full bloom. After seeds were freeze-dried, gibberellins (GAs) were extracted, purified and fractionated via C₁₈ reversed-phase high-performance liquid chromatography (HPLC). Gibberellins A₁, A₃, A₄, A₇, A₈, A₉, A₁₅, A₁₇, A₁₉, A₂₀, A₂₄, A₃₄, A₃₅, A₄₄, A₅₁, A₅₃, A₅₄, A₆₁, A₆₂, A₆₃ and A₆₈ were identified by using C₁₈ HPLC, gas chromatography-selected ion monitoring and Kovats retention indices. Eight of the GAs identified were also quantified by using deuterated internal standards. The paclobutrazol applications caused a 55% reduction of vegetative shoot elongation in 1989, but both treated and untreated trees had developed a biennial bearing pattern by that time (heavy bloom or “on year’in 1989). Levels of early 13-hydroxylation pathway GAs, viz. GA₅₃, GA₁₉, GA₂₀, GA₁ and also GA₃, were not altered by treatment. However, GA₄, GA₇ and GA₉ were increased 13.4, 6.5 and 3.8 times, respectively, in seeds of fruit from treated compared to untreated trees.     

Interaction of jasmonic acid with some plant growth regulators in the control of apple (Malus domestica) embryo germination

Embryos isolated from dormant apple seeds were treated with jasmonic acid (JA), gibberellin A₃ (GA₃), abscisic acid (ABA) and hydrogen cyanide in darkness and in light. The chemicals were present in the culture medium continuously and simultaneously or applied for 2 days and in different sequences. All treatments stimulated embryo germination except ABA, which was strongly inhibitory. Additive effects of JA with light and with GA₃ on embryo germination were observed, whereas ABA interacted synergically with JA, HCN and light. ABA and GA₃ were most effective when applied early during embryo incubation, but the late JA treatment was more stimulatory. It is concluded that JA does not act on the regulatory pathway that is initiated by light and which leads to embryo germination through gibberellin accumulation and alkaline lipase activation. ABA and HCN appear to be involved in the control of this pathway. JA and ABA may be involved in the control of alkaline lipase activity, independently of this regulatory chain.Abbreviations ABA abscisic acid - GA₃ gibberellin A₃ - JA jasmonic acid     

Abscisic acid promotes lectin biosynthesis in developing and germinating rice embryos

Immature rice (Oryza sativa, L) embryos isolated about 12 days post anthesis are fully able to develop into young seedlings when cultured in vitro. Concomitantly, they rapidly loose their lectin synthesis activity. Abscisic acid added to the nutrient medium prevents precocious germination of the immature embryos and simultaneously strongly promotes lectin biosynthesis activity. Similarly, abscisic acid keeps mature embryos grown in a nutrient medium in a dormant state and maintains their lectin synthesis activity, whereas control embryos rapidly germinate but also quickly loose their lectin synthesis activity. It appears, therefore, that rice lectin is typically synthesized in embryos which are kept in a dornant state.Abbreviations ABA abscisic acid - GA₃ gibberellic acid - WGA wheat germ agglutinin - DPA days post anthesis     

Dormancy of <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis> seeds can be broken by different compounds

The influence of after-ripening, sodium nitroprusside, potassium ferricyanide, cyanide, paclobutrazol and nitrite on germination of seeds of Nicotiana benthamiana was investigated as well as the influence of plant hormones such as gibberellins and abscisic acid. Dormancy of N. benthamiana seeds was broken by all treatments except treatments with abscisic acid, paclobutrazol and gibberellic acid (GA₃). Gibberellins had an interesting effect on dormancy breakage of studied seeds which was dependent on use of particular gibberellin: GA₃ or GA₄₊₇. Unlike GA₃, GA₄₊₇ had broken seed dormancy.     

Free Radicals Developed in the Amino-Garbonyl Reaction of Sugars with Amino Acids

GA_l5, GA₃, GA₅, GA₁₉, GA₂₀ and GA₂₃ were identified by GC-MS in the acidic ethyl acetate-soluble fraction from the seeds of sweet potato (Ipomoea batatas Lam.). GA₁₉ and GA₂₃ were major GAs in the mature seeds, their contents being about 200 and 160 μg/kg fresh weight, respectively, while those of GA₁₉ and GA₂₃ in immature seeds were below 100 μg/kg fr. wt. The occurrence of glycosyl conjugates of GA₃, GA₅, GA₈, GA₁₇, GA₁₉, GA₂₀, GA₂₃ and GA₄₄ in the butanol fraction from mature seeds was shown by GC/MS analysis after enzymatic hydrolysis.

Besides the endogenous GAs in sweet potato, those in immature seeds of several other Convolvulaceae plants were investigated. The species of endogenous GAs were discussed in terms of chemotaxonomy.     

Identification of gibberellin A20, abscisic acid,and phaseic acid from flowering Bryophyllum daigremontianum by combined gas chromatography-mass spectrometry

Summary The presence of abscisic and phaseic acid in a purified acidic extract from flowering plants of the long-short-day plant Bryophyllum daigremontianum [(R. Hamet and Perr.) Berg.] was conclusively established by combined gas chromatography-mass spectrometry (GC-MS) of their methyl esters. Gibberellin A₂₀ (GA₂₀) was identified by GC-MS of the methyl ester and the trimethylsilyl ether of the methyl ester. The following levels of the 3 compounds per kg fresh weight were estimated: Abscisic acid, 5.5 g; phaseic acid, 9.4g; gibberellin A₂₀, 0.8 g. When GA₂₀ and four other GAs were applied to Bryophyllum under shortday conditions, the order of effectiveness for induction of flower formation was: GA₂>GA₁>GA₅=GA₇>GA₂₀. The low biological activity of the native GA₂₀ is discussed.     

Evidence that ethylene, light and abscisic acid interact to inhibit germination in the recalcitrant seeds of Quercus robur L.

The effects of ethylene, light and abscisic acid (ABA) on thegermination of prematurely-harvested and mature Q. robur seedswere studied. There was evidence of active biosynthesis of ethylene,but not of ABA in seeds from both harvests. Addition of ABA,the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid(ACC) or the ethylene-releasing compound Ethephon (E) to germinationmedia and exposure to light individually inhibited germinationand their effects were additive. The ethylene biosynthesis inhibitoraminoethoxyvinylglycine (AVG) tended to promote germinationof premature seeds only. Key words: Quercus robur, germination, ethylene, light, abscisic acid, recalcitrant seed     

Metabolism of Tritiated Gibberellin A(20) in Immature Seeds of Dwarf Pea, cv. Meteor

Tritium-labeled gibberellin A₂₀ ([³H]GA₂₀) 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 [³H]GA₂₀, the three in highest yield were GA₂₉, an α,β-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 [³H]GA₂₉ in both seeds and pods was highest at day 5 and declined to its lowest level at maturity. The amount of the [³H]ketone 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 GA₂₉, a 2β-hydroxylated GA of low biological activity, and that the ketone is not metabolized, or only slowly metabolized, during seed maturation.     

Effect of Abscisic Acid on Uptake and Metabolism of [H]Gibberellin A(1) and [H]Pseudogibberellin A(1) by Barley Half-seeds

Uptake and metabolism of 1,2-[³H]gibberellin A₁ ([³H]GA₁, I) and its 3-hydroxy epimer ([³H]pseudoGA₁, II) by barley (Hordeum vulgare L.) half-seeds were measured after 24 hours of incubation, in the presence or absence of abscisic acid in the media. Uptake of both compounds was enhanced by abscisic acid, and abscisic acid enhanced the extent of metabolism of [³H]GA₁. However, [³H]pseudoGA₁ was not metabolized, even in the presence of abscisic acid. The significance of the stereo-chemistry of the 3-hydroxyl position is discussed.     

The effect of seed drying and gibberellin treatment on the germination performance of developing carrot seed

In a succession of seed harvests of carrot, the highest percentage and most rapid germination was obtained from seed harvested 51 days after anthesis (DAA) when dried by conditioning at 25°C and 60% RH for one week and from seeeds harvested 65 or 79 DAA with or without conditioning treatment. Seed from these harvests had reached maximum weight when dried, had embryos of maximum length and were considered mature. The germinaton of seed from these treatments was unaffected by a mixture of the gibberellins A₄ and A₇ (GA_4/7) applied in the incubation medium. Seed harvested 37 DAA also gave maximum percentage germination when it was both conditioned and incubated in GA_4/7 solution. Seed harvested earlier than this germinated poorly. Germination times of both mature and immature seed were reduced after storage for 18 months but there was no response to GA_4/7.Abbreviations ABA abscisic acid - DAA days after anthesis - GA gibberellin     

Effects of Kinetin, Gibberellins and (±) Abscisic acid on the Germination of Lettuce (Lactuca sativa)

Germination responses of achenes of lettuce (Lactuca sativa L, cv. Arctic King) to treatment with kinetin, gibbe-rellins and abscisic acid were examined over a range of temperatures: in both light and dark. Kinetin (0.1–10 mg/l) strongly promoted germination at temperatures above 27±C in continuous light or after short periods of illumination during the early stages of imbibition. It also relieved the inhibitory affects of abscisic acid in these conditions. In total darkness however kinetin treatment resulted in only a minor promotive effect. Treatment with gibberellic acid (A₃) or a mixture of gibberellins A₄ and A₇ were much less effective in promoting germination at higher temperatures of lettuce achenes exposed to light but were strongly promotive in the dark.     

Synthetic Studies on Nephritogenic Glycosides. Synthesis of β-Glc-(1→6)-α-Glc-(l→6)-α-Glc-(1→Asn)

Gibberellins (GAs) A₉, A₁₅, A₁₉, A₂₀, A₂₉, A₃₅, A₄₄, A₅₀ and A₆₁ were identified by capillary gas chromatography/selected ion monitoring (GC/SIM) in immature seeds of loquat (Eriobotrya japonica Lindl). Furthermore, five unknown GA-like compounds with apparent parent ions of m/z 418, 504 or 506 (as methyl ester trimethylsilyl ether derivatives) were found by GC/mass spectrometry (GC/MS) in the biologically active fractions. The m/z 418 and 504 compounds may have been C-11β hydroxylated GA₉ and dehydro-GA₃₅, respectively. The bioassay and GC/MS results suggest that the major GAs were GA₅₀ and the five unknown GA-like compounds. In the immature seeds, at least two GA metabolic pathways may thus exist, one being the non-hydroxylation pathway of GA₁₅→GA₂₄→GA₉, and the other, the early C-13 hydroxylation pathway of GA₄₄→GA₁₉→GA₂₀→GA₂₉. A late C-11β hydroxylation pathway is also possible.     

Evidence for the accumulation of a germination inhibitor during progressive thermoinhibition of seeds of celery (Apium graveolens L.)

The germination of seeds of celery (Apium graveolens L.) becomes progressively thermoinhibited on incubation in the dark at high temperatures, the inhibitory temperature being dependent on the cultivar used. In two high-dormancy cultivars of celery, the production of germination inhibitors in seeds incubated in the dark at 26°C gradually increased over a 7-day period. Inhibitor production was measured by incubating seeds of the low-dormancy cultivar Florida 683 in homogenates of the thermoinhibited seeds of the high-dormancy cultivars and recording germination either in the light or with the gibberellins A₄ and A₇ (GA_4/7) in the dark. Most Florida 683 seeds which failed to germinate in the homogenates after 15 days were induced to germinate by addition of N⁶-benzyladenine (BA). The presence of BA in addition to GA_4/7 throughout incubation in the dark completely overcame the inhibitory effects of homogenates. This indicates that thermoinhibition of celery seeds is associated with the accumulation of a germination inhibitor which interacts with cytokinins. This does not appear to be abscisic acid (ABA) since ABA levels in thermoinhibited seeds were lower than in untreated seeds and did not increase with duration of high temperature treatment.Abbreviations ABA Abscisic acid - BA N⁶-benzyladenine - GA_4/7 a mixture of the gibberellins A₄ and A₇ - HTP high-temperature pretreatment     

A rapid, simple synthesis and purification of abscisic Acid glucose ester

A simple and rapid technique was developed to synthesize abscisic acid glucose ester. The free acid of abscisic acid (ABA) was combined with CsHCO₃ to form the Cs salt of ABA. The Cs salt of ABA was then combined with acetobromo-α-d-glucose tetraacetate, and the tetraacetate derivative of abscisic acid glucose ester was formed. Acetate groups were selectively removed from the glucose moiety with a crude enzyme preparation derived from Helianthus annuus seeds. Abscisic acid glucose ester was purified via silica gel column chromatography and identified by micro NMR.     

Metabolism of gibberellins by immature barley grain

Gibberellins A₁, A₄, A₉, A₁₂-aldehyde, A₂₀ and A₅₁, each labelled with both a radioactive and stable isotope were fed to immature barley grain by injection into the endosperm. After 7 d, extensive metabolism of all substrates had occurred, and metabolites were identified by combined capillary gas chromatography-mass spectrometry. A proposed scheme of gibberellin metabolism in immature barley grain is presented.Abbreviations GA_n gibberellin A_n - GC-MS combined gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography     

Biochemical Characteristics Associated with the Development of the Desiccation-sensitive Seeds of Machilus thunbergii Sieb. & Zucc.

Biochemical properties, i.e. endogenous abscisic acid, proline,sugars, respiration, adenosine phosphates and adenylate energycharge, and growth and moisture content were measured duringthe development of seeds of Machilus thunbergii. As dry matteraccumulated in the embryo during development, moisture content,ABA, proline, respiration and sugars all declined. At maturity,the dry mass of the seeds failed to attain a plateau beforethe period of natural seed shedding; the axis and cotyledonsreached moisture contents of 58 and 45%, respectively. Dryingof immature seeds at 73% relative humidity and 25 °C for30 d resulted in a complete loss of viability at all developmentalstages tested with the exception of mature seeds that were ableto tolerate a 5% decrease in moisture content before germinationdeclined. ABA was detected in all embryos tested, with a maximum value16.·16 µg g^-1 d. wt about midway through development.Although the presence of ABA induced no tolerance to desiccationof mature seeds, it did coincide with decreased content of waterin the developing seeds and decreased respiration. Desiccationdamage of M. thunbergii seeds occurred when moisture contentwas still high (45%) and this damage was not related to theabsence of oligosaccharides in the mature seeds. We concludethat developing embryos and mature seeds of M. thunbergii haveproperties common to many recalcitrant seeds, with seeds beingsensitive to desiccation at all stages, having a prominent ABApeak, little proline, lacking oligosaccharides, and specifically,little dormancy and a moderate rate of respiration of matureseeds (0·9 µmol O₂ min^-1 g^-1 f. wt). Adenosinetriphosphate content and energy charge decreased from stagefour to stage eight of seed development, then increased againto 103 nmol g^-1 d. wt and 0·73, respectively, in matureseeds. The moderate energy charge observed in mature seeds indicatesthat continuous metabolism is also a characteristic of recalcitrantseeds.Copyright 1995, 1999 Academic Press Machilus thunbergii, seed development, recalcitrant seed, abscisic acid, energy charge