Gibberellins inCitrus sinensis: A comparison between seeded and seedless varieties

The gibberellin (GA) content of the reproductive organs ofCitrus sinensis (L.) Osb., cv. Bianca Comuna and the seedless variety, Salustiana, were examined by combined gas chromatography-mass spectrometry (GC/MS) at different stages of development. Gibberellins A₁, A₂₀, and A₂₉ were identified in the reproductive buds of both cultivars 21 days prior to anthesis and in fruits 35 days after anthesis by comparison of their mass spectra and Kovats retention indices with those of standards. In addition, three uncharacterized isomers of GA₁ were detected, one in buds and two in fruits. The presence of GA₄ in both tissues, and of GA₈ in the reproductive buds, was indicated by the occurrence of characteristic ions at the expected retention times, although their spectra were too weak for full identification. Vegetative shoots of cv. Salustiana contained gibberellins A₁, A₁₉, A₂₀, and A₂₉, and the unidentified isomer of GA₁ present in reproductive buds. The presence of trace amounts of gibberellins A₈ and A₁₇ was also indicated. Although the two varieties did not differ qualitatively in the GAs present during flower and fruit development, the seedless variety contained slightly greater amounts. The concentrations of gibberellins A₁, A₄, and A₂₀ were determined by gas chromatography-selected ion monitoring (GC/SIM) throughout ovary development and early fruit growth. In both varieties, the maximum GA₁ concentration occurred at anthesis. Highest concentrations of gibberellins A₂₀ and A₄ were found in fruit 35 days after anthesis, although the GA₁ concentration at this stage remained low.     

Hormonal changes associated with fruit set and development in mandarins differing in their parthenocarpic ability

Satsuma [Citrus unshiu (Mak) Marc.] and Clementine [Citrus reticulata (Hort.) Ex. Tanaka, cv. Oroval] are two related species of seedless mandarins which differ in their tendency to set parthenocarpic fruits. Satsuma fruits naturally set parthenocarpically whereas Clementine mandarins show very low ability to set fruit in the absence of cross-pollination. The endogenous levels of gibberellins (GAs) and free and conjugated indole-acetic acid (IAA) and abscisic acid (ABA) throughout early stages of fruit development were investigated in seedless cultivars of both species. Analyses performed by full-scan combined gas chromatography-mass spectrometry (GC-MS) of extracts from ovaries at anthesis demonstrated the presence of GA₁₉, GA₂₀, GA₂₉, GA₁, GA₈, GA₃ and iso-GA₃ in Satsuma mandarin, whereas only GA₂₉, GA₃ and trace levels of GA₈ were detected in Clementine. At this developmental stage GA-like substances, as estimated by bioassay, reached their highest levels in Satsuma, while Clementine mandarins contained relatively lower levels. In both species the highest levels of free IAA were found at petal-fall stage at which time free ABA levels also peaked. Developing fruits of Clementine had higher amounts of both free IAA and ABA. In Satsuma, levels of conjugated IAA remained low throughout reproductive development whereas in Clementine they increased as the free form declined. In contrast, conjugated ABA was at low levels in Clementine but reached higher concentrations in Satsuma. These results suggest that in these mandarins the potential for setting parthenocarpic fruits is mainly influenced by the hormonal status of the fruit during the later stages of cell division and early stages of cell enlargement. Thus, the condition of low ability to set parthenocarpic fruits appears to be associated with lower levels of active GAs, lower capability to catabolize ABA to conjugated ABA and higher ability to conjugate IAA during this period.     

Gibberellins and parthenocarpic ability in developing ovaries of seedless mandarins

Satsuma (Citrus unshiu [Mak] Marc.) and Clementine (Citrus reticulata [Hort.] Ex. Tanaka, cv Oroval) are two species of seedless mandarins differing in their tendency to develop parthenocarpic fruits. Satsuma is a male-sterile cultivar that shows a high degree of natural parthenocarpy and a high fruit set. Seedless Clementine varieties are self-incompatible, and in the absence of cross-pollination show a very low ability to set fruit. The gibberellins (GAs) GA53, putative 17-OH-GA₅₃, GA₄₄, GA₁₇, GA₁₉, GA₂₀, GA₂₉, GA₁, 3-epi-GA₁, GA₈, GA₂₄, GA₉, and GA₄ have been identified from developing fruits of both species by full-scan combined gas chromatography-mass spectrometry. Using selected ion monitoring with [²H₂]- and [¹³C]-labeled internal standards, the levels of GA₅₃, GA₄₄, GA₁₉, GA₂₀, GA₁, GA₈, GA₄, and GA₉ were determined in developing ovaries at anthesis and 7 days before and after anthesis, from both species. Except for GA8, levels of the 13-hydroxy-GAs were higher in Satsuma than in Clementine, and these differences were more prominent for developing young fruits. At petal fall, Satsuma had, on a nanograms per gram dry weight basis, higher levels of GA₅₃ (10.4x), GA₄₄ (13.9x), GA₁₉ (3.0x), GA₂₀ (11.2x), and GA₁ (2.0x). By contrast, levels of GA₈ were always higher in Clementine, whereas levels of GA₄ did not differ greatly. Levels of GA₉ were very low in both species. At petal fall, fruitlets of Satsuma and Clementine contained 65 and 13 picograms of GA₁, respectively. At this time, the application of 25 micrograms of paclobutrazol to fruits increased fruit abscission in both varieties. This effect was reversed by the simultaneous applications of 1 microgram of GA₃. GA₃ alone improved the set in Clementine (13x), but had little influence on Satsuma. Thus, seedless fruits of the self-incompatible Clementine mandarin may not have adequate GA levels for fruit set. Collectively, these results suggest that endogenous GA content in developing ovaries is the limiting factor controlling the parthenocarpic development of the fruits.     

Gibberellic acid causes earlier flowering and synchronizes fruit ripening of coffee

The effect of 100 mgl^–1 gibberellic acid (GA₃) on flowering and fruit ripening synchrony, fruit set, fruit fresh weight, and vegetative growth were studied for different size classes of coffee (Coffea arabica L. cv. Guatemalan) flower buds. Flower buds that were > 4 mm, but not developed to the candle stage at the time of GA₃ treatment, reached anthesis 20 days earlier than the controls, and their development was independent of precipitation, unlike the controls. Fruit from buds that were treated with GA₃ at the candle stage showed earlier and more synchronous ripening than the control, although no differences in flowering were found during anthesis. Buds that were smaller than 4 mm at the time of treatment did not respond to GA₃ applications. Treatment with GA₃ did not affect fruit set, fresh weight of fruits, or vegetative shoot growth.     

The role of gibberellins A1 and A3 in fruit growth of Pisum sativum L. and the identification of gibberellins A4 and A7 in young seeds

Gibberellins A₁ and A₃ are the major physiologically active gibberellins (GAs) present in young fruit of pea (Pisum sativum L.). The relative importance of these GAs in controlling fruit growth and their biosynthetic origins were investigated in cv. Alaska. In addition, the non-13-hydroxylated active GAs, GA₄ and GA₇, were identified for the first time in young seeds harvested 4 d after anthesis, although they are minor components and are not expected to play major physiological roles. The GA₁ content is maximal in seeds and pods at 6 d after anthesis, the time of highest growth-rate of the pod (Garcia-Martinez et al. 1991, Planta 184: 53–60), whereas gibberellic acid (GA₃), which is present at high levels in seeds 4–8 d after anthesis, has very low abundance in pods. Gibberellins A₁₉, A₂₀ and A₂₉ are most concentrated in seeds at, or shortly after, anthesis and their abundance declines rapidly with development, concomitant with the sharp increase in GA₁ and GA₃ content. Application of GA₁ or GA₃ to the leaf subtending an emasculated flower stimulated parthenocarpic fruit development. Measurement of the GA content of the pods at 4 d after anthesis indicated that only 0.002–0.5% of the applied GA was transported to the fruit, depending on dose. There was a linear relationship between GA₁ content and pod weight up to about 2 ng · (g FW)⁻¹, whereas no such correlation existed for GA₃ content. The concentration of endogenous GA₁ in pods from pollinated ovaries is just sufficient to give the maximum growth response. It is concluded that GA₁, but not GA₃, controls pod growth in pea; GA₃ may be involved in early seed development. The distribution of GAs within the seeds at 4 d post anthesis was also investigated. Most of the GA₁, GA₈, GA₁₉, GA₂₀ and GA₂₉ was present in the testa, whereas GA₃ was distributed equally between testa and endosperm and GA₄ was localised mainly in the endosperm. Of the GAs analysed, only GA₃ and GA₂₀ were detected in the embryo. Metabolism experiments with intact tissues and cell-free fractions indicated compartmentation of GA biosynthesis within the seed. Using ¹⁴C-labelled GA₁₂, GA₉, 2,3-didehydroGA₉ and GA₂₀ as substrates, the testa was shown to contain 13-hydroxylase and 20-oxidase activities, the endosperm, 3β-hydroxylase and 20-oxidase activities. Both tissues also produced 16,17-dihydrodiols. However, GA₁ and GA₃ were not obtained as products and it is unlikely that they are formed via the early 13-hydroxylation pathway. [¹⁴C]gibberellin A₁₂, applied to the inside surface of pods in situ, was metabolised to GA₁₉, GA₂₀, GA₂₉, GA₂₉-catabolite, GA₈₁ and GA₉₇, but GA₁ was not detected. Gibberellin A₂₀ was metabolised by this tissue to GA₂₉ and GA₂₉-catabolite. Received: 23 July 1996 / Accepted: 2 September 1996     

Gibberellins in developing fruits of Pisum sativum cv. Alaska: Studies on their role in pod growth and seed development

Gibberellins A₁, A₈, A₂₀ and A₂₉ were identified by capillary gas chromatography-mass spectrometry in the pods and seeds from 5-d-old pollinated ovaries of pea (Pisum sativum cv. Alaska). These gibberellins were also identified in 4-d-old non-developing, parthenocarpic and pollinated ovaries. The level of gibberellin A₁ within these ovary types was correlated with pod size. Gibberellin A₁, applied to emasculated ovaries cultured in vitro, was three to five times more active than gibberellin A₂₀. Using pollinated ovary explants cultured in vitro, the effects of inhibitors of gibberellin biosynthesis on pod growth and seed development were examined. The inhibitors retarded pod growth during the first 7 d after anthesis, and this inhibition was reversed by simultaneous application of gibberellin A₃. In contrast, the inhibitors, when supplied to 4-d-old pollinated ovaries for 16 d, had little effect on seed fresh weight although they reduced the levels of endogenous gibberellins A₂₀ and A₂₉ in the enlarging seeds to almost zero. Paclobutrazol, which was one of the inhibitors used, is xylem-mobile and it efficiently reduced the level of seed gibberellins without being taken up into the seed. In intact fruits the pod may therefore be a source of precursors for gibberellin biosynthesis in the seed. Overall, the results indicate that gibberellin A₁, present in parthenocarpic and pollinated fruits early in development, regulates pod growth. In contrast the high levels of gibberellins A₂₀ and A₂₉, which accumulate during seed enlargement, appear to be unnecessary for normal seed development or for subsequent germination.Abbreviations GA_(a) gibberellin A_n - GC-MS combined gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - PFK perfluorokerosene - PVP polyvinylpyrrolidone     

Identification and quantitative analysis of gibberellins inCitrus

Nine gibberellins (GAs) have been identified from tissues of Valencia orange (Citrus sinensis Osbeck) using gas chromatography—mass spectrometry and gas chromatography-selected ion monitoring of high-performance liquid chromatography (HPLC)-fractionated extracts. These GAs are GA₁, GA₃, GA₈, GA₁₉, GA₂₀, GA₂₉, 3-epi-GA₁, 2-epi-GA₂₉, and iso-GA₃. Selected-ion monitoring and stable-isotope dilution assays have been used to estimate levels of some of these GAs in vegetative and reproductive tissues. GA₂₉ was found to be the most abundant GA measured. GA₁ was found in all samples examined, and there was always less 3-epi-GA₁ than GA₁. GA₂₀ was present in most extracts. Leaves of developing inflorescence shoots contained six times more GA₂₉ than did leaves of comparable vegetative shoots. Levels of GA₂₉ increased during the early stages of fruit development. GA₂₀ may be more abundant in growing fruitlets than in those about to abscise; however, there were no consistent differences in the relative amounts of the other GAs. No major differences were found between tissues of immature seeded and seedless fruit, and developing seeds did not contain high levels of any of the GAs measured. It is concluded that seed-produced GAs are not essential for normal fruit development in Valencia orange.     

Quantification of GA1, GA3, GA4, GA7, GA9, and GA20 in vegetative and male cone buds from juvenile and mature trees of Pinus radiata

The gibberellins GA₁, GA₃, GA₄, GA₇, GA₉ and GA₂₀ were quantified in vegetative and pollen cone buds of juvenile and mature trees of Pinus radiata by combined gas chromatography-mass spectrometry and selected ion monitoring (GC-MS-SIM) using deuterated GAs as internal standards. Higher levels of GA₇ and GA₉ and lower levels of GA₄ were detected in juvenile vegetative buds compared to mature buds, and there were no differences in relation to age for GA₁, GA₃ and GA₂₀. Conversely, when differences between vegetative and pollen cone buds from a mature tree were studied, the highest levels of GA₁ and GA₄ were found in pollen cone buds, similar levels of GA₃, GA₇ and GA₉ were observed in both, and ten fold lower levels of GA₂₀ were found in pollen cone buds as compared with vegetative buds. These results indicate a difference in GA metabolism in relation to both the tree age as well as the physiological status of buds: vegetative or reproductive in this conifer.     

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.     

The source of gibberellins in the parthenocarpic development of ovaries on topped pea plants

The role and source of gibberellins (GAs) involved in the development of parthenocarpic fruits of Pisum sativum L. has been investigated. Gibberellins applied to the leaf adjacent to an emasculated ovary induced parthenocarpic fruit development on intact plants. The application of gibberellic acid (GA₃) had to be done within 1 d of anthesis to be fully effective and the response was concentration-dependent. Gibberellin A₁ and GA₃ worked equally well and GA₂₀ was less efficient. [³H]Gibberellin A₁ applied to the leaf accumulated in the ovary and the accumulation was related to the growth response. These experiments show that GA applied to the leaf in high enough concentration is translocated to the ovary. Emasculated ovaries on decapitated pea plants develop without application of growth hormones. When [³H] GA₁ was applied to the leaf adjacent to the ovary a substantial amount of radioactivity accumulated in the growing shoot of intact plants. In decapitated plants, however, this radioactivity was mainly found in the ovary. There it caused growth proportional to the accumulation of CA₁. Application of LAB 150978, an inhibitor of GA biosynthesis, to decapitated plants inhibited parthenocarpic fruit development and this inhibition was counteracted by the application of GA₃ (either to the fruit, or the leaf adjacent to the ovary, or through the lower cut end of the stem). All evidence taken together supports the view that parthenocarpic pea fruit development on topped plants depends on the import of gibberellins or their precursors, probably from the vegetative aerial parts of the plant.Abbreviations FW flesh weight - GA_n gibberellin A_n - HPLC high-performance liquid chromatography     

Hormonal regulation of fruit set, parthenogenesis induction and fruit expansion in Japanese pear

The effects of applied gibberellins (GAs), GA₁, GA₃, GA₄ and GA₇ with a cytokinin, N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) and indole-3-acetic acid (IAA) on fruit set, parthenogenesis induction and fruit expansion of a number of Rosaceae species were assessed. These included Japanese pear cv. ‘Akibae’ (self-compatible) and cv. ‘Iwate yamanashi’ (a seedless cultivar). Other Rosaceae species (Pyrus communis, Chaenomeles sinensis, Cydonia oblonga, and Malus pumila) were also investigated. GA₄, GA₇ and CPPU are very effective in inducing parthenocarpic fruit growth, whereas GA₁, GA₃ and IAA, have no ability to induce parthenogenesis in Japanese pear. GA₄- and GA₇-induced parthenocarpic fruit tended to be smaller in size, higher in flesh hardness, and showed advanced fruit ripening in comparison to pollinated fruit and to parthenocarpic fruit induced by CPPU. GA₄- and GA₇-induced parthenocarpic fruit also had an increased pedicel length and fruit shape index and also showed a slight protrusion of the calyx end. CPPU, GA₄ and GA₇ alone or combination with uniconazole were also active in inducing parthenogenesis in three other Rosaceae species, although final fruit set was extremely low. GA₁ was essentially inactive in promoting fruit expansion unlike the other bioactive GAs, whose effectiveness in promoting fruit cell expansion was as follow: GA₄ ≈ GA₇ > GA₃ > GA₁.     

Delay of early fruitlet abscission by branch girdling in citrus coincides with previous increases in carbohydrate and gibberellin concentrations

Current evidence in citrus indicates that gibberellins (GAs) are main determinants of early fruit set while subsequent growth of developing fruits is mostly dependent upon carbohydrate availability. In this work, branch girdling performed at anthesis in Satsuma mandarin (Citrus unshiu (Mak.) Marc.) cv. Okitsu transitorily reduced early abscission rates (12–32 days after anthesis, DAA) delaying initially the process of natural fruitlet drop. The effects of girdling on growth, gibberellin (GA) and carbohydrate concentrations in developing ovaries and fruitlets were assessed during this initial growth stage (0–69 DAA). In girdled branches, abscission rate reduction was preceded by elevated concentrations of carbohydrate and GA in developing ovaries and fruitlets. Girdling at anthesis stimulated higher hexose (21 DAA) and starch (6–20 DAA) concentrations and also higher GA₁ (6 DAA), GA₁₉ (13–20 DAA) and GA₂₀ (6–20 DAA). The results established a relationship between the reduction of early abscission rates and higher concentrations of carbohydrates and GAs induced by girdling in developing fruitlets. These findings revealed that girdling certainly increased GA concentration and strongly suggested that its effect on early fruitlet abscission delay is likely mediated by both GA and carbohydrates.     

Strong synergistic effects of gibberellins with the synthetic cytokinin N-(2-chloro-4-pyridyl)-N-phenylurea on parthenocarpic fruit set and some other fruit characteristics of apple

The induction of parthenocarpic fruit set was investigated using the apple cvs. Golden Delicious and Jonagold. The gibberellins GA₃, GA₄, GA₅ and GA₇ and the synthetic phenylurea-type cytokinin CPPU (N-(2-chloro-4-pyridyl)-N-phenylurea), were applied alone and in combination to unpollinated flowers at the end of petal fall. Gibberellins induced only a marginal final set of parthenocarpic fruits. CPPU sprays were more effective, particularly in the first year. When applied in combination, CPPU and gibberellins had a positive synergistic effect on parthenocarpic fruit set and fruit size, but a negative effect on flower induction the next year. After CPPU + GA sprays, percent fruit set was similar, or greater, compared to natural pollinated trees. The parthenocarpic fruits induced by CPPU + GA had an increased length to diameter ratio. CPPU stimulated, and GA₄ and GA₇ reduced, the russeting of the parthenocarpic fruits. The internal quality of the fruits was hardly affected, but Ca-deficiency symptoms occurred more frequently in parthenocarpic fruits.     

Dormancy in Peach (Prunus persica L.) Flower Buds : I. Floral Morphogenesis and Endogenous Gibberellins at the End of the Dormancy Period

Flower buds of peach (Prunus persica L.) trees, cv Novedad de Cordoba (Argentina), were collected near the end of the dormant period and immediately before anthesis. After removal of scale leaves, morphological observations of representative buds, made on transverse and longitudinal microtome sections, showed that all verticils making up the flower are present in an undifferentiated form during the dormant period (June). Flower buds collected at the end of dormant period (August) showed additional growth and differentiation, at which time formation of two ovules was beginning in the unicarpelar gynoecium. Dehiscence of anthers had not yet occurred 10 days before full bloom, and the ovules were still developing. Free endogenous gibberellin (GA)-like substances were quantified by bioassay (Tan-ginbozu dwarf rice microdrop) after SiO₂ partition column chromatography, reversed phase C18-high performance liquid chromatography, and finally Nucleosil [N(CH₃)₂]high performance liquid chromatography. Bioactive fractions were then subjected to capillary gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM). Gibberellins A₁, A₃, and A₈ were tentatively identified in peach flower buds using GC-SIM and Kovat's retention indices, and relative amounts approximated by GC-SIM (2:8:6 for GA₁, GA₃, and GA₈, respectively). The highest concentration (330 nanograms per gram dry weight) of free GA₁/GA₃ was found in dormant buds (June) and diminished thereafter. The concentration free of GA₁/GA₃ did not increase immediately prior to bud break. However, high GA₁/GA₃ concentrations occurred during stages where rate of growth and cellular differentiation of (mainly fertile) verticils can be influenced.     

Induction of fruit set and development in pea ovary explants by gibberellic acid

The response of unpollinated ovary explants ofPisum sativum L. cv. Alaska No. 7 to several plant growth regulators and nutrients has been studied. Explants consisted of a segment of stem and an emasculated flower with or without the adjacent leaf. They were made on the day equivalent to anthesis and were cultured in a liquid medium. Growth regulators were applied either in the solution or directly to the ovaries. Giberellic acid (GA₃) in the presence of sucrose, but not indole-3-acetic acid or N⁶-(Δ²-isopentenyl)-adenine (2iP), induced fruit set and development of parthenocarpic fruits, the final length of these being a function of the intensity of the GA₃ treatment. The capacity of ovaries to respond fully to GA₃ was not lost after incubation of explants in water or 50 mM sucrose for 1 day and was similar in explants made between the day of anthesis and 3 days later. Limited growth was obtained with 100 mM sucrose alone but this effect was counteracted by 2′-isopropyl-4′-(trimethyl ammonium chloride)-5′-methylphenyl piperidine-1-carboxylate (AMO-1618). This inhibitor was ineffective when GA₃ was applied to the ovary. The development of the fruit was proportional to the length of the segment of stem up to 5 cm. The presence of the leaf in the explant enhanced the development of the fruit. These results indicate that a gibberellin is necessary for setting and development of fruits from cultured ovaries and that this effect depends on an appropriate source of nutrients. The course of development of parthenocarpic fruits on explants was similar to that of seeded fruits on the intact plant. The cultured pea ovary systemoffers convenient means to investigate the role of gibberellins and nutrients in fruit set and development.     

Transport and metabolism of exogenously applied gibberellins to Malus domestica Borkh. cv. Jonagold

The radio-labeled gibberellins GA₁, GA₃,GA₄, and GA₇ were applied to intact developing applefruits (Malus domestica Borkh. cv. Jonagold) during theperiod when GAs are suggested to inhibit flower bud induction for the followingyear. Radioactivity from these compounds was found to be transported intoadjacent tissues as there are pedicels and bourses (4%). Application topedicels, after removal of the fruits, enhanced the transport into adjacentbourses up to 11%. The bud-carrying lateral bourse shoots contained onlyminor amounts of radioactivity on average 0.4% in both cases. Theseexport rates were identical, 1 or 5 days after application.After application of the corresponding deuterium-labeled GAs and analyses bymass spectrometry the specific metabolization of GA₁ toGA₁ 13-O-glucoside and of GA₃ to GA₃13-O-glucoside was demonstrated. Additional metabolites of GA₁ andGA₃ were not detected. After fruit application of GA₃ theratio of GA₃ to GA₃ 13-O-glucoside was found to be 1:2 inthe fruit. Pedicel application led to ratios of 1:4 and 1:5, respectively, inthe pedicel and in the adjacent bourse. After the application of GA₄and GA₇, neither glucosylation products nor other GA-like metabolitescould be identified.This is the first report of the metabolism of GAs to GA 13-O-glucosides indeveloping apple fruits. The possible function of the GAs as a signal in flowerbud formation for the following year is discussed.     

The Structure of Gibberellin A23 and the Biological Properties of 3,13-Dihydroxy C20-Gibberellins

Two aldehydic C₂₀-gibberellins, L-2 and L-4, were isolated from the immature fruits of yellow lupine (Lupinus luteus L.). L-2 was shown to have the structure II and named gibberellin A₂₃. L-4 was identified as gibberellin A₁₉(VI). Two new C₂₀-gibberellins, tentatively called 3,13-dihydroxy GA₁₅(IV) and 13-hydroxy GA₁₅(VIII), were derived from gibberellins, A₂₃ and A₁₉, respectively. The biological activities of four 3,13-dihydroxy C₂₀-gibberellins-GA₁₈(I), GA₂₃(II), GA₂₈(III) and 3,13-dihydroxy GA₁₅(IV), which were isolated from the fruits except for 3,13-dihydroxy GA₁₅—were compared in six gibberellin bioassays.     

Identification,quantitation and distribution of gibberellins in fruits of Pisum sativum L. cv. Alaska during pod development

In addition to the previously-reported gibberellins: GA₁; GA₈, GA₂₀ and GA₂₉ (García-Martínez et al., 1987, Planta 170, 130–137), GA₃ and GA₁₉ were identified by combined gas chromatography-mass spectrometry in pods and ovules of 4-d-old pollinated pea (Pisum sativum cv. Alaska) ovaries. Pods contained additionally GA₁₇, GA₈₁ (2-hydroxy GA₂₀) and GA₂₉-catabolite. The concentrations of GA₁, GA₃, GA₈, GA₁₉, GA₂₀ and GA₂₉ were higher in the ovules than in the pod, although, with the exception of GA₃, the total content of these GAs in the pod exceeded that in the seeds. About 80% of the GA₃ content of the ovary was present in the seeds. The concentrations of GA₁₉ and GA₂₀ in pollinated ovaries remained fairly constant for the first 12 ds after an thesis, after which they increased sharply. In contrast, GA₁ and GA₃ concentrations were maximal at 7 d and 4–6 d, respectively, after anthesis, at about the time of maximum pod growth rate, and declined thereafter. Emasculated ovaries at anthesis contained GA₈, GA₁₉ and GA₂₀ at concentrations comparable with pollinated fruit, but they decreased rapidly. Gibberellins a₁ and A₃ were present in only trace amounts in emasculated ovaries at any stage. Parthenocarpic fruit, produced by decapitating plants immediately above an emasculated flower, or by treating such flowers with 2,4-dichlorophenoxyacetic acid or GA₇, contained GA₁₉ and GA₂₀ at similar concentrations to seeded fruit, but very low amounts of GA₁ and GA₃ Thus, it appears that the presence of fertilised ovules is necessary for the synthesis of these last two GAs. Mature leaves and leaf diffusates contained GA₁, GA₈, GA₁₉ and GA₂₀ as determined by combined gas chromatography-mass spectrometry using selected ion monitoring. This provides further evidence that vegetative tissues are a possible alternative source of GAs for fruit-set, particularly in decapitated plants.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - FW fresh weight - GA_n gibberellin A_n - GC-MS combined gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - KRI Kovats retention index - m/z mass to charge ratio We thank Mr M.J. Lewis for qualitative GC-MS analyses and Ms M.V. Cuthbert (LARS), R. Martinez Pardo and T. Sabater (IATA) for technical assistance. We are also grateful to Professor B.O. Phinney, University of California, Los Angeles, for gifts of [17-¹³C]GA₈ and -GA₂₉ and to Mr Paul Gaskin, University of Bristol, for the mass spectrum of GA₂₉-catabolite and for a sample of GA₈₁ The work in Spain was supported by Dirección General de Investigación Cientifica y Técnica (grant PB87-0402 to J.L.G.-M.). We also acknowledge the British Council and Ministerio de Educacion y Ciencia for travel grants through Accion Integrada Hispano-Britanica 56/142 (J.L.G.-M. and P.H.).     

Gibberellin Structure-Dependent Interaction between Gibberellins and Deoxygibberellin C in the Growth of Dwarf Maize Seedlings

The effects of 3-deoxygibberellin C (DGC) on the growth-promoting actions of gibberellins A₁, A₂, A₃, A₄, A₅, A₇, A₈, A₉, A₁₃, A₁₈, A₁₉, A₂₀, and A₂₃ (GAn) as well as 13-deoxygibberellin A₅ (deoxy-GA₅) were tested with seedlings of gibberellin-deficient dwarf mutants (d₂ and d₅) of maize (Zea mays L.). It was found that DGC promoted the actions of gibberellins having both C-1 double bond and C-3 axial hydroxyl group, and it inhibited the action of gibberellins having the saturated ring A and lacking the C-3 axial hydroxyl group, whereas it did not affect that of the ones having the hydroxyl group. The presence of C-2 double bond, as in GA₅ and deoxy-GA₅, diminished the inhibitory action of DGC. The DGC inhibition was alleviated by raising the doses of the relevant GAs, suggesting that it is a competitive inhibition. These results and the finding that the growth of normal maize and rice seedlings are inhibited by DGC indicate that GA₉, GA₁₉, GA₂₀ or other gibberellins having ring A of the same structure are involved in the growth of these plants as active form(s) or as intermediate(s) leading to the active form(s).     

Self‐pollination and parthenocarpic ability in developing ovaries of self‐incompatible Clementine mandarins (Citrus clementina)

This study aimed to determine if self‐pollination is needed to trigger facultative parthenocarpy in self‐incompatible Clementine mandarins (Citrus clementina Hort. ex Tan.). ‘Marisol’ and ‘Clemenules’ mandarins were selected, and self‐pollinated and un‐pollinated flowers from both cultivars were used for comparison. These mandarins are always seedless after self‐pollination and show high and low ability to develop substantial parthenocarpic fruits, respectively. The time‐course for pollen grain germination, tube growth and ovule abortion was analyzed as well as that for carbohydrates, active gibberellins (GA₁ and GA₄), auxin (IAA) and abscisic acid (ABA) content in the ovary. ‘Clemenules’ showed higher pollen grain germination, but pollen tube development was arrested in the upper style 9 days after pollination in both cultivars. Self‐pollination did not stimulate parthenocarpy, whereas both un‐pollinated and self‐pollinated ovaries set fruit regardless of the cultivar. On the other hand, ‘Marisol’ un‐pollinated flowers showed greater parthenocarpic ovary growth than ‘Clemenules’ un‐pollinated flowers, i.e. higher ovule abortion rate (+21%), higher fruit set (+44%) and higher fruit weight (+50%). Further, the greater parthenocarpic ability of ‘Marisol’ paralleled higher levels of GA₁ in the ovary (+34% at anthesis). ‘Marisol’ ovary also showed higher hexoses and starch mobilization, but lower ABA levels (?64% at anthesis). Self‐pollination did not modify carbohydrates or GA content in the ovary compared to un‐pollination. Results indicate that parthenocarpy in the Clementine mandarin is pollination‐independent with its ability to set depending on the ovary hormone levels. These findings suggest that parthenocarpy in fertile self‐incompatible mandarins is constitutively regulated.