Helminthosporic acid functions as an agonist for gibberellin receptor

Helminthosporol was isolated from a fungus, Helminthosporium sativum, as a natural plant growth regulator in 1963. It showed gibberellin-like bioactivity that stimulated the growth of the second leaf sheath of rice. After studying the structure–activity relationship between the compound and some synthesized analogs, it was found that helminthosporic acid (H-acid) has higher gibberellin-like activity and chemical stability than helminthosporol. In this study, we showed that (1) H-acid displays gibberellin-like activities not only in rice but also in Arabidopsis, (2) it regulates the expression of gibberellin-related genes, (3) it induces DELLA degradation through binding with a gibberellin receptor (GID1), and (4) it forms the GID1-(H-acid)-DELLA complex to transduce the gibberellin signal in the same manner as gibberellin. This work shows that the H-acid mode of action acts as an agonist for gibberellin receptor.     

Dormancy and Flowering Are Regulated by the Reciprocal Interaction Between Cytokinin and Gibberellin in <Emphasis Type="Italic">Zantedeschia</Emphasis>

Floral productivity of Zantedeschia is dependent on the conversion rate of buds to shoots, which is controlled by varying intensities of para- (apical dominance), endo- (dormancy), and ecodormancy. We present evidence of cross-talk between cytokinin and gibberellin in their complementary roles to alleviate bud dormancy and enhance flowering in a perennial geophyte. We assessed the impact of cytokinin and gibberellin, applied alone and in sequential combinations, on bud fate during three phases along the ontogeny of growth, which coincide with the progressive transition of buds from apical dominance to dormancy. Given that cytokinin can stimulate branching and gibberellin can induce flowering in Zantedeschia, we measured these phenotypic responses as parameters of bud commitment. The efficacy of cytokinin alone to stimulate branching declined with the transition to dormancy (phase 1 = 3.8 ± 0.2 shoots; phase 3 = 1.0 ± 0.3 shoots). To sustain branching during this transition, a sequential application of gibberellin was necessary. Gibberellin alone failed to stimulate branching. The efficacy of gibberellin alone to stimulate flowering diminished with the transition to dormancy. Any flowering during this transition occurred only after the sequential application of cytokinin. Cytokinin alone failed to stimulate flowering. Alleviating bud dormancy and enhancing flowering in Zantedeschia, achieved by the reciprocal cross-talk between cytokinin and gibberellin, contributes to the pool of evidence drawing common mechanisms between dormancy and flowering and may have commercial implications.     

Comparative effects of some Amo-1618 analogs on gibberellin production in Fusarium moniliforme and on growth in higher plants

Summary The plant growth retardant 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride (Amo-1618) and three analogs (Carvadan, XII, XIII) were tested for ability to inhibit gibberellin production in Fusarium moniliforme and to suppress stem elongation in three species of higher plants.Amo-1618 and compound XII were highly effective in suppressing GA production in Fusarium whereas Carvadan and XIII were inactive. These inactive analogs were not degraded to any appreciable extent by Fusarium cultures.All four compounds suppressed stem growth although the relative effectiveness varied with the species tested. Carvadan was most active in dwarfing Phaseolus vulgaris and Pharbitis nil; compound XII was most active in Helianthus annuus.The lack of correlation between the ability to inhibit gibberellin production in Fusarium and to cause dwarfed growth in higher plants is interpreted to indicate that enzymes involved in gibberellin biosynthesis in different organisms exhibit certain variations which make them more or less sensitive to inhibitors such as the four growth retardants tested.     

α-Amylase Production in the Endosperm flinders of Ungerminated Barley treated with Gibberellin The Relation with their Respiration

When the barley endosperm flinders are incubated with the gibberellin solution under aerobic condition, α-amylase is produced. But it is not clear whether this aerobic condition is necessary for the biosynthesis of α-amylase or for the absoption of gibberellin into the endosperm tissue. The authors found that the rate of the α-amylase production did not decrease when the incubation medium containing gibberellin was flung away after 48 hours incubation and then this incubation was continued with the fresh medium free from gibberellin. When, at the begining of this second incubation, DNP, KCN or chloramphenicol was added to the fresh medium or anaerobic condition was made, this α-amylase production was found to be inhibited to a considerable extent. As at this period (i.e. the first 48 hours incubation) gibberellin seems to be fully absorbed into the tissue, the authors estimated that the aerobic condition is necessary for this α-amylase production.     

Persistence of [14C] gibberellin A3 and [3H] gibberellin A1 in senescing,ethylene treated Citrus and tomato fruit

The fate of [¹⁴C] gibberellin A₃ and [³H] gibberellin A₁ was examined in senescing fruit of Shamouti orange (Citrus sinensis L. Osbeck) and tomato (Lycopersicon esculentum Mill.). Gibberellin A₃ was highly persistent in Citrus peel (t 1/2=18 days) and to a lesser degree in tomato (t 1/2=5.5 days). Ethylene and ethephon caused a slight enhancement of gibberellin A₃ metabolism in Citrus and tomato fruit, respectively. Gibberellin A₁ was metabolized by Citrus peel at a relatively high rate (t 1/2 < 24 h) and ethylene slightly reduced this rate. It is concluded that the ethylene-induced enhancement of senescence does not involve major effects on the deactivation of applied gibberellins.Abbreviations GA₃ gibberellin A₃ - GA₁ gibberellin A₁     

Growth substances and the relation between phenotype and genotype in Pisum sativum

Summary Reciprocal grafts between plants of the tall variety Alaska and the dwarf Progress No. 9 show that neither roots nor mature leaves determine shoot phenotype. It is demonstrated that differences in stem growth between the two varieties are essentially controlled by a single Mendelian factor, and the effect of this Le locus is not graft transmissible. Combined with published data for gibberellin content this confirms that the Le locus does not control shoot phenotype by regulating gibberellin synthesis. Growth of slender plants (Le la cry ^s ) and early growth of microcryptodwarfs (le la cry ^c lm) is not inhibited by AMO-1618 at concentrations which greatly reduce growth of tall plants. This is consistent with the suggestion that rapid growth in these varieties, in the absence of the inhibitory effect of La and Cry, is not dependent on endogenous gibberellin.The work was supported by United States Atomic Energy Commission Contract No AT (11-1)-1338 wilhe Dr. A.J.McComb was on leave from the University of Western Australia.     

Structure Elucidation of Gibberellin A32 and Its Acetonide

The structures of two gibberellin-like substances isolated from the immature seeds of Prunus persica, tentatively named PG–I and PG–II, were elucidated. PG–I was an ammonium salt of a novel gibberellin, ent-3α,10,12β,13,15α-pentahydroxy-20-norgibberella-l,16-diene-7,19-dioic-19,10-lactone (1), to which gibberellin number A₃₂ was allocated. PG–II was shown to be gibberellin A₃₂ acetonide (7), and concluded to be an artifact produced from gibberellin A₃₂ in the isolation process.     

Biochemical Studies on “Bakana” Fungus. Part 69. Synthesis of Substances related to Gibberellins

Degradation of gibberellin C afforded methyl 1,7-dimethyl-2α-hydroxy-8-oxo-7α-gibb-4a(4b)-ene-1,10-dicarboxylate, from which gibberellin C was reconstructed. Reactions of desoxogibberellin C are also described.     

Inhibition of growth by ancymidol and tetcyclacis in the gibberellin-deficientdwarf-5 mutant ofZea mays L. and its prevention by exogenous gibberellin

Leaf sheath length and shoot dry matter of the gibberellin-deficientdwarf-5 mutant ofZea mays L. were further reduced by micromolar concentrations of two putative gibberellin biosynthesis inhibitors, ancymidol [-cyclopropyl--(p-methoxyphenyl)-5-pyrimidine methyl alcohol] and tetcyclacis [5-(4-chlorophenyl)-3,4,5,9,10-pentaazatetracyclo-5,4,1,0^2,6,0^8,11-dodeca-3,9-diene]. Growth retardant action was prevented by the subsequent application of gibberellin (GA₄₊₇). Plants treated with both gibberellin and growth retardants were identical in all outward respects to those treated with gibberellin alone. Although thedwarf-5 mutant is blocked in the synthesis ofent-kaurene and does not contain detectable quantities of gibberellin, the above results are consistent with the interpretation that biologically active levels of endogenous gibberellin are present in the dwarf which can be decreased by biosynthesis inhibitors.Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the United States Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

Free and Bound Gibberellin Activities and Ent-kaurene Synthesis during Induction of Cold Hardiness in Black Locust Seedlings

The role of gibberellin in the development of cold hardiness in black locust (Robinia pseudoacacia L.) seedlings was investigated. Free and bound gibberellin activities were followed during induction of cold hardiness using ethyl acetate partitioning and pH variation, with subsequent paper chromatographic fractionation and gibberellin bioassay. While total gibberellin activity decreased during the induction of cold hardiness in black locust seedlings, no convincing evidence was found to support conversion of free gibberellin to a bound form. However, bound gibberellin activity did appear to be more stable than did free gibberellin activity during the final stages of cold hardening at freezing temperatures. Gibberellin synthesis was followed using ¹⁴C-mevalonate conversion to ent-kaurene in a cell-free extract of the tissue. Ent-kaurene synthesis decreased during cold hardening with no detectable synthesis in fully hardened seedlings. However, since growth cessation precedes development of cold hardiness, decreased gibberellin synthesis and corresponding trends in free and bound fractions might have been expected, and a cause and effect relationship is difficult to establish. Even so, a decline in one step in gibberellin synthesis and a greater stability of bound than free gibberelin activity are associated with induction of cold hardiness in black locust seedlings.     

Endogenous gibberellin transport and biosynthesis in relation to geotropic induction of excised sunflower shoot-tips

Summary Surgical experiments on Helianthus annuus and Phaseolus multiflorus seedlings involving the application of auxin and gibberellin to decapitated plants, suggested that internode extension growth occurs under the controlling influence of apically synthesised gibberellin rather than auxin. Studies were made of diffusible gibberellins from sunflower apical buds in relation to geotropic stimulation. Approximately ten times as much gibberellin was obtained from lower than from upper tissues of horizontal shoot-tips, whereas approximately equal quantities were obtained from the two halves of upright tips. Evidence was obtained suggesting both lateral transport of gibberellin in the young internode, and also enhanced gibberellin synthesis in buds maintained in a horizontal position during the collection of diffusible gibberellins into agar. The results are discussed in relation to current concepts of the role of auxin in geotropism.     

Increase in the gibberellin response by low temperature pretreatment of azuki bean epicotyls

Epicotyl segments cut from azuki bean (Vigna angularis) seedlings grown at 27 C for 5 days and then treated at 15 C for 2 days were compared with those cut from seedlings not receiving the 15-C treatment, for their response to auxin and auxin plus gibberellin. The segments cut from 15C-treated seedlings showed greater elongation than the segments cut from untreated seedlings both in the absence and in the presence of gibberellin. The 15-C treatment increased the elongation caused by auxin plus gibberellin to a greater extent than it did the elongation caused by auxin alone; the gibberellin response of epicotyl was increased by the treatment. Diffusates from leaves and buds inhibited both the elongation caused by auxin and that caused by auxin plus gibberellin. The diffusates inhibited the latter more strongly than the former; the gibberellin response was decreased by the diffusates. Leaves or buds appear to supply epicotyls with a substance which decreases the gibberellin response. The supply of this substance was found to be temperature dependent. The diffusates obtained at 15 C caused no inhibition, while those obtained at 27 C decreased the gibberellin response. The lack of the supply of this substance at 15 C may account for the increase in the gibberellin response by the 15-C treatment.     

Biochemical Studies on “Bakanae” Fungus

The production of gibberellin A₇ by Gibberella fujikuroi was studied by using newly devised assay method. Gibberellin A₇ increased at preferable temperature range between 32°C and 34°C at the controlled pH(6.0~7.5). The improved isolation process by using column chromatography composed of granular charcoal was found to be extremely convenient, because of its quick elution with satisfactory separation from gibberellic acid which is always accompanied by gibberellin A₇ in culture medium.     

Interactions between Abscisic Acid, Gibberellins and Cytokinins in Grand Rapids Lettuce Seeds Germination

Experiments with Grand Rapids lettuce seeds (Lactuca sativa L.) maintained in darkness or irradiated with red light have shown that the inhibition of germination induced by low concentrations of ABA (2, 4, 6 μM) could be overcome by gibberellins (GA₃ or GA₄). The same results were obtained, although to a lesser extent, under the influence of two out of the four cytokinins tested (K and BAP) for seeds maintained in darkness. To suppress the block induced by higher concentrations of ABA (for example 8 μM), it was necessary to apply a cytokinin (K, BAP, Z or 2iP) and a gibberellin (GA₄ or GA₃) simultaneously, or a cytokinin following a red light treatment. Experiments conducted in darkness in which ABA (8 μM) was applied together with a cytokinin (BAP) and a gibberellin (GA₄) showed that the gibberellin and the cytokinin played similar roles towards each other and towards ABA.     

THE ROLES OF PLANT HORMONES IN THE GROWTH OF THE COROLLA OF GAILLARDIA GRANDIFLORA (ASTERACEAE) RAY FLOWERS

Corolla elongation and the roles of plant hormones in this process in Gaillardia grandiflora Van Houtte ray flowers were examined. The sterile ray flowers elongated during a 2-day period, and corolla growth was accompanied by fresh and dry weight increases and epidermal cell elongation (greatest near the base of the corolla) but not by cell division. Corollas excised from young ray flowers were measured during treatment in vitro with solutions of plant growth regulators. They elongated in response to gibberellins and fusicoccin but did not respond to auxins, cytokinins, abscisic acid, ethylene, or inhibitors of ethylene biosynthesis. Sequential and simultaneous hormone applications indicated no additive or synergistic effects between hormones, but auxin did reduce gibberellin-promoted growth. Analyses of endogenous auxins showed no significant variation, and ethylene production decreased prior to elongation, while a 20-fold increase in endogenous gibberellin activity was observed just prior to rapid corolla elongation. It appears that corolla growth in Gaillardia is accomplished by an increase in gibberellin activity alone, that multiple hormone interactions are not important in the control of corolla growth, and that part of the mode of action of gibberellin is acid-induced growth.     

Enhancement by Low pH of Gibberellin Effects on Dormant Celery Seeds and Embryoless Half-Seeds of Barley

The action of gibberellins on celery (Apium graveolens L.) seed germination and the release of reducing sugars from barley (Hordeum vulgare L.) seed endosperm were enhanced by decreasing the pH of the incubation solution to below the pKa point. In most cases, low pH was obtained by mixing the solution with weak acids such as succinic acid 2.2 dimethylhydrazide (SADH), 2-chloroethylphosphonic acid (CEPA), or citric acid. However, lowering the pH of the gibberellin solution with strong acid (HCl) also increased markedly the activity of low concentrations of GA_4/7. The synergistic action of ethylenediaminetetraacetic acid (EDTA) with gibberellin was not dependent on the pH level of the incubation solution. The response of celery seeds to gibberellins was increased when their distal ends were removed: solution pH and EDTA had no effect on this response. The possible explanations of the synergism between low pH compounds and gibberellins are discussed.     

Control of tuberisation in potato by gibberellins and phytochrome B

Phytochrome B-deficient plants exhibit increased gibberellin (GA) levels or responsiveness, which may contribute to their elongated growth and reduced chlorophyll levels. We have investigated the effects of applications of gibberellic acid and an inhibitor of gibberellin biosynthesis, ancymidol, on wild-type and phytochrome B-antisense potato (Solanum tuberosum ssp. andigena) plants. The results showed that some phenotypes of the phytochrome B-antisense plants, i.e. increased stem length and reduced chlorophyll, can be mimicked by treating wild-type plants with gibberellic acid. However, another phenotype, i.e. tuberisation response in long days, is mimicked by application of a GA biosynthesis inhibitor ancymidol, thus appearing to be the result of a reduction in the gibberellin levels. A simple increase in gibberellin levels or sensitivity is, therefore, not sufficient to explain the phenotype of the antisense plants.     

Gibberellin Biosynthetic Pathway inGibberella fujikuroi:Evidence for a Gene Cluster

Differential screening of aGibberella fujikuroicDNA library was used to successfully clone and identify genes involved in the pathway of gibberellin biosynthesis. Several cDNA clones that hybridized preferentially to a cDNA probe prepared from mycelium induced for gibberellin production were isolated and characterized. The deduced amino acid sequences of two (identical) clones contained the conserved heme-binding motif of cytochrome P450 monooxygenases (FXXGXXXCXG). One of these cDNA fragments was used as a homologous probe for the screening of a genomic library. A hybridizing 6.7-kb genomicSalI fragment was cloned into pUC19. The sequencing of this clone revealed that a second cytochrome P450 monooxygenase gene was closely linked to the first one. Since at least four cytochrome P450 monooxygenase-catalyzed steps are involved in the synthesis of gibberellins, chromosome walking was performed to find a further gene of this family or other genes involved in gibberellin pathway. Next to the two P450 monooxygenase genes, a putative geranylgeranyl diphosphate synthase gene, the copalyl diphosphate synthase gene, which is the first specific gene of the gibberellin pathway, and a third P450 monooxygenase gene were identified. These results suggest that at least some of the genes involved in the biosynthesis of gibberellins are closely linked in a gene cluster inG. fujikuroi,as has been recently found for other “dispensable” pathways in fungi.     

Genetic variation in yield under hot ambient temperatures spotlights a role for cytokinin in protection of developing floral primordia

Unusually hot ambient temperatures (HAT) can cause pre‐anthesis abortion of flowers in many diverse species, limiting crop production. This limitation is becoming more substantial with climate change. Flower primordia of passion fruit (Passiflora edulis Sims) vines exposed to HAT summers, normally abort. Flower abortion can also be triggered by gibberellin application. We screened for, and identified a genotype capable of reaching anthesis during summer as well as controlled HAT conditions, and also more resistant to gibberellin. Leaves of this genotype contained higher levels of endogenous cytokinin. We investigated a possible connection between higher cytokinin levels and response to gibberellin. Indeed, the effects of gibberellin application were partially suppressed in plants pretreated with cytokinin. Can higher cytokinin levels protect flowers from aborting under HAT conditions? In passion fruit, flowers at a specific stage showed more resistance in response to HAT after cytokinin application. We further tested this hypothesis in Arabidopsis. Transgenic lines with high or low cytokinin levels and cytokinin applications to wild‐type plants supported a protective role for cytokinin on developing flowers exposed to HAT. Such findings may have important implications in future breeding programmes as well as field application of growth regulators.