Promotive and inhibitory effects of uniconazole and prohexadione on the sprouting of bulbils of Chinese yam,Dioscorea opposita

Gibberellin A₁ (GA₁), GA₃ and GA₄ inhibited the sprouting of nondormant bulbils of Chinese yam, Dioscorea opposita, where the effectiveness of the GAs was as follows: GA₄>GA₁+GA₃. Uniconazole and prohexadione, plant growth retardants, promoted the sprouting of half-dormant bulbils. By contrast, these retardants inhibited the sprouting of nondormant bulbils. Gibberellin A₃ (GA₃) and A₄ (GA₄) which were applied to the stems of the sprouted bulbils, promoted stem elongation, but GAs applied to the bulbous parts inhibited this process. The effectiveness of the GAs on stem elongation was as follows: GA₃+GA₄ for the promotion and GA₄ > GA₃ for the inhibition. Uniconazole applied to the stem inhibited the stem elongation of the sprouted bulbils. These results suggest the possible involvement of endogenous GAs in the induction and maintenance of bulbil dormancy of D. opposita, as well as in the bulbil sprouting and subsequent stem elongation.     

Effects of gibberellic acid,ethephon, and 1-aminocyclopropane-1-carboxylic acid on germination ofAmaranthus caudatus seeds inhibited by paclobutrazol

The specific inhibitor of gibberellin biosynthesis, (2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol (paclobutrazol), inhibited germination ofAmaranthus caudatus L. seeds. Addition of gibberellic acid (GA₃), 2-chloroethylphosphonic acid (ethephon), or 1-aminocyclopropane-1-carboxylic acid (ACC) effectively antagonized inhibition. Ethephon was found to be the most efficient antagonist. The transfer of seeds after 1 day's incubation in paclobutrazol to solutions of GA₃ or ethephon reversed the inhibition, the effect increasing with increasing concentration of GA₃ or ethephon. Seeds incubated in paclobutrazol for 5 days decreased sensitivity to GA₃ and ethephon.     

Changes in responsiveness to ethylene and gibberellin during corolla expansion ofIpomoea nil

Corolla expansion inIpomoea nil appears to be triggered by changes in gibberellin concentration and ethylene production during development. We investigated the role of responsiveness to GA and ethylene in corolla expansion. The effects of growth regulators applied in vitro were measured as a change in area of corolla segments from younger (15–17 mm) and older (18–20 mm) whole corollas. Applied gibberellic acid (GA₃) significantly (p < 0.05) promoted growth in the younger segments but was less effective in the older segments. Moreover, applications of the GA biosynthesis inhibitors, PP333 (paclobutrazol) AMO₁₆₁₈ (2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride), chlorocholine chloride, and tetcyclasis had little effect on younger segments but inhibited growth of older segments. The older corollas have apparently synthesized and accumulated enough GA-like substances to become less responsive to additional applied GA₃. The amount of growth induced by applied or endogenous GA depended on the amount of ethylene simultaneously produced in the tissue. The younger corollas rapidly produced ethylene from endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) and did not respond to applied ACC whereas the older corollas naturally produced much less ethylene and were significantly (p < 0.05) inhibited by applied ACC. When ethylene production was inhibited by applying aminoethoxyvinylglycine (AVG), growth was promoted in all segments. However, only the growth of the younger segments was further stimulated by simultaneously applied AVG and GA₃ over the GA₃ control. Thus the differential responses of segments from 15- to 20-mm long corollas to applied growth regulators reflect developmental changes in responsiveness of the developing corolla. The change in responsiveness is attributed in part to the changes in production of endogenous growth regulators and to the effect of one endogenous plant growth regulator (PGR) on the responsiveness of the corolla to another PGR.     

Growth Responses and Allocation of Assimilates of Rice Seedlings by Paclobutrazol and Gibberellin Treatment

Paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)methyl-4,4-dimethyl-2-(1h-1,2,4-trizol-1-yl)penten-3-ol] effectively decreased vegetative growth of rice (Oryza sativa L.) seedlings and increased the chlorophyll content. The number of veins in a leaf, the calculated number of stomata per leaf, and the length of guard cells were not altered by the paclobutrazol treatment, suggesting an effect on cell elongation. The allocation pattern of carbohydrates was changed by either gibberellin (GA) or paclobutrazol treatment. GA₃ induced more shoot growth and less accumulation of starch than the control and paclobutrazol-treated seedlings. Photosynthetic ability was not affected by either paclobutrazol or GA₃ treatment. Paclobutrazol-treated plants allocated a smaller amount of photosynthates for vegetative shoot growth and stored more as starch in the crowns than the control and GA₃-treated plants. The same starch degrading activity in the crown tissue of paclobutrazol-treated seedlings as in control plants suggests that the accumulated starch is utilized in a normal activity for growth including leaf emergence, tiller formation, and root production, resulting in improved seedling quality. Received May 30, 1996; accepted December 10, 1996     

Reversing the inhibitory effect of paclobutrazol on seed germination of Amaranthus paniculatus by GA3, ethephon or ACC

The germination of Amaranthus paniculatus seeds was inhibited by applying paclobutrazol, a specific inhibitor of gibberellin biosynthesis. This inhibition was markedly counteracted by gibberellin A₃ (GA₃), suggesting that endogenous gibberellins are required for germination in this species. The inhibitory effect of paclobutrazol was also overcome by ethephon (2-chloroethylphosphonic acid) or the precursor of ethylene biosynthesis, ACC (1-aminocyclopropane-l-carboxylic acid). Thus the physiological effect of gibberellin can be mimicked by ethylene released from ethephon or synthesised from exogenous ACC. It is suggested, that endogenous gibberellins are involved in germination of Amaranthus paniculatus seeds and that action of GA₃ can be substituted by ethylene.Abbreviations ACC 1-aminocyclopropane-l-carboxylic acid - AMO-1618 (2-isopropyl-5methyl-4-trimethylammoniumchloride)-phenyl-l-piperidinium-carboxylate - ancymidol -cyclopropyl--(4-methoxyphenyl)-5-pyrimidine methanol - chloromequat chloride (2-chloroethyl)trimethylammoniumchloride - ethephon 2-chloroethylphosphonic acid - GA gibberellin A₃ - paclobutrazol (2RS, 3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-lyl)pentan-3-ol - Phosphon D 2,4,dichlorobenzyl-tributhylphosphoniumchloride - tetcyclacis 5,(4-chlorophenyl)-3,4,5,9,10-pentaaza-tetracyclo)5,4,1,0,Z,6,0^8,11 dodeca-3,9-diene     

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.     

Germination of Tagetes minuta L. II. Role of gibberellins

The effect of the application of gibberellins to Tagetes minuta L. achenes (seeds) was determined at both 25°C, the optimal germination temperature, and 35°C, at which temperature the achenes are thermoinhibited. Both GA₃ and GA₄₊₇ accelerated germination at 25°C. Seed germination at 25°C was inhibited by paclobutrazol, but on subsequent application of GA₄₊₇ rapid germination was induced. Following application of GA₃ or GA₄₊₇ to thermoinhibited seeds, a significantly higher final germination percentage was observed than in the distilled water control. However, endogenous gibberellin levels in germinating and thermoinhibited seeds did not differ significantly.     

Gibberellin physiology of safflower: endogenous gibberellins and response to gibberellic acid

Endogenous gibberellins (GAs) were extracted from safflower (Carthamus tinctorius L.) stems and detected by capillary gas chromatography-mass spectrometry from which GA₁, GA₃, GA₁₉,, GA₂₀, GA₂₉, and probably, GA₄₄ were detected. The detection of these GAs suggests that the early 13-OH biosynthetic pathway is prevalent in safflower shoots. Deuterated GAs were used as internal standards and GA concentrations were determined in stems harvested at weekly intervals. GA₁ and GA₁₉ levels per stem increased but concentrations per gram dry weight decreased over time. GA₂₀ was only detected in young stem tissue.Gibberellic acid (GA₃) was also applied in field trials and both GA₃ and the GA biosynthetic inhibitor, paclobutrazol, were applied in growth chamber tests. GA₃ increased epidermal cell size, internode length, and increased internode cell number causing stem elongation. Conversely, paclobutrazol reduced stem height, internode and cell size, cell number and overall shoot weight. In field tests, GA₃ increased total stem weight, but decreased leaf weight, flower bud number and seed yield. Thus, GA₃ promoted vegetative growth at the expense of reproductive commitment. These studies collectively indicate a promotory role of GAs in the control of shoot growth in safflower, and are generally consistent with gibberellin studies of related crop plants. Author for correspondence     

The effects of gibberellic acid,fluridone, abscisic acid and paclobutrazol on anther culture of Brussels sprouts

Both gibberellic acid (GA₃) and fluridone, a non-specific inhibitor of ABA biosynthesis, promoted embryo production in anther cultures of Brussels sprouts cv. Hal, but not in cv. Gower. Abscisic acid (ABA) and the gibberellin-biosynthesis inhibitor paclobutrazol inhibited embryo production in both cultivars.     

Application of paclobutrazol and GA3 to adult peach trees: Effects on nutritional status and photosynthetic pigments

Soil applications of 1 or 2 g paclobutrazol (Pbz) and foliar sprays of 1000 mg L^-1 GA₃ to adult peach trees [Prunus persica (L.) Batsch] had opposite effects: vegetative growth was inhibited by paclobutrazol and promoted by GA₃, whereas mean fruit yield was greater in Pbz-treated trees and less in GA₃-sprayed trees. Compared to controls, differences in the concentration of different mineral nutrients occurred in leaves collected shortly after fruit harvest: in Pbz-treated trees, the concentration of N and K significantly decreased, whereas that of Mg and Mn increased. Leaves of trees sprayed with GA₃ had a significantly lower concentration of N and Ca and Mn, and a slightly greater concentration of K. In spite of these changes, element concentrations were within accepted ranges and therefore the nutritional status of peach trees did not change with the application of both growth regulators. Pbz did not alter the concentrations of chlorophylls, whereas GA₃ significantly reduced the concentrations of chlorophylls a and b. Neoxanthin, violaxanthin, antheraxanthin, lutein, zeaxanthin, and ß-carotene concentrations were unaffected by all treatments.Mention of trademark, warranty, proprietary product, or vendor does not constitute a guarantee by the C.S.I.C. (Ministry of Education, Spain) and does not imply its approval to the exclusion of other products or vendors that may be suitable.     

Stamens and gibberellin in the regulation of corolla pigmentation and growth in Petunia hybrida

Removal of stamens, or even of only the anthers, at an early stage of corolla development, before the start of main anthocyanin production, inhibited both growth and pigmentation of attached corollas of Petunia. When only one or two stamens were removed from one side, the inhibition was restricted to the corolla side adjacent to the detached stamens. Application of gibberellic acid (GA₃) substituted for the stamens in its effect on both growth and pigmentation. In detached corollas, isolated at the early-green stage and grown in vitro in sucrose medium, GA₃ promoted growth and was essential for anthocyanin synthesis. A marked enhancement of anthocyanin production was observed 48 h before the increase in corolla growth rate. Corollas detached at later stages were able to continue their growth and pigmentation in sucrose without GA₃. When Paclobutrazol (-[(4-chlorophenyl)-ethyl]-(1,1-dimethylethyl)-H-1,2,4-triazol-1-ethanol), an inhibitor of gibberellin biosynthesis, was added to the growth medium of in-vitro-grown corollas, pigmentation was inhibited but there was no effect on corolla growth. Low levels of GA₃ counteracted the Paclobutrazol effect on pigmentation but did not affect growth. The above results indicate that the effect of GA₃ (and probably that of the stamens) on corolla growth is independent of its effect on pigmentation. Gibberellic acid and paclobutrazol had no effect on [¹⁴C]sucrose uptake by in-vitro-grown corollas. The activity of phenylalanine ammonialyase was correlated with the effect of stamens and GA₃ on pigmentation in corollas grown in vivo and in vitro.Abbreviations GA gibberellin - GA₃ gibberellic acid - PAC Paclobutrazol - PAL phenylalanine ammonia-lyase     

Elicitation of secondary metabolism in Echinacea purpurea L. by gibberellic acid and triazoles

In vitro bioreactor production of Echinacea purpurea L. was used to facilitate the screening of compounds capable of eliciting increased secondary metabolite production. Based on previous experience with various bioreactors, the Southern Sun Liquid Lab Rocker was selected for this study as it produces healthy, vigorous, whole plants. The focus of the present study was to quantify the concentration of the medicinally important secondary metabolites caftaric acid and cichoric acid and to study the effect of growth regulators on the production of these compounds in this system. Both marker compounds were produced at levels that compared favorably to field grown plants. Application of gibberellic acid (GA₃), paclobutrazol, uniconazole and a combined treatment of GA₃ with paclobutrazol to the in vitro plants generally increased the concentration of caftaric acid in the roots with little effect on the concentration in the shoots. The concentration of cichoric acid was higher in the roots of treated plants than in the roots of control plants, but lower in corresponding shoots. The present study highlights the use of in vitro production of whole plants as a model system for studying the regulation of plant secondary metabolism in a controlled environment.     

The control of flower initiation by gibberellin in Helianthus annuus L. (sunflower), a non-photoperiodic plant

The involvement of gibberellins in the control of flowering of sunflower was studied by direct application of GA₃ to the apex of the plants, analysis of the endogenous levels of gibberellin-like substances at different plant ages, and indirectly by the application of paclobutrazol, an inhibitor of gibberellin synthesis. GA₃ speeded-up flower initiation and floral apex development. The time of GA₃ application was more critical than the amount of GA₃ applied. The endogenous levels of gibberellin-like compounds increased significantly by day 15 after sowing. The application of paclobutrazol markedly delayed floral initiation and this effect was also depedent on plant age. Both GA₃ and paclobutrazol had their greatest effects between 10 and 20 days after sowing suggesting that an increase in gibberellins in that time period plays a role in floral initiation.     

Promotion of asparagus shoot and root growth by growth retardants

Plantlets regenerated from shoot-tip culture of Asparagus officinalis L. possessed weak shoots and roots. Various combinations of auxins and cytokinins did not improve the plantlets. Incorporation of a number of growth retardants, viz. ancymidol, B-995, phosfon, Amo 1618, cycocel and paclobutrazol, promoted growth of stronger shoots and roots. The effectiveness of the growth retardants varied, with ancymidol being most effective and cycocel least effective.The response to ancymidol was prevented by exogenous GA₃ and GA_4/7. GA_1/3 and GA_4/7-like activities were detected in asparagus shoot-tip culture and these activities were reduced by the presence of the growth retardants ancymidol, Amo-1618, and cycocel.     

Different inhibitors of the gibberellin biosynthesis pathway elicit varied responses during in vitro culture of aspen (<Emphasis Type="Italic">Populus tremula</Emphasis> L.)

Several synthetic plant growth regulators (PGRs), including prohexadione-calcium (ProCa), paclobutrazol (PBZ), and chlormequat chloride (CCC), known for their ability to inhibit gibberellin (GA) biosynthesis, were investigated for their influence on Populus tremula L. (aspen) shoots grown in vitro. Changes in plant growth induced by these inhibitors were compared to the effects of exogenous gibberellins (GA₃ and GA_4/7). All PGRs were added to the nutrient medium at concentrations of either 1 or 5 μM. Stem segments with and without apical buds were excised from in vitro-grown shoot culture, and these explants were incubated either in test tubes or Petri dishes. In the presence of 5 μM ProCa, shoot growth and rooting were inhibited when grown in test tubes, while shoots grown in Petri dishes exhibited strongly enhanced shoot and root growth. PBZ suppressed shoot development both in test tubes and Petri dishes, although 1 μM PBZ promoted adventitious root formation when shoots were grown in test tubes. Five micromolars CCC suppressed shoot and root development in test tubes, but promoted shoot growth in Petri dishes.     

Necessity of gibberellin for stimulatory effect of KAR1 on germination of dormant Avena fatua L. caryopses

Avena fatua L. florets (caryopses enclosed by lemma and palea) were partially dormant at 10–20 °C and did not germinate at temperatures outside this range. After-ripening florets at 25 °C for 12 weeks completely removed dormancy. Caryopses (florets without lemma and palea) were able to germinate totally at 20 °C. Karrikinolide (KAR₁) and gibberellic acid (GA₃) applied at 10–25 °C partially or markedly induced germination of dormant florets and caryopses, respectively. Both florets and caryopses were more sensitive to KAR₁ than to GA₃. To obtain similar effects, 1,000 to 10,000 times lower concentrations of KAR₁ than GA₃ were required. After-ripening with time gradually increased sensitivity of caryopses to these regulators. Likewise, after-ripened, non-dormant caryopses were sensitive to KAR₁ and GA₃. Inhibitors of gibberellin biosynthesis, ancymidol, paclobutrazol and flurprimidol inhibited the effect of KAR₁. This inhibition was reversed by GA₃. Caryopses pre-incubated in water with ancymidol or paclobutrazol in the presence or absence of KAR₁ germinated completely but with different rates after transfer to GA₃. KAR₁ probably requires gibberellin biosynthesis to stimulate germination of dormant Avena fatua L. caryopses. Both KAR₁ and GA₃ increased α-amylase, β-amylase and dehydrogenases activities during imbibition before visible germination occurred.     

Growth and organogenesis in moth bean callus cultures as influenced by triazole growth regulators and gibberellic acid

The triazole plant growth regulators, paclobutrazol and uniconazole, reduced in vitro growth of moth bean callus by 20–25% when added to the culture medium at 1 mg/L (3.4 μM). The addition of 10 mg/L (29 μM) gibberellic acid (GA₃) to the culture medium in combination with the triazoles restored callus growth to a level equivalent to that of the untreated control. GA₃ alone had little effect on callus growth. When added to a regeneration medium at 1 mg/L both paclobutrazol and uniconazole reduced the percentage of cultures that formed roots, as well as the mean number of roots per culture. In contrast, GA₃ increased root formation and counteracted the inhibitory effects of the triazoles on rooting. The addition of triazoles or GA₃ to the regeneration medium reduced the formation of green meristematic nodules, which are precursors of shoots in moth bean callus. When callus was grown in the presence of either paclobutrazol or uniconazole, subsequent root and green meristematic nodule formation were reduced upon transfer to a growth regulator-free regeneration medium. The results of this study indicate that exposure of moth bean callus tissue to micromolar concentrations of triazoles or GA₃ can significantly alter in vitro growth and differentiation.     

Gibberellin content of immature apple seeds from paclobutrazol-treated trees over three seasons

Seeds from heavily fruiting (on-year), mature untreated, and paclobutrazol-treated apple trees (Malus domestica Borkh. cv. Spartan) were sampled in mid-June 1987, mid-July 1987, and mid-July 1990. After seeds were freeze-dried, gibberellins (GAs) were extracted, purified, and fractionated via C₁₈ reversed-phase high-performance liquid chromatography (HPLC). Nine GAs (GA₁, GA₃, GA₄, GA₇, GA₈, GA₉, GA₁₉, GA₂₀, and GA₅₃) were quantified by the use of deuterated GA internal standards. Paclobutrazol trunk drench treatments reduced vegetative shoot elongation in the seasons that seeds were sampled by 55% or more. Between June 17, 1987 and July 15, 1987, the dry weight of seeds from both untreated and treated trees increased about 2.5 times and there were reductions, on a per seed basis, of GA₄ in seeds from both untreated and treated trees, of GA₇ in seeds from treated trees, and of GA₉ in seeds from untreated trees. However, GA₉ increased in seeds from treated trees. Changes in levels of some of the early-13-hydroxylation pathway GAs (GA₁₅ GA₃, GA₈, GA₁₉, GA₂₀, and GA₅₃) also occurred during the month. For mid-July harvested seeds, the pattern, with some exceptions, was that 2 years after paclobutrazol treatment (1987), levels of early-13-hydroxylation pathway GAs in seeds from treated trees were lower compared to controls but after 5 years (1990) their levels tended to increase. For the non-13-hydroxylated GAs (GA₄, GA₇, and GA₉), 2 years after paclobutrazol treatment, GA₄ levels were equal in seeds from untreated and treated trees, GA₇ decreased in seeds from treated trees compared with controls, but GA₉ levels increased. Levels of these three GAs were higher in seeds from treated trees 5 years after treatment (1990) but levels of GA₄, GA₇, and GA₉ dramatically increased in seeds from treated trees 4 years after treatment (1989), as we previously reported.     

Responses of ivy (Hedera helix L.) to combinations of gibberellic acid,paclobutrazol and abscisic acid

A reduction in concentration of gibberellins has been implicated in the phase change from juvenile to mature forms of ivy (Hedera helix L.). Attempts were made to increase the effective internal concentration of gibberellins by exogenous application of GA₃, and to decrease them by various applications of abscisic acid (ABA) and paclobutrazol (PP333), alone or in combination with GA₃. ABA and GA₃ were fed directly into the xylem of ivy plants by a wick system (a less drastic procedure than the defoliation or decapitation used by earlier workers) whereas PP333 was applied as a soil drench.Mature ivy responded to the application of GA₃ by reversion to the juvenile form, although this reversion was incomplete with respect to leaf lobing and red (anthocyanin) pigmentation and could occur spontaneously without the application of GA₃. Contrary to expectation, applications of ABA and PP333 caused the stimulation of growth in juvenile ivy. No adult characteristics were induced. As similar concentrations of ABA and PP333 produced severe retardation of growth (which could be alleviated by the application of GA₃) in other species, it is suggested that ivy may be an unsuitable model system for the investigation of phase change in woody plants.     

The Response to Gibberellin in Pisum sativum Grown under Alternating Day and Night Temperature

The application of gibberellins (GA) reduces the difference in stem elongation observed under a low day (DT) and high night temperature (NT) combination (negative DIF) compared with the opposite regime, a high DT/low NT (positive DIF). The aim of this work was to investigate possible thermoperiodic effects on GA metabolism and tissue sensitivity to GA by comparing the response to exogenously applied GA (in particular, GA₁ and GA₃) in pea plants (Pisum sativum cv. Torsdag) grown under contrasting DIF. Control plants not treated with growth inhibitors or additional GA were 38% shorter under negative (DT/NT 13/21°C) than positive DIF (DT/NT 21/13°C) because of shorter internodes. Additional GA₁ or GA₃ decreased the difference between positive and negative DIF. In pea plants dwarfed with paclobutrazol, which inhibits GA biosynthesis at an early step, the response to GA₁ was reduced more strongly by negative compared with positive DIF than the response to GA₃. The induced stem elongation by GA₁₉ and GA₂₀ did not deviate significantly from the response to GA₁. Plants treated with prohexadione-calcium, an inhibitor of both the production and the inactivation of GA₁, grew equally tall under the two temperature regimes in response to both GA₁ and GA₃. We hypothesize that the reduced response to GA₁ compared with GA₃ in paclobutrazol-treated plants grown under negative DIF is caused by a higher rate of 2β-hydroxylation of GA₁ into GA₈ under negative than positive DIF. This contributes to lower levels of GA₁ and consequently shorter stems and internodes in pea plants grown under negative than positive DIF. Differences in tissue sensitivity to GA alone cannot account for this specific thermoperiodic effect on stem elongation. Received May 28, 1998; accepted May 29, 1998