Effects of Exogenously Applied Gibberellins and Thidiazuron on Phytohormone Profiles of Long-Shoot Buds and Cone Gender Determination in Lodgepole Pine

In long-shoot buds of lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm), cone-bud initiation and gender differentiation occur in a site-specific manner: female cone buds are normally restricted to the distal portion, whereas male cone buds are located in the proximal portion. Application of a paste containing two plant growth regulators gibberellins A₄ + A₇ (GA_4/7) combined with thidiazuron (TDZ) to long-shoot buds prior to cone-bud gender determination altered endogenous phytohormone profiles and induced female cone-bud formation in the proximal portion of the long-shoot bud, where male cone buds normally occur. Induced cone clusters observed in the following spring were either entirely female or a mixture of both female and male cones. Endogenous phytohormones in the long-shoot bud tissues were quantified by the stable isotope dilution method using high performance liquid chromatography-electrospray ionization tandem mass spectrometry in multiple-reaction monitoring mode. Applied GA_4/7 + TDZ led to increased concentrations of endogenous zeatin-type cytokinins, that is, trans-zeatin riboside and dihydrozeatin riboside, whereas concentrations of abscisic acid (ABA) and its catabolite, ABA glucose ester, were decreased, all relative to control, in untreated long-shoot bud tissue. Concentrations of extractable GA₄ and GA₇ declined in long-shoot bud tissues over 4 weeks following treatment with exogenous GA_4/7. This study demonstrates that high levels of endogenous zeatin-type cytokinins, together with reduced levels of ABA, both induced by applied GA_4/7 + TDZ, are positively associated with an increased female cone-bud formation in long-shoot buds.     

Gibberellin A(4/7) and the Promotion of Flowering in Pinus radiata: Effects on Partitioning of Photoassimilate within the Bud during Primordia Diferentiation

Gibberellin A_4/7 mixture (GA_4/7), a highly effective promoter of early and enhanced flowering in the Pinaceae, caused a significant reallocation of dry matter and ¹⁴C-photosynthate within terminal buds of Pinus radiata D. Don within 8 days of hormone treatment. Treatment with GA_4/7 to terminal shoots of vigorous, potentially flowering mature grafted propagules reduced the flow of photoassimilated ¹⁴C and dry matter into the terminal bud as a whole, but significantly increased the dry matter and ¹⁴C allocated within the bud to developing long-shoot primordia (potential seed-cone buds). This was accomplished at the expense of the structural tissues, the apical dome region, and the vegetative branch buds. Although GA₃ caused a similar reallocation of dry matter within the terminal bud, it was significantly less effective than GA_4/7 thus appears to have, in addition to any nutrient diversion abilities, a distinct morphogenic function in sexual differentiation.     

Promotion of flowering in the Pinaceae by gibberellins

Significant female flowering of 6- to 11-year-old seedlings and grafted ramets of sexually mature scions of lodgepole pine (Pinus contorta Dougl.) was promoted by both topical and spray applications of a gibberellin (GA) A_4/7 mixture (1.6 to c. 5 mg per plant in total) during that period (June to September) when sexual differentiation of lateral primordia would be expected to take place. Girdling was used in most experiments to enhance the GA_4/7 effect, as was the auxin, naphthaleneacetic acid (NAA). Average frequency of flowering branches on treated plants over all experiments ranged from 27 to 59% (control ranged from 0 to 36%) and average number of female strobili was increased from 2- to 6-fold by growth regulator treatment, relative to controls. Within an experiment, clonal or family frequency of flowering for treated plants ranged from 11 to 67% (controls were 0 to 28%), and number of female strobili was increased from 2- to 14-fold by growth regulator treatment, relative to controls. Movement of the flowering stimulus from the point of application was apparent in several experiments, the response in adjacent branches being correlated positively with increasing dosage of GA_4/7. Significant male flowering occurred only in one experiment, girdling and GA_4/7 treatment being promotive factors. The use of spray applications of GA_4/7+ NAA is warranted to induce early and enhanced flowering in lodgepole pine seedlings and vegetative propagules for genetic improvement programs.     

Effect of Gibberellic Acid and Monophenols on Flowering of lmpatiens balsamina in Relation to the Number of Inductive and Non-inductive Photoperiodic Cycles

A single treatment of plants with GA₃ (gibberellic acid) is not adequate to cause induction under LD (long day: 24-h photo-period) condition, but its effect is added to the sub-threshold induction caused by one SD (short day: 8-h photoperiod) cycle. Floral bud initiation is hastened, and the number of floral buds and flowers per flowering plant increases in plants receiving a single treatment with the combination GA₃+ SA (salicylic acid) accompanying a single SD cycle. However, the increase on 10 replicate basis is more marked in plants receiving three treatments with the combination GA₃+β-N (β-naphthol) and five treatments with the combination GA₃+ SA accompanying six and 10 SD cycles, respectively. The number of floral buds and flowers decreases with an increase hi the number of SD cycles, but it is higher in plants treated with GA₃, SA or GA₃+β-N than in the water-treated controls. — Under long days, treatment of plants with the combinations GA₃+ SA or GA₃+β-N accelerates the initiation as well as increases the number of floral buds. While a minimum of five treatments with GA₃ or of 25 with SA or β-N alone is needed for floral bud initiation under a 24-h photoperiod, three treatments are adequate to induce floral buds with the combination GA₃+ SA or GA₃+β-N under continuous illumination. Ten or more treatments with these combinations under a 24-h photoperiod produce more flowers than the same treatments under an 8-h photoperiod.     

Gibberellins and bud break,vegetative shoot growth and flowering in Metrosideros collina cv. Tahiti

Applications of the growth promotive gibberellins (GAs) GA₄ and 2,2-dimethyl GA₄, and of C-16,17 endo-dihydro GA₅, which is known to promote flowering while inhibiting stem growth in the long-day grass Lolium temulentum, were made to micropropagated plants of Metrosideros collina cv. Tahiti, a highly ornamental cultivar with an intermittent flowering pattern. Gibberellin A₄ and 2,2-dimethyl GA₄ stimulated vegetative growth both in elongating shoots, and internodes of shoots developing from buds that were quiescent at the time of GA application. Abscission of the apices of expanding shoots, a feature of mature Metrosideros plants, was inhibited by these GAs, the rejuvenation of micropropagated plantlets being enhanced. However, C-16,17 endo-dihydro GA₅ differed from GA₄ and 2,2-dimethyl GA₄ by having no promotive effects on vegetative growth, and no inhibition of apical abscission. Notwithstanding this contrasting effect on vegetative growth, high doses of GA₄ or C-16,17 endo-dihydro GA₅ similarly reduced flowering on shoots to which either GA was applied. Reduced flowering in response to applied GAs is common in many woody angiosperms, and in this instance was probably the combined result of abortion of developing floral structures in quiescent buds, and a preferential inhibition of bud break for floral buds relative to vegetative buds, particularly by GA₄. Finally, both C-16,17 endo-dihydro GA₅ and GA₄ strongly inhibited bud break in this woody angiosperm, although GA₄ could initially stimulate bud break when applied to vegetative buds close to the expansion stage. The above findings, in toto, highlight the sensitivity of Metrosideros to both classes of GA in a variety of growth and development processes.     

Effect of Gibberellic Acid and Some Phenols on Flowering of Impatiens balsamina, a Qualitative Short-day Plant

GA₃ as well as SA (salicylic acid) and β-N (β-naphthol) induce floral buds in Impatiens balsamina under strictly non-inductive photoperiods. The floral bud initiation is accelerated when 1 mg/1 SA is used in combination with 100 mg/1 GA₃. 100 mg/1 GA₃+ 1 mg/1 SA and 100 mg/1 GA₃+ 100 mg/1 β-N increase the number of floral buds as compared with 100 mg/1 GA₃ alone.     

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.     

A Developmental Pattern of Flowering in Colored <Emphasis Type="Italic">Zantedeschia</Emphasis> spp.: Effects of Bud Position and Gibberellin

The restricted flowering of colored cultivars ofZantedeschia is a consequence of developmental constraints imposed by apical dominance of the primary bud on secondary buds in the tuber, and by the sympodial growth of individual shoots. GA₃ enhances flowering inZantedeschia by increasing the number of flowering shoots per tuber and inflorescences per shoot. The effects of gibberellin on the pattern of flowering and on the developmental fate of differentiated inflorescences along the tuber axis and individual shoot axes were studied in GA₃ and Uniconazole-treated tubers. Inflorescence primordia and fully developed (emerged) floral stems produced during tuber storage and the plant growth period were recorded. Days to flowering, percent of flowering shoots and floral stem length decreased basipetally along the shoot and tuber axes. GA₃ prolonged the flowering period and increased both the number of flowering shoots per tuber and the differentiated inflorescences per shoot. Activated buds were GA₃ responsive regardless of meristem size or age. Uniconazole did not inhibit inflorescence differentiation but inhibited floral stem elongation. The results suggest that GA₃ has a dual action in the flowering process: induction of inflorescence differentiation and promotion of floral stem elongation. The flowering pattern could be a result of a gradient in the distribution of endogenous factors involved in inflorescence differentialtion (possibly GAs) and in floral stem growth. This gradient along the tuber and shoot axes is probably controlled by apical dominance of the primary bud. Online publication: 7 April 2005     

Effect of Gibberellic Acid and Phosfon on the Critical Dark Period Reqnirement for Flowering of Impatiens balsamina cv. Rose

The critical dark period requirement for flowering of Impatiens balsamina L. cv. Rose, an obligate short day plant, is about 8.5 hours. While GA₃ completely substituted for the dark period requirement, Phosfon prolonged it to 9.5 hours. GA₃ hastened and Phosfon delayed the initiation of floral buds under all photoperiods. Floral buds opened into flowers only during 8 and 14 hour photoperiods in control and Phosfon-treated plants but during all photoperiods in GA₃-treated ones. The delay in floral bud initiation and flowering was correlated with shifting up of the node bearing the first floral bud and flower respectively. While GA₃ increased the numher of floral buds and flowers in all photoperiods except 8-hour, Phosfon increased their number in the 14-hour photoperiod only. The number of flowering plants decreased with increasing photoperiod regardless of GA₃ and Phosfon application. The effect of Phosfon was completely or partially overcome, depending upon the photoperiod, by simultaneous application of GA₃.     

Effects of Growth Regulators on Direct Flowering of Isolated Ginseng Buds <Emphasis Type="Italic">in vitro</Emphasis>

An in vitro method for obtaining gingseng inflorescences directly from explants of gingseng (Panax ginseng) is reported. Isolated shoot-buds of somatic embryo-derived plantlets ginseng were used as explants and incubated in B5 medium supplemented with 1 mg l⁻¹ benzyladenine (BA) and 1 mg l⁻¹ gibberellic acid (GA₃). About 15% of the buds flowered directly without developing vegetative organs. Cytokinin was found to be the key factor for inducing these isolated buds to proliferate and flower, but both these processes also occurred when benzyladenine (BA) was replaced by thidiazuron (TDZ). The optimal concentration of TDZ for obtaining the best ratios of bud proliferation and total flowering was 0.1 mg l⁻¹, while the highest number of vegetative shoots was obtained in medium supplemented with 1 mg l⁻¹ GA₃ and 0.1 mg l⁻¹ TDZ. The explant elongated abnormally in the presence of 10 mg l⁻¹ GA₃. Although a low concentration (1 mg l⁻¹) of NAA increased the bud proliferation ratio in the medium supplemented with 0.1 mg l⁻¹ TDZ and 1 mg l⁻¹ GA₃, a high concentration (5 mg l⁻¹) of NAA reduced the bud proliferation ratio and inhibited the flowering.     

Promotion of Flowering in the Pinaceae by Gibberellins

Flowering was significantly promoted in 4-year-old grafts of mature coastal Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) clones by exogenous gibberellins (GAs) A₄ and A₇ (as a mixture) applied alone and in combination with A₅ and A₉. Biweekly applications of 400 μg GA_4/7 per branch between late March and late June gave a 5-fold increase in ovulate and 3-fold increase in staminate strobilus production over untreated controls. ⁶N-benzyladenine and 2,3,5-triiodobenzoic acid applied in combination with GAs had no consistent effect on strobilus production. Non-destructive branch girdling, ineffective by itself as a cultural treatment, enhanced the GA benefit to flowering. Exogenous application of GA_4/7 is effective and appears to be a practical method for promotion of early and enhanced flowering in grafted Douglas-fir seed orchards.     

In vitro flowering and pod formation from cotyledons of groundnut (Arachis hypogaea L.)

Summary The response of groundnut cotyledons to the presence of various growth regulators in concentrations from 0.1 to 5 mg/l has been studied in detail using several genotypes of groundnut on two different media. Cotyledons with embryo axis, cultured on Blaydes' medium with cytokinins, produced shoots, in the axils of which 2–7 flower buds could be seen. The frequency of flower bud induction in general increased with increasing concentrations of cytokinins, the optimal levels being 3 mg/l of KN or 4 mg/l of BAP. Cotyledons without embryo axis, cultured on Blaydes' medium with BAP (0.5 mg/l), produced a cluster of flower buds directly, ranging in number from 8–28, without any vegetative growth. Excised embryo axes cultured on the same medium gave plantlets without flower buds. The growth regulators IAA, NAA, GA₃ and ABA failed to induce flower buds in independent treatments. However, lower concentrations of IAA and NAA in combination with cytokinins exerted a positive influence on flowering. The blooming of the flower buds was facilitated on media supplemented with low concentrations of cytokinins. Six percent of the induced flowers resulted in gynophore development and ultimately formed pods when cultured under complete dark conditions in modified MS medium supplemented with kinetin.     

Non-Reversibility of Gibberellin-Induced Inhibition of Regeneration in Begonia Leaves

With applied to the petioles of detached Begonia x cheimantha leaves before planting, Gibberellic acid (GA₃) inhibited the formation of adventitious buds and roots ill an apparently irreversible manner. Bud formation was entirely suppressed by 10^?6M and higher concentrations and a significant inhibition was still present at 10^?9M the lowest concentration tested. Root formation was not affected by GA₃ below 10^?7M and was possible even at 10^?4 M GA₃. Petiole elongation was stimulated by GA₃ with an optimum at 10^?5M. GA₃ also blocked the action of 6-benzyiamino-purine (BAP) and 1-naphthaleneacetic acid (NAA), compounds which are potent stimulators of bud and root formation, respectively. When applied simultaneously with GA₃ they were, at their optimal concentrations, devoid of any effect in counteracting or reversing the gibberellin-induced inhibitions. Abscisic acid and the growth retardants CCC and Phosfon also were unable to restore bud and root formation. In leaves initially treated with water or 10^?5M BAP, endogenous bud and root formation as well as BAP-induced bud formation were entirety suppressed when 10^?5M GA₃ was applied 8 days after the initial treatments. Even when delayed for 14 days GA₃ treatment inhibited BAP-induced bud formation, while treatment after 21 days bad little effect on bud and root formation. Development of pre-existing, visible bud primordia was not inhibited by GA₃. BAP and NAA competitively inhibited the action of GA₃ in petiole extension growth. The results are discussed in relation to results obtained in other plant systems. It is suggested that GA₃ acts by blocking of the organized cell divisions initiating the formation of bud and root primordia.     

Within-crown distribution of male and female cones on clonal Sitka spruce trees

The within-crown distribution of cones (strobili) was mapped on 48 mature grafts of Sitka spruce [Picea schensis (Bong.) Carr.]. The total number of cones per tree was increased by a mainstem injection of 20 mg GA_4/7 and 8 mm wide bark ring. The cones and buds were classified as either lateral or terminal on each individual branch. The distribution of lateral female, terminal female, lateral male and terminal male cones, showed a general progression from the upper distal to lower proximal regions of the tree crown in all 12 clones. Evidence for preferential allocation of cones to particular branch types is presented. The region of the tree with the greates number of cones varied with treatment and reflected differences in the sex ratio of the individual trees. There is evidence for a relationship between branch length, position within the crown and the type of cone produced.     

Culture of Shoot Apices of Theobroma cacao

Surface sterilized buds of young cocoa plants (Theobroma cacao L.) taken at particular stages of the flush cycle were placed in Linsmaier and Skoog agar medium supplemented with a range of growth regulators. Only buds taken at the 1–2 (dormant) stage of the flush cycle and treated with gibberellic acid (GA₃) alone and GA₃plus kinetin (KN) supplement showed bud opening. In liquid Linsmaier and Skoog medium buds isolated at the 1–2 stage also responded to GA₃ and KN. In this case addition of KN caused bud opening, while GA₃ either initiated bud opening only or opening followed by leaf expansion depending on the concentration of GA₃ supplied. Bud development was inhibited when ABA was included in the medium hut this was overcome by the presence of GA₃ but not KN. Since a hormonal supplement was required for any response from the excised buds, it is suggested that the intermittent growth of the shoot apex in the intact plant may be determined by hormonal stimuli derived from other parts of the plant. The findings also indicate that the bud apices could be maintained in culture for long periods which may provide a basis for the development of a micropropagation procedure for cocoa.     

Hormonal Profile in Vegetative and Floral Buds of Azalea: Levels of Polyamines, Gibberellins, and Cytokinins

The floral transition includes a complex system of factors that interact and involve various biochemical signals, including plant growth regulators. The physiological signals involved in the control of the floral transition have been sparsely studied and mainly in plant species whose genetics are poorly known. In this work, the role of polyamines, gibberellins, and cytokinins was investigated by analyzing their endogenous content in vegetative and floral buds of azalea. The results showed that there is a clear distinction between floral and vegetative buds with respect to the levels of these plant hormones, with floral buds containing higher amounts of conjugated polyamines, gibberellins (GAs) from the non-13-hydroxylation pathway (GA₉, GA₇, and GA₄), and cytokinins (particularly isopentenyl-type species), and vegetative buds containing higher amounts of free polyamines and gibberellins from the early 13-hydroxylation pathway and fewer cytokinins. In conclusion, there is a specific pattern of endogenous hormone profiles in both vegetative and floral bud development in azalea, which may be relevant for future research on the control of flowering by exogenous hormone applications.     

Promotion of Flowering in the Pinaceae by Gibberellins

Gibberellins (GAs) were effective in promoting flowering in sexually mature (45-year-old scions) grafts of loblolly pine (Pinus taeda L.). Seed-cone production was increased 12-fold in field-grown, grafted ramets by bi-weekly, May-September applications of 500 μg per branch of GA_4/7. Gibberellin A₃ was equally effective at 500 μg but not at 100 μg per branch, while GA₅ was ineffective at either concentration. A second study using potted, less-sexually mature (8-to 10-year-old scions) grafts gave a reduced level of seed cones in response to GAs. However, even on these younger grafts GA_4/7 was a significant promotive treatment, GA₃ being considerably less effective. Branch girdling, tested as an adjunct treatment, was ineffective. It is now apparent that exogenous applications of GA_4/7 are effective on a number of Pinaceae species, and their use to promote earlier and more abundant flowering in breeding orchards of grafted ramets for at least two species, loblolly pine and Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] is practical.     

Effects of external ABA,GA<Subscript>3</Subscript> and NAA on the tiller bud outgrowth of rice is related to changes in endogenous hormones

Hormones play an important role in regulating the growth of rice tiller buds. However, little is known about the hormonal changes that occur during tiller bud growth and the mechanism of hormonal regulation of tiller bud growth. Here, two rice cultivars, Yangdao 6 (Indica) and Nanjing 44 (Japonica), were used to investigate the changes in plant hormones during tiller bud growth and the mechanism that underlies the hormonal regulation of tiller bud growth. In the present study, panicles were removed after heading to stimulate the growth of dormant tiller buds located at the elongated upper internodes. At the same time, external abscisic acid (ABA), gibberellic acid (GA₃) and α-naphthalene acetic acid (NAA) were applied. The results demonstrated that auxin and cytokinin (CTK) play important and different roles in the regulation of tiller bud growth. Auxin in the nodes inhibits tiller bud growth, while CTK is transferred to the tiller buds to promote growth. The inhibitory effects of GA₃ and NAA on tiller bud growth are mainly due to the control of the indole-3-acetic acid (IAA) or CTK contents in plants. As opposed to auxin and CTK, the ABA contents in nodes and tiller buds remained unchanged before tiller bud growth after panicle removal. Meanwhile, external ABA application only slightly slowed the growth of the tiller buds, suggesting that ABA may not be a key regulator of tiller bud growth. These results indicate that auxin, CTK and ABA together likely play roles in the regulation of tiller bud growth.     

Flowering of Chrysanthemum under non-inductive long days by gibberellins and N6-benzyladenine

Summary The flowering and inflorescence development of Chrysanthemum morifolium cv. Pink Champagne under non-inductive long days was promoted by exogenous application of GA₅, GA₃, GA₄+GA₇ or GA₉ in combination with the cytokinin, BA. The combination of BA and GA₅ was highly effective, BA and GA₃ moderately effective. Applications of the GAs alone or BA alone also resulted in some flowering, with GA₃ and GA₅ being most effective. In general, the effects of GA and BA were synergistic, and the concentration of both growth substances was a limiting factor with regard to the number of plants flowering under long days. Only the concentration of GA was a limiting factor for inflorescence development, however. Simultaneous application of indole-3-acetic acid reduced inflorescence development in most treatments. Development to the stage of anthesis was in no case effected.Abbreviations BA N⁶-benzyladenine - GA gibberellin - IAA indole-3-acetic acid This research was supported by National Research Council of Canada Grant No. 2585.     

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.