Interaction between Pfr and growth substances in the germination of light-requiring Kalanchoë seeds

Seeds of Kalanchoë blossfeldiana Poelln. cv. Feucrblüte, incubated on gibberellic acid, become very light-sensitive through a synergism between the far-red absorbing form of phytochrome and the growth substance, which results in high physiological activity of short far-red (FR) exposures. On 2 × 10^-3 M gibberellic acid (GA₃), one saturating FR pulse is somewhat more effective than one saturating red light (R) irradiation. Fluence-response curves for R and FR confirm this observation. At lower GA₃ concentrations, this difference disappears and the effects of one saturating R and FR pulse decrease in an identical way with the GA₃ concentration. When two saturating irradiations, separated by 24 h are given, the effect of FR falls off faster than that of R at low GA₃ concentrations. Consequently, the second irradiation must have a different impact in comparison with the first one. Of the other growth substances tested, only a mixture of gibberellins A4 and A₇ had an analogous, still more pronounced effect than GA₃. Abscisic acid (ABA) inhibits the phytochrome-mediated germination of Kalanchoë , both in the absence and presence of GA₃. An antagonism between ABA and GA₃ was demonstrated.     

Interactions between hydrogen cyanide, gibberellin, abscisic acid and red light in germination of lettuce seeds

Germination of lettuce ( Lactuce sativa L. cy. Grand Rapids) seeds was promoted by red light and by pulse treatments with gibbercllie acid (GA₃) or hydrogen cyanide, whereas it was inhibited by short exposure of seeds to absusic acid (ABA). The eflects of unsalLirating red light and of 10 μ M GA₃ on lettuce germination were completely reversed the effect of ABA (100 μ M ). In contrast, hydrogen cyanide did not reverse the effect of 100 μ M ABA and only partly eliminated the effect of 10μ M ABA, independently of the sequence of treatments. Possible interactions between HCN GA₃, ABA and red light were discussed. It was concluded thai light GA₃ and HCN affect different mechanisms involved in lettuce germination: ABA counteracts the stimulatory action of all these faclors. being the most effective against cvanide Additional key words - Lactuca sativa, pholodormancy, phylohormoncs.     

Gibberellic acid decreases anthocyanin accumulation in wild carrot cell suspension cultures but does not alter 3'-nucleotidase activity

Gibberellic acid (GA₃) applied at different times during the growth of wild carrot ( Daucus carota ssp. Carota ) cell suspension cultures inhibited anthocyanin accumulation. Application of 3 × 10^–6 M GA₃ to cultures on day 0 or day 4 gave, respectively, 10 or 35% of anthocyanin accumulation relative to levels occurring when GA₃ was applied at the end of the growth period. Endogenous GAs were separated by high pressure liquid chromatography, and identified and quantified by gas chromatography-selected ion monitoring. Gibberellins GA₁, GA₃ and traces of GA₈. GA₁₉ and GA₂₀ were identified in carrot cell suspension cultures of both high and low anthocyanin-accumulating clones. The concentrations of GA₁. GA₃ and GA₈ in the two clones were similar and were not significantly different after the application of uniconazole which promoted anthocyanin accumulation. This suggests that these endogenous GAs are not the sole factors controlling the accumulation of anthocyanin in these different clones. Exogenous GA₃ and uniconazole had no effect on 3'-nucleotidase and 5'-nucleotidase activity in the carrot cell suspension cultures. Thus 3'-nucleotidase does not appear to play a role in the inhibition of anthocyanin accumulation by exogenous GA₃.     

Effect of plant growth hormones and polyamines on ornithine decarboxylase activity during the germination of barley seeds

Gibberellic acid (GA₃) and β-indolylacetic acid (IAA), two of the well known growth hormones, induce four fold the activity of ornithine decarboxylase (ODC) during the germination of barley seeds ( Hordeum vulgare L. var. Beca). The optimal concentration for induction of ODC was 10^–5 M for GA₃ and 10^–3 M for IAA. When 10^–3 M of a polyamine, putrescine or spermidine, is added to the growth medium, ODC activity is significantly inhibited. This inhibition is due to the induction of a protein inhibitor of ODC (antizyme), whose apparent molecular weight is 16 000 ± 2 000 daltons. Addition of GA₃ to cultures which have been grown for 50 or 98 h in the presence of polyamines, abolishes the observed inhibition of ODC activity, while in the reverse experiment, addition of polyamines at 50 or 98 h does not affect the ODC activity induced by GA₃. Cadaverine, a physiological plant diamine, enhances ODC activity; whereas 1,8-diaminooctane (the alkyl analogue of spermidine) does not have any effect.     

Effects of the triazole plant growth retardant BAS 111¨W on gibberellin levels in oilseed rape, Brassica napus

Three-week-old shoots of the spring oilseed rape cv. Petranova ( Brassica napus L. ssp. napus ) were found by combined gas chromatography-mass spectrometry to contain GA₁, GA₈, GA₁₅, GA₁₇, GA₁₉, GA₂₀, GA₂₄, GA₂₉, 3-epi-GA₁ and a previously uncharacterised C₁₉ dicarboxylic acid that is probably structurally related to GA₂₄. Shoots of the winter cultivar Belinda, harvested at the early flowering stage, contained the same GAs with the exception of the C₁₉ dicarboxylic acid and, in addition, GA₃₄ and GA₅₁ were identified. All material contained higher levels of GA₂₀ than of GA₁; the ratio of GA₁ to GA₂₀ was highest in shoots containing the largest proportion of young immature tissues. Soil treatment of cv. Petranova seedlings with the growth retardant BAS 111¨W [1-phenoxy-5,5-dimethyl-3-(1,2,4-triazol-1-yl)-hexan-4-ol] caused 80% reduction in height 18 days after treatment and the levels of all GAs were 20% or less that of control plants. Foliar treatment at the same dosage reduced height by 50% and caused an 85% or greater reduction in the concentrations of the GA₁ precursors GA₂₀, GA₁₉ and GA₄₄. However, the levels of GA₁, GA₈ and GA₂₉ were affected to a much smaller extent. Foliar application of BAS 111¨W to cv. Belinda 1 month after sowing resulted in only a 20% height reduction at flowering, but no uniform decrease in the concentrations of endogenous GAs at this stage.     

Effects of exogenously applied growth regulators on shoot growth of inbred lines of Plantago major differing in relative growth rate: differential response to gibberellic acid and (2-chloroethyl)-trimethyl-ammonium chloride

The possibility of modulating shoot growth charaeteristics of seedlings of two inbred lines of Plantago major L., differing in relative growth rate (RGR), by exogenously applied 6-benzylaminopurine (BA), α-naphthalene acetic acid (NAA), (gibberellic acid (GA₃) and (2-chloroethyl)-trimethyl-ammonium chloride (CCC) was investigated. BA completely inhibited growth of the shoot at a concentration of 1 m M , while lower concentrations had no effect. NAA reduced growth of the shoot at 10 ü M , while 1 m M completely inhibited growth. Addition of 10 μ M GA₃ or higher stimulated shoot fresh weight up to 20% and leaf area up to 30% for the slow growing inbred line (W9), but less for the fast growing line (A4). Application of 1 m M CCC, an inhibitor of gibberellin metabolism, reduced growth of both inbred lines, but to a larger extent in the fast growing seedlings.
The lower shoot growth of W9 was associated with a lower specific leaf area (SLA) and a higher dry matter percentage of the shoot, as compared with A4. NAA reduced growth by reducing SLA and increasing leaf thickness, but the percentage dry matter of the leaves was unaffected. Stimulation of the shoot growth by GA₃ application was associated with higher SLA and lower dry matter percentage. Application of CCC had opposite effects on SLA and dry matter percentage as compared with GA₃. GA seems to be involved in the regulation of at least part of the genetic difference in RGR in Plantago major .     

Influence of Plant Growth Regulators on Growth and Virulence of Rhizoctonia solani (Kühn) Pathogenic to Groundnut Seedlings (Arachis hypogaea L. var. TMV 2)

Growth of R. solani was markedly influenced by GA₃, kinetin but not by lAA. The mycelium used for inoculation of groundnut hypocotyls was obtained from cultures grown on solid media supplemented with various concentrations of GA₃, kinetin and IAA. The virulence was evaluated by measuring the lesion size 48 h after inoculation. Gibberellic acid followed by kinetin caused enhanced lesion size at 0.5 μg/ml in the medium. A correlation was observed between the infection cushions produced and the virulence of the pathogen.     

Role of endogenous gibberellins in germination of melon (Cucumis melo) seeds

The effect of the plant growth retardants ancymidol. mefluidide and uniconazole on germination of two melon accessions differing in their ability to germinate at 14°C was examined. The accessions were the cold sensitive Noy Yizre'el and the cold tolerant Persia 202. The three growth retardants were able to delay the germination of intact Noy Yizre'el seeds, but did not affect that of intact Persia 202 seeds. On the other hand germination of decoated seeds of both accessions was unaffected by these inhibitors at normal oxygen concentration, but was inhibited at 5% oxygen. When gibberellin-like activity was measured by a dwarf rice biological assay following HPLC fractionation, it was found that seeds of Persia 202 contained much more gibberellin-like activity than Noy Yizre'el seeds. Among the extracted compounds several endogenous gibberellins were identified by combined gas chromatography-mass spectrometry (GC-MS). They included GA₄, GA₂₀, GA₁ and GA₃ in Noy Yizre'el and GA₃₄, GA₂₀, GA₁ and GA₈ in Persia 202. It is suggested that the better germination of intact Persia 202 seeds, compared to Noy Yizre'el seeds at low temperature and low oxygen concentration, is due to a higher endogenous level of GA and a better seed coat permeability to oxygen.     

Germination of annual celery (Apium graveolens) seeds: Inhibition by paclobutrazol and its reversal by gibberellins and benzyladenine

Plants of annual celery ( Apium graveolens L.) were treated with paclobutrazol during anthesis. Seeds collected from the treated plants showed a marked reduction in germination in light and failed to germinate in the dark. Application of GA_4/7 to the imbibition solution reversed the inhibitory effect of paclobutrazol while gibberellic acid (GA₃) was ineffective. Benzyladenine (BA) interaction with GA_4/7 was light and concentration-dependent. At relatively low concentrations in the dark there was a synergistic effect, but at higher concentrations, especially in the light, BA, antagonized the GA_4/7 effect. Seedlings emerging from the seeds from paclobutrazol-treated plants were only slightly shortened. It is suggested that paclobutrazol applied to the mother plants inhibited the biosynthesis of endogenous GAs, which normally enable the germination of annual seeds under unfavorable conditions. Exogenously applied GA_4/7 fulfills the function of the absent endogenous GAs.     

The effect of light and exogenous gibberellic acid on respiration pathways during germination of tomato seeds

Tomato (Lycopersicon esculenlum Mill. cv. Taiwan Red) seeds are typical lightsensitive seeds where light requirement can be substituted by gibberellic acid (GA₃). During the initial stage of germination, the hexose monophosphate pathway (HMP) in seeds incubated for 24 h under white light or 12 h in 0.3 mW GA₃ solution acidified with 1 M HCI for 1 h in the dark (HCl→GA₃) was much greater than in seeds incubated for 24 h in the dark. The results were obtained from measurements of the respiration rate by man metric method of Warburg with or without iodoacetic acid, and from activities of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and aldolase (EC 4.1.2.13). Although the HMP tended to decrease at the later stage (72 h under white light or in the dark, or 36 h in HCl→GA₃), it remained greater in light- than in dark-incubated seeds. CN-resistant respiration increased from 27–30% to 57–64% of the total respiration in seeds incubated under white light or in HCI→GA₃, while respiration of non-germinating, dark-incubated seeds remained zero. Benzohydrox–amic acid (BHAM), an inhibitor of the alternative pathway, inhibited both respiration and seed germination. It is concluded that the enhancement of HMP and the CN–resistant pathways are both controlled by P_fr, but there is no direct connection between them.     

Effects of salicylhydroxamate on respiration, seed germination and seedling growth in Avena fatua

The effects of inhibitors of alternative respiration [salicylhydroxamate (SHAM) and propyl gallate (PG)] on germination, seedling growth and O₂ uptake in Avena fatua L. (wild oats) were studied. SHAM did not inhibit germination or O₂ uptake prior to germination. SHAM-sensitive (alternative) respiration, therefore, cannot be a pre-requisite for germination. Following germination, both chemicals inhibited seedling growth with the root being more susceptible than the shoot. SHAM concentrations that inhibited root growth by 90 to 95%, inhibited O₂ uptake of 1 cm root apices by less than 15%. While sodium azide (a cytochrome-oxidase inhibitor; 1 m M ) alone inhibited O₂ uptake by only 40 to 50%, in the simultaneous presence of SHAM (or PG), O₂ uptake was inhibited by 90 to 99%. Thus: 1) respiration of wild oat seedling root apices is predominantly cytochrome-mediated and incomplete inhibition of O₂ uptake in the presence of azide alone is due to diversion of electrons to the alternative pathway and 2) even though these roots have little alternative respiration, they maintain the capacity to support a much greater flux of electrons via this path way. SHAM and PG at concentrations (0.05 to 0.4 m M ) which inhibited O₂ uptake significantly in the presence (but not in the absence) of azide had little effect on root growth suggesting that an effect(s) other than that on respiration is involved in the inhibition of root growth at higher concentrations. The effect of SHAM on wild oat root growth is not selective as it also inhibits growth of a number of crop species.     

The effect of some growth regulators on light-induced germination of Paulownia tomentosa seeds

Grubišié, D., Konjevié, R. and Neškovié, M. 1988. The effect of some growth regulators on light-induced germination of Paulownia tomentosa seeds. - Physiol. Plant. 72: 525–528.
The germination of Paulownia tomentosa Steud. (Empress tree) seeds can be induced either by red light (R) or by exogenously applied gibberellic acid (GA₃). The R induced germination is completely suppressed by far red irradiation, abscisic acid (ABA) or by growth retardants such as ancymidol, tetcyclacis and paclobutrazol, though not by AMO 1618 or chloro-choline chloride (CCC). The inhibition caused by far red light (FR) and growth retardants can be overcome by GA₃, while the ABA-induced inhibition can be reversed by fusicoccin (FC), which does not reverse the inhibition caused by FR or growth retardants. It is noteworthy that the germination of light insensitive wheat, corn, alfalfa and mung bean seeds is not inhibited by growth retardants.     

Free and conjugated gibberellins in dormancy and germination of apple seeds

Halińska, A. and Lewak, St. 1987. Free and conjugated gibberellins in dormancy and germination of apple seeds.
The presence of gibberellin A₄ (GA₄) was confirmed in partly stratified seeds of apple ( Malus domestica Borb., cv. Antonówka) by mass spectrometry of the methyl ester. Levels of free and conjugated gibberellins A₄₊₇ and A₉ changed during drying of mature seeds, during cold and warm stratification, as well as during germination of dormant and non-dormant embryos. The temporary rise in GA₄₊₇ during cold stratification and during the culture of dormant embryos as well as the lack of it under conditions of warm stratification, allowed us to postulate a role for GA₄₊₇ in the removal of dormancy. In addition, GA₉ was absent in dormant embryos and increased during cold stratification and during the culture of non-dormant embryos. This suggests the involvement of GA₉, in induction of normal development of the seedling. The equivalence between changes in free and conjugated GAs suggests that formation and hydrolysis of conjugates are involved in the control of the physiologically active levels of free GA₄₊₇ and GA₉.     

Effects of prohexadione (BX-112) and gibberellins on shoot growth in seedlings of Salix pentandra

Effects of gibberellins A₁, A_4/7, A₉, A₁₉ and A₂₀ and growth retardants were studied on shoot elongation in seedlings of Salix pentandra L. The growth-retarding effects of CCC and ancymidol were antagonized by all the gibberellins tested. The novel plant growth regulator prohexadione (free acid of BX-112), which is suggested to block 3β-hydroxylation of gibberellins, effectively prevented shoot elongation in seedlings grown under long photoperiod. Initiation of new leaves was only slightly reduced. GA₁, but not GA₁₉ and GA₂₀, was active in overcoming the inhibition of stem elongation of seedlings, treated with prohexadione, GA₁₉, GA₂₀ and GA₁ are native in S. pentandra , and the results are compatible with the hypothesis that GA₁ is active per se in shoot elongation, and that the effect of GA₁₉ and GA₂₀ is dependent on their conversion to GA₁.
A mixture of GA₄ and GA₇ was as active as GA₁ in promoting shoot elongation in seedlings treated with prohexadione, while GA₉ showed slight activity only when applied at high doses.     

Interaction of gibberellic acid and photoperiod on leaf sheath elongation in normal and gibberellin-insensitive genotype of wheat

Effect of gibberellic acid (GA₃) on leaf sheath elongation in a normal (cv. Møystad) and a gibberellin(GA)-insensitive (cv. Siete Cerros) genotype of wheat ( Triticum aestivum L.) were studied at 18 and 12°C under short (SD, 12 h) or long (LD, 24 h) photoperiod. Leaf sheath length in cv. Møystad was signficantly increased by exogenous GA₃ both under SD and LD. LD alone stimulated leaf sheath elongation and the combined effect of LD and GA₃ was additive, and there was no statistically signficant interaction between photoperiod and GA₃ concentrations. Leaf sheath length in cv. Siete Cerros was not significantly affected by GA₃ under any conditions. However, there was a highly significant stimulation of leaf sheath elongation by LD in cv. Siete Cerros as well. These results indicate that stimulation of elongation growth in wheat leaves by LD is not mediated by gibberellin.     

Ethylene-dependent action of gibberellin in seed germination of Amaranthus caudatus

The role of gibberellins in the germination of seeds of Amaranthus caudatus L. was examined. Tetcyclacis (BAS 106), an inhibitor of gibberellin biosynthesis, inhibited germination of the seeds. The inhibition caused by BAS 106 was antagonised by gibberellin A₄₊₇ (GA₄₊₇). Ethephon (2-chloroethylphosphonic acid) and 1-aminocyclopropane-1-carboxylic acid (ACC) could replace GA₄₊₇. Ethephon and ACC counteracted also the side effects of BAS 106 that are not reversible by GA₄₊₇. The rate of seed germination was not increased by gibberellin in the presence of aminoethoxyvinylglycine (AVG). AVG increased the effect of BAS 106. GA₄₊₇ could not reverse the effect of BAS 106 when AVG was simultaneously applied. The results indicate that the biosynthesis of endogenous gibberellins may be required for germination of A. caudatus seeds and that main physiological effects of GA₄₊₇ on seed germination may depend on ethylene biosynthesis.     

BIOASSAY OF ANTHERIDIOGEN IN LYGODIUM JAPONICUM

Antheridia were induced by exogenously applied GA₃ at concentrations between 10⁻⁶ and 3 × 10⁻⁴ M in very young filamentous protonemata of Lygodium japonicum grown in darkness; the longer the dark preculture of protonemata, the lower was the sensitivity of the protonemata to GA₃. Antheridial initials were discernible after 36 hr of GA₃ treatment in the most sensitive protonemata, and the timing of antheridial initiation was delayed with increasing protonemal age.
This quantitative response of the protonemata provided the basis for a new method of assaying gibberellins in terms of the degree of antheridial formation. According to this method, all the gibberellins tested and one of their precursors were active in inducing antheridia in the protonemata, and the activity spectrum of the gibberellins was as follows: GA₇>GA₄>GA₉>GA₃>GA₅>GA₁>GA₈.
The amounts of antheridiogen contained in conditioned media were measured by the present bioassay. A semi-logarithmic relation was shown between the percentage of antheridial formation and the concentration of conditioned medium within a certain dilution range. The amounts of antheridiogen secreted by the prothallia were quantitatively compared by transferring samples onto fresh media for a short period of time.     

Interaction between the effects of phytochrome and gibberellic acid on the senescence of Alstroemeria pelegrina leaves

Chlorophyll loss in the leaves of cut flowering branches of Alstroemeria pelegrina L. cv. Stajello, placed in water in darkness at 20°, was inhibited by irradiation with red light and by the inclusion of gibberellic acid (GA₃) in the water. The effects of red light were abolished when it was followed by far-red light. Effects of GA₃ and red light were additive over a range of GA₃ concentrations (0. 01–1 μ M ). Chlorophyll breakdown was increased by the inclusion of AMO-1618, ancymidol, or tetcyclasis in the water. The effect of these inhibitors of gibberellin synthesis was fully reversed by GA₃. The inhibition of chlorophyll breakdown by red light was absent when AMO-1618, ancymidol or tetcyclasis were included in the water. The results indicate that leaf yellowing is controlled by endogenous gibberellins and that the effect of phytochrome is mediated by gibberellin synthesis.     

Induction of phenylalanine ammonia-lyase (PAL) in germinating lettuce seeds (Lactuca sativa)

Dry lettuce seeds (achenes of Lactuca sativa L. cv. Grand Rapids) contain no detectable phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity. Enzyme activity could be detected in these seeds within 4 h of imbibition under white light. The specific activity of PAL increased rapidly during the next 12–16 h of imbibition. Far-red light completely suppressed germination as well as the development of PAL. Gibberellic acid (GA₃, 0.1 m M ), although effective in causing almost 100% germination in dark, did not induce proportionate increases in PAL. Seed germination as well as PAL activity were substantially inhibited by cis -4-cyclohexene-l, 2-dicarboximide (CHDC, 1.0 m M ) both in light and dark. Both GA₃ and benzyladenine (BA, 0.1 m M ) retarded radicle elongation in light. Concomitantly, a decrease in PAL activity was observed. Benzyladenine was able to reverse the effects of CHDC on germination but PAL activity was still highly reduced, probably due to the inhibitory effects of BA on elongation of the radicle. More than 95% of the extractable PAL was found to be present in the radicle. When seeds incubated in white light for 10 h were transferred to FR, further increases in PAL activity as well as the growth of the radicle were severely inhibited. It is suggested that the induction of PAL in light-sensitive lettuce seeds is coincidental with the germination of seeds, and the amount of PAL per germinated seed is related to the extent of elongation of the embryonic axes.     

Interaction of photoperiod and gibberellin on growth and photosynthesis of high-latitude Poa pratensis

The relative growth rate of pot-grown plants of Poa pratensis L. cv. Holt, origin 69s°N, was increased by 20–40% by photoperiod extension with low intensity incandescent light from 8 to 24 h at 9–21°C. The main increase occurred over the 14 to 18 h photoperiod range. The true photoperiodic nature of the response was demonstrated by the effectiveness of night interruption in stimulating growth. Fortnightly sprayings with gibberellic acid (GA₃) (3 × 10^-6 to 3 × 10^-5 M ) mimicked all the effects of long days, whereas (2-chloroethyl)-trimethylammonium chloride (CCC) counteracted the effects of long days. Both growth substances exhibited pronounced interactions with photoperiod, GA₃ being most effective in short days and CCC in long days. The growth stimulation, whether caused by long days or GA₃, was exerted mainly through increases in individual and total leaf area. This was associated with a reduction in CO₂, exchange rate and a parallel fall in specific leaf weight. Proportionally, however, the increase in leaf area was greater than the fall in CO₂ exchange rate, resulting in a 38 to 118% increase in photosynthesis per leaf. No evidence was found of any direct and promotive effect of transition to long days on the CO₂ exchange rate of already expanded leaves.