Photocontrol of germination in Amaranthus caudatus

Summary Germination of Amaranthus caudatus is inhibited by light, far-red being the most effective part of the spectrum. At temperatures of 25° and below there is a low final germination percentage under continuous far-red whereas above 25° there is only a delaying effect. In the presence of a saturating concentration of gibberellic acid (GA₃) at 25° seeds germinate under continuous far-red although they are delayed. At 25° seeds exposed to 48 hr far-red fail to germinate when transferred to darkness. This induced dormancy can be broken by a single short exposure to red light given at any time after the far-red illumination. This effect of short red can be reversed by a subsequent short period of far-red indicating that the seeds are phytochrome controlled. Although most seeds have escaped from the reversing effect of short far-red after an intervening dark period of 5 hours, germination is greatly reduced by continuous far-red at this time. Results of exposing seeds to varying periods of far-red before and after dark imbibition are interpreted in terms of a continual production of phytochrome in its active P _fr form and a requirement for P _fr action over a long period of time. Effects of intermittent and continuous low intensity far-red on the inhibition of germination provides further evidence for a low energy photoreaction involving phytochrome. Effects on Germination Index of continuous illumination with various light sources maintaining different P _fr /P _total ratios have been investigated. The results suggest that the proportion of phytochrome in the P _fr form is the most important factor in the regulation of germination. A scheme for the phytochrome control of germination in Amaranthus caudatus is presented and possible explanations for the dependence on P _fr /P _total ratio are discussed.Holder of a Science Research Council Studentship.     

Responsiveness of Amaranthus retroflexus seeds to ethephon, 1-aminocyclopropane 1-carboxylic acid and gibberellic acid in relation to temperature and dormancy

Dormant Amaranthus retroflexus seeds do not germinate in the dark at temperatures below 35°C. Fully dormant seeds germinate only at 35–40°C whereas non-dormant ones germinate within a wider range of temperatures (15 to 40°C). Germination of non-dormant seeds requires at least 10% oxygen, but the sensitivity of seeds to oxygen deprivation increases with increasing depth of dormancy. 10^–6 to 10^–4 M ethephon, 10^–3 M 1-aminocyclopropane 1-carboxylic acid (ACC) and 10^–3 M gibberellic acid (GA₃) break this dormancy. In the presence of 10^–3 M GA₃ dormant seeds are able to germinate in the same range of temperatures as non-dormant seeds. The stimulatory effect of GA₃ is less dependent on temperature than that of ethephon, while ACC stimulates germination only at relatively high temperatures (25–30°C). The results obtained are discussed in relation to the possible involvement of endogenous ethylene in the regulation of germination of A. retroflexus seeds.Abbreviations ACC 1-aminocyclopropane 1-carboxylic acid - GA₃ gibberellic acid - SD standard deviation     

Biphasic fluence-response curves for phytochrome-mediated kalanchoë seed germination : sensitization by gibberellic Acid

The fluence-response curves for the effect of two red pulses separated by 24 hours on the germination of Kalanchoe blossfeldiana Poelln. cv Vesuv seeds, incubated on gibberellic acid (GA₃) are biphasic for suboptimal concentrations. The response in the low fluence range corresponds with a classical red/far-red reversible phytochrome mediated reaction. GA₃ induces an additional response in the very low fluence range, which is also phytochrome mediated. The sensitivity to phytochrome-far-red absorbing form (Pfr), however, is increased about 20,000-fold, so that even far-red fluences become saturating. Both in the very low and low fluence response range, the maximal responses induced by saturating fluences are modulated by the GA₃ concentration. GA₃ having no direct influence on the phytochrome phototransformations, alters the Pfr requirement and determines the responding seed population fraction in the very low and low fluence range. The effet of GA₃ appears to be on the transduction chain of the phytochrome signal.     

Floral initiation in sorghum hastened by gibberellic acid and far-red light

Combinations of far-red light (FR) (4 min) and gibberellic acid (GA₃), given at the beginning of a daily 12-h dark period in a growth room, were used to study floral induction in four maturity genotypes of the milo group of sorghum (Sorghum bicolor (L.) Moench). The 12-h dark period without GA₃ application or FR induced flowering in only the early genotype; FR hastened initiation in the early genotype, while GA₃ hastened floral initiation in the two intermidiate-flowering genotypes. GA₃ and FR together had a strong synergistic effect, hastening floral initiation by 30 to more than 80 d in the early and intermediate genotypes. Red light (R) did not hasten flowering; FR preceded by R gave the same effect as FR alone. GA₃ promoted stem elongation equally whether floral initiation occurred or not; thus, its effect on stem elongation was independent of floral initiation. The capacity of GA₃ to induce flowering in sorghum, a short-day plant, seems to be enhanced by phytochrome being in the P_R form at the beginning of the night when GA₃ was applied.Abbreviations FR far-red light - GA(s) gibberellin(s) - GA₃ gibberellic acid - R red light     

Changes in endogenous abscisic acid levels during dormancy release and maintenance of mature seeds: studies with the Cape Verde Islands ecotype,the dormant model of<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

Mature seeds of the Cape Verde Islands (Cvi) ecotype of Arabidopsis thaliana (L.) Heynh. show a very marked dormancy. Dormant (D) seeds completely fail to germinate in conditions that are favourable for germination whereas non-dormant (ND) seeds germinate easily. Cvi seed dormancy is alleviated by after-ripening, stratification, and also by nitrate or fluridone treatment. Addition of gibberellins to D seeds does not suppress dormancy efficiently, suggesting that gibberellins are not directly involved in the breaking of dormancy. Dormancy expression of Cvi seeds is strongly dependent on temperature: D seeds do not germinate at warm temperatures (20–27°C) but do so easily at a low temperature (13°C) or when a fluridone treatment is given to D seeds sown at high temperature. To investigate the role of abscisic acid (ABA) in dormancy release and maintenance, we measured the ABA content in both ND and D seeds imbibed using various dormancy-breaking conditions. It was found that dry D seeds contained higher amounts of ABA than dry ND after-ripened seeds. During early imbibition in standard conditions, there was a decrease in ABA content in both seeds, the rate of which was slower in D seeds. Three days after sowing, the ABA content in D seeds increased specifically and then remained at a high level. When imbibed with fluridone, nitrate or stratified, the ABA content of D seeds decreased and reached a level very near to that of ND seeds. In contrast, gibberellic acid (GA₃) treatment caused a transient increase in ABA content. When D seeds were sown at low optimal temperature their ABA content also decreased to the level observed in ND seeds. The present study indicates that Cvi D and ND seeds can be easily distinguished by their ability to synthesize ABA following imbibition. Treatments used here to break dormancy reduced the ABA level in imbibed D seeds to the level observed in ND seeds, with the exception of GA₃ treatment, which was active in promoting germination only when ABA synthesis was inhibited.Abbreviations ABA Abscisic acid - Cvi Cape Verde Islands - D Dormant - GA Gibberellin - GA₃ Gibberellic acid - ND Non dormant     

Responsivencess of Intact and Scarified Seeds of Barbarea vulgaris to Gibberellic Acid and Changes in Phytochrome

Either red light or millimolar levels of gibberellic acid promoted germination of seeds of yellow rocket (Barbarea vulgaris R. Br.), and their effects were generally additive. Buffering the substrate pH at 3.0, addition of 20 mM nitrate to the substrate, preliminary incubation at 10 or 30°C, and brief scarification of the seeds increased responsiveness of the seeds to gibberellic acid. Scarification increased several thousandfold the sensitivity of the seeds to gibberellic acid. Pretreatment of the seeds in darkness or far-red light did not lower their responsiveness to gibberellic acid. These results suggest that uptake of gibberellic acid is a limiting factor in the stimulation of germination in intact seeds and that there is only a minimal requirement for active phytochrome to express gibberellic acid action.     

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.     

Dormancy of the barley grain is correlated with gibberellic Acid responsiveness of the isolated aleurone layer

The relationship between barley grain dormancy and gibberellic acid (GA₃) responsiveness of aleurone layers has been investigated. Barley (Hordeum distichum L. cvs Triumph and Kristina) grains were matured under defined conditions in a phytotron. Grains of Triumph plants grown under long-day/warm conditions had lower dormancy levels than grains of plants grown under short-day/cool conditions. Aleurone layers isolated from grains of long-day Triumph plants secreted more α-amylase and had a higher responsiveness to GA₃ as measured by α-amylase secretion. Storage of the grains increased both the percentage of germination and the responsiveness of the aleurone to GA₃. Use of different sterilization methods to break dormancy confirmed the correlation between germination percentage and aleurone layer GA₃ responsiveness. The response of embryoless Triumph grains to GA₃ was lower than that of the isolated aleurone layers, suggesting a role of the starchy endosperm in regulating the GA₃ response of the aleurone layer. Grains of the cultivar Kristina harvested from short day- and long day-grown plants lacked dormancy, and their isolated aleurone layers had a similar responsiveness to GA₃ as measured by α-amylase secretion. The data indicate that the physiological state of the aleurone layers contributes to the percentage germination of the grains.     

Interaction between a plant-derived smoke extract,light and phytohormones on the germination of light-sensitive lettuce seeds

Plant-derived smoke extracts mimics the effect of red light on germination in light-sensitive lettuce seeds and partially overcomes the inhibitory effect of far-red light. Interaction between a smoke extract and gibberellins, cytokinins, abscisic acid and ethephon was investigated. Smoke acted synergistically with GA₃ and increased the sensitivity of the lettuce seeds to ABA. It seems likely that smoke affects membrane permeability or receptor sensitivity rather than influencing the phytochrome system of these seeds.Abbreviations R red light - FR far-red light - SM smoke extract     

Dormancy and germination in Stachys germanica L. subsp. bithynica (Boiss.) Bhattacharjee seeds: Effects of short-time moist chilling and plant growth regulators

We investigated the germination requirements of the species Stachys germanica L. subsp. bithynica (Boiss.) Bhattacharjee (Lamiaceae). We studied the effects of scarification, short-time moist chilling (+4 °C) for 15 and 30 days, and various doses of gibberellic acid (GA₃; 0, 100, 150 and 250 ppm), Kinetin (KIN; 50 ppm) and a combination of 250 ppm GA₃ and 50 ppm KIN. The hormone and moist chilling treatments were carried out under both continuous darkness (20 °C) and photoperiodic (20/10 °C; 12/12 h, respectively) conditions. Seeds failed to germinate in response to short-time moist chilling treatments with distilled water under both continuous darkness and photoperiodic conditions. Seeds were found to have dormancy. Treatments with GA₃ or a combination of GA₃ and KIN were successful at breaking seed dormancy. A maximum of 37% of the seeds germinated after GA₃ application in all series. When only KIN was applied at a 50 ppm concentration, germination (12%) was found only with moist chilling for 30 days under continuous darkness. The highest germination rates were found in seeds treated with combination of 250 ppm GA₃ and 50 ppm KIN. In the combination treatments, while the moist chilling treatments for 15 days resulted in 68 and 73% germination, respectively, these rates were up to 95% in the moist chilling treatments for 30 days under continuous darkness and photoperiodic conditions. Mean germination time (MGT) in GA₃ and KIN combinations was lower than in other treatments. Scarification with 80% sulphuric acid did not promote germination. The characteristics of physiological dormancy of S. germanica ssp. bithynica seeds are consistent with conditions of existence in the in alpine habitat of this species.     

The influence of plant growth regulators on the growth of the embryonic axes of red- and far-red-treated lettuce seeds

The influence of several plant growth regulators on the growth of the embryonic axes from red- and far-red-(R- and FR-)treated lettuce (Lactuca sativa L., cv. Grand Rapids) seeds was examined; as shown previously, the water potential of the axes from R-treated seeds has been lowered by 3.5–5.6 bars compared to that in axes from FR-treated ones. Kinetin and abscisic acid (ABA), when included in the incubation medium, reduced the elongation of the axes whereas fusicoccin stimulated it; however, these effects were the same in axes of both R- and FR-treated seeds. In contrast, elongation of axes from FR-treated seeds was stimulated by gibberellic acid (GA₃, but elongation of axes from R-treated ones was not affected by this hormone. This latter result indicates that gibberellins may be involved in the phytochrome-mediated growth responses in lettuce axes.When the root caps of the embryos were removed prior to light treatment, R was still able to induce a water-potential decrease in the embryonic axes, indicating that at least a portion of the active Pfr resides in the axis and not the root cap.Abbreviations ABA abscisic acid - FR far red light - GA₃ gibberellic acid - PEG polyethylene glycol - Pfr far-red-absorbing form of phytochrome - R red light     

Control of seed dormancy in Nicotiana plumbaginifolia: post-imbibition abscisic acid synthesis imposes dormancy maintenance

The physiological characteristics of seed dormancy in Nicotiana plumbaginifolia Viv. are described. The level of seed dormancy is defined by the delay in seed germination (i.e the time required prior to germination) under favourable environmental conditions. A wild-type line shows a clear primary dormancy, which is suppressed by afterripening, whereas an abscisic acid (ABA)-deficient mutant shows a non-dormant phenotype. We have investigated the role of ABA and gibberellic acid (GA₃) in the control of dormancy maintenance or breakage during imbibition in suitable conditions. It was found that fluridone, a carotenoid biosynthesis inhibitor, is almost as efficient as GA₃ in breaking dormancy. Dry dormant seeds contained more ABA than dry afterripened seeds and, during early imbibition, there was an accumulation of ABA in dormant seeds, but not in afterripened seeds. In addition, fluridone and exogenous GA₃ inhibited the accumulation of ABA in imbibed dormant seeds. This reveals an important role for ABA synthesis in dormancy maintenance in imbibed seeds. Received: 31 December 1998 / Accepted: 9 July 1999     

Effect of chilling and gibberellic acid on growth potential of excised embryos of Ruellia humilis

Summary Dormancy in intact seeds of Ruellia humilis is broken by chilling or by treatment with gibberellic acid (GA₃). Embryos are nondormant and will grow when the seed coat is removed completely. Embryos from chilled or GA₃-treated seeds have more growth potential than embryos from nontreated seeds. Dormancy is ascribed to the mechanical restriction of the embryo by the seed coat. Chilling and treatment with GA₃ break dormancy by increasing the expansive force of the embryo; thus, chilled or GA₃-treated embryos exert enough expansive force to break through the seed coat, whereas nontreated embryos do not.This work was supported by an NIH Biomedical Sciences Support Grant to the University of Kentucky.     

Comparative effects of gibberellins and an N-substituted phthalimide on seed germination and extension growth of celery (Apium graveolens L.)

The N-substituted phthalimide AC 94377 (1-(3-chlorophthalimido)-cyclohexanecarboxamide) was equally effective as a mixture of the gibberellins A₄ and A₇ (GA_4/7) in breaking dormancy and stimulating germination of celery seeds when either was used in combination with ethephon or daminozide as a seed soak. Whereas seedlings emerging from GA_4/7-treated seeds became etiolated in comparison with those from untreated seeds, those from AC 94377-treated seeds showed normal development. Preharvest sprays of gibberellic acid (GA₃) increased the height of mature plants in comparison with untreated controls by about 16 per cent whereas AC 94377 was ineffective. The yield from GA₃-treated plots was about 10 per cent greater than that from AC 94377-treated plots.     

Hormones are no causal links in phytochrome-mediated adventitious root formation in mustard seedlings (Sinapis alba L.)

The question of whether or not hormones are causal links in the realization of phytochrome control during photomorphogenesis was investigated using the phytochrome-dependent formation of adventitious roots in hypocotyl cuttings excised from mustard seedlings as a test system. Histological examination of regenerating rest seedlings revealed that phytochrome (operationally, continuous far-red light) mediates the de novo formation of root primordia in the pericycle region of the hypocotyl near the cutting surface withing 12–24 h after excision.Auxin (IAA), gibberellin (GA₃), Cytokinin (kinetin), abscisic acid (ABA), and ethylene had no promotive effect on primordium formation in dark-grown or far-red irradiated rest seedlings. Depending on concentration, the application of these hormones was either ineffective or inhibitory in the rooting response. It is concluded that phytochrome does not operate through changes of hormone (auxin, gibberellin, cytokinin, ABA, ethylene) levels.While externally applied ethylene had no specific effect on primordium formation, the number of primordia produced in darkness could be increased to the far-red light level by removing the endogenously formed ethylene. Since the stimulatory effect of light could not be related to a lower ethylene level, it is concluded that ethylene interferes with primordium formation by modulating the susceptibility of this process to phytochrome control. This ethylene effect takes place in a concentration range below the range that can be manipulated by external application of the hormone.Abbreviations ABA abscisic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - P_r P_fr red and far-red absorbing forms of phytochrome     

Effects of Repetitive Acid Immersion, Red Light, and Gibberellin A3 Treatments on Phytochrome-mediated Germination Control in Skotodormant Lettuce Seeds

Dry lettuce seeds (Lactuca sativa L. cv. Grand Rapids), whichreceived 5 min far-red light (FR) 0.5 h after the onset of waterimbibition, showed 17% and 50% germination without and withacid immersion treatment (pH 0.1) for 1 h and rinsing with water,respectively. The acid treatment caused only 6% germinationor less in FR-treated seeds held for 10 to 30 d in dark storage.The 10 to 30 d skotodormant seeds did not respond to red light(R) or gibberellin A₃ (GA₃) singly, but showed 84% or higherpercentage germination if 1 h acid immersion was given beforeR or GA₃. The 20 d skotodormant seeds, which received R treatmentat day 10 but remained dormant showed 89% germination with onlyacid treatment. Similar values were obtained with 30 d skotodormantseeds which received one or two R treatments at day 10 or 20,i.e. the only requirement for these R-treated dormant seedswas an acid immersion. This releases the skotodormancy and rendersthe seeds more sensitive to R or GA₃, but the skotodormancywas initiated again if no light or hormone treatments were givenimmediately. The repetitive R or GA₃ treatments, which did notcause skotodormant seeds to germinate, lessened the degree ofskotodormancy. The germination of these skotodormant seeds canonly be induced by the synergistic action of R and GA₃. In thisstudy, GA₃ caused higher germination percentages in R-treatedskotodormant seeds than R stimulated in GA3-treated seeds. Itis suggested that (i) repetitive R or Ga₃ treatments maintaina high endogenous level of the far-red-absorbing form of phytochrome(P_fr) and GA activity, respectively, (ii) the accumulated stableintermediates of phytochrome persist in fully-imbibed skotodormantseeds for up to 20 d, without phytochrome expressing its functionuntil the seeds are acidified and (iii) a model is formulatedto interpret the results of acidification, growth promotersand R effects on germination of light-sensitive lettuce seeds. Key words: Phytochrome, Latuca saliva, seed germination, dark reversion of phytochrome, gibberellin A₃, acidification, skotodormancy     

Response of Amaranthus retroflexus L. seeds to gibberellic acid, ethylene and abscisic acid depending on duration of stratification and burial

Freshly harvested, dormant seeds of Amaranthus retroflexus were unable to germinate at 25 and 35 °C. To release their dormancy at the above temperatures, the seeds were stratified at a constant temperature (4 °C) under laboratory conditions or at fluctuating temperatures in soil or by outdoor burial in soil. Fully dormant, or seeds stratified or buried (2006/2007 and 2007/2008) for various periods were treated with exogenous gibberellic acid (GA₃), ethephon and abscisic acid (ABA). Likewise, the effects of these regulators, applied during stratification, on seed germination were determined. The results indicate that A. retroflexus seed dormancy can be released either by stratification or by autumn–winter burial. The effect of GA₃ and ethylene, liberated from ethephon, applied after various periods of stratification or during stratification, depends on dormancy level. GA₃ did not affect or only slightly stimulated the germination of non-stratified, fully dormant seeds at 25 and 35 °C respectively. Ethylene increased germination at both temperatures. Seed response to GA₃ and ethylene at 25 °C was increased when dormancy was partially removed by stratification at constant or fluctuating temperatures or autumn–winter burial. The response to GA₃ and ethylene increased with increasing time of stratification. The presence of GA₃ and ethephon during stratification may stimulate germination at 35 °C. Thus, both GA₃ and ethylene can partially substitute the requirement for stratification or autumn–winter burial. Both hormones may also stimulate germination of secondary dormant seeds, exhumed in September. The response to ABA decreased in parallel with an increasing time of stratification and burial up to May 2007 or March 2008. Endogenous GA_n, ethylene and ABA may be involved in the control of dormancy state and germination of A. retroflexus. It is possible that releasing dormancy by stratification or partial burial is associated with changes in ABA/GA and ethylene balance and/or sensitivity to these hormones.     

Far-red Sensitive Dark Processes Essential for Light- and Gibberellin-induced Germination of Lettuce Seed

The action of prolonged far-red on seed germination was studied in Lactuca sativa L. var. Grand Rapids. Exposure of imbibed seeds to 6 hours far-red before the application of gibberellic acid (GA₃) and thiourea completely prevented germination. Using GA₃, this far-red was effective after the sixth hour of imbibition. At 6, 12, and 18 hours of imbibition equal durations of far-red had equal effects. The kinetics of far-red action was investigated: it was found that although far-red for several hours, irrespective of the energy level, was needed for maximum inhibition, shorter durations (15 and 30 mins) were also appreciably effective provided they were followed by several hours darkness before the supply of GA₃. This is taken to indicate the existence of labile product(s) of the action of a far-red sensitive pigment. Evidence is provided for the existence of promotive dark processes controlled by this pigment, which are essential for germination whether triggered by GA₃, thiourea or red-light. A model for the operation of the pigment system is proposed and its role in the germination mechanism of this seed is discussed.     

Interaction of light and temperature on seed germination of Rumex obtusifolius L.

Germination of Rumex obtusifolius L. seeds (nutlets) is low in darkness at 25° C. Germination is stimulated by exposure to 10 min red light (R) and also by a 10-min elevation of temperature to 35° C. A 10-min exposure to far-red light (FR) can reverse the effect of both R (indicating phytochrome control) and 35° C treatment. Fluence-response curves for this reversal of the effect of R and 35° C treatments are quantitatively identical. Treatment for 10 min with light of wavelenght 680, 700, 710 and 730 nm, after R and 35° C treatment, demonstrates that germination induced by 35° C treatment results from increased sensitivity to a pre-existing, active, far-red-absorbing form of phytochrome (Pfr) in the seeds.Abbreviations FR far-red light - P phytochrome - Pr red-absorbing form of P - Pfr far-red-absorbing form of P - R red light     

Effect of Temperature, Oxygen, and Gibberellic Acid on the Development of Photosensitivity in Oldenlandia corymbosa L. Seeds during Their Incubation in Darkness

Two successive phases can be distinguished in the development of the responsiveness to light in Oldenlandia corymbosa L. seeds during their incubation in darkness. During phase I, the responsiveness to light increases with time if there is sufficient O₂, and the higher the temperature, the faster the increase. This phase is stimulated by gibberellic acid. During the following phase (II), seeds remain responsive to light at 10 or 20°C, but lose their responsiveness at higher temperature (≥30°C). This second phase depends on O₂: loss of responsiveness is accelerated at lower O₂ concentration. Phase II is only slightly affected by gibberellic acid. The results are discussed in terms of variation of phytochrome and of a reaction along the transduction chain initiated by phototransformation of this pigment, which is finally expressed in germination.