Thermodormancy in Celery Seeds and its Removal by Cytokinins and Gibberellins

Imbibition of celery (Apium graveolens L.) seeds at 32°C for up to 96 h lowered the upper temperature limit for germination. If this high temperature treatment was given in the light, these seeds germinated slightly earlier than those treated in the dark although the final percentage germination was similar for both treatments. The inhibitory effect of the high temperature treatment was completely removed by allowing the seeds to imbibe in a mixture of the gibberellins A₄ and A₇ (GA_4/7) and partially removed by the cytokinin N⁶-benzylaminopurine (BA). GA_4/7 was less effective when added before rather than after the high temperature treatment, whereas the opposite was true of BA. At constant temperatures more GA_4/7 was required to promote germination as the temperature was raised but addition of BA reduced the concentration of GA_4/7 required. A model is proposed for the control of celery seed germination by light and temperature through the action of endogenous cytokinins and gibberellins.     

Evidence for the accumulation of a germination inhibitor during progressive thermoinhibition of seeds of celery (Apium graveolens L.)

The germination of seeds of celery (Apium graveolens L.) becomes progressively thermoinhibited on incubation in the dark at high temperatures, the inhibitory temperature being dependent on the cultivar used. In two high-dormancy cultivars of celery, the production of germination inhibitors in seeds incubated in the dark at 26°C gradually increased over a 7-day period. Inhibitor production was measured by incubating seeds of the low-dormancy cultivar Florida 683 in homogenates of the thermoinhibited seeds of the high-dormancy cultivars and recording germination either in the light or with the gibberellins A₄ and A₇ (GA_4/7) in the dark. Most Florida 683 seeds which failed to germinate in the homogenates after 15 days were induced to germinate by addition of N⁶-benzyladenine (BA). The presence of BA in addition to GA_4/7 throughout incubation in the dark completely overcame the inhibitory effects of homogenates. This indicates that thermoinhibition of celery seeds is associated with the accumulation of a germination inhibitor which interacts with cytokinins. This does not appear to be abscisic acid (ABA) since ABA levels in thermoinhibited seeds were lower than in untreated seeds and did not increase with duration of high temperature treatment.Abbreviations ABA Abscisic acid - BA N⁶-benzyladenine - GA_4/7 a mixture of the gibberellins A₄ and A₇ - HTP high-temperature pretreatment     

Thermodormancy Release of Celery Seed by Gibberellins, 6-Benzylaminopurine, and Ethephon Applied in Organic Solvent to Dry Seeds

The emergence of celery (Apium graveolens L. cv. Utah 52–70)seeds was promoted by growth regulators when exposed to hightemperatures during the germination period. The growth regulatorswere applied to dry seeds prior to sowing, by means of the organicsolvent dichloromethane (DCM). A mixture of gibberellins A₄and A₇ (GA_4/7) strongly enhanced emergence at a high day-timetemperature of 35°C alternating with night temperaturesof 20°C and 25°C; however, emergence was very poor whenthe night temperature was raised to 30°C. Under the latterregime, only mixtures of GA_4/7 with 6-benzylaminopurine (BA)or with 2-chlorophosphonic acid (ethephon) promoted seed emergence.However, BA and ethephon applied separately or in combinationwere much less effective in enhancing seed emergence withoutthe addition of GA_4/7, under all the temperature regimes.     

Implication of Ethylene in the Release of Secondary Dormancy in Amaranthus caudatus L. Seeds by Gibberellins or Cytokinin

Ethephon (Eth), gibberellin A₃, A_{4 + 7} (GA₃, GA_{4 + 7}), and 6-benzyladenine (BA) removed secondary dormancy of Amaranthus caudatus seeds. The GAs and BA potentiated the effect of ethephon or 1-aminocyclopropane-1-carboxylic acid (ACC), an ethylene biosynthesis precursor, in terms of the rate or final percent of germination. Aminoethoxyvinylglycine (AVG), an ACC synthase activity inhibitor, was observed to simultaneously inhibit the release from dormancy effected by GA₃ or BA as well as the ethylene production stimulated by these regulators. Breaking of secondary dormancy by GA₃, GA_{4 + 7} or BA was prevented by 2,5-norbornadiene (NBD), an inhibitor of ethylene binding. Ethylene completely or markedly reversed the inhibitory effect of NBD. We thus conclude that the removal of secondary dormancy in Amaranthus caudatus seeds by gibberellin or benzyladenine involves ethylene biosynthesis and action.     

Growth Regulators and the Phytochrome-Mediated Dormancy of Celery Seeds

Seeds of five celery (Apium graveolens L.) cultivars germinated at 15°C in the light or dark but at 22°C only in the light. This light requirement was overcome by treatment with a mixture of the gibberellins GA₄ and GA₇ (GA_4/7) but interactions of cytokinins, daminozide, ethephon, EDTA and N-phenyl-N′-4-pyridylurea (NC5392) with GA_4/7 were observed. Varietal differences in response to GA_4/7 concentration and the requirement for cytokinins were related to the upper temperature limits for germination of the different cultivars. Seeds of cultivars responding to low concentrations of GA_4/7 appeared to contain less natural inhibitor than those requiring either high concentrations of GA_4/7 or cytokinin in addition to low GA_4/7. The cytokinin requirement for germination was partially removed by leaching the seeds with water. Interaction studies with applied hormones indicated that in seeds incubated in the light inhibition by abscisic acid was partially alleviated by N⁶-benzyladenine but not by GA_4/7 application. The implications of these results are discussed in relation to the involvement of natural plant hormones in the dormancy mechanism of celery seeds.     

Endogenous ethylene and reversing methyl jasmonate inhibition of Amaranthus caudatus seed germination by benzyladenine or gibberellin

BA at 10^–5 M, GA₃ at 3×10^–4 M or GA₄₊₇ at 3×10^–5 M partially or largely reversed the inhibition of Amaranthus caudatus seed germination due to JA-Me. BA or GA₃ did not affect ethylene production and ACC oxidase activity in vivo in the presence of JA-Me before radicle protrusion. However, both increased ethylene production after 72 h of incubation, when the reversal of the JA-Me inhibition of seed germination was observed. AVG at 3×10^–4 M decreased ethylene production when it was applied simultaneously with BA and JA-Me or GA₃ and JA-Me, but it had no effect on seed germination. NBD almost completely reversed the stimulatory effect of BA, GA₃ or GA₄₊₇ on the germination of seeds in the presence of JA-Me. Exogenous ethylene reversed the inhibitory effect of NBD. The results indicate that action of endogenous ethylene is involved in the response of JA-Me inhibited seeds to BA or GA_s.     

Germination behaviour of seeds of Withania somnifera (L.) Dunal: a high value medicinal plant

In order to evolve a quick method for smooth and optimum germination for Withania somnifera- a medicinally efficacious multipurpose plant, present investigation was carried to study the effect of physico-chemical treatments, storage, temperature, photoperiod and growth regulators (GA₃, IAA, IBA, 2–4 D and BA) on germinability. The most effective treatment is GA₃ at 150 μg/ml concentration at 25 °C. The optimal temperature for germination is 25 °C and continuous light favored germination showing that photoperiod has a significant role. The seedlings derived from seeds performed well when grown in a glasshouse. The data have implications for conservation and cultivation of the species studied.     

Dormancy break of celery (Apium graveolens L.) seeds by plant derived smoke extract

Seed dormancy of a highly-dormant cultivar of celery (Apium graveolens L.) was broken by combinations of plant-derived smoke extract or N⁶-benzyladenine (BA) and gibberellins A_4/7 (GA_4/7) in the dark at temperatures between 18 and 26°C. A less dormant cultivar which responded to GA_4/7 alone showed no additional response to smoke extract or BA. Neither smoke extract nor BA affected either cultivar in the dark in the absence of GA_4/7. The partial dormancy-breaking effect of short exposures to red-light was also enhanced by smoke extracts in this highly-dormant cultivar. The results suggest that smoke extracts act in a similar way to cytokinins, by enhancing gibberellin activity in the celery seed system.Abbreviations BA N⁶-benzyladenine - GA_4/7 A₄ and A₇ gibberellin mixture     

Germination response of black and yellow seed coated canola (<Emphasis Type="Italic">Brassica napus</Emphasis>) lines to chemical treatments under cold temperature conditions

Seed quality is a key critical component to produce well established and vigorous seedlings under cool soil (<10°C) conditions experienced in Western Canada. A simple, relatively quick germination assay is required to separate small differences in seed germination which can have a significant impact on seedling growth. It has long been established that phytohormones regulate seed germination: abscisic acid inhibits germination whereas gibberellins enhance germination. We investigated the effects of ABA, GA, ethylene and inhibitors of these phytohormones alone and in combination on the germination rate of a black and a yellow seed canola (Brassica napus) imbibed at 8°C. The effects of either saline solutions, osmotic solutions, fusicoccin or testa on the germination of canola seeds imbibed at 8°C were also investigated. This temperature is representative of the soil temperatures experienced in the early spring of Western Canada. The two canola seed lines, especially the yellow seed line, were very sensitive to increasing concentration of saline solutions at 8°C, but not at 23°C; however, iso-osmotic solutions that reduced water potential were more inhibitory. The seed coat (testa) including the endosperm was a major factor affecting the germination rate of the yellow seed line at 8°C, however, GA₄₊₇ overcame the inhibitory effect of the testa, whereas ABA exacerbated it. Fusicoccin was more stimulatory to germination than GA₄₊₇, however, unlike GA₄₊₇, it was unable to overcome the inhibitory effect of paclobutrazol, a GA biosynthesis inhibitor. Fluridone, an ABA biosynthesis inhibitor, was unable to overcome the inhibitory effects of a saline solution suggesting that the inhibitory effect was not due to elevated ABA levels. Ethylene, a stimulator of germination did not appear to be involved in the germination of these two lines. Controlled deterioration at 35°C, 85% RH could be either partially or completely overcome by exogenous GA₄₊₇. This study demonstrated the effect of hormones, salinity and testa on the germination of canola seeds under less than ideal environmental conditions.     

Low temperature, gibberellin and acid lipase activity in removal of apple seed dormancy

The activity of acid lipase and the level of gibberellin A₄ (GA₄) were determined in apple embryos excised from seeds after different time periods of stratification and subsequently cultured in darkness at 4°C or at 25°C. Enzyme activity and GA₄ content were higher at 4°C. Exogenous gibberellin stimulated lipase activity, while AMO-1618, an inhibitor of gibberellin biosynthesis, inhibited, to the same degree, both the enzyme activity and the GA₄ accumulation. The involvement of GA₄ and lipolytic enzymes in cold-mediated removal of embryonal dormancy has been discussed and compared with the role of these two factors in light-stimulated germination of dormant apple embryos, described earlier (Smoleńska and Lewak 1974).     

Germination responses to GA3 and short‐time chilling of three endemic species: Tripleurospermum pichleri,Cirsium leucopsis and Senecio olympicus (Asteraceae)

In the present study, the germination characteristics of three endemic species from Turkey, Tripleurospermum pichleri (Boiss.) Bornm., Cirsium leucopsis D.C and Senecio olympicus Boiss. (Asteraceae), were investigated. Germination was studied for fresh seeds, for seeds subjected to short‐time chilling (15 days, moist +4°C), to GA₃ (100, 150 and 250 ppm) and a combination of chilling and GA₃; in all cases seeds were incubated either at 20/10°C day/night with light daytime or at 20°C in darkness with daily short‐time dim light (DSDL). In C. leucopsis seeds, all GA₃ treatments enhanced the final germination percentages. The mean germination time (MGT) of C. leucopsis was lower under DSDL than with photoperiod. The chilling treatment with GA₃ in DSDL significantly increased germination in S. olympicus seeds (from 45 to 87%). Germination increased to 55% in T. pichleri by chilling under photoperiod compared with 32% by chilling followed by DSDL. In conclusion, these three co‐existing endemic Asteraceae species have different germination behaviours; something that should be taken into account for ex situ propagation. However, an efficient way to germinate all species is to use 250 ppm GA₃ and 20/10°C with photoperiod.     

Gibberellins and the photoperiodic control of leaf growth in Poa pratensis

The role of gibberellins (GAs) in photoperiodic control of leaf elongation in Poa pratensis was studied by both application of exogenous GAs and analysis of endogenous GAs. Leaf elongation was strongly increased under long day (LD, 24 h) conditions at both 9 and 21°C, leaf length at 9°C LD being similar to that in plants grown in short days (SD, 8 h) at 21°C. However, even at 21°C leaf elongation was enhanced by LD. Exogenous GA₁ could completely compensate for LD at both 9 and 21°C. Gibberellins A₂₀, A₁₉ and A₄₄ could also partly replace LD, but they were significantly less active than GA₁, GA₅₃ was inactive when applied to plants grown at 9°C in SD and exhibited only marginal activity at 9°C LD and 21°C SD. The total level of GAs of the early 13-hydroxylation pathway (A₅₃, A₄₄, A₁₉, A₂₀ and A₁) increased rapidly when plants were transferred from SD to LD at 9°C. After transfer from 9 to 21°C, there was an increase in GA levels at both LD and SD, followed by a decrease under LD conditions. In all cases, GA₁₉ was the predominant GA, accounting for 60 to 80% of the analysed GAs. Levels of the bioactive GA₁ were low and increased transiently by LD four days after transfer from SD to LD. At both temperatures, the ratio GA₁₉ to GA₂₀ and GA₂₀ to GA₁ at 9°C was enhanced by LD compared with SD. Taken together, these results support the hypothesis that photoperiodic regulation of leaf elongation in Poa pratensis is GA-mediated, and they indicate a photoperiodic control of oxidation of GA₅₃ to GA₄₄ and GA₁₉ to GA₂₀, and perhaps also of 3β-hydroxylation of GA₂₀ to GA₁.     

Abscisic acid regulates seed germination of Vellozia species in response to temperature

• The relationship between the phytohormones, gibberellin (GA) and abscisic acid (ABA) and light and temperature on seed germination is still not well understood. We aimed to investigate the role of the ABA and GA on seed germination of Vellozia caruncularis, V. intermedia and V. alutacea in response to light/dark conditions on different temperature.
• Seeds were incubated in GA (GA₃ or GA₄) or ABA and their respective biosynthesis inhibitors (paclobutrazol – PAC, and fluridone – FLU) solutions at two contrasting temperatures (25 and 40 °C). Furthermore, endogenous concentrations of active GAs and those of ABA were measured in seeds of V. intermedia and V. alutacea during imbibition/germination.
• Exogenous ABA inhibited the germination of Vellozia species under all conditions tested. GA, FLU and FLU + GA₃ stimulated germination in the dark at 25 °C (GA₄ being more effective than GA₃). PAC reduced seed germination in V. caruncularis and V. alutacea, but did not affect germination of V. intermedia at 40 °C either under light or dark conditions. During imbibition in the dark, levels of active GAs decreased in the seeds of V. intermedia, but were not altered in those of V. alutacea. Incubation at 40 °C decreased ABA levels during imbibition in both V. caruncularis and V. alutacea.
• We conclude that the seeds of Vellozia species studied here require light or high temperature to germinate and ABA has a major role in the regulation of Vellozia seed germination in response to light and temperature.

     

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.     

Effect of gibberellic acid- and water-based pre-soaking treatments under different temperatures and photoperiods on the seed germination of Allium stracheyi Baker: An endangered alpine species of Central Himalaya,India

Allium stracheyi Baker (Alliaceae, 2600–3000 m asl), an endangered species of Central Himalaya, India, has low seed germination in its natural habitat. This study is an attempt to improve seed germination by determining the seed viability with a low mean germination time (MGT) and germination index (GI) under optimum temperature, light, and pre-soaking treatments. The seeds were pre-soaked in hot water (80°C), cold water (10°C), and gibberellic acid (GA₃ at 50 and 100 mg/l) for 24 h and subjected to light (12 h light and 12 h dark) and continuous dark (24 h) conditions with different temperature regimes (10, 15, 20, 25, and 30°C). The viability varied between 66.0% and 69.67% and declined rapidly after 12 months of storage. Our studies suggest that the 100 mg/l GA₃ treatment was beneficial for seed germination and seedling growth. Pre-soaking in a 100 mg/l GA₃ solution and incubation at 20°C under light conditions enhanced the germination significantly (p < 0.05) and resulted in the highest (97.3%) germination with the lowest MGT = 5.7 days, with GI = 8.11. The recommendations of this study support the conservation of alpine A. stracheyi via simple and cost-effective techniques for optimal seed germination.     

Allelopathy in Heracleum laciniatum: Inhibition of Lettuce Seed Germination and Root Growth

Both dark and red light germination of lettuce seeds (cv. “Maikönig”) as well as their root and hypocotol elongation were inhibited when the seeds were sown in petri dishes together with a few seeds of Heracleum laciniatum Horn. This inhibition was not significantly counteracted by the presence of gibberellic acid (GA₃) or/and 6-benzylaminopurine (BA). However, a large proportion of the lettuce seeds germinated abnormally (only cotyledons emerged) when treated with BA in the presence of Heracleum seeds. GA₃ had alone no significant effect on abnormal germination, but it counteracted the effect of BA to some extent. The inhibitory effect of Heracleum seeds gradually disappeared during a moist incubation period of one to seven days in darkness at 25°C. When lettuce seeds were pre-incubated together with Heracleum seeds for one to five days the remaining, non-germinated lettuce seeds had lost their ability for subsequent germination in darkness in distilled water. This induced dark dormancy was to a great extent broken by red light, but not by GA₃ or/and BA. H. laciniatum seeds inhibited the germination of Salix pentandra seeds and to some extent also the germination of radish but had no effect on the germination of spruce.     

The germination characteristics of <Emphasis Type="Italic">Scrophularia marilandica</Emphasis> L. (Scrophulariaceae) seeds

Investigations on seeds of Scrophularia marilandica L. were undertaken to determine their germination requirements. Seeds were collected from three naturally occurring sites and one greenhouse-grown population in London, Ontario in September and October of 1997. Some were set to germinate immediately after collection; others were stored in or on soil outside and/or under controlled laboratory conditions before testing. Germination was assessed under two light/temperature regimes (35°C 14 h light, 20°C 10 h dark and 25°C 14 h light, 10°C 10 h dark), in continuous darkness, and in the presence of two germination-promoting chemicals (GA₃ and KNO₃). Fresh seeds germinated best at 35/20°C, while stored seeds germinated best at 25/10°C. No differences in percent germination were found among three seed-maturity stages. All chemical treatments, except 0.01 M KNO_3, increased percent germination. Significant differences were found both among and within sites for most chemical treatments, but exposure to 3 × 10⁻⁴ M GA₃ caused almost every seed to germinate. When compared to the control, both the gibberellic acid and the soil-storage treatments contributed to faster germination. Exposure of seeds to naturally prevailing conditions on the soil surface followed by testing under the 25/10°C regime produced the highest percent germination. No seeds germinated in the dark. In summary, seeds of S. marilandica exhibit physiological dormancy, which can be alleviated by exposure to light, after-ripening and/or cold stratification. It is probable that the differences in germination response among sites can be attributed to differences in environmental conditions during seed production. These experiments indicate that the seeds of S. marilandica must be buried shortly after dispersal in order to form a persistent seed bank.     

Relationships between position on the parent plant and germination characteristics of seeds of parsley (Petroselinium crispum Nym.)

The percentage germination of seeds of parsley cv. Imperial Curled was higher in the light than in the dark, the high temperature limits for germination being 30 and 28°C for light and dark respectively. At the higher temperatures, the germination rate was slower in the dark. At 30°C, treatment with a gibberellin A_4/7 mixture at 2 × 10^–4 M partially alleviated the inhibiting effect of darkness on the germination percentage. Pre-incubation of parsley seeds at 35°C in the dark for 30 h increased the rate, but decreased the percentage, of germination of seeds incubated at 15°C in the light. Germination and seedling emergence studies were made on seed harvested from four different umbel positions. Although heavier seeds were produced from primary umbels than from other umbel orders, they were less viable as measured by seedling emergence in the glasshouse. The rate of emergence was decreased with increasing umbel order i.e. with later seed development: this was reflected in subsequent seedling weights, with seedlings from quarternary umbel seeds being about half the weight of those from primary umbel seeds. The upper temperature limit for dark germination was only slightly affected by umbel order, with quarternary umbel seeds being the most thermo-inhibited.Abbreviations BA N⁶-benzyladenine - GA_4/7 a mixture of gibberellins A₄ and A₇ - SD8339 6-benzyl-amino-9-(tetrahydropyran-2-yl)-9H-purine     

Deep and intermediate complex morphophysiological dormancy in seeds of Ferula gummosa (Apiaceae)

We tested the hypothesis that seeds of the monocarpic perennial Ferula gummosa from the Mediterranean area and central Asia have deep complex morphophysiological dormancy. We determined the water permeability of seeds, embryo morphology, temperature requirements for embryo growth and seed germination and responses of seeds to warm and cold stratification and to different concentrations of GA₃. The embryo has differentiated organs, but it is small (underdeveloped) and must grow inside the seed, reaching a critical embryo length, seed length ratio of 0.65–0.7, before the seed can germinate. Seeds required 9 weeks of cold stratification at <10°C for embryo growth, dormancy break and germination to occur. Thus, seeds have morphophysiological dormancy (MPD). Furthermore, GA₃ improved the germination percentage and rate at 5°C and promoted 20 and 5% germination of seeds incubated at 15 and 20°C, respectively. Thus, about 20% of the seeds had intermediate complex MPD. For the other seeds in the seed lot, cold stratification (5°C) was the only requirement for dormancy break and germination and GA₃ could not substitute for cold stratification. Thus, about 80% of the seeds had deep complex MPD.     

Responses of florence fennel (Foeniculum vulgare azoricum) seeds to light,temperature and gibberellin A4/7

The percentage and the rate of germination of seeds of three varieties of Florence fennel was higher in the dark than in the light, the high temperature cut-off points being between 27.2 and 29.4°C. The optimum temperature for germination was between 20 and 25°C. Seeds of all three varieties responded to incubation in solutions containing gibberellin A_4/7 mixture (GA_4/7; Regulex), giving higher germination in the light at temperatures from 20 to 30°C. Seeds steeped for 4 h at 25°C or for 24 h at 5°C in GA_4/7 solutions gave a higher percentage and increased rate of emergence as compared with untreated dry seeds, when sown in compost at 25°C; steeping in water alone was also beneficial. In general, drying the treated seeds before sowing reduced the rate but sometimes increased the percentage of germination as compared with seeds sown when still moist. Seeds harvested from secondary umbels of var. Zefa fino germinated better both in the light and dark than those taken from primary umbels.Abbreviations GA_4/7 a commercial mixture of the gibberellins A₄ and A₇ (trade name Regulex)