Dormancy and Impotency of Cocklebur Seeds: VII. Inability of Dormant Cotyledons to Form Chlorophyll

Dormant seeds of cocklebur (Xanthium pennsylvanicum Wallr.) were characterized by the lack of ability to form chlorophyll. Such an inability of cotyledons of the dormant seeds was improved by the application of various factors and reagents which were capable of breaking the dormancy and of increasing cotyledon enlargement. Of these, ethylene, benzyladenine, and high temperature treatments were particularly effective, and, in turn, oxygen enrichment, gibberellic acid, thiourea, carbon doxide, and potassium nitrate were also promotive to the greening of the dormant cotyledons. The effects of benzyladenine, oxygen enrichment, and high temperature were reduced in the presence of absorbents for endogenously evolve carbon dioxide and ethylene. δ-Aminolevulinic acid could not restore their greening ability.     

Germination Promotion of Loblolly Pine and Baldcypress Seeds by Stratification and Chemical Treatments

The effects of 4 chemicals on the germination promotion of stratified and unstratified seeds of loblolly pine (Pinus taeda) and baldcypress (Taxodium distichum) were studied. The chemicals used were gibberellic acid, kinetin, potassium nitrate and thiourea, each at 3 different concentrations. Stratification promoted the germination of both seed species. Certain concentrations of gibberellic acid, potassium nitrate and thiourea improved the germination of unstratified loblolly pine and baldcypress seeds while kinetin had no stimulatory effect. All 4 chemicals at specific concentrations promoted the germination of loblolly pine seeds stratified for a short period of time. Considering both speed and completeness of germination, best results were obtained when 21-day stratified seeds were treated with either gibberellic acid (100 mg/1) or kinetin (10 mg/1). In baldcypress, on the other hand, none of these chemicals had any stimulatory effect on the germination of stratified seeds. Germination of both species of seeds was either partially or completely inhibited by the highest concentration of thiourea (30,000 mg/1) used.     

The Effects of Gibberellic Acid and Scarification on the Seed Dormancy and Germination in Luzula spicata

Luzula spicata L. seeds are completely dormant at maturity. A germination inhibitor is present at the micropylar end. Normally, the only effective means of eliciting germination is a precise scarification of the micropylar end which inactivates the inhibitor. Exogenous application of gibberellic acid, kinetin, KNO₃, and thiourea have no affect on the dormancy of unscarified seeds. Scarification of the hilar end of the seed does not elicit germination, but when gibberellic acid is applied to the hilar scarified seeds moderate germination results. Presumably, these seeds are dormant due to a deficit of endogenous gibberellin; a condition which can be overcome by the application of gibberellic acid to seeds scarified at a site in itself ineffective in producing germination. Apparently the gibberellic acid serves to initiate amylase activity in the endosperm, overcoming the inhibitor block. Luzula spicata seed dormancy is apparently unique in that a germination inhibitor is operative in conjunction with the commonly recognized gibberellin-amylase mechanism.     

Ethylene, seed germination, and epinasty

Ethylene activity in lettuce seed (Lactuca satina) germination and tomato (Lycopersicon esculentum) petiole epinasty has been characterized by using heat to inhibit ethylene synthesis. This procedure enabled a separation of the production of ethylene from the effect of ethylene. Ethylene was required in tomato petioles to produce the epinastic response and auxin was found to be active in producing epinasty through a stimulation of ethylene synthesis with the resulting ethylene being responsible for the epinasty. In the same manner, it was shown that gibberellic acid stimulated ethylene synthesis in lettuce seeds. The ethylene produced then in turn stimulated the seeds to germinate. It was hypothesized that ethylene was the intermediate which caused epinasty or seed germination. Auxin and gibberellin primarily induced their response by stimulating ethylene production.     

Possible Involvement of the Alternative Respiration System in the Ethylene-stimulated Germination of Cocklebur Seeds

Respiration of nondormant upper cocklebur (Xanthium pensylvanicum Wallr.) seeds was enhanced by exogenous C₂H₄, proportionally to the concentration of C₂H₄ and the duration of presoaking of the seeds. Benzohydroxamic acid (BHM) and salicylhydroxamic acid (SHM), inhibitors of alternative respiration, inhibited both the germination of nondormant lower cocklebur seeds and the respiration of the upper seeds presoaked for periods of 12 to 30 hours. Both the growth and respiration of axial and cotyledonary tissues were also inhibited by BHM. Moreover, BHM inhibited both the C₂H₄-induced germination of the upper seeds and their C₂H₄-stimulated respiration; the inhibition occurred only with concomitant addition of C₂H₄ and BHM. The respiration of seeds with a secondary dormancy induced by presoaking for prolonged periods was markedly stimulated by C₂H₄ but not suppressed by BHM. It was suggested that the alternative respiration system may be involved in the normal germination process of cocklebur seeds, secondary dormancy may result from its inactivation, and C₂H₄ may exert its germination-promoting action by stimulating the alternative respiration. The effects of BHM and SHM can suggest but not prove the involvement of the alternative respiration in seed germination.     

Cytokinin-Inhibitor Antagonism in the Hormonal Control of α-Amylase Synthesis and Growth in Barley Seed

The effect of cytokinins and gibberellic acid on the inhibition of growth and α-amylase synthesis by germination inhibitors was investigated in intact and embryoless seed halves. The cytokinins, kinetin and benzyladenine, effectively reversed the inhibition of coleoptile growth and α-amylase synthesis by abscisic acid and courmarin in barley seed. An antagonism between cytokinins, kinetin and benzyladenine, effectively reversed the inhibition of coleoptile growth and α-amylase synthesis by abscisic acid and coumarins in barley seed. An antagonism between cytokinins and germination inhibitors was also shown in root growth. Abscisic acid inhibited coleoptile growth to a greater extent than the root growth while the opposite held true in the case of coumarin. The apparent increase in coleoptile growth and α-amylase synthesis by gibberellic acid plus abscisic acid (or coumarins) over abscisic acid (or coumarin) appears to be a result of the overall stimulation of growth and metabolism by exogenous gibberellic acid and probably does not involve an interaction of gibberellic acid with the inhibitors. Gibberellic acid reversed root inhibition to some extent. Abscisic acid inhibition of gibberellic acid induced α-amylase synthesis in the embryoless endosperm was not reversed by excess gibberellic acid or kinetin Cytokinin reversal of inhibition of growth and enzyme synthesis probably depends on some factor(s) in the embryo. Cytokinin reversal of inhibitor action leading to enzymen synthesis and growth may be at the level of genome or at the site protein assembly.     

Effects of Germination-promoting Substances Given in Conjunction with Red Light on the Phytochrome-mediated Germination of Dormant Lettuce Seeds (Lactuca sativa L.)

Ethylene or thiourea can substitute for gibberellic acid but not for red light in breaking the secondary dormancy induced by extended dark storage of fully hydrated lettuce seeds (Lactuca sativa cv. Grand Rapids). After 10 days of storage, ethylene, thiourea, or gibberellic acid applied either separately or in any combination in conjunction with red light induced near maximal germination. When applied separately without red light, none of the substances promoted germination of seeds stored 10 days. Combinations of any two or all three of the substances in the absence of red light induced some germination but no combination was as effective as any single substance given with red light.     

Inhibition of cottonseed germination with abscisic Acid and its reversal

Germination of cottonseed (Gossypium hirsutum L.) was inhibited by abscisic acid. Inhibition was greater when seeds were soaked in abscisic acid for 5 hours and dried prior to germination than when abscisic acid was applied in the germination medium. (2-Chloroethyl)phosphonic acid, gibberellic acid, and kinetin partially overcame the inhibitory action of abscisic acid. Combinations of (2-chloroethyl)phosphonic acid with gibberellic acid or kinetin were more effective than the individual substances. Germination also was partially restored by removal of seed coats. Fusicoccin completely restored germination of abscisic acidtreated seeds.     

β-Glucosidase activities and HCN liberation in unimbibed and imbibed seeds, and the induction of cocklebur seed germination by cyanogenic glycosides

In many seed species, the major source of HCN evolved during water imbibition is cyanogenic glycosides. The present investigation was performed to elucidate the role of endogenous cyanogenic glycosides in the control of seed germination and to examine the involvment of β-glucosidase in this process. All seed species used here contained some activities of β-glucosidase already in the dry state before imbibition. in the decreasing order of Malus pumila, Daucus carota, Hordeum vulgare, Chenopodium album and so on. β-Gluosidase activity in upper and lower seeds of cocklebur (Xanthium pennsylvanicum Wallr.) decreased with imbibition, and in lower seeds the activity disappeared when they germinated. On the contrary, in caryopses of rice (Oryza sativa L. cv. Sasanishiki) β-glucosidase increased during imbibition, and this increase continued even after germination. β-Glucosidase in cocklebur seeds was more active in the axial than in the cotyledonary tissue. Amygdalin, prunasin and linamarin could all serve as substrattes for the β-glucosidase(s) from both cocklebur and rice. Amygdalin, prunasin and linamarin as well as KCN, were effective in stimulating the germination of upper cocklebur seeds. The seeds evolved much more free HCN gas when they were exposed to the cyanogenic glycosides than when the glycosides were absent. Moreover, the application of the cyanogenic glycosides or of KCN caused accumulation of bound HCN in the seeds. Carbon monoxide, which stimulated cocklebur seed germination only slightly, did not cause accumulation of bound HCN. We suggest that a balance between the cytochrome and the alternative respiration pathways, which is adequate for germination (Esashi et al. 1987. Plant Cell Physiol. 28: 141–150), may be brought about by the action of endogenous HCN; a large portion of which is liberated from cyanogenic glycosides via the action of β-glucosidase. In addition to the partial suppression of the cytochrome path and unlike carbon monoxide, the HCN thus produced may act to supply cyanide group(s) to unknown compounds necessary for germination.     

The effects of time of seed production on the germination response of Spergularia marina

Germination studies were carried out with seeds of Spergularia marina L. Griseb produced over an interval of six months (June-November). The response of the seeds to light and dark, various constant and alternating temperature regimes, and salinity were determined. In addition, the effects of soil moisture status at the time of seed production on the subsequent germination response of seeds were also determined. Light was an absolute requirement for germination. While a constant temperature regime did not generally favour germination of seed of any month, alternating temperature greatly enhanced germination with an optimum at 5/15°C in all seeds. When imbibed in solutions of different salinities, seeds collected in July and October behaved like true halophyte seeds whereas those collected in June. August, September and November behaved like glycophyte seeds.
High concentration of gibberellic acid (3 000 μ M ) stimulated dark germination in the June and November seed lots, but in light, low GA3 concentration (300 μ M ) stimulated germination most. The addition of kinetin (30 μ M ) plus gibberellic acid enhanced germination in the dark in contrast to GA3 alone; kinetin alone stimulated a very low percentage germination.
The moisture status of the soil at the time seeds were produced did not affect the germination response of an early seed crop (July) but affected that of the later seeds (August).
Judging from the different germination responses, it appears that the seeds belong to at least two physiological groups, one which appears to need either a dark-wet or cold-wet pretreatmem for high germination to occur; and the other group which does not need pretreatmem. The ecological significance of these varied responses is discussed in relation to the survival of the species in its habitat.     

Promoting Effect of Fusicoccin on Seed Germination

The effects of fusicoccin on the germination of dormant, light-requiring or abscisic acid-inhibited seeds has been investigated. (1) Fusicoccin (10^?6M) induces germination in dormant wheat seeds (Triticum durum cv. Cappelli; 1972 crop) and stimulates it in seeds already relieved from dormancy (1971 crop), with an effect similar to that of gibberellic acid. (2) Fusicoccin (1.5 × 10^?6M) is more active than the two phytohormones gibberellic acid and benzyladenine and than white light in stimulating light-requiring lettuce seeds (Lactuca sativa cv. Grand Rapids) to germinate. Germination of radish seeds (Raphanus sativus) is also accelerated by fusicoccin, while benzyladenine and gibberellic acid are less active in this material. (3) Fusicoccin (1.5 × 10^?5M) removes almost completely the inhibitory effect of abscisic acid on germination of radish and lettuce seeds, whereas benzyladenine (10^?4M) and gibberellic acid (3 × 10^?4M) remove the inhibition only partially. The possible relationship between these results and previous information on growth by cell enlargement is discussed in terms of the mechanism of action of fusicoccin as compared with natural hormones.     

The role of hormones in regulating the germination of polymorphic seeds and early seedling growth of Atriplex triangularis under saline conditions

Germination of the polymorphic seeds and seedling growth of Atriplex triangularis under various salinity, gibberellic acid and kinetin treatments were determined. Gibberellic acid (GA₃; 2.9 m M ) promoted germination and growth at high NaCl concentrations (345 m M ). Kinetin (4.7 μ M ) stimulated germination at all salinities and seed sizes tested. GA₃ and kinetin generally increased seedling growth at all concentrations of salinity studied. Higher concentrations of kinetin were found to be inhibitory.     

The role of darkness, GA and fusicoccin (FC) in breaking photodormancy in Phacelia tanacetifolia seeds

The germination of the negatively photoblastic seeds of Phacelia tanacetifolia Benth. (cv. Bleu Clair) is promoted by gibberellic acid and fusicoccin. In the dark, or in the light in the presence of fusicoccin, seed germination is accompanied by an increase of gibberellic acid-like substances. In these conditions, the inhibition of the synthesis of gibberellic acid-like substances does not prevent seed germination, but it affects the growth and the survival of the seedlings. Seed germination, growth, and survival of seedlings are discussed in relation to phytochrome, fusicoccin, and gibberellic acid-iite substances.     

Secondary dormancy (skotodormancy) in seeds of lettuce (Lactuca sativa cv. Grand Rapids) and its release by light, gibberellic acid and benzyladenine

Lettuce seeds (Lactuca sativa L. cv. Grand Rapids) imbibed in darkness at supra-optimal temperatures (23 ± 1°C) develop a secondary dormancy, termed skotodormancy. The seeds first lose their ability to be promoted to germinate by gibberellic acid, and then lose their ability to be promoted by red light. A combination of red light and gibberellic acid will break skotodormancy for longer than either alone, but red light and benzyladenine together are much more effective. Desiccation of skotodormant seeds does not diminish their dormancy. Embryos dissected from skotodormant seeds will germinate, and are as capable of radicle expansion in the osmoticum polyethylene glycol as are newly-imbibed seeds. Hence skotodormancy is a whole seed dormancy and does not reside within the embryo as an inherent block to germination processes, but as an inability to respond to the stimulation of red light or to hormone.     

Germination characteristics of some plant species from calcareous fens in southern Germany and their implications for the seed bank

Seeds of 25 plant species from calcareous fen hay meadows were exposed to different experimental conditions and their germination was characterised. Constant temperature inhibited germination especially in Cyperaceae . Both gibberellic acid and potassium nitrate failed to terminate dormancy. Increased germination rates were found in dicot species after treatment with gibberellic acid. Temperature fluctuations increased germination of Cyperaceae as well as dicotyledons. Treatment with gibberellic acid removed the chilling requirement in some of the species. Dormancy of small seeds with thin seed coats was broken by the application of gibberellic acid or fluctuating temperature; large thick-coated seeds were unaffected by gibberellic acid. No obligatory darkness requirement was found in any species; three species germinated irrespective of light treatments. All other species achieved higher percentage germination in daylight or in red (670 nm) light. Permanent darkness and far-red light (730 nm) reduced germination drastically. The results indicate that germination characteristics of the species investigated can be related to their seed bank types.     

Effect of water stress and gibberellic acid on germination of flax,sesame and onion seeds

The effects of different osmotic stresses (from 0 to –8× 10⁵ Pa) obtained with NaC or polyethylene glycol 6000 solutions on the germination of flax, sesame and onion seeds were investigated. The effect of presoaking with gibberellic acid (GA³) on the germination of the above mentioned seeds was also studied. It was found that the rate of seed germination and the final germination percentages as well as the amount of water absorbed by the seeds were considerably lowered with the rise of osmotic stress levels whatever the stress agent used, more considerable reduction was obtained under polyethylene glycol 6000 than under NaCl. Presoaking with gibberellic acid increased the rate and the final germination percentage of osmotically stressed flax and sesame seeds, while those of stressed onion seeds were slightly retarded.     

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.     

Synergism between gibberellic acid and low Pfr levels inducing germination of Kalanchoë seeds

Sown on water, seeds of Kalanchoëbiossfetdiana Poelln. cv. Feuerblute are absolutely light-requiring and show full red/far-red reversibility. In seeds, sown on 2 ×10^-3 M gibberellic acid, red/far-red reversibility disappears and both short red and far-red irradiations induce germination. Gibberellic acid alone does not induce germination, but it increases the physiological activity of P_fr to the extent, that the low P_fr level obtained by far-red irradiation becomes very effective. The synergism between gibberellic acid and far-red light appears after a two-day incubation; period. The nature of this lag phase was examined by measuring both germination and uptake of labelled gibberellic acid in intact seeds and seeds with a punctured seed coat. The lag phase was shown to be independent of the uptake kinetics of gibberellic acid and allows development to a specific stage, necessary for germination after phytochrome-phototransformation. The kinetics of the uptake of gibberellic acid by intact seeds and embryos of intact seeds are different. In intact seeds most of the gibberellic acid is retained in the seed coat; only a small fraction actually penetrates to the embryo where it can exert its physiological activity.     

Precocious Germination during In Vitro Growth of Soybean Seeds

Immature Glycine max (L.) Merrill seeds were grown and matured in liquid medium at 25°C under fluorescent light. In standard medium containing minerals, 146 millimolar sucrose and 62.5 millimolar glutamine (osmolality 0.24), precocious germination seldom occurred with a starting seed size of less than 300 milligrams fresh weight. Frequency of precocious germination increased with increased starting seed size. Sucrose concentration strongly affected precocious germination while glutamine concentration had no effect. Starting with 300 to 350 milligrams fresh weight seeds, treatments which reduced the sucrose concentration or lowered the osmolality of the culture medium stimulated precocious germination, and increased the fresh weight growth but not the dry weight growth of seeds. Increasing the osmolality to 0.38 with sucrose or mannitol prevented precocious germination without reducing dry weight accumulation in seeds. In medium with initially low osmolality, precocious germination was inhibited by addition of 1 to 100 micromolar abscisic acid to the medium without a reduction in seed growth. During growth and maturation of large soybean seeds in vitro, precocious germination and other abnormal tissue growth can be prevented by high sucrose or mannitol concentrations in the medium or by addition of abscisic acid.     

Inhibitory Effects of Methyl Jasmonate on the Germination and Ethylene Production in Cocklebur Seeds

Methyl jasmonate (JA-Me) inhibited the germination of cocklebur (Xanthium pennsylvanicum Wallr.) seeds. The inhibition of the germination of cocklebur seeds treated with JA-Me at concentrations less than 300 μm was nullified by ethylene applied exogenously, although the inhibitory effect of 1,000 μm JA-Me was not recovered completely even by high concentrations of ethylene (10,000 μL/liter). JA-Me inhibited ethylene production before seed germination. The level of 1-aminocyclopropane-1-carboxylic acid (ACC) in the cotyledonary tissues treated with JA-Me decreased but not the level of 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC). JA-Me inhibited the conversion of ACC to ethylene in the tissues. These results suggested that JA-Me inhibits ethylene production by prevention of ACC oxidation in addition to ACC synthesis. We believe that the inhibition of ethylene production by JA-Me results in the retardation of the germination of cocklebur seeds. Received June 4, 1997; accepted October 23, 1997