Respiration of Grand Rapids lettuce (Lactuca sativa L.) seeds in relation to chemical and photocontrol of germination

A 10-min irradiation with red light (R) of ‘Grand Rapids’lettuce (Lactuca sativa L.) seeds increased respiratory rates(QO₂ and QCO₂) over those of far red (FR)-irradiated seeds within2 to 3 hr. Differences in respiration between R- and FR-irradiatedseeds became more pronounced with time. The respiration, indarkness at 25?C, of seeds air dried after germination promotingimpregnation treatments with 1.0 mM GA₃+0.5 mM kinetin in acetonewas stimulated slightly by the 6th hour after planting and markedlyby the 12th and 24th hour. Results with seeds in GA₃+kinetindissolved in water were similar. ABA, which inhibits germinationand seedling growth, prevented stimulation in respiration oflight-induced seeds. ABA also repressed respiration when usedwith GA₃+kinetin. The germination and growth retardant, isopropylN-(3-chlorophenyl) carbamate (CIPC), did not affect the respirationof either R- or FR-treated seeds. A comparison of the effectsof chemical and light treatments on respiration indicate thatchemical and photocontrol mechanisms are not identical. (Received May 17, 1976; )     

Changes in Ultrastructure and Abscisic Acid Level, and Response to Applied Gibberellins inTaxus maireiSeeds Treated With Warm and Cold Stratification

Seeds ofTaxus maireiare known for their deep dormancy whichcan only be broken by a procedure involving warm stratificationfollowed by cold stratification. Treatments with alternatingtemperatures of 25/15 or 23/11 °C (12 h light) for 6 monthsfollowed by 5 °C for 3 months were successful in overcomingseed dormancy. After 6 months of warm stratification, cytologicalchanges observed included: enlargement of the embryo; a decreasein the number of lipid bodies; appearance of ER; and increasesin mitochondria, plastids, dictyosomes, vacuoles and microbodiesin the shoot apical meristem. Cold stratification followingthe warm treatment induced cell division, and one or two distinctnucleoli in the shoot apical meristem cells were observed. Bothwarm and cold stratification reduced endogenous ABA concentrationsfrom the original 8888 pg per freshly harvested seed to 392and 536 pg, respectively. Treatment with exogenous gibberellinsafter seeds had been warm-stratified showed that GA₄and GA₇wereeffective at promoting seed germination, but GA₃was not. Theseresults suggest that the strong seed dormancy ofT. maireicouldbe caused by a high ABA content and underdevelopment of theembryos in freshly shed seeds. We conclude that warm stratificationwith alternating temperatures increases the growth of embryosby cell expansion and enlargement and decreases ABA content,but seeds still remain ungerminated. Cold stratification mayinduce the response to GAs and initiate cell division resultingin release from physiological dormancy and subsequent germinationofT. maireiseeds.Copyright 1998 Annals of Botany Company Taxus mairei; ultrastructure; abscisic acid; gibberellin; seed dormancy; stratification; germination.     

Germination of seeds in Jussiaea suffruticosa

Seeds of Jussiaea suffruticosa reach high germination percentagesonly when exposed to long periods of continuous illumination.The light reaction may be repeatedly reversed by short exposuresto red and far red light, thus being mediated by the phytochromesystem. Seeds also germinate at high percentages if exposedto various cycles of 1 hr light and 24 hr of darkness at 20°C.If the temperature in the periods of darkness is raised up to30°C or lowered to 10°C the promotive effect of lightis inhibited. High temperatures (35°C) during imbibitionhave a promotive effect, whereas a pure O₂ atmosphere decreasesthe response to light. KNO₃ and kinetin enhance the responseto light but do not provoke germination in the dark. Only ifseed coats are punctured or removed does germination in thedark occur. (Received January 14, 1969; )     

Induction of Secondary Dormancy in Chenopodium bonus-henricus L. Seeds by Osmotic and High Temperature Treatments and Its Prevention by Light and Growth Regulators

Factors controlling the establishment and removal of secondary dormancy in Chenopodium bonus-henricus L. seeds were investigated. Unchilled seeds required light for germination. A moist-chilling treatment at 4 C for 28 to 30 days removed this primary dormancy. Chilled seeds now germinated in the dark. When chilled seeds were held in the dark in −8.6 bars polyethylene glycol 6000 solution at 15 C or in water at 29 C a secondary dormancy was induced which increased progressively with time as determined by subsequent germination. These seeds now failed to germinate under the condition (darkness) which previously allowed their germination. Continuous light or daily brief red light irradiations during prolonged imbibition in polyethylene glycol solution at 15 C or in water at 29 C prevented the establishment of the secondary dormancy and caused an advancement of subsequent germination. Far red irradiations immediately following red irradiation reestablished the secondary dormancy indicating phytochrome participation in “pregerminative” processes. The growth regulator combination, kinetin + ethephon + gibberellin A₄+A₇ (GA₄₊₇), and to a relatively lesser extent GA₄₊₇, was effective in preventing the establishment of the secondary dormancy and in advancing the germination or emergence time. Following the establishment of the secondary dormancy by osmotic or high temperature treatments the regulator combination was relatively more active than light or GA₄₊₇ in removing the dormancy. Prolonged dark treatment at 29 C seemed to induce changes that were partially independent of light or GA₄₊₇ control. The data presented here indicate that changes during germination preventing dark treatment determine whether the seed will germinate, show an advancement effect, or will become secondarily dormant. These changes appear to be modulated by light and hormones.     

A Model of the Effects of a Wide Range of Constant and Alternating Temperatures on Seed Germination of FourOrobanche Species

Seeds of the obligate parasitic plants, Orobanche spp., wereconditioned in water or GA₃for 2 or 12 weeks and then stimulatedto germinate by the synthetic stimulant GR24. Temperature treatmentsduring the germination tests comprised 169 different constantand alternating temperature regimes on a two-dimensional gradientplate. Optimum temperatures for germination of seeds of O. aegyptiacaand O. crenata were 18–21 °C and 18 °C, respectively.However, longer conditioning periods slightly lowered the optimain both species, and the maximum germination percentage wasalso reduced due to an induction of secondary dormancy. At agiven mean temperature, more seeds germinated at constant thanat alternating temperatures. Results were analysed in termsof characteristics of alternating temperatures that appearedto control germination, i.e. mean temperature, maximum temperature,amplitude (difference between daily maximum and minimum temperatures)and thermoperiod (the time spent at the maximum temperatureeach day). Final germination was modelled on the basis of therebeing two prerequisites for germination: a minimum mean temperaturewhich must be exceeded and a maximum temperature above whichthe seed will not germinate. These two requirements were assumedto be independent and to be normally distributed in the seedpopulation so that final germination could be described by amultiplicative probability model. Because of the response tomaximum temperature, inhibitory effects were more evident atalternating temperatures. Amplitude and thermoperiod influencedthis effect of maximum temperature. The implications of thedetrimental effect of alternating temperatures for germinationofOrobanche spp. in the field are discussed. Copyright 1999Annals of Botany Company Orobanche aegyptiaca, O. crenata, O. cernua, O. minor, broomrape, seed germination, temperature, germination model, secondary dormancy.     

INTERACTION OF GIBBERELLIN A3, AND INORGANIC PHOSPHATE IN TOBACCO SEED GERMINATION

1. Investigation was made on the influence of inorganic phosphateupon the germination of positively photoblastic tobacco seed(Nicotiana tabacum L. var. uirginica (AGDH.) COM. "Bright Yellow")induced by GA₃, GA₃M, kinetin, red light, and ammonium saltsof various organic acids.
2. Inorganic phosphate increases theGAs-induced germination, andinhibits the germination causedby ammonium citrate, while itdoes not influence the germinationbrought about by GA₃M, kinetin,and red light.
3. The optimumpH for the GA₃-induced germination lies in the acidicpH range,indicating that the undissociated form of GA₃ is operative.The stimulatory effect of phosphate is, however, not ascribedmerely to the pH control in the mediurr. Phosphate exerts somespecific influence for which the presence of the free carboxylgroup of GAs is required.
4. The observed contrasting effectsof phosphate on the GA₃-inducedgermination (i.e., acceleration),on the one hand, and on theammonium citrate-induced germination(i.e., inhibition), onthe other, were explained by assumingthat the phosphate effectsultimately consist in acceleratingthe uptake of the carboxylicacid into the seeds.
5. GA₃M alsohas an activity of inducing the germination of tobaccoseedwithout light.

¹Present address: Department of Vegetable Crops, Universityof California, Davis, California, U.S.A. (Received March 12, 1962; )     

Germination and Dormancy in Immature and Fresh-mature Lettuce Seeds

The effects of red light, far-red light, Gibberellin A₃, andethephon were studied on the germination of lettuce seeds cv.Grand Rapids harvested at different stages of development. Seeds did not become capable of germination until 8 days afteranthesis. Red light promoted seed germination from the age of8–9 days following anthesis up to the newly mature stage.Ten or 11 days following anthesis, a large percentage of seedsbecame capable of germination in the dark and therefore couldbe considered not dormant. They were affected by far-red light,but less so than the mature seeds. The effect of light on the germination of developing seeds appearedto be similar to the known light effect on mature lettuce seedgermination. Gibberellin A₃ and ethephon had no effect on immatureand fresh seed germination. Lactuca sativa L., Lettuce, germination, dormancy, red light, far-red light, gibberellin A₃, ethephon     

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.     

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

Ten-30 d imbibed skotodormant lettuce seeds (Lactuca sativaL. cv. Grand Rapids) showed no germination with water alone.However, following a single treatment of red light (R), gibberellinA₃ (GA₃) or 1 h acid immersion (pH 0–1) plus water rinse,7% germinated. These imbibed skotodormant seeds germinated 85%or higher if acid immersion was carried out before R or GA₃.Similar values were obtained with imbibed skotodormant seedsunder acid immersion plus drying treatment applied at day 10or 20 plus R or GA3 treatment applied 10 d later. One or twodrying treatments alone reduced the degree of skotodormancyand made seeds more responsive to R but not GA₃. Seeds withone R plus drying treatment at day 10 or 20 germinated about50% with or without an additional R, and 80% or more with GA₃on day 20 or 30. The 20 or 30 d skotodormant seeds having R(with or without drying) or acid plus R and drying treatmenton day 10 or 20 and additional dark incubation in water for10d showed 85 to 100% germination with only acid immersion.The skotodormancy was eliminated by the acid immersion but itwas initiated again if R or GA₃ treatment was not given immediately.It is concluded that the drying treatment, after eliminationof skotodormancy by acid or acid + R pretreatment, preventsthese seeds re-entering skotodormancy and maintains a high germinationpotential under dark storage for up to 20 d. Key words: Dark reversion of phytochrome, gibberellin A₃, acidification, skotodormancy, induction and breakage of seed dormancy     

Quantal Response of Seed Germination in Seven Genera of Cruciferae to White Light of Varying Photon Flux Density and Photoperiod

The response of the germination of seeds of Barbarea vema (Mill.)Aschers, Brassica chinensis L., Brassica juncea (L.) Czern.& Coss., Brassica oleracea L. var. gongylodes L., Camelinasaliva (L.) Crantz, Eruca saliva Mill., Lepidium sativum L.,Nasturtium officinale R. Br., and Rorippa palustris (L.) Besserto white fluorescent light of different photon flux densitiesapplied for different daily durations in a diurnal alternatingtemperature regime of 20 °C/30 °C (16 h/8 h) was quantifiedby linear relations between probit percentage germination andthe logarithm of photon dose, the product of photon flux densityand duration. The low energy reaction, in which increasing dosepromotes germination, was detected in all the seed populationsbut in Barbarea vema and Brassica Juncea the lowest photon doseapplied (10^–5–2 and 10^{–5 7} mol m^–2 d^–1,respectively) was sufficient to saturate the response. Comparisons,where possible, between photoperiods demonstrated reciprocity,i.e. germination was proportional to photon dose irrespectiveof photoperiod, for the low energy reaction in Brassica oleracea(1 min d^–1 to 1 h d^–1), Camelina saliva (1 min d^–1to 8 h d^–1), Eruca saliva (1 min d^–1 to 24 h d^–1),Lepidium sativum (I min d^–1 to 8 h d^–1) and Rorippapalustris (1 min d^–1 to 8 h d^–1), but not in Brassicachinensis and Nasturtium officinale. The high irradiance reaction,in which increasing dose inhibits germination, was detectedin Barbarea vema, Brassica chinensis, Brassica juncea, Brassicaoleracea, and Camelina saliva. The minimum dose at which inhibitionwas detected was lO^–0–3 mol m^–2 d^–1.These results are discussed in the context of devising optimallight regimes for laboratory tests intended to maximize germination The response of germination to photon dose was also quantifiedwith 3 x 10^–4 M GA₂, co-applied (Brassica chinensis, Camelinasaliva, and Lepidium sativum) and with 2 x 10^–2 M potassiumnitrate co-applied (Brassica chinensis). In the latter casepotassium nitrate had no effect in the dark and inhibited germinationin the light, but GA₂, promoted germination substantially inall three species. Variation amongst seeds in the minimum photondose required to stimulate germination was not affected by co-applicationof GA₂, in Brassica chinensis and Camelina saliva, whereas seedsof Lepidium salivum showed a narrower distribution of sensitivitiesto the low energy reaction in the presence of GA₂ Barbarea vema (Mill.) Aschers, Brassica chinensis L., Brassica juncea (L.) Czern. & Coss., Brassica oleracea L. var. gongylodes L., Camelina saliva (L.) Crantz, Eruca saliva Mill., Lepidium satiaum L., Nasturtium officinale R. Br., Rorippa palustris (L.) Besser, Cruciferae, light, gibberellic acid, seed germination, seed dormancy     

Effects of Temperature and Various Red or Far-red Irradiations on Phytochrome- and Gibberellin A3-mediated Germination Control in Partially Hydrated Lettuce Seeds

Dark reversion of phytochrome in partially hydrated lettuceseeds (Lactuca sativa cv. Grand Rapids) is temperature dependent.After initial red irradiation (R) the higher the storage temperature,the higher the dark reversion rate. Following dark moist storage(DMS) at 30 ?C for 15 d none of the seeds receiving initialR germinated, whereas seeds stored at 0 ?C germinated nearlyas well (about 80%) as unstored controls. The half-time fordark reversion at 20 ?C and 30 ?C is 9 d and 3 d respectively.Repeated R treatments given at 5 d intervals during DMS at 20?C and 30 ?C maintained a high germination capacity. With threeor more R treatments the effect of high temperature largelydisappeared. Dark reversion of phytochrome was not observed in partiallyhydrated lettuce seeds receiving continuous red irradiation(cont R) for two or more days. The promotive effect of contR could be reversed at any time with a brief far-red irradiation(FR), indicating that the phytochrome system remained fullyphototransformable. With continuous far-red light (cont FR)the ability of gibberellin A₃ (GA₃) to stimulate germinationdisappeared and response to GA₃ also diminished in cont R followedby FR but at a slower rate indicating the induction of secondarydormancy in these partially hydrated seeds. This induction ofdormancy was retarded by repetitive or cont R but was enhancedby cont FR. The results of this study suggest a role for theaccumulated stable intermediates of phytochrome transformationin partially hydrated seeds with repeated or continuous R treatmentsand different effects of GA₃ and R in the regulation of germination. Key words: Phytochrome, Lactuca sativa, Seed germination, Temperature, Dark reversion of phytochrome, Seed water content     

Light Actions in the Germination of Cocklebur Seeds: V. EFFECTS OF ETHYLENE, CARBON DIOXIDE AND OXYGEN ON GERMINATION IN RELATION TO LIGHT

Esashi, Y., Hase, S. and Kojima, K. 1987. Light actions in thegermination of cocklebur seeds. V. Effects of ethylene, carbondioxide and oxygen on germination in relation to light.–J.exp. Bot. 38: 702–710. Effects of ethylene, CO² and O² on the germination of after-ripenedupper cocklebur (Xanthium pennsylvanicum Wallr.) seeds wereexamined in relation to pre-irradiation by red (R) or far-red(FR) light In order to remove the pre-existing Pfr, seeds weresoaked in the dark for various periods prior to light irradiationand gas treatments. Regardless of light, 0.3 Pa C₂H₄ promotedgermination at 23 ?C, but it strongly inhibited germinationwhen applied at 33 ?C, the optimal temperature for the germinationof this seed. However, delayed application of C₂H₄ during 33?C incubation stimulated germination independently of lightin a similar manner to that seen at 23 ?C. It is, therefore,suggested that the germination-regulating action of C2H4 iscompletely independent of phytochrome. In contrast, the germination-promoting effect of 3–0 kPaCO₂ was pronounced only when the seeds were previously irradiatedby R, regardless of temperature, suggesting that CO₂ actionto promote germination depends upon Pfr. A synergism betweenCO₂ and C₂H₄ at 23 ?C was observed only in the germination ofseeds pre-irradiated by R, while at 33 ?C an antagonism occurredindependently of light. The stimulation of C₂H₄ production byCO₂ was most striking in the cotyledonary tissue pre-irradiatedby R. However, the R-dependent enhancement of CO₂-stimulatedC₂H₄ production was negated by the subsequent FR and it wasnot found in the presence of 1-aminocyclopropane-1-carboxylicacid (ACC). Moreover, the R dependency of the germination-promotingCO₂ effect disappeared in the presence of C₂H₄. The R-dependentC₂H₄ production enhanced by CO₂ may thus be involved, at leastpartially, in some step of conversion from methionine to ACC. The germination-promoting effect of C₂H₄, but not CO₂, was enhancedby O₂ enrichment regardless of light. However, the germination-promotingeffect of pure O₂ itself appeared to depend upon pre-irradiationwith R Key words: Carbon dioxide, cocklebur seed, ethylene, far-red light, germination, oxygen, red light, Xanthium pennsyloanicum     

Seed germination behavior of five Resedaceae species from arid Arabian desert: gibberellic acid and light effects

Abstract

Light and growth-promoting compounds, such as gibberellic acids (GA₃), are among the most important factors that can break physiological seed dormancy. Here, we investigate the effects of GA₃ and light on germination of five species of Resedaceae that are known to have different levels of physiological dormancy. Seeds were incubated at 20/30?°C in both 12-hr photoperiod and complete darkness. To study the effect of growth hormone on germination, seeds were soaked for 24?h in different concentrations of GA₃ before sowing. The annuals (Reseda aucheri and Oligomeris linifolia) and the perennial Ochradenus arabicus had deep physiological dormancy, and exogenous application of GA₃ enhanced their germination in the light, but not in darkness; few or no seeds germinated in the dark in these species. Ochradenus aucheri and O. baccatus had intermediate and non-deep dormancy, respectively, and application of GA₃ enhanced their germination in both light and darkness. Germination of the annual species was much slower than that of the shrubby perennials. Overall, these results indicate that conditions under which seed developed, matured and stored on maternal plants as well as incubation conditions should be taken into consideration when assessing germination behavior of the perennial species of Resedaceae.     

Variation in seed dormancy and germination among populations of Silybum marianum (Asteraceae)

Milk thistle (Silybum marianum) is a medicinal plant; however, lack of consistency in past dormancy studies has hindered propagation of this species from seeds. We tested the germination responses of freshly harvested and after-ripened (stored for 2 and 7 months; 25°C at 50% relative humidity) seeds from three populations (P1, P2 and P3) in Iran at varying constant or alternating temperatures, with or without GA₃ and in light and continuous darkness. No germination occurred in freshly harvested seeds incubated at any condition without GA₃ application, indicating that all the seeds were dormant. Seeds from P1 and P2, which developed under relatively dry, warm conditions, germinated over a wider range of temperatures after 2 months of dry storage, indicating type 6 of non-deep physiological dormancy (PD). Seeds from P3, which developed under relatively wet, cool conditions, incubated at constant temperatures (especially on GA₃), exhibited an increase in maximum temperature for germination, indicating type 1 of non-deep PD. Light improved germination of after-ripened seeds, and GA₃ application substituted for the light requirement for germination. This is the first report that environmental conditions during seed development may be correlated with differences in the type of non-deep PD. We conclude that milk thistle seeds are positively photoblastic and photodormant and the germination responses of after-ripened seeds from different populations are different under darkness. Therefore, the impacts of genetic differences and maternal effects on the induction of dormancy during seed development should be considered in attempts to domesticate this medicinal plant.     

Analysis of the Effect of Cold Stratification on the Germination Response to Light and Alternating Temperatures using Selected Seed Populations of Ranunculus sceleratus L.

From a single population of achenes (seeds) of Ranunculus sceleratusL. sub-populations were selected on the basis of the sensitivityof individuals to an increasing number of daily light/temperatureshift cycles. Each cycle comprised a 4 h pulse of red lightfollowed by a 4 h temperature shift from 16 to 31 ?C. Selectionfor low dormancy (NND and ND) and high dormancy (D and DD) populationsresulted in a > 5-fold difference in the number of cyclesrequired for 50% germination. Despite a shift in the mean levelof dormancy the distributions of sensitivity (slopes of dose-responsecurves) were similar in all four selected populations. Differentialeffects of cold stratification on the germination response tolight and alternating temperatures were related to the depthof primary dormancy. The proportion of individuals that respondedpositively to the dormancy breaking effects of cold stratificationfollowed the trend DD < D < ND < NND. In high dormancypopulations (D and DD) the rate and uniformity of germinationof some individuals was reduced by cold stratification, indicatinga dormancy inducing effect. Over the range 2 to 11 ?C the effectivenessof dormancy release or dormancy induction was inversely relatedto temperature. The effects of cold stratification on the expressionof dormancy in R. sceleratus are discussed in relation to areproductive strategy involving winter and summer annual behaviour. Key words: Cold stratification, selection, dormancy, light, alternating temperatures, germination     

Mode of action of gibberellic Acid and light on lettuce seed germination

The seeds of lettuce (Lactuca sativa L. cv. Grand Rapids) germinate in darkness at 25 C when treated by gibberellic acid (GA₃) for 1 hour following 2 hours of imbibition. The time of GA₃ application influences the rate and the final percentage of seeds that germinate. In contrast, red light illumination given at different times affects only the rate and not the final germination percentage. The early process(es) of germination initiated by GA₃ or light treatment can be arrested by subjecting the treated seeds to a nongerminative temperature of 35 C. The results suggest differences in the mode of action of light and GA₃ during germination. They indicate that different kinds of processes are involved in the biochemical control of germination.     

Germination responses of Discopodium penninervium Hochst. to temperature and light

The germination responses of Discopodium penninervium were tested at different constant and alternating temperature regimes as well as under various light conditions both in the laboratory and glasshouse. Seeds incubated at 10, 15, 20, 25 and 30 °C failed to germinate. When the seeds were incubated at alternating temperatures of 20/12 °C and 30/12 °C under continuous light, germination was 89 and 61%, indicating that the species requires alternating temperatures as a cue for germination. However, germination declined as the amplitude of alternating temperatures increased from 8 °C and was completely inhibited at an amplitude of 23 °C, suggesting that the optimum amplitude is around 8 °C. Germination was less than 10% in light and nil in darkness at 20 °C in the laboratory. In contrast, seeds incubated at 20/12 °C germinated to 96 and 86% in light and darkness, respectively. Seeds incubated under leaf shade in the glasshouse failed to germinate whereas those incubated under direct daylight and darkness germinated to 44 and 50%, respectively, 30 days after sowing. When seeds incubated under leaf shade and in darkness were exposed afterwards to light, final percent germination was 83% from seeds incubated initially under direct daylight, 79% from those incubated under leaf shade and 86% from those incubated in darkness. The requirement for alternating temperatures and light rich in red:far red ratio to break the dormancy of seeds of D. penninervium could restrict germination to gaps in the vegetation. The results conform with the ecology of the species.     

Effects of Gibberellins on Seed Germination of Phytochrome-Deficient Mutants of Arabidopsis thaliana

Experiments were carried out to explore the involvement of gibberellins(GAs) in the light-induced germination of Arabidopsis thaliana(L.) Heynh, using wild type (WT) and phytochrome-deficient mutants(phyA, phyB and phyAphyB deficient in phytochrome A, B and Aplus B, respectively). Seed germination of WT and phytochrome-deficientmutants was inhibited by uniconazole (an inhibitor of an earlystep in biosynthesis of GA, the oxidation of ent-kaurene) andprohexadione (an inhibitor of late steps, namely, 2rß-and 3rß-hydroxylation). This inhibition was overcomeby simultaneous application of 10^-5 M GA₄. The relative activityof GAs for promoting germination of uniconazole-treated seedswas GA₄>GA₁=GA₉>GA₂₀. The wild type and the phyA and phyBmutants had an increased response to a red light pulse in thepresence of GA₁, GA₄, GA₉, GA₂₀ and GA₂₄ but there were no significantdifferences in activity of each GA between the mutants. Therefore,neither phytochrome A nor hytochrome B appears to regulate GAbiosynthesis from GA₁₂ to GA₄ during seed germination, sincethe conversion of GA₁₂ to GA₉ is regulated by one enzyme (GA20-oxidase). However, GA responsiveness appears to be regulatedby phytochromes other than phytochromes A and B, since the phyAphyBdouble mutant retains the photoreversible increased responseto GAs after a red light pulse. (Received February 13, 1995; Accepted July 11, 1995)     

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