Effects of long-term storage on phytochrome-mediated germination in lettuce seeds

The effects of long-term seed storage on the physiological properties of phytochrome-mediated germination including water uptake, the temperature and light flunnce dependencies of germination and dark germination were studied. The fluenceresponse relationships of the brief irradiation with monochromatic red (660 nm, 7.5 W m⁻²) and far-red (750 nm, 6.6 W m⁻²) light at various times after sowing were also studied. The samples used consisted of three lots of seeds ofLactuca sativa L. cv. MSU-16, which had been harvested in 1976, 1979 and 1985 and stored dry for 9, 6 and 0 years, respectively, in darkness at 23±2 C until the experiments were carried out in July–August, 1985. Seeds with the longer storage periods showed the higher ability to germinate in both continuous darkness and continuous white fluorescent light at 20–30 C. In the seeds stored for 6 or 9 years, red light irradiation for 20 sec given at 15 min or more after sowing at 25 C induced as high a percent germination (85–95%) as those under continuous white fluorescent light. In the freshly harvested seeds, however, germination under continuous white fluorescent light (46%) was considerably lower than the germination induced by the red pulse (97%). Germination of the seeds decreased when the intervals between sowing and a far-red irradiation for 20 sec increased up to 100 min (or 30 min in the freshly harvested seeds). The far-red pulse given later than 100 min (or 6 hr in the freshly harvested seeds) after sowing resulted in an increased germination up to the dark-germination levels with increasing intervals between sowing and the pulse irradiation. Before or at 3 min after sowing, the seeds stored for 6 or 9 years were responsive to the far-red pulse although they were not or hardly responsive to the red pulse, while the freshly harvested seeds were responsive to both the far-red and the red pulses. These data indicate that normal functions of phytochrome completely survived in the dry seeds during storage at 25 C for as long as 6 or 9 years and that these functions are restored into full operation by means of imbibition. The differences in the dependence of germination on the time and fluence of a single pulse of red or far-red light seems to be related to the smaller water content throughout the imbibition in the seeds with the longer storage periods. The greater ability to germinate in the dark indicates the greater amounts of P_FR or the greater responsivity to P_FR, in the seeds with the longer storage periods.     

THE EFFECTS OF TEMPERATURE ON THE GERMINATION AND RADICLE GROWTH OF PHOTOSENSITIVE LETTUCE SEED

1. The time course of germination of Grand Rapids lettuce seedshas heen followed with different combinations of temperature(3°–35°) and irradiation (red or far-red light).For each set of conditions the following three parameters weredetermined: (i) the time required for half maximum germination,(ii) the rate of germination during the actively germinatingphase, and (iii) the maximum germination attained. In general,as the temperature was lowered, with dark-imbibed seeds, (i)became longer, (ii) became lower, but (iii) became progressivelyhigher. The effect of red light at any temperature was to shorten(i) and increase (ii) and (iii) over the values dark controls.Far-red light exerted an effect opposite to that of red light.Temperatures higher than 25° inhibited (ii) and (iii) underany light conditions. The optimum temperature to the actionof red and far-red light is 25°, at which the stimulatoryeffect of red light and the inhibition of this effect by far-redlight are both maximal. 2. The growth of the radicles of de-coated seeds of Grand Rapidslettuce shows two phases at all temperatures studied. PhaseI is characterized by slow but linear growth which continuesuntil shortly after visible differentiation of the radicle intothe hypocotyl and the root. Phase II is a phase of active growthin which the total length reflects mainly the length of theroot. The optimum temperature for Phase I is 25°-35°,and that, for Phase II is 25°. In neither phase, and atnone of the temperatures studied, is there any effect of redor far-red radiation on the growth of the radicle. The firstvisible sign of radicle elongation in red light induced seeds,however, takes place at exactly the same time as that of germination. 3. Similarities and dissimilarities between the germinationand the growth are pointed out, and it is concluded that thetwo phenomena are different, but proceed at sites closely associatedin the embryo. ¹Present address: Johnson Foundation for Medical Physics, Universityof Pennsylvania, Philadelphia, Pa., U.S.A.     

The Effects of Red, Far-red, and Blue Light on the Geotropic Response of Coleoptiles of Zea mays

The effects of red, far-red, and blue light on the geotropicresponse of excised coleoptiles of Zea mays have been investigated.Seedlings were grown in darkness for 5 or 6 days, exposed tovarious light treatments, and then returned to darkness fordetermination of the geotropic response. The rate of response of the coleoptiles is decreased after theyhave been exposed to red light (620–700 mµ, 560ergs cm^–2sec^–1 for the 24 hrs, but not for the 4hrs, preceding stimulation by gravity. Furthermore, their rateof response is greatly reduced if they are exposed to red lightfor 10 min and then returned to darkness for 20 hrs before geotropicstimulation. At 25° C an interval of 6 to 8 hrs elapses between a 10-minexposure to red light and the first detectable decrease in thegeotropic response of the coleoptile. This interval can be lengthenedby exposing the seedlings to low temperatures (0° to 2°C) after the light treatment but cannot be greatly shortenedby increasing the duration of exposure to red light. Using a standard procedure of exposing 5-day-old etiolated seedlingsto light for various times, replacing them in darkness for 20hrs and then determining the response of the coleoptiles to4 hrs geotropic stimulation, it has been found that: (a) Exposureto red light for 15 sec significantly decreases the geotropiccurvature of the coleoptiles and that further reduction occurson increasing the length of the light treatment to 2 and 5 min.(b) Far-red light has no effect on the geotropic response ofthe coleoptiles but it can completely reverse the effect ofred light. After repeated alternate exposure to red and far-redlight the geotropic response of the coleoptile is determinedby the nature of the last exposure, (c) Complete reversal ofthe effect of red light by far-red radiation only occurs whenexposure to far-red follows immediately after exposure to red.The reversing effect of far-red radiation is reduced if a periodof darkness intervenes between the red and far-red light treatments,and is lost after a dark interval of approximately 2 hrs. The effect of red light on the rate of geotropic response ofthe coleoptiles is independent of their age and length at thetime of excision. Blue light acts in a similar way to red light, but the seedlingsare less sensitive to blue than to red light. Coleoptiles grown throughout in a mixture of continuous, weak,red, and far-red light have a lower rate of geotropic responsethan etiolated coleoptiles.     

Blue light induced inhibition of seed germination: The necessity of the fruit coats for the blue light response

The seeds (achenes) of Laportea bulbifera require a chilling to break their dormancy and are negatively photoblastic. Their germination is inhibited by both continuous blue light and continuous or prolonged far-red radiation. The germination of de-coated seeds, prepared by removing the fruit coats, however, was strongly inhibited by continuous far-red, but not by continuous blue light. Photoreversible germination by a brief irradiation with red light occurred when the chilled seeds were exposed to prolonged far-red light. These results suggest that far-red light may regulate the germination of L. bulbifera seeds through the phytochrome system which exists in the regions other than fruit coats and that the blue light reaction may be governed by other photoreceptor system(s).     

An unusual effect of the far-red absorbing form of phytochrome: Photoinhibition of seed germination inBromus sterilis L.

Seeds ofBromus sterilis L. germinated between 80–100% in darkness at 15° C but were inhibited by exposure to white or red light for 8 h per day. Exposure to far-red light resulted in germination similar to, or less than, that of seeds maintained in darkness. Germination is not permanently inhibited by light as seeds attain maximal germination when transferred back to darkness. Germination can be markedly delayed by exposure to a single pulse of red light following 4 h inhibition in darkness. The effect of the red light can be reversed by a single pulse of far-red light indicating that the photoreversible pigment phytochrome is involved in the response. The response ofB. sterilis seeds to light appears to be unique; the far-red-absorbing form of phytochrome (Pfr) actually inhibiting germination.Abbreviations Pr red absorbing form of phytochrome - Pfr far-red absorbing form of phytochrome     

Primary dormancy in tomato (Lycopersicon esculentum cv. Moneymaker): studies with the sitiens mutant

Intact wild-type tomato (Lycopersicon esculentum cv. Moneymaker)seeds do not complete germination to the same percentage orat the same speed as intact ABA-deficient sitiens (sit^w) mutantseeds when seeds of both genotypes are imbibed on polyethyleneglycol (PEG) solutions of –0.3 to –1.5 MPa osmoticpotential. However, if the thicker testas of wild-type seedsare removed (stripped) from the micropyle without damaging theendosperm, both the percentage and speed of germination at lowexternal water potential are similar to that of sit^w mutantseeds. Removing the micropylar end of the testa from sit^w seedsdid not enhance either the speed or percentage of germinationon PEG solution. Despite similar germination percentage and speed between strippedwild-type seeds and either stripped or intact sit^w seeds underosmotic stress, some differences in seed metabolism are evidentbetween genotypes. The activity of endo-ß-mannanasewas greater in the endosperm of sit^w mutant seeds compared tothe endosperm of wild-type seeds when seeds were exposed toosmotic stress. Although     

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     

Variations in Tiller Dynamics and Morphology in Lolium multiflorum Lam.Vegetative and Reproductive Plants as affected by Differences in Red/Far-Red Irradiation

Lolium multiflorum Lam, plants were grown in a growth room undertwo light sources with red/far-red ratios of 1·62 and0·84 but similar photosynthetically active radiation.In both situations the capacity to produce new tillers and thelight available per tiller decreased with canopy growth. Tilleringwas further reduced by the low red/far-red ratio while lightinterception and plant dry weight were unaffected by this treatment.In both reproductive and vegetative plants under the lower red/far-redratio the time between leaf expansion and the appearance ofa tiller in its axil was increased and the proportion of ‘maturebuds’ that developed was reduced. Irradiation with lowred/far-red advanced the reproductive development and increasedthe number of fertile tillers per plant. It also caused longerleaf sheaths, blades and reproductive shoots. The results suggestthat as canopy density increases the lower light interceptionper tiller and the photomorphogenic effect of low red/far-redratios may reduce the capacity to produce new tillers. Lolium multiflorum, Lam., annual ryegrass, tillering, tiller growth, leaf growth, flowering, light quality.     

Abscission: Response to Red and Far-Red Light

Craker, L. E., Zhao, S. Y. and Decoteau, D. R. 1987. Abscission:response to red and far-red light.—J. exp. Bot. 38: 883–888. The dose-response and time relationship of red and far-red lightin the inhibition and promotion, respectively, of dark-inducedleaf abscission was quantified using cuttings of coleus (ColeusBlumei Benth.). A continuous photon flux of approximately 15nM m^–2 s^–1 of red light was sufficient to preventleaf abscission. Abscission was promoted by exposure to a photonflux of approximately 10 nM m^–2 s^–1 of far-red lightThe inhibition of abscission by red light could be reversedby treatment with far-red and the promotion of abscission byfar-red light could be reversed by treatment with red lightThe data were consistent with a phytochrome receptor systemlocated in the leaves that controlled the presence of an abscission-inhibitingsubstance in the abscission zones. Key words: Abscission, Coleus Blumei, far-red light red light     

Effects on Phytochrome Controlled Germination Produced by Far-Red Irradiation of Seeds Before and During Rehydration

Seeds irradiated with red light and then re-dried will respondto this light treatment on subsequent rehydration in the dark.If such high-Pfr seeds are irradiated in the dry state withfar-red light immediately before rehydration the percentagegermination is significantly reduced in the case of Plantagomajor and Sinapis arvensis but increased in Bromus steriliswhere Pfr inhibits germination. This effect of far-red lightcan be reversed by red light despite the fact that red lightalone has no effect on dry seed. This is due to the interconversionof Pfr and the red light absorbing phytochrome intermediatecomplex meta-Fa. If there is a delay between far-red irradiationand rehydration of Sinapis seeds, the inhibitory effect of thefar-red irradiation becomes progressively less the longer thedelay. This reduction in effectiveness of far-red is interpretedin terms of a dark reversal of meta-Fa to Pfr with a half-lifeof about 4–6 h. The reappearance of Pfr is either veryslow or docs not occur in dehydrated Plantago seeds, as far-redtight given 96 h prior to hydration is just as inhibitory asfar-red light given immediately before hydration. Meta-Fa doesappear to revert to Pfr in darkness in Bromus seeds, but onlyvery slowly. The rapid increase in effectiveness of red irradiationduring rehydration of high-Pfr Plantago seeds suggests that,in this species, the pre-treatment used in preparation of high-Pfrseeds may increase the receptivity or amount of the Pfr reactionpartner. Key words: Phytochrome intermediates, Seeds, Germination     

Seed Dormancy in Acer pseudoplatanus L.: the Role of the Covering Structures

The present studies with Acer pseudoplatanus L. suggest thatthe covering structures play an important and multiple rolein the dormancy of the fruit. Whole fruits and seeds with thetesta intact required a period of chilling at 5 °C beforedormancy was broken whereas bare embryos germinated immediatelyat 20 °C without pretreatment. This suggested that dormancywas coat-imposed and that the testa was responsible for thiseffect. Germination of dormant seeds was inhibited by lightwhereas the non-dormant bare embryos showed little response.Studies on the manner in which the testa imposed dormancy onthe embryo indicated that restriction on oxygen uptake, wateruptake, mechanical restriction to embryo enlargement, and thepresence of germination inhibitors in the testa were not limitingfactors at this stage of dormancy. Results from leaching experimentssuggest that dormancy was the result of the restriction by thetesta of the outward diffusion of a germination inhibitor(s)present in the embryo. In seeds that had nearly completed theirstratification requirements, the covering structures seemedto act in a manner other than by preventing the leaching ofan inhibitor from the embryo. At this point the physical propertiesof the covering structures seem to determine any further delaysin germination by the mechanical restriction of embryo enlargementby the testa and by restriction of oxygen uptake by the pericarp.     

The Role of the Red/Far-Red Ratio in the Response of Tropical Tree Seedlings to Shade

The West African species Khaya senegalensis and Terminalia ivorensiswere grown in a controlled environment, varying the photon fluxdensity in the range 18–610 µmol m^–2 s^–1and the red/far-red ratio over an appropriate range to simulatethe shade of a tree canopy versus unattenuated daylight. Theshade tolerant seedlings of Khaya were relatively insensitiveto the red/far-red ratio. The light demanding Terminalia wasconsiderably affected: when the ratio was low the specific leafarea was increased and the leaves produced were very much largerin area. Thus, the Leaf Area Ratio was enhanced and the plantsdisplayed an increase in Relative Growth Rate. Khaya, Terminalia, tropical trees, shade, red/far-red ratio     

Light Actions in the Germination of Cocklebur Seeds: IV. DISAPPEARANCE OF RED LIGHT-REQUIREMENT FOR THE GERMINATION OF UPPER SEEDS SUBJECT TO ANOXIA, CHILLING, CYANIDE OR AZIDEPRETREATMENT

Esashi, Y., Fuwa, Nn Kojima, K. and Hase, S. 1986. Light actionsin the germination of cocklebur seeds. IV. Disappearance ofred light-requirement for the germination of upper seeds subjectto anoxia, chilling, cyanide or azide pretreatmenL—J.exp. Bot. 37: 1652–1662. The effects on the germination of positively photoblastic uppercocklebur (X anthium pennsylvanicum Wallr.) seeds by pretreatingwith anoxia, chilling, cyanide or azide, which stimulates theirdark germination, were examined in relation to light actions.Prior to experiments, seeds were pre-soaked at 23 °C inthe dark for 1 or 2 weeks to remove the pre-existing Pfr. Whenthe prctreatment conditions were suboptimal for germinationinduction, the stimulating effects of the pretreatments on germinationduring a subsequent dark period at 23 °C were manifest onlywhen seeds were irradiated with red light before or after thepretreatment Red light promotion was reversed by blue or far-redlight treatment. However, both prc-chilling for 6 d at 8 °Cand prctreatment with 1· 5 mol m ^{– 3} NaN³ for 2d could induce full germination without red light exposure.On the other hand, both pre-exposure to anoxia for 8 d and pretreatmentwith 30 mol m^–3 KCN could induce the dark germinationonly when germination occurred at 33 °C which is known toaugment the ratio of an alternative respiration flux to a cytochromeone. Moreover, the dark germination in response to these inductionswere strongly inhibited by the inhibitors of alternative respiration,propyl gallate and benzohydroxamic acid, applied during a subsequentdark period. It was thus suggested that Pfr has some relationto the operation of two respiration systems of cocklebur seeds,but it is not indispensable to germination of this positivelyphotoblastic seed. Key words: Anoxia, azide, blue light, chilling cyanide, dark germination, far-red light, red light, seed germination, X anthium pennsylvanicum     

The Role of ABA in the Control of Apple Seed Dormancy Re-appraised by Combined Gas Chromatography-Mass Spectrometry

Measured by GC—MS²—SIR³, endogenous ABA⁴ in embryonicaxes of seeds of Malus pumila L. cv. Golden Delicious decreased8-fold and cotyledon ABA by only 60%, during 10–50 d ofstratification at 5 ?C, after ABA leaching during an initial24 h soaking. During stratification, the percentage germinationof embryos transferred to 17?C showed a significant linear dependenceon log_e of ABA levels in the axes at transfer. Between 50 and70 d, ABA levels increased markedly in axes and testa both ofstratified seeds and seeds allowed to re-dry at 17 ?C afterinitial soaking. The ability of fully stratified axes with elevatedendogenous ABA to germinate indicated that stratification haddecreased their ABA sensitivity. Changes in cotyledon ABA couldnot account for the promotory effect of cotyledons on germinationduring the first 30 d of stratification. Loss of testa inhibitionof germination during stratification was not linked with changesin testa ABA. Stratification markedly increased the sequestrationin the axes of exogenous ABA supplied via the cotyledons. Changesboth in axis ABA levels and sensitivity were thus correlatedwith dormancy release, but subject to modifying control by thecotyledons and testa not involving ABA. Rehydration of driedseeds affected axis ABA later during storage via mechanismsunconnected with dormancy. Key words: ABA, seed dormancy, stratification     

Evidence of Phytochrome Mediation in the Low-pH-Induced Root Hair Formation Process in Lettuce (Lactuca sativa L. cv. Grand Rapids) Seedlings

−2 . The inductive effect of 100 Jm⁻² red light could be partially reversed by subsequent far-red light only one time. On the other hand, the inductive effect of 1,000 Jm⁻² red light was partially reversed by subsequent far-red light irradiation at least twice. These results indicate the involvement of phytochrome in this response. The inductive effect of blue light was repeatedly reversed by subsequent far-red light irradiation, suggesting that the blue-light induction was mainly mediated by phytochrome. Received 13 August 1999/ Accepted in revised form 22 December 1999     

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.     

Stimulation of lettuce seed germination by ethylene

Ethylene increased the germination of freshly imbibed lettuce (Lactuca sativa L. var. Grand Rapids) seeds. Seeds receiving either red or far-red light or darkness all showed a positive response to the gas. However, ethylene was apparently without effect on dormant seeds, those which failed to germinate after an initial red or far-red treatment. Carbon dioxide, which often acts as a competitive inhibitor of ethylene, failed to clearly reverse ethylene-enhanced seed germination. While light doubled ethylene production from the lettuce seeds, its effect was not mediated by the phytochrome system since both red and far-red light had a similar effect.     

PHOTOCONTROL AND TEMPERATURE DEPENDENCE OF GERMINATION OF RUMEX SEEDS

1. Light is not obligatory for the germination of the seed ofRumex obtusifolius L. subsp.agrestis DANSER, which has beenregarded as being a typical light sensitive seed. Even in continuousdarkness, a short period of high (30°) or low temperature(5°) treatment evokes germination very readily. 2. Germination is markedly promoted by 1 min exposure to a redlight and this red light effect is completely removed by 1-hrexposure to a far-red light. Alternations of the red and far-redradiation bring about an alternate promotion and inhihibitionof germination. 3. When a dark interval is inserted between the red and thefar-red treatments, inhibition of germination becomes less distinctas the duration of darkness increases. When the seeds are irradiatedwith far-red prior to red, with an inserted darkness, germinationpromotion due to the red light also decreases with the durationof inserted darkness. 4. Complicated interdependence between the light and temperatureeffects are demonstrated. This suggests a participation of somereactants besides pigments in the photoreaction. 5. The observed interdependence between the light and temperatureeffects on the germination of Rumex seeds implies that, if,as BORTHWICK has assumed, two forms of pigment, viz., a far-red-absorbingform and a red-absorbing one, are participating in the photoreaction,they should be presumed to coexist from the start of imbibition. (Received September 27, 1960; )     

Photoinduction of Spore Germination in a Fern, Mohria caffrorum

Irradiation of spores of the fern Mohria caffrorum Sw. witheither red light (67.4 µW cm^–2) or far-red light(63.2 µW cm^–2) for a period of 24 h induced maximumlevels of germination. Brief irradiations with blue light (127.6µW cm^–2) administered before or after photoinductioncompletely nullified the effects of red or far-red light; however,with prolonged exposure to blue light, germination levels roseto near maximum. The similar effects of red and far-red lightin promoting spore germination makes the involvement of phytochromein this process questionable. Based on energy requirements,the promotive and inhibitory phases of blue light appear toinvolve independent modes of action. Mohria caffrorum, ferns, spore germination, photoinduction, phytochrome