Accumulation of Storage Materials,Precocious Germination and Development of Desiccation Tolerance During Seed Maturation in Mustard (Sinapis alba L.)

Under defined environmental conditions (20°C, continuous light of 15 klx) development of mustard seeds from artificial pollination to maturity takes about 60 d. After surpassing the period of embryo cell division and histodifferentiation (12–14d after pollination = dap), the seed enters into a maturation period. The time courses of various physiological, biochemical, and structural changes of embryo and testa during seed maturation were analyzed in detail (dry and fresh mass changes, osmotic and water potential changes, respiration, DNA amplification by endomitosis, total ribosome and polysome formation, storage protein synthesis and accumulation, storage lipid accumulation). In addition to the final storage products protein and lipid, embryo and testa accumulate transiently large amounts of starch within the chloroplasts during early maturation. Concomitantly with the subsequent total breakdown of the starch, the plastids lose most of their internal structure and chlorophyll and shrink into proplastids, typical for the mature seed. At about 30 dap the seeds shift from a desiccation-sensitive to a desiccation-tolerant state and are able then to germinate rapidly upon drying and reimbibition. If isolated from the immature fruit and sown directly on water, the seeds demonstrate precocious germination from about 13 dap onwards. Young seeds (isolated ≦ 38 dap) germinate only after surpassing a lag-phase of several days (after-ripening) during which the embryo continues to accumulate storage protein and lipid at the expense of the surrounding seed tissues. We conclude from these results that the maturing seed represents a rather closed developmental system which is able to continue its development up to successful germination without any specific regulatory influence from the mother plant. Immature seeds are able to germinate without a preceding dehydration treatment, which means that partial or full desiccation does not serve as an environmental signal for reprogramming seed development from maturation to germination. Instead, it is argued that the water relations of the seed are a critical element in the control of maturation and germination: during maturation on the mother plant the embryo is subject to a considerable turgor pressure (of the order of 12 bar) accompanied by a low water potential (of the order of ?12 bar). This turgor permits maturation growth but is subcritical for germination growth. However, upon imbibition in water, the low water potential provides a driving force for a burst of water uptake overcoming the critical turgor threshold and thereby inducing germination.     

Water Relations of Tomato Seeds During Imbibition and Osmotic Priming

Water uptake of tomato (Lycopersicon esculentum Mill. cv. Moneymaker) seeds during germination was obviously triphasic. The completion of the first phase of water uptake by whole seed could not be realized until 10~12 h later after sowing though varies in different parts of seed. The mechanical resistance of endosperm and seed coat restricted water uptake of the embryo envoleped by the endosperm. Water potential of the intact embryo was still 0. 6~0. 9 Mpa lower than the whole seed when the equilibrium between seed and imbibing solution was established. GA and ABA had no direct effects on the water uptake of tomato seeds. The water potential of embryo was positively correlated with its moisture content. The osmotic potential of tomato embryos decreased slowly during imbibition in water and osmotic solution as well.     

After-ripening and phytochrome action in seeds of dormant lines of wild oat (Avena fatua)

The effects of a short exposure to red, far-red or alternate red/far-red light on the germination of seeds after-ripened for different periods of time were studied in dormant lines of wild oat ( Avena fatua L.). Three stages were distinguishable in the after-ripening period in the response of germination to light. Seeds stayed dormant and showed no response to light during stage I. Phytochrome-mediated germination was observed in seeds during stage II. The phytochrome action disappeared during stage III, i.e. seeds fully germinated following treatments of all light qualities. When the seeds were imbibed in polyethylene glycol solutions, dark germination was reduced and phytochrome again had an effect, which suggested the involvement of phytochrome in water uptake of the seed.     

Release of reactive oxygen intermediates (superoxide radicals, hydrogen peroxide, and hydroxyl radicals) and peroxidase in germinating radish seeds controlled by light, gibberellin, and abscisic acid

Germination of radish (Raphanus sativus cv Eterna) seeds can be inhibited by far-red light (high-irradiance reaction of phytochrome) or abscisic acid (ABA). Gibberellic acid (GA3) restores full germination under far-red light. This experimental system was used to investigate the release of reactive oxygen intermediates (ROI) by seed coats and embryos during germination, utilizing the apoplastic oxidation of 2',7'-dichlorofluorescin to fluorescent 2',7'-dichlorofluorescein as an in vivo assay. Germination in darkness is accompanied by a steep rise in ROI release originating from the seed coat (living aleurone layer) as well as the embryo. At the same time as the inhibition of germination, far-red light and ABA inhibit ROI release in both seed parts and GA3 reverses this inhibition when initiating germination under far-red light. During the later stage of germination the seed coat also releases peroxidase with a time course affected by far-red light, ABA, and GA3. The participation of superoxide radicals, hydrogen peroxide, and hydroxyl radicals in ROI metabolism was demonstrated with specific in vivo assays. ROI production by germinating seeds represents an active, developmentally controlled physiological function, presumably for protecting the emerging seedling against attack by pathogens.     

Phytochrome-mediated germination control of maize caryopses

Maize caryopses sown in water germinate equally well either in darkness or under any light regime. However, when they are imbibed in mannitol solutions, continuous far-red light proves to be strongly inhibitory on the final germination as compared to darkness. Similar but less pronounced inhibition is also exhibited by continuous red or blue light. Intermittent far-red light can partially substitute for continuous far-red light in inhibiting maize caryopsis germination, and its effect is reversed to the intermittent red light level when red light is given immediately after each far-red illumination. These results are interpreted as a proof of existence and involvement of phytochrome in the germination control of maize caryopses, though its manifestation is realized only under osmotic stress.Abbreviations D darkness - FR far-red - R red - B blue - c-FR, c-R, c-B continuous FR, R, B, resp. - i-FR, i-R intermittent FR, R, resp.     

A model of the effects of water stress on seed advancement and germination

A model of seed germination is proposed which uses a variable with the units of an osmotic potential (virtual osmotic potential) to integrate the effect of a constant or a varying water potential. This differs from existing models that describe the effects of fixed water potentials on germination, or the effects of fixed priming water potentials on the subsequent germination at a fixed water potential. When a seed is sown, the virtual osmotic potential is assumed to fall at a rate that depends on the ambient water potential, and on the difference between its current and a minimum value. Radicle growth is assumed to initiate when the difference between the ambient water potential and the virtual osmotic potential exceeds a threshold. The germination of carrot and onion seeds at various fixed potentials below 0 MPa was well described by the virtual osmotic potential model. The model was also used to simulate the results of experiments in which seeds were given a single step change in water potential.     

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     

Effects of 2,6-dichlorobenzonitrile on differentiation to tracheary elements of isolated mesophyll cells of Zinnia elegans and formation of secondary cell walls

Seeds of Caesulia axillaris Roxb. displayed an absolute light requirement for germination throughout the period of dry storage at 28°C. The seeds were found to show a gradual increase in percent germination with storage time - reaching a maximum value between 8-14 months and then a sharp decline. Percent water uptake and photosensitivity were at maximum after a 5-day imbibition period in the dark in both seedlots studied. Seedlot I, which was only marginally responsive to far-red light, showed a nearly complete red-far-red reversal effect in contrast to seedlot II. The latter also displayed a considerable promotion of germination in far-red light. Interestingly, a noticeable degree of heterogeneity, besides the one observed in both seedlots with reference to red light, was found to exist in seedlot II for far-red light. Exogenous application of nitrate and ammonium, at the levels occurring in soil during seed germination/seedling emergence phase of the plant in nature, promoted a considerable proportion of high Ø-requiring seeds to germinate under irradiation conditions establishing low Ø-value. The probable ecological implication of this reponse has been discussed. Little correlation was found between the requirement for an exogenous supply of nitrate and the endogenous nitrate level in the seeds in their response to far-red light.     

Effect of light on seed germination of eight wetland Carex species

BACKGROUND AND AIMS: In wetland plant communities, species-specific responses to pulses of white light and to red : far-red light ratios can vary widely and influence plant emergence from the seed bank. Carex species are the characteristic plants of sedge meadows of natural prairie wetlands in mid-continental USA but are not returning to restored wetlands. Little is known about how light affects seed germination in these species-information which is necessary to predict seed bank emergence and to develop optimal revegetation practices. The effects of light on germination in eight Carex species from prairie wetlands were investigated. METHODS: Non-dormant seeds of eight Carex species were used to determine the influence of light on germination by examining: (a) the ability of Carex seeds to germinate in the dark; (b) the effect of different lengths of exposures to white light on germination; (c) whether the effect of white light can be replaced by red light; and (d) whether the germination response of Carex seeds to white or red light is photoreversible by far-red light. KEY RESULTS: Seeds of C. brevior and C. stipata germinated >25 % in continuous darkness. Germination responses after exposure to different lengths of white light varied widely across the eight species. Carex brevior required <15 min of white light for > or =50 % germination, while C. hystericina, C. comosa, C. granularis and C. vulpinoidea required > or =8 h. The effect of white light was replaced by red light in all species. The induction of germination after exposure to white or red light was reversed by far-red light in all species, except C. stipata. CONCLUSIONS: The species-specific responses to simulated field light conditions suggest that (a) the light requirements for germination contribute to the formation of persistent seed banks in these species and (b) in revegetation efforts, timing seed sowing to plant community development and avoiding cover crops will improve Carex seed germination.     

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.     

Antagonistic action of low-fluence and high-irradiance modes of response of phytochrome on germination and beta-mannanase activity in Datura ferox seeds

Seed germination is often induced by a pulse of red light perceived by phytochrome and cancelled by a subsequent pulse of far-red light. When the pulse of red light is followed by several hours of darkness, a pulse of far-red light is no longer effective and prolonged far-red is necessary to block germination. The aim was to investigate whether the red light pulse and prolonged far-red light act on the same or different processes during germination of Datura ferox seeds. Forty-five hours after the inductive red light pulse, germination could not be blocked by one pulse or six hourly pulses of far-red light, but was significantly reduced by 6 h of continuous far-red light. The pulse of red light increased embryo growth potential and the activities of beta-mannanase and beta-mannosidase extracted from the micropylar region of the endosperm. Continuous far-red light had no effect on embryo growth potential or beta-mannosidase activity, but severely reduced the activity of beta-mannanase. The effect of far-red light had the features of a high-irradiance response of phytochrome. Both germination and beta-mannanase activity were restored by a pulse of red light given after the end of the continuous far-red treatment. It is concluded that the low-fluence response and the high-irradiance response modes of phytochrome have antagonistic effects on seed germination and that the control of beta-mannanase activity is one process where this antagonism is established.     

Significance of inhibition of seed germination under the leaf canopy in ash coppice

Abstract. The effect on seed germination of the red/far-red ratio of light (ζ) as modified by a leaf canopy was studied. The artificial light sources used and daylight filtered through leaves appeared to have identical effects in tests with lettuce seed.
The sensitivity of the seeds to ζ varied with the environmental conditions during pretreatment and during the actual germination test. Freshly harvested and dry-stored seeds of Cirsium palustre were strongly inhibited at low ζ; after stratification, the inhibition was much less. The fact that C. palustre emerges mainly after the coppice is felled could not be explained from these results combined with field data on ζ and the response of germination to temperature.
The ecological significance of inhibition at low ζ is to be found directly after dispersal. Seed germination will then be delayed until the following spring or later.
Geum urbanum showed primary dormancy, which was released by a stratification treatment. No inhibition of germination at low ζ was found in the spring and seeds germinated in light and darkness at low temperatures. These results explain why the species emerges early in spring regardless of the presence of the ash canopy.     

GERMINATION OF LIGHT-INHIBITED SEED OF NEMOPHILA INSIGNIS

Germination of Nemophila insignis seed is inhibited by light over a wide range of temperatures. At low temperatures the light intensity required for inhibition is higher. At 25 C there is little germination (in darkness as well as in light); at 27.5 C germination is inhibited altogether. Virtually complete germination in light is obtained when the endosperm directly covering the radicle is removed. This operation also improves germination in darkness at 25 C. Mechanical scarification performed elsewhere in the seed is without effect. As with Phacelia tanacetifolia, Nemophila seed apparently fails to germinate in light because the endosperm restrains the expansive growth of the embryo. The embryo of dark-imbibed seed develops a force which enables it to overcome the physical resistance of the endosperm. Light inhibits the process which leads to generation of “expansive force.” Gibberellic acid at 5 × 10^–4 m stimulates germination in light and in the dark. Abscisic acid at 10^-4 m inhibits germination; at 10^-6 m it inhibits only root growth. The inhibition of germination or root growth caused by abscisic acid cannot be reversed by gibberellic acid. Eighty per cent oxygen under certain conditions promotes germination. The rate of O₂ uptake is enhanced in oxygen-enriched atmosphere; however, there is no corresponding increase in the rate of CO₂ output. Thus high oxygen tension enhances an oxidative process other than respiration. This reaction is favorable to seed germination.     

Triadimefon reduces transpiration and increases yield in water stressed plants

The total free amino acid pools in radicles of watermelon seeds, investigated during imbibition of water at 25°C, were higher under the most (darkness) than under the least (continuous broad spectrum far-red light) favourable light regime for germination. When seeds were imbibed in an appropriate osmotic solution of PEG-6000 (fully suppressing germination), in darkness or under continuous red or far-red light, the biochemical analyses of the radicles after 1,2,3 and 4 days from the onset of imbibition show that while the total soluble sugar content remains rather constant in all treatments, significant changes are observed in the total free amino acid pools. After the first day, a considerable increase characterizes the "darkness" pool in contrast to a moderate one under red, while the "far-red" pool remains constant. Ultimately, at 4 days, the three pools are 190,142 and 123% of the 0 day radicle one. The qualitative free amino acid determination of the 4 day darkness and far-red pools shows a considerably increased percentage contribution of glutamic acid, arginine and citrulline in the "darkness" pool. The free amino acid increase in non-illuminated radicles may be correlated to germinability; moreover, it is evidently a phytochrome-mediated, pre-germinatory event, probably due to the hydrolysis of proteins (known to be rich in glutamic acid and arginine), stored in the radicle.     

Photocontrol of seed germination in the hemiparasite Buchnera hispida (Scrophulariaceae)

Abstract Buchnera hispida, a facultative root parasite of grasses and graminaceous crops, has a light requirement for germination. Studies were carried out on the effects of varying photoperiods with or without preceding dark incubation, on seed germination. Buchnera seeds showed long-day behaviour, since they germinated at all photoperiods including continuous light, and longer photoperiods were more effective in triggering seed germination than shorter photoperiods. Also, effects of red and far-red light indicated that the phytochrome system is operative in the light-induced germination of Buchnera. Although dark incubation in water before illumination was not absolutely necessary for germination, it caused the seeds to respond more rapidly to light. The longer the time of the dark incubation the more responsive the seeds were to photoperiod except when 15 min light was given. The effectiveness of a preceding dark incubation in making Buchnera seeds sensitive to rapid light action was completely inhibited at 4°C. This is in agreement with the hypothesis that a reaction partner of the far-red absorbing form of phytochrome is produced during dark incubation of Buchnera seeds. Such an intermediate has also been reported in some positively photoblastic seeds of non-parasitic flowering plants.     

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.     

Water uptake and distribution in non-dormant and dormant wild oat (Avena fatua L.) caryopses

The influence of seed coat modification and light quality onwater uptake and distribution in caryopses of dormant and non-dormantlines of wild oat (Avena fatua L.) was determined using NMRmicroimaging. Non-dormant seeds absorbed water more rapidlythan dormant seeds during imbibition on distilled water. Thiseffect was detected first in the embryo-scutellar region (8h) and later in the proximal endosperm (12 h). Cutting the testaand pericarp close to the embryo or scarification with KOH promotedrapid embryo/scutellum hydration and germination. Cutting atthe middle part of the caryopsis did not enhance embryo hydrationnor did it greatly improve germination. The sensitivity of waterdistribution to the phytochrome germination effect was examined.Significant differences in imbibitional water uptake by embryos-scutellumtissue were detected by 18 h following red-light (germinationpromoter) compared with far-red (germination inhibitor) treatment.The results indicated that both the rate and the sequence ofembryo/scutellum hydration were important in initiating germinationin dormant seeds. A refinement of the model that describes waterimbibition in wild oat seeds during the early stages of germinationis discussed. Key words: Water uptake, water distribution, Avena fatua, seed coat modification, light quality, dormant and non-dormant seeds     

Possible significance of changes in the light requirement of Cirsium palustre seeds after dispersal in ash coppice

Abstract. It is demonstrated that after the feelling of ash coppice Cirsium palustre becomes established from seed that was produced during the previous coppice cycle and has lain underground for several years. These seeds are light-requiring and it is probable that their germination is triggered by exposure to light during coppicing in winter.
It is assumed that the seeds become light-requiring because phytochrome in the far-red absorbing form high. This facilitates their incorporation in the seed bank in summer. However, the disappearance of Pfr is inhibited at low temperatures; this means that the seeds can store the light stimulus in winter and will not germinate until they experience higher temperatures in spring. The significance of the inhibition of seed germination under a leaf canopy is that it enhances the seeds chance of being incorporated in the soil seed bank.     

The growth physics and water relations of red-light-induced germination in lettuce seeds

Using lettuce (Lactuca sativa L., cv. Grand Rapids) embryos in osmotica, we have demonstrated that when the growth rates of the embryonic axes of seeds treated with red (R) or far-red (FR) light are equalized, the axes of R-treated seeds develop a 3.4-bar decrease in water potential (paper No. III).As axial growth begins, reserve protein and phytin decrease rapidly, concomitant with increases in reducing sugars, -amino nitrogen, and inorganic and esterified soluble phosphates. However, no differences between the axes of R-and FR-treated seeds are found with respect to the changes in these compounds, indicating that these changes arise as a result of growth and are not under immediate phytochrome control. Little change in the total lipid content is found in either treatment. The axes of FR-treated seeds hydrolyze endogenous sucrose at a greater rate thant those of R-treated seeds. Axes of R-treated seeds accumulate K⁺ and Na⁺ to a greater extent than those of FR-treated seeds. When potassium salts are added to the incubation medium, R induces increased K⁺ uptake by the axis and greater medium acidification by the axis. Malate and other organic acids and acidic amino acids increase at equal rates in both treatments, indicating that inorganic anions may also be taken up to balance the ionic charges. The results are compatible with the assumption that changes in the osmotic and pressure potentials of the embryonic axes of R-treated seeds are the result of a phytochrome-stimulated proton pump which, in whole dormant seeds, would initiate water-potential changes allowing the embryos to overcome the mechanical restraint of the surrounding seed layers, resulting in germination.Abbreviations FR far-red light - PEG polyethylene glyeol 4000 - Pfr far-red-absorbing form of phytochrome - R red light III=Carpita et al. 1979     

Light and nitrate interaction in phytochrome-controlled germination ofPaulownia tomentosa seeds

Pulsed light and nitrate exhibit an interactive effect on the germination ofPaulownia tomentosa Steud. seeds that require long periods of light irradiation. Two pulses of red light (R), separated by an adequately long dark interval, substitute for continuous prolonged irradiation. A far-red (FR) pulse given at the beginning of the dark interval inhibits germination, while it has no effect if given at the end. The requirement for certain ratios of the far-red-absorbing form of phytochrome/total phytochrome (P_fr/P_tot) differs when a FR+R-pulse is given as the first or second of two pulses (FR+R or R) separated by a dark interval. An equal decrease of the P_fr/P_tot ratio leads to a more pronounced decrease in germination when the pulse of the same FR+R ratio is given as the second pulse at the end of the dark interval. The length of dark interval between light pulses needed for maximal germination, differed in (i) seeds with a natural requirement for long periods of light irradiation from that in (ii) seeds with their long light requirement imposed by two weeks of imbibition in darkness or by (iii) imbibition in 40% heavy water. However, a single R pulse was sufficient to induce a high percentage of germination if the seeds were supplied with KNO₃ (10 mM) from the onset of imbibition up to the onset of light. This effect decreased with a delayed time of application, and was prevented if FR preceded the KNO₃ application. We dedicate this paper to Professor Hans Mohr on the occasion of his 60th birthday