Ethylene in seed dormancy and germination

The role of ethylene in the release of primary and secondary dormancy and the germination of non-dormant seeds under normal and stressed conditions is considered. In many species, exogenous ethylene, or ethephon – an ethylene-releasing compound - stimulates seed germination that may be inhibited because of embryo or coat dormancy, adverse environmental conditions or inhibitors (e.g. abscisic acid, jasmonate). Ethylene can either act alone, or synergistically or additively with other factors. The immediate precursor of ethylene biosynthesis, 1-aminocyclopropane-1-carboxylic acid (ACC), may also improve seed germination, but usually less effectively. Dormant or non-dormant inhibited seeds have a lower ethylene production ability, and ACC and ACC oxidase activity than non-dormant, uninhibited seeds. Aminoethoxyvinyl-glycine (AVG) partially or markedly inhibits ethylene biosynthesis in dormant or non-dormant seeds, but does not affect seed germination. Ethylene binding is required in seeds of many species for dormancy release or germination under optimal or adverse conditions. There are examples where induction of seed germination by some stimulators requires ethylene action. However, the mechanism of ethylene action is almost unknown.
The evidence presented here shows that ethylene performs a relatively vital role in dormancy release and seed germination of most plant species studied.     

On the role of abscisic acid in seed dormancy of red rice

Abscisic acid (ABA) is commonly assumed to be the primary effector of seed dormancy, but conclusive evidence for this role is lacking. This paper reports on the relationships occurring in red rice between ABA and seed dormancy. Content of free ABA in dry and imbibed caryopses, both dormant and after-ripened, the effects of inhibitors, and the ability of applied ABA to revert dormancy breakage were considered. The results indicate: (i) no direct correlation of ABA content with the dormancy status of the seed, either dry or imbibed; (ii) different sensitivity to ABA of non-dormant seed and seed that was forced to germinate by fluridone; and (iii) an inability of exogenous ABA to reinstate dormancy in fluridone-treated seed, even though applied at a pH which favoured high ABA accumulation. These considerations suggest that ABA is involved in regulating the first steps of germination, but unidentified developmental effectors that are specific to dormancy appear to stimulate ABA synthesis and to enforce the responsiveness to this phytohormone. These primary effectors appear physiologically to modulate dormancy and via ABA they effect the growth of the embryo. Therefore, it is suggested that ABA plays a key role in integrating the dormancy-specific developmental signals with the control of growth.     

Dormancy in Rice Seed II: THE INFLUENCE OF COVERING STRUCTURES

Most of the dormancy in rice seed can be accounted for by theinhibitory influence of the husk, and most of the residual dormancyafter dehusking can be attributed to the inhibitory influenceof other covering structures—either the pericarp or testa,or both. It is shown that the rate of water absorption is thesame in dormant and non-dormant seeds and that dormant seedsare capable of absorbing sufficient water for germination. Thecovering structures therefore do not cause dormancy by restrictingthe entry of water. Removal of a small area of the husk breaks the dormancy of alarge proportion of the seeds; but for some seeds this treatmentis ineffective whereas removal of the entire husk would breakdormancy. The site of the excision of a small area of the huskcan alter the effectiveness of the treatment: removal of a portionof husk immediately over the embryo is no more effective thanexcising a similar portion nearby, but the removal of part ofthe husk some distance from the embryo is not as effective.Sealing the perforations with paraffin wax has little effectexcept when carried out as soon as possible after the excisionis made, and then only in positions distant from the embryo. Attempts to extract a water-soluble or ether-soluble germinationinhibitor from the husk and other parts of dormant seed or todemonstrate the presence of inhibitors by indirect methods havenot been successful. Nor has it been found possible to extracta water-soluble germination stimulator from seed which has brokendormancy. The implications of these results are discussed.     

Studies in Wild Oat Seed Dormancy: I. THE ROLE OF ETHYLENE IN DORMANCY BREAKAGE AND GERMINATION OF WILD OAT SEEDS (AVENA FATUA L.)

Seed of Avena fatua were shown to exhibit a characteristic loss of dormancy during dry storage at 25 C, whereas similar seed stored at 5 C maintained dormancy. 2-Chloroethylphosphonic acid was shown to increase germination of partly dormant seed imbibed under certain temperature regimes; a similar effect could not be established for fully dormant or fully nondormant seed. Using gas-liquid chromatography, natural ethylene levels were followed during imbibition of fully dormant and nondormant seed. A large peak in production was observed in the period prior to radicle emergence in the case of the nondormant seed. Measurements of ethylene production taken at 15 C, following periods of after-ripening in moist soil at either 5 or 25 C, indicated that endogenous production was unlikely to be a main cause of dormancy breakage in this species. The possibility that endogenous ethylene could play a role in natural dormancy breakage in aged seeds is discussed. The practical possibilities of 2-chloroethylphosphonic acid as a dormancy breaking agent in a field situation are outlined.     

Interaction of karrikinolide and ethylene in controlling germination of dormant <Emphasis Type="Italic">Avena fatua</Emphasis> L. caryopses

Caryopses of Avena fatua L. are dormant after harvest and germinate poorly at 20 °C. Dormancy was released by after-ripening the dry caryopses in the dark at 25 °C for 3 months. Karrikinolide (butenolide, 3-methyl-2H-furo[2,3-c]pyran-2-one, KAR₁), in contrast to exogenous ethylene and the precursor of ethylene biosynthesis 1-aminocyclopropane-1-carboxylic acid (ACC), completely overcame dormancy. The effect of KAR₁ was not affected by aminoethoxyvinylglycine (AVG), α-aminoisobutyric acid (AIB) and CoCl₂, inhibitors of ACC synthase and oxidase, respectively. 2,5-Norbornadiene (NBD), a reversible inhibitor of ethylene binding to its receptor, counteracted the stimulatory effect of KAR₁. Ethylene, ethephon and ACC counteracted and AVG reinforced inhibition caused by norbornadiene. Inhibition due to norbornadiene, applied during the first 3 days of imbibition in the presence of KAR₁, disappeared after transfer to air or ethylene. The obtained results confirm that KAR₁ breaks dormancy and indicate that ethylene alone plays no role in releasing dormancy of Avena fatua caryopses. KAR₁ probably did not relieve dormancy via the stimulation of ethylene biosynthesis. Some level of endogenous ethylene is probably required for ethylene action, which might be required for releasing dormancy by KAR₁ or for subsequent germination of caryopses after removing dormancy.     

Ethylene as a Component of the Emanations From Germinating Peanut Seeds and Its Effect on Dormant Virginia-type Seeds

The embryonic axes of Spanish-type peanut seeds that do not exhibit dormancy to any extent were found to produce ethylene during germination. Virginia-type peanut seeds of the extremely dormant variety NC-13 produced low levels of ethylene when imbibed but not germinating. Treatments that released dormancy of NC-13 peanut seeds resulted in increased ethylene production by the embryonic axis. The estimated internal concentration of ethylene in Virginia-type peanut seeds was 0.4 ppm at 24 hr of germination. Fumigation with an external concentration of 3.0 to 3.5 ppm for 6 hr was sufficient to break dormancy of Virginia-type peanut seeds. These results suggest that ethylene is associated with the germination processes of non-dormant seeds and participates in the breaking of seed dormancy of dormant peanut varieties.     

Soil burial induced dormancy in weedy rice seeds through hormone level changes: Implications in adaptive evolution and weed control

Seed dormancy plays a key role in preventing seeds of higher plants from random germination under adverse environmental conditions. Previous studies suggested that a critical temperature could regulate germination of weedy rice seeds without primary dormancy at seed dispersion. However, what will happen to the non-dormant seeds after shattering in the soil seed banks when temperature fluctuates to exceed the critical temperature remains an interesting question to be investigated. To determine whether or not soil burial can change the status of dormancy in weedy rice seeds, we examined germination ratios of weedy rice seeds after soil-burial treatments. In addition, we compared hormone levels in the untreated seeds and viable but ungerminated seeds after soil burial. Results showed that soil burial induced a proportion of 41%–72% dormant seeds in the initially non-dormant weedy rice seeds. Also, the induction of seed dormancy is associated with the change of hormone levels in the seeds treated by soil burial, suggesting that soil burial can significantly activate the control of hormone production in seeds. Together, the previously reported mechanism of critical temperature-inhibited seed germination and the newly found phenomenon of soil burial-induced seed dormancy provide a “double-security” strategy to ensure germination of weedy rice seeds under a favorable condition in agricultural ecosystems. The findings not only reveal the important role of rapid evolution of adaptive functions in weeds, such as weedy rice, in adapting to changing agricultural environments, but also facilitate the design of strategies for effective weedy rice control practices.     

Control of seed dormancy in Nicotiana plumbaginifolia: post-imbibition abscisic acid synthesis imposes dormancy maintenance

The physiological characteristics of seed dormancy in Nicotiana plumbaginifolia Viv. are described. The level of seed dormancy is defined by the delay in seed germination (i.e the time required prior to germination) under favourable environmental conditions. A wild-type line shows a clear primary dormancy, which is suppressed by afterripening, whereas an abscisic acid (ABA)-deficient mutant shows a non-dormant phenotype. We have investigated the role of ABA and gibberellic acid (GA₃) in the control of dormancy maintenance or breakage during imbibition in suitable conditions. It was found that fluridone, a carotenoid biosynthesis inhibitor, is almost as efficient as GA₃ in breaking dormancy. Dry dormant seeds contained more ABA than dry afterripened seeds and, during early imbibition, there was an accumulation of ABA in dormant seeds, but not in afterripened seeds. In addition, fluridone and exogenous GA₃ inhibited the accumulation of ABA in imbibed dormant seeds. This reveals an important role for ABA synthesis in dormancy maintenance in imbibed seeds. Received: 31 December 1998 / Accepted: 9 July 1999     

Seed Dormancy in Red Rice (Oryza sativa) (IX. Embryo Fructose-2,6-Bisphosphate during Dormancy Breaking and Subsequent Germination)

Fructose-2,6-bisphosphate (Fru-2,6-bisP) was evaluated as a potential marker for the dormancy-breaking phase or the germination phase before pericarp splitting in red rice (Oryza sativa). During 4 h of imbibition at 30[deg]C, Fru-2,6-bisP of dehulled dormant and nondormant seeds increased to 0.26 and 0.38 pmol embryo-1, respectively. In nondormant seeds, embryo Fru-2,6-bisP content remained stable until the onset of pericarp splitting (12 h) and increased rapidly thereafter. In dormant seeds, Fru-2,6-bisP declined to 0.09 pmol embryo-1 at 24 h. Embryo Fru-2,6-bisP was correlated with O2 uptake of dormant and nondormant seeds. A 24-h exposure of dehulled, water-imbibed, dormant seeds to treatments yielding >90% germination (sodium nitrite [4 mM], propionic acid [22 mM], methyl propionate [32 mM], propanol [75 mM], and propionaldehyde [40 mM]) led to changes in embryo Fru-2,6-bisP that were unrelated to the final germination percentages. Furthermore, a 2-h pulse of propionaldehyde increased Fru-2,6-bisP 4-fold but did not break dormancy. Whereas nitrite and propionaldehyde increased Fru-2,6-bisP to 0.33 pmol embryo-1 after 2 h of contact, propionic acid and methyl propionate did not increase Fru-2,6-bisP above the untreated control. In all cases, further increases in Fru-2,6-bisP occurred after pericarp splitting. However, the plateau Fru-2,6-bisP attained during chemical contact was inversely correlated with elapsed time to 30% germination (r = -0.978). Therefore, although Fru-2,6-bisP is not a universal marker for dormancy release, its rapid increase during nitrite and propionaldehyde treatments suggests that events associated with dormancy breaking can occur within 2 h of chemical treatment.     

Action of ferric and aluminium ions on the dormancy breakage of <Emphasis Type="Italic">Stylosanthes humilis</Emphasis> seeds

Dormancy of scarified seeds of Stylosanthes humilis was broken by acidic Al³⁺ and Fe³⁺ solutions. Fe⁺³-stimulated seeds exhibited a high activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and produced great amounts of ethylene, which showed correlated with the germination process. In addition, specific inhibitors of ethylene biosynthesis and action largely depressed the Fe³⁺-stimulated germination, leading to the conclusion that the ion broke dormancy by triggering ethylene production by the seeds. By contrast, inhibitors of ethylene biosynthesis and action did not impair germination of Al³⁺-stimulated dormant seeds. Moreover, ethylene production and activity of ACC oxidase of Al³⁺-treated seeds was substantially decreased by inhibitors of ethylene biosynthesis, but germination kept large. Together these data suggest that ethylene biosynthesis was not required in the chain of events triggered by Al³⁺ leading to dormancy breakage. Methyl viologen (MV), a reactive oxygen species-generating compound, broke dormancy of seeds to the same extent as Al³⁺ did. Germination of both Al³⁺- and MV-stimulated dormant seeds was inhibited by sodium selenate, an antioxidant compound; selenate, however had no effect on germination of Fe³⁺-stimulated seeds. Together these data indicate that the mechanisms underlying the germination of Al³⁺- and Fe³⁺-treated seeds are not the same.     

ACC conversion to ethylene by sunflower seeds in relation to maturation,germination and thermodormancy

Conversion of exogenous 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene was studied in sunflower (Helianthus annuus L., cv. Mirasol) seeds in relation to germinability. Ethylene production from ACC decreased during seed maturation, and non-dormant mature seeds were practically unable to synthesize ethylene until germination and growth occurred, indicating that ethylene forming enzyme (EFE) activity developed during tissue imbibition and growth. ACC conversion to ethylene was reduced by the presence of pericarp, and in young seedlings it was less in cotyledons than in growing axes.ACC conversion to ethylene by cotyledons from young seedlings was optimal at c. 30°C, and was strongly inhibited at 45°C. Pretreatment of imbibed seeds at high temperature (45°C) induced a thermodormancy and a progressive decrease in EFE activity.Abscisic acid and methyl-jasmonate, two growth regulators which inhibit seed germination and seedling growth, and cycloheximide were also shown to inhibit ACC conversion to ethylene by cotyledons of 3-day-old seedlings and by inbibed seeds.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - CH cycloheximide - EFE ethylene forming enzyme - IAA indole-3-acetic acid - Me-Ja methyl-jasmonate     

Barley (Hordeum vulgare) seed dormancy as related to glumella characteristics

Dormancy of freshly harvested barley ( Hordeum vulgare L. cv. Sonja) caryopses results mainly from glumellae which fix oxygen by polyphenol oxidase (EC 1.14.18.1)-mediated oxidation of phenolic compounds present in high amounts. The breaking of dormancy during dry storage is not due to qualitative or quantitative modifications of the phenols or polyphenol oxidases. Glumellae of dormant caryopses start to take up oxygen at the beginning of inbibition, whereas those of non-dormant caryopses start to take up oxygen only after about 10 h. That delay should allow germination.     

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     

Hormonal and environmental regulation of seed germination in flixweed (<Emphasis Type="Italic">Descurainia sophia</Emphasis>)

Flixweed is one of the most abundant weeds in North America and China, and causes a reduction in crop yields. Dormancy of flixweed seeds is deep at maturity and is maintained in soil for several months. To identify regulators of seed dormancy and germination of flixweed, the effect of environmental and hormonal signals were examined using dormant and non-dormant seeds. The level of dormancy was decreased during after-ripening and stratification, but long imbibition (over 5 days) at 4 °C in the dark resulted in the introduction of secondary dormancy. The strict requirement of duration of cold treatment for the break of dormancy may play a role in the seasonal regulation of germination. The germination of non-dormant flixweed seeds was critically regulated by red (R) and far-red (FR) light in a photoreversible manner. Sodium nitroprusside, a donor of nitric oxide (NO), promoted germination of half-dormant seeds, suggesting that NO reduced the level of seed dormancy. As has been shown in other related species, light elevated sensitivity to GA₄ in dark-imbibied flixweed seeds, but cold treatment did not affect GA₄-sensitivity unlike in Arabidopsis. Taken together, our results indicate that seed germination in flixweed and its close relative Arabidopsis is controlled by similar as well as distinct mechanisms in response to various endogenous and environmental signals.     

Participation of the tightly-bound (putative cytoskeleton-bound) polysomes in translation during germination of dormant and non-dormant cereal caryopses

Research was done on dormant and non-dormant barley cv. Ars caryopses and triticale cv. Grado caryopses treated and non-treated with abscisic acid (ABA). During germination higher participation of populations of so-called tightly-bound polysomes (TBP) in embryos of dormant barley caryopses was observed, as well as their high metabolic activity. In embryos of triticale caryopses of which dormancy was imposed in an artificial way by ABA (100 microM), the strongest incorporation of 14C-amino acids into nascent polypeptide chains in vivo was found in population of TBP, as well as the highest participation among three of the studied fractions (free polysomes, membrane-bound polysomes and tightly-bound polysomes). These results may indicate the significant role of TBP (putative cytoskeleton-bound polysomes--CBP) in maintaining dormancy during imbibition of cereal caryopses.     

Role of reactive oxygen species in the regulation of Arabidopsis seed dormancy

Freshly harvested seeds of Arabidopsis thaliana, Columbia (Col) accession were dormant when imbibed at 25°C in the dark. Their dormancy was alleviated by continuous light during imbibition or by 5 weeks of storage at 20°C (after-ripening). We investigated the possible role of reactive oxygen species (ROS) in the regulation of Col seed dormancy. After 24 h of imbibition at 25°C, non-dormant seeds produced more ROS than dormant seeds, and their catalase activity was lower. In situ ROS localization revealed that germination was associated with an accumulation of superoxide and hydrogen peroxide in the radicle. ROS production was temporally and spatially regulated: ROS were first localized within the cytoplasm upon imbibition of non-dormant seeds, then in the nucleus and finally in the cell wall, which suggests that ROS play different roles during germination. Imbibition of dormant and non-dormant seeds in the presence of ROS scavengers or donors, which inhibited or stimulated germination, respectively, confirmed the role of ROS in germination. Freshly harvested seeds of the mutants defective in catalase (cat2-1) and vitamin E (vte1-1) did not display dormancy; however, seeds of the NADPH oxidase mutants (rbohD) were deeply dormant. Expression of a set of genes related to dormancy upon imbibition in the cat2-1 and vet1-1 seeds revealed that their non-dormant phenotype was probably not related to ABA or gibberellin metabolism, but suggested that ROS could trigger germination through gibberellin signaling activation.     

A meta-analysis of the effects of frugivory (endozoochory) on seed germination: role of seed size and kind of dormancy

Heretofore, no study has determined how germination of ingested seeds is affected by the kind (class) of dormancy nor by seed dormancy x seed size interaction. Thus, we aimed to determine the effects of seed size, kind of dormancy and their interaction on germination of defecated seeds using a meta-analysis. We collected data for 366 plant species in 97 plant families from 76 publications. In general, gut passage significantly increased germination percentage of defecated seeds by 5% compared with that of control seeds. Germination percentages of non-dormant, physiologically dormant, and morphologically/morphophysiologically dormant seeds (all water-permeable) significantly decreased after gut passage by 40, 18, and 14%, respectively, compared with control seeds (non-gut-passed). Changes in germination percentage of seeds with physical dormancy (water-impermeable) were positive, and gut passage increased germination by 69% compared with control seeds. Germination of small seeds decreased 8% after gut passage, whereas germination of both medium and large seeds increased by 18%. However, changes in germination percentage differed between categories of seed size in each class of dormancy. In physically dormant seeds, germination of all seed sizes improved after gut passage, and the magnitude of increase was higher for large than for medium and small seeds. Thus, gut passage increased germination of medium-size water-permeable seeds (physiologically dormant and morphologically/morphophysiologically dormant) more than it did for large and small seeds. However, gut-passage decreased or did not change the germination percentage of non-dormant seeds. Seed size and kind of dormancy should be included in studies on the effect of gut passage on germination.