The timing of the germination of seeds is highly responsive to inputs from the environment. Temperature plays a key role in the control of germination, with low temperatures acting to stimulate this developmental transition in many species. In Arabidopsis, extensive gene expression changes have been reported at the whole seed level in response to cold, while much less is known about their spatial distribution across the diverse cell types of the embryo. In this study we examined the spatiotemporal patterns of promoter activity and protein abundance for key gibberellic acid (GA) and abscisic acid (ABA) factors which regulate the decision to germinate both during a time course of germination and in response to cold. Low temperature stimulated the spatial relocalization of these factors to the vasculature. The response of these seeds to dormancy-breaking stratification treatments therefore stimulates the distribution of both positive (GA) and negatively acting (ABA) components to this same cell type. This altered spatial pattern persisted following the transfer of seeds to 22°C, as well as after their rehydration, indicating that this alteration is persistent. These observations suggest that the vasculature plays a role in the low temperature-mediated stimulation of germination in this species, while novel cell types are recruited to promote germination in response to stratification.

The wheat bZip transcription factor TaABF1 mediates both abscisic acid (ABA)-induced and ABA-suppressed gene expression. As levels of TaABF1 protein do not change in response to ABA, and TaABF1 is in a phosphorylated state in vivo, we investigated whether TaABF1 could be regulated at the post-translational level. In bombarded aleurone cells, a TaABF1 protein carrying phosphomimetic mutations (serine to aspartate) at four sites (S36D, S37D, S113D, S115D) was three to five times more potent than wild-type TaABF1 in activating HVA1, an ABA-responsive gene. The phosphomimetic mutations also increased the ability of TaABF1 to downregulate the ABA-suppressed gene Amy32b. These findings strongly suggest that phosphorylation at these sites increases the transcriptional regulatory activity of TaABF1. In contrast to the activation observed by the quadruple serine to aspartate mutation, a single S113D mutation completely eliminated the ability of TaABF1 to upregulate HVA1 or downregulate Amy32b. Thus phosphorylation of TaABF1 can either stimulate or inhibit the activity of TaABF1 in regulating downstream genes, depending on the site and pattern of phosphorylation. Mutation of S318 and S322 (in the bZIP domain) eliminated the ability of TaABF1 to activate HVA1, but had no effect on the ability of TaABF1 to downregulate Amy32b, suggesting that TaABF1 represses Amy32b expression through a mechanism other than direct DNA binding. An important step towards understanding how ABA and gibberellin (GA) signals are integrated through TaABF1 phosphorylation to regulate downstream gene expression is to clarify the effects of those hormones on the expression of specific genes. In contrast to some other ABA-induced genes, we found that HVA1 induction by ABA or TaABF1 is not inhibited by GA.

The influence of acid scarification on the responsiveness of curly dock (Rumex crispus L. ♯ RUMCR) seeds to physical and chemical factors was studied. Scarification made the seeds responsive to GA4+7 (a mixture of gibberellin A4 + A7) and markedly improved the responsiveness to light, moist-chilling, and a 1-h 40-C temperature shift. Scarification reduced the mechanical restraint of the seedcoat by 0.4 to 0.8 MPa. Removal of the entire pericarp induced 69% germination in the dark. Moist-chilling treatment of seeds reduced the soaking period needed for maximum response to the high-temperature shift from 24 to 6 h. The embryo growth potential generated by moist-chilling was less in magnitude than that by the reduction in seed coat restraint by scarification. Induction of secondary dormancy was prevented to a marked degree in scarified (chilled or unchilled) seeds during polyethylene glycol-6000 (PEG) treatment in light or in the presence of GA4+7 while intact seeds rapidly became secondarily dormant in light. The ability of scarified seeds to remain germinable or nondormant for a longer period than the intact seed in low water potential imbibing medium may be due to a net reduction (by 0.4 to 0.8 MPa) in the impact on the embryo of germination-preventing forces (low water potential plus mechanical restraint of the seed coat).

Medusahead is an invasive annual grass that, once established, severely affects range-land productivity and stability. Medusahead builds large seedbanks in the litter and on the soil surface. Effective weed control of medusahead involves either inhibiting germination from the seedbank, eliminating the seedbank, or enhancing germination so that plants are available for control. The purpose of this study was to determine the influence of nitrogen enrichment, immobilization, and nitrification inhibition treatments in the field on the size and germination status of medusahead seedbanks. The germination status of medusahead seeds in seedbanks was determined by periodically collecting field samples of surface soil and litter and bioassaying them in greenhouse emergence tests. Control seedbanks had increased seedling emergence with KNO3 or GA3 enrichment of the bioassay substrate. The combination of these two materials increased emergence. Nitrogen enrichment increased seedling establishment in the field. Carbon enrichment in the field decreased seedling establishment and increased medusahead seeds in seedbanks. Nitrapyrin treatment decreased medusahead in the field similar to carbon enrichment. In comparison to the control or other treatments, GA3 enrichment was not as effective in increasing emergence from nitrapyrin-treated bioassay samples. The combination of carbon and nitrapyrin treatments was very effective in eliminating medusahead emergence in the field, but in wetter years, it never completely eliminated medusahead seedling recruitment and subsequent reproduction. These treatments have promise for influencing succession in medusahead infestations if an adapted perennial species, capable of competing under low nitrogen levels, becomes available.

Winter chilling in the field removed the primary dormancy in common ragweed (Ambrosia artemisiifolia L.) seeds and allowed them to germinate in both light and darkness. A secondary dormancy was induced in the spring and seeds again failed to germinate in light or darkness. Kinetin (K) + ethephon (E) + gibberellin A4 + A7 (GA4+7) applied to the soil in December prevented secondary dormancy. The following May only 20% of the seeds at the 15-cm depth remained viable. The rest had germinated or decayed. In contrast, 89% of the untreated seeds remained viable. Application of K + E + GA4+7 on April 22 and May 2 also broke the secondary dormancy. Gibberellin A3 (GA3) was less effective than GA4+7 in preventing or breaking secondary dormancy.

Effects of ethylene, gibberellins, kinetin, nitrate, light, and temperature, each alone or combined with other factors, were studied on the germination of dormant seed of field pennycress (Thlaspi arvense L. # THLAR). Treated seed were incubated at a range of constant temperatures, at temperatures alternating daily between 12 and 22 C (12 h each), or at 12 C followed by transfer to 22 C. No germination ocurrred in water under any conditions. When applied in the dark KNO3 had no effect on germination under any of the temperature regimes. A combination of KNO3, light, and a shift in temperature from 12 to 22 C induced 100% germination. Four days of incubation at 12 C induced maximal seed sensitivity towards KNO3 + light, and the sensitivity remained unchanged if the incubation at 12 C was extended to 15 days. Upon transfer to 22 C, the effect of low-temperature incubation was lost within 3 days. Of the two seed lots studied, one had a relatively lower chilling requirement, but both had similar requirements for light and KNO3. Gibberellic acids4+7 (GA4+7) induced 100% germination in the dark or light at all temperature regimes. Gibberellic acid3 (GA3) was promotive under isothermal conditions, but its effectiveness was enhanced by a shift in temperature from 12 to 22 C. Kinetin or ethephon, each applied alone, had little effect on germination. Ethephon and GA3 together induced 100% germination.

Germination of dormant wild oat (Avena fatua L. # AVEFA) seed was examined in response to various concentrations of ethephon (2-chloroethylphosphonic acid), KNO3, or both together in the dark at temperatures ranging from 7 to 32 C. Both chemicals significantly (P = 0.05) promoted germination and their effects were additive. Maximum stimulation of germination in response to all treatments occurred at 7 C, with a gradual decrease in reponse as temperature or duration of after-ripening, the seed that retained dormancy during storage remained responsive to ethephon also occurred during storage of mature seed at 3 to 32 C. The rate and extent of this dormancy loss increased with an increase in storage temperature. Regardless of the temperature or duration of after-ripening, the seed that retained dormancy during storage remained responsive to ethephon and KNO3 treatments. Suitable combinations of after-ripening, ethephon, and KNO3 induced over 90% germination of seed that was dormant at maturity. Experimental evidence has been obtained that suggests that this percentage may represent the total number of viable seed in the population. While ethephon promoted seed germination, it severely stunted root and shoot growth of the resulting seedlings. It appears that strategies could be developed to combine the use of ethephon and nitrate with the natural process of after-ripening to enhance and synchronize wild oat germination.

Buthidazole {3-[5-(1,1-dimethylethyl)-1,3,4-thiadiazol-2-yl]-4-hydroxy-1-methyl-2-imidazolidinone} may be useful for selective weed control in corn (Zea mays L.) but injures corn seedlings at high rates. Greenhouse studies were initiated to test potential antidotes for buthidazole injury to corn seedlings. NA (1,8-naphthalic anhydride) and CDAA (N,N-diallyl-2-chloroacetamide) were the most effective of six chemicals evaluated. A ratio of 1:3 (buthidazole: CDAA) appeared to be optimal for the protection effect. CDAA appeared to be more effective than NA. Since both CDAA and NA offered limited protection but R-25788 did not, this action appears to be through a different mechanism than the one proposed for the thiocarbamate or acetanilide herbicides. The other chemicals tested were R-25788 (2,2-dichloro-N,N-diallylacetamide), R-29148 (2,2-dimethyl-5-methyl-dichloroace-tyloxazolidine), carboxin (2,3-dihydro-5-carboxanilido-6-methyl-1,4-oxanthiin) and gibberellin (GA3). Seven herbicides, members of the acetanilide, thiocarbamate, and dinitroaniline groups, were also tested for their antagonistic interactions but none offered protection.

Laboratory and field experiments were conducted to determine the effects of several environmental factors on seed germination and seedling emergence of the invasive weed, apple of Peru. The anatomy of the seed was also investigated in this study. The anatomy of the seed is similar to that of an eggplant (Solanum melongena L.) seed. The seed is albuminous, and consists of a seed coat, endosperm, and an embryo. The seed coat, which consists of both outer and inner integuments, is relatively thick. Fresh and dry stored seeds exhibited strong dormancy. Hot water treatment did not affect the seed germination in the light, but significantly (P < 0.05) reduced the germination rate in the dark. Dry-heat treatment had no significant effect on seed germination in the dark, but it significantly (P < 0.05) increased the germination rate in the light. Cold stratification alone did not influence seed germination under constant temperature (25 C) but affected it under alternating temperature (25–15 C). However, a combination of warm and cold stratification produced significant (P < 0.05) germination in the dark as well as in the light. In addition, the effect of warm stratification followed by cold stratification was more pronounced in the dark than in the light. Gibberellin A3 (GA3) (10–3 and 10–4 M) treatments significantly (P < 0.05) increased the seed germination rate compared with that of the control. Emergence rate was maximum for seeds placed on the soil surfaces; no seedling emergence occurred when seeds were placed at a depth of 5 cm.

The dwarfing wheat (Triticum aestivum L.) breeding has promoted a dramatic increase in yields. Since the utilized dwarfing genes in wheat are very limited, identification of novel dwarfing genes is necessary for improving the genetic diversity of wheat. In this study, more than 300 dwarfing wheat lines from induced mutation were screened by kompetitive allele-specific PCR and gibberellin (GA) treatment. The 17.49% of Rht-D1b and 1.37% of Rht-B1b dwarfing mutants were identified in this mutant group. Additionally, Rht-D1b mutants showed more effective in reduction of plant height and higher 1000-grain weight comparing with that of Rht-B1b mutants. By combing with comparison of yield components and expression profile of GA biosynthetic genes with wild-type, the GA-responsive mutant dm15 without directly involvement in GA metabolism, significantly increased 1000-grain weight but no change of other yield components in two locations of field experiments. Meanwhile, another elite mutant dm11 with change of GA biosynthetic genes expression was also identified. These mutants will be promising candidates for dwarfing wheat breeding.

Knowledge about the hormonal control of grain dormancy and dormancy loss is essential in wheat, because low grain dormancy at maturity is associated with the problem of pre-harvest sprouting (PHS) when cool and rainy conditions occur before harvest. Low GA (gibberellin A) hormone sensitivity and high ABA (abscisic acid) sensitivity were associated with higher wheat grain dormancy and PHS tolerance. Grains of two PHS-tolerant cultivars were very dormant at maturity, and insensitive to GA stimulation of germination. More PHS-susceptible cultivars were less sensitive to ABA inhibition of germination, and were either more GA sensitive or germinated efficiently without GA at maturity. As grain dormancy was lost through dry afterripening or cold imbibition, grains first gained GA sensitivity and then lost ABA sensitivity. These changes in GA and ABA sensitivity can serve as landmarks defining stages of dormancy loss that cannot be discerned without hormone treatment. These dormancy stages can be used to compare different cultivars, seed lots and studies. Previous work showed that wheat afterripening is associated with decreasing ABA levels in imbibing seeds. Wheat grain dormancy loss through cold imbibition also led to decreased endogenous ABA levels, suggesting that reduced ABA signalling is a general mechanism triggering dormancy loss.

The wheat transcription factor TaABF1 physically interacts with the protein kinase PKABA1 and mediates both abscisic acid (ABA)-induced and ABA-suppressed gene expression. In bombarded aleurone cells of imbibing grains, the effect of TaABF1 in down-regulating the gibberellin (GA)-induced Amy32b promoter was stronger in the presence of exogenous ABA. As these grains contained low levels of endogenous ABA, the effect of TaABF1 may also be mediated by ABA-induced activation even in the absence of exogenous ABA. Levels of TaABF1 protein decreased slightly during imbibition of afterripened grains. However, TaABF1 levels (especially in aleurone layers) were not substantially affected by exogenous ABA or GA, indicating that changes in TaABF1 protein level are not an important part of regulating its role in hormone signalling. We found that TaABF1 was phosphorylated in vivo in aleurone cells, suggesting a role for post-translational modification in regulating TaABF1 activity. Induction of Amy32b by overexpression of the transcription factor GAMyb could not be prevented by TaABF1, indicating that TaABF1 acts upstream of GAMyb transcription in the signalling pathway. Supporting this view, knockdown of TaABF1 by RNA interference resulted in increased expression from the GAMyb promoter. These results are consistent with a model in which TaABF1 is constitutively present in aleurone cells, while its ability to down-regulate GAMyb is regulated in response to ABA.

The expression patterns of β-1,3-glucanases (βGlu) and chitinases (Chn) were investigated during the seed germination of members of the Cestroideae (three Nicotiana species, Petunia hybrida) and the Solanoideae (Capsicum annuum, Physalis peruviana) subgroups of Solanaceous species. Rupture of the micropylar testa (seed coat) and rupture of the micropylar endosperm, i.e. radicle emergence, were distinct and temporally separate events during the germination of Cestroideae-type seeds. βGlu accumulation in imbibed Cestroideae-type seeds, occurring after testa rupture but prior to endosperm rupture, was inhibited by abscisic acid (ABA) and promoted by gibberellins (GA) and light, in strict association with germination, and appeared to be caused by transcriptional regulation of the class I βGlu genes. The micropylar cap of Solanoideae-type seeds does not allow a distinction between testa and endosperm rupture, but βGlu accumulation occurred prior to radicle emergence of pepper and P. peruviana seeds. ABA inhibited endosperm rupture and βGlu accumulation in the micropylar cap of pepper seeds. In contrast to tomato, βGlu accumulation in pepper seeds was not only confined to the micropylar cap, was due to distinct, tissue-specific βGlu isoforms, and was not accompanied by Chn accumulation. In conclusion, ABA inhibition of germination and βGlu accumulation in the micropylar endosperm appears to be a widespread event during the seed germination of Solanaceous species. In contrast, accumulation of Chn and distinct βGlu isoforms in the embryo, prior to germination, appears to be a species-specific phenomenon within the Solanaceae. In addition, a post-germination co-induction of βGlu and Chn in the root of the emerged seedling was found in endospermic and non-endospermic species and could represent an evolutionarily conserved event during dicot seedling development.

Cereal grain germination and early seedling growth involve the co-ordinated action of endosperm and embryo tissues to mobilize the storage reserves of the starchy endosperm. This mobilization is accomplished by hydrolases secreted from the aleurone and scutellar tissues. The breakdown products are then transported to the growing seedling by the scutellum. This resource-harvesting system is regulated at multiple levels. One well-defined aspect of control is brought about by the hormone gibberellin (GA). Gibberellin is released from the embryo upon imbibition and activates the aleurone cells. The secretory apparatus of the aleurone then proliferates, supporting increased hydrolase synthesis and secretion to degrade the starchy endosperm. The molecules that regulate this response to GA are now being increasingly characterized. Elements such as cGMP, calcium, calmodulin and protein kinases are well known as regulators in other eukaryotic cell types and are emerging as key control factors in the aleurone hormone response. However, superimposed upon this molecular regulatory system is another level of control, the structural pattern of tissues and stored macro- molecules that was laid down during grain development. It is the interaction of these structural motifs combined with the molecular regulatory mechanisms that ensure the appropriate timing and positioning of hydrolase production and endosperm reserve mobilization. This integrated control system ensures an extended release of nutrients to fuel early seedling growth.

Grains of indica rice (Oryza sativa cv. Peiza 67) exhibit an increase in endo-β-mannanase activity, mostly after the completion of germination. According to tissue blots, the initial increase occurs in association with the embryo, and possibly the scutellum, although the largest sustained increase in activity is in the peripheral regions of the endosperm. The aleurone layer, being the only living region of the endosperm, is presumably the site of synthesis and secretion of the enzyme into the non-living, starch-laden region. β-Mannosidase activity is low throughout germination and subsequent seedling growth, particularly in the endosperm regions. Its activity profile does not mimic that of endo-β-mannanase. In the intact grain, gibberellin (GA) causes a relatively small increase in endo-β-mannanase activity, while abscisic acid (ABA) causes a large decrease; this inhibition is overcome to a considerable extent when GA is supplied along with ABA. β-Mannosidase activity is little affected by either GA or ABA. Embryoless half-grains imbibed in water exhibit only a small increase in endo-β-mannanase activity with time of imbibition, showing the necessity for a stimulus from the embryo for this to occur. Incubating half-grains in the presence of GA results in a large increase in enzyme activity; ABA reduces the amount of activity compared to the water controls. GA is capable of reversing the inhibitory effect of ABA with respect to endo-β-mannanase activity. As in the intact grains, β-mannosidase activity in the half-grains is unaffected by either GA or ABA. It is concluded that the major site for the production of endo-β-mannanase activity is the aleurone layer, and this event is influenced by the presence of the embryo; in the absence of the latter, the increase in enzyme activity is stimulated by GA. β-Mannosidase activity is low throughout germination and post-germination, it is not influenced by GA and ABA, and thus its activity is not regulated in a coordinated manner with that of endo-β-mannanase.

This review focuses mainly on eudicot seeds, and on the interactions between abscisic acid (ABA), gibberellins (GA), ethylene, brassinosteroids (BR), auxin and cytokinins in regulating the interconnected molecular processes that control dormancy release and germination. Signal transduction pathways, mediated by environmental and hormonal signals, regulate gene expression in seeds. Seed dormancy release and germination of species with coat dormancy is determined by the balance of forces between the growth potential of the embryo and the constraint exerted by the covering layers, e.g. testa and endosperm. Recent progress in the field of seed biology has been greatly aided by molecular approaches utilizing mutant and transgenic seeds of Arabidopsis thaliana and the Solanaceae model systems, tomato and tobacco, which are altered in hormone biology. ABA is a positive regulator of dormancy induction and most likely also maintenance, while it is a negative regulator of germination. GA releases dormancy, promotes germination and counteracts ABA effects. Ethylene and BR promote seed germination and also counteract ABA effects. We present an integrated view of the molecular genetics, physiology and biochemistry used to unravel how hormones control seed dormancy release and germination.

As seed dormancy is released within a seed population, both the rate and percentage of germination increase progressively with increasing dose of a dormancy-breaking treatment or condition. Population-based models can account for this behaviour on the basis of shifting response thresholds as dormancy is alleviated. In particular, hydrothermal time analysis of germination sensitivity to water potential (Ψ) and temperature (T) can describe these features of seed behaviour. We used the hydrothermal time model to analyse the effects of dormancy-breaking treatments on germination of dormant true (botanical) potato (Solanum tuberosum L.) seeds (TPS). After-ripening (37°C and 4% seed moisture content) of TPS for 7 or 30 days partially or fully alleviated primary dormancy. The median base water potential required to prevent germination [Ψb(50)] decreased from –0.25 MPa in control seeds to –0.87 MPa and –1.83 MPa after 7 and 30 days of after-ripening, respectively. In contrast, the base temperature for germination (Tb) was relatively unaffected (0–3.3°C). Fluridone (50 μM), an inhibitor of abscisic acid (ABA) biosynthesis, also promoted germination of dormant TPS and lowered Ψb(50), indicating a role for de novo synthesis of ABA during dormancy maintenance. Moist chilling (3 days at 4°C) or gibberellin (100 μM) alleviated secondary dormancy and lowered Ψb(50) values from –0.08 MPa to –0.36 and –0.87 MPa, respectively. The hydrothermal time model allows quantification of dormancy levels and explains why changes in germination speed and percentage are closely correlated during dormancy alleviation.

β-1,3-Glucanase (βGlu) expression in seeds plays important roles in the regulation of seed germination, dormancy and in the defence against seed pathogens. A thick β-1,3-glucan layer is typical for the seed envelope of cucurbitaceous species, confers seed semipermeability and is degraded during germination. In many species with coat-imposed dormancy, the seed envelope confers a physical constraint to radicle emergence. In the solanaceous species, the micropylar endosperm and testa have this function, and endosperm weakening appears to be a prerequisite for germination. Class I βGlu is transcriptionally induced in the micropylar endosperm of tobacco, tomato and other solanaceous seeds just prior to radicle emergence. βGlu induction and germination are tightly linked in response to plant hormones and environmental factors, e.g. they are both promoted by gibberellins and inhibited by abscisic acid (ABA). Sense and antisense transformation of tobacco reveals two sites of βGlu action: after-ripening-mediated release of testa-imposed dormancy and endosperm rupture during germination. The use of an ABA-inducible chimeric sense-transgene resulted in overexpression of class I βGlu in seeds and provided direct evidence that βGlu contributes to endosperm rupture. A model integrating βGlu, seed dormancy, after-ripening and germination is presented, and possible mechanisms for βGlu action are discussed. It is proposed that βGlu not only helps defend seeds against pathogens, but is also a key factor in regulating coat-imposed dormancy and germination of seeds in response to environmental and hormonal cues.

As a member of the tribe Brassiceae, Eruca sativa, although a minor crop worldwide, is considered a valuable genetic resource for cabbage, rapeseed and other Brassica crops. Self-incompatibility (SI) in Brassica has been extensively studied, but information on SI in E. sativa is limited. Of six chemicals used to treat the stigmas to overcome SI in five E. sativa lines, gibberellin was the most effective. As gibberellin is well known for its ability to break dormancy and to promote cell elongation, its effectiveness may help to understand the mechanisms of SI. Urea and ammonium sulphate were also effective. These two chemicals are known to affect protein stability, which may help explain their effects on SI. Although table salt has been reported as being effective in overcoming SI in B. rapa, B. oleracea and B. napu, it was not effective in E. sativa. Sucrose and alcohol also had negligible effect. There was significant variation among the genotypes in SI intensity and response to chemicals, but the genotype–chemical interaction was not significant. The data presented in this paper add to our understanding of SI in E. sativa and may lead to a better use of this genetic resource.

The range of processes regulated by gibberellins (GAs) covers all aspects of the life history of the plant from seed germination to vegetative growth and flowering. In seeds there has been an intensive search, using the techniques of both biochemistry and cell biology, for the regulatory molecules linking GA perception to gene regulation and the events of germination. Although a GA receptor has yet to be identified, the site of perception has been localized to the plasma membrane. Calmodulin, Ca2+ and cGMP have also been identified as elements of the GA signal transduction pathway. These regulators parallel many of the signalling elements identified in the transduction of other signals such as phytochrome and ABA. Studies of GA-regulated gene expression, principally of the α-amylases of cereal aleurone, have identified core GA-responsive promoter elements, such as the gibberellin response element (GARE), box-1 and pyrimidine boxes, as well as elements that may lend specificity to GA-regulated expression, such as the Opaque-2-similar element (O2S), and TRE and CRE motifs. One of the most striking features of all of these studies of the molecular basis of GA action is the interaction of GA-dependent regulatory elements with those of other factors such as ABA. GA-response elements also appear to be conserved between disparate GA-response systems. For example, Myb transcription factors appear to regulate a multitude of GA-induced genes in cereal aleurone as well as to alter GA responses when expressed in Arabidopsis. Thus the study of GA signal transduction and response systems is highlighting the conservation of regulatory elements used by plants. These common factors, used by distinct signal transduction systems, provide a molecular basis for the integration of the GA signal with other growth regulators that is the hallmark of plant growth and development.