Abscisic acid (ABA) is a plant hormone that regulates seed dormancy and germination. Seeds undergo changes in both ABA content and sensitivity during seed development and germination in response to internal and external cues. Recent advances in functional genomics have revealed the integral components involved in ABA metabolism (biosynthesis and catabolism) and perception, the core signalling pathway, as well as the factors that trigger ABA-mediated transcription. These allow for comparative studies to be conducted on seeds under different environmental conditions and from different genetic backgrounds. This review summarizes our understanding of the control of ABA content and the responsiveness of seeds to afterripening, light, high temperature and nitrate, with a focus on which tissues are involved in its metabolism and signalling. Also described are the regulators of ABA metabolism and signalling, which potentially act as the node for hormone crosstalk. Integration of such knowledge into the complex and diverse events occurring during seed germination will be the next challenge, which will allow for a clearer understanding of the role of ABA.

The ability of crop seeds to retain their viability over extended periods of uncontrolled temperature and/or relative humidity conditions has not been widely investigated, although this is an important issue for genebank management. We report here the response of 18 crop species to storage for up to 26 years at 20.3 ± 2.3°C and 50.5 ± 6.3% relative humidity. Germination rates decreased in a sigmoid fashion, but the curve parameters were species characteristic. Pea, common bean and maize seeds retained their viability over the longest period (23, 21 and 19 years, respectively). In contrast, chive seeds survived for only 5 years and lettuce for 7 years. In addition to this interspecific variability, there were also indices for intraspecific variability, particularly in bean and chive seeds, just as in collard, lupin, poppy, wheat and maize seeds. A significant correlation was obtained between germination performance in the laboratory and seedling emergence following autumn sowing. Seeds in which oil was the major seed storage component were more short lived, whereas carbohydrates or proteins did not show an effect on seed longevity.

The mechanism of starch metabolism in the endosperm of germinating rice (Oryza sativa L.) seed (caryopsis) has been well studied; however, little is known about the occurrence and function of transitory starch in germinating rice embryos. During rice seed germination and seedling establishment, starch in the endosperm is hydrolysed to glucose, which is taken up by scutellar epithelial cells, converted to sucrose, and transported through vascular bundles to other embryonic tissues, such as growing shoots and roots. In this study, we found that soluble sugar was converted to starch in the scutellum 6 d after imbibition (DAI). Starch appeared primarily in cells surrounding the vascular bundles of the embryonic axis 12 DAI. The removal of growing shoots caused hyper-accumulation of starch in embryonic tissues, including the scutellum. In contrast, placing seedlings in the dark, which reduced photosynthetic efficiency, lowered starch levels in the embryonic axis cells. The disappearance of transitory starch from embryonic tissues of dark-grown seedlings was accompanied by a reduction of soluble sugar content and the down-regulation of the expression and activity of starch biosynthesis enzymes. These results suggest that the amount of transitory starch in embryonic tissues was dependent on the demand of growing sink tissues.

Consider the following four quotations concerning the distinction between breaking dormancy and stimulating germination.

Physiological dormancy of scarified seeds of Townsville stylo (Stylosanthes humilis HBK) was released by acidic aluminium (Al3+) solution. Antiethylenic substances inhibited germination of low-pH-stimulated dormant seeds, with a correspondingly low ethylene production and low activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase in seeds. On the other hand, antiethylenic substances did not decrease the germination of Al3+-stimulated seeds, but ACC oxidase activity and ethylene production by the seeds was decreased to a large extent. These data provide evidence that dormancy breakage by Al3+ differs from that caused by low pH and is not associated with ethylene production. Similarly to Al3+ action, methyl viologen (MV), a reactive oxygen species-generating compound, broke dormancy of Townsville stylo seeds. Sodium selenate and N-acetyl cysteine, antioxidant compounds, largely decreased germination of MV- and Al3+-stimulated dormant seeds. Altogether these data point to oxidative radicals constituting key molecules in the chain of events triggered by Al3+ leading to dormancy breakage.

Feeding stem–leaf–pod explants with d-chiro-inositol and d-pinitol was used as a method to modify α-d-galactosides in developing pea (Pisum sativum) seeds. Four genotypes differing in the composition of raffinose, stachyose and verbascose (raffinose family oligosaccharides or RFOs) in seeds – high RFOs (cv. Tiny), low RFOs (SZD175) and low verbascose (cv. Hubal and cv. Wt 506) – were studied. Although seeds of all examined pea lines were able to take up both d-chiro-inositol and d-pinitol, only d-chiro-inositol was effectively converted into its galactosides: mainly fagopyritol B1 (O-α-d-galactopyranosyl-(1 → 2)-d-chiro-inositol) and fagopyritol B2 (O-α-d-galactopyranosyl-(1 → 6)-O-α-d-galactopyranosyl-(1 → 2)-d-chiro-inositol). In seeds of pea lines naturally containing low levels of verbascose (cv. Hubal) and low RFOs (SZD175), the enhanced accumulation of fagopyritols depressed the RFO level by c. 64 and 20%, respectively. Moreover, in both genotypes, about 25 and 30% of total galactose bound in α-d-galactosides occurred in fagopyritols. d-Pinitol present in the pea seeds was converted into monogalactosides, but their accumulation was several-fold lower than that of fagopyritols and did not significantly influence the accumulation of RFOs. Pea seeds with the composition of soluble carbohydrates modified by feeding with either of the cyclitols were able to complete germination.

Dormant barley grains cannot germinate at 30°C and this inability to germinate is imposed mostly by the glumellae which have been suggested to limit oxygen supply to the embryo. Hypoxia imposed either artificially or by the glumellae to embryos from dormant grains, increases embryo sensitivity to abscisic acid (ABA) and promotes the accumulation of ABA during the first hours after imbibition. Expression of candidate genes involved in ABA synthesis (HvNCED), catabolism (HvABA8OH1) and signalling (HvABI5, HvVP1 and HvPKABA) was analysed in embryos isolated from dormant whole or de-hulled grains incubated in air or in hypoxia (5% oxygen). The presence of the glumellae enhanced the expression of genes involved in ABA metabolism and signalling with respect to that observed in de-hulled grains incubated in air. These results suggest that at least part of the observed physiological responses to the presence of the glumellae are regulated at the level of gene expression. However, hypoxia imposed on dormant de-hulled grains did not mimic the presence of the glumellae in terms of expression of candidate genes. Hypoxia mimics the presence of the glumellae in terms of dormancy maintenance and ABA accumulation and sensitivity, but its effects appear to operate through different mechanisms.

The forest seed bank has been demonstrated to vary spatially at scales from 2 to 10 m. To our knowledge, the fine-scale spatial structure, i.e. < 2 m, has not been studied before. This study aims to make a thorough investigation of fine-scale spatial structure. Soil samples (128) were collected from each of five 2.1 m × 2.1 m plots, using a combined systematic (64) and random design (64). This allowed investigation of the fine-scale spatial structure of individual species–plot combinations using indicator-variograms. Our results indicated that over half of all species recorded in a particular plot were spatially structured. Remarkably, the presence of spatial structure seemed independent of species frequency. Visualization of the spatial structure showed an irregular spatial pattern, i.e. seed clusters that were randomly distributed in space. Spatial dependence occurred over small distances, possibly suggesting that a significant proportion of seeds was deposited near the mother plant. We conclude by presenting the relevance and implications of small-scale spatial seed-bank patterning for seed-bank sampling.

Timing of seed germination influences plant lifetime fitness and can affect the ability of plant populations to colonize and persist in changing environments. However, the genetic variation of the seed germination response remains poorly understood. The amplified restriction fragment polymorphism (AFLP) technique was applied to characterize the genetic variation of germinated seeds collected from three Festuca hallii populations in the Canadian prairie. Three subpopulations with early, intermediate and late germination were identified from each population, based on germination tests at 10, 15 and 20°C in controlled growth chambers. Three AFLP primer pairs were employed to screen a total of 540 assayed seedling samples and 188 polymorphic AFLP bands were scored for each sample. None of the assayed AFLP bands were significantly associated with seed germination, but marked differences in estimates of mean band frequency were observed for various groups of germinating seeds under different test temperatures. Partitioning of the total AFLP variation showed that 5.9% AFLP variation was present among seeds of the three populations, 0.3% among seeds of three germination subpopulations, and 0.5% among seeds grouped for germination temperature. Genetic differentiation was significant among 27 groups of seeds representing population, germination timing and test temperature. Subpopulations with early and intermediate germination shared similar genetic backgrounds and were genetically differentiated from the late germination subpopulation. These results indicate that seed genotypes respond slightly differently to environmental variation, resulting in significant but weak genetic differentiation in the germination of F. hallii seeds. Implications for plant establishment and fescue restoration are discussed.

Seed banks are often crucial for the survival of plant species, especially short-lived ones. Nevertheless, empirical studies about the fate of seeds under natural conditions are scarce, particularly for parasitic plants. Therefore, fresh seeds of the holoparasitic Cuscuta epithymum were buried in the field under natural conditions in Belgium or kept at room temperature for up to 31 months, and germination was tested periodically by incubating seeds in light at 23°C. Buried C. epithymum seeds had cyclic changes in their germinability, while those stored dry at room temperature never germinated without scarification. Buried seeds exhibited a seasonal cycle of physiological dormancy, the first to be reported for a species with combinational (i.e. physical+physiological) dormancy. The physiological dormancy of the embryo was broken during winter ( ≤ 5°C) but induced during exposure to late spring temperatures (>10°C). Therefore, germination of C. epithymum seeds is fine tuned to seasonal temperature changes. Each year only a portion of the viable seeds could germinate. The portion of seeds buried in the soil that remained available for delayed germination, i.e. in the seed bank, decreased rather quickly, with only 8.5% of them being viable after 31 months of burial; all seeds stored in the laboratory remained viable. Nonetheless, we concluded that some C. epithymum seeds are capable of surviving for several years in the soil. These data indicate that the parasite is well adapted to survive in a dynamic heathland landscape where conditions for survival change constantly, but it is still sensitive to local extinction.

MicroRNAs (miRNAs) are involved in developmental programmes of plants, including seed germination and post-germination. Here, we provide evidence that two different miRNA pathways, miR156 and miR172, interact during the post-germination stages in Arabidopsis. Mutant seedlings expressing miR156-resistant SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE13 (mSPL13), which has silent mutations in the miR156 complementary sequence, over-accumulated SPL13 mRNA and exhibited a delay in seedling development. Microarray analysis indicated that SCHNARCHZAPFEN (SNZ), an AP2-like gene targeted by miR172, was down-regulated in these mutants. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and miRNA gel blot analyses showed that the MIR172 genes were up-regulated in mSPL13 mutants. These results suggest that the miRNA regulation cascades (miR156⊣SPL13 → miR172⊣SNZ) play a critical role during the post-germination developmental stages in Arabidopsis.

Raffinose, stachyose and phytin are undesirable compounds for soybean food and animal feed products. In seeds, raffinose and stachyose are believed to contribute to desiccation and cold stress tolerance. Thus, removal of these compounds from soybean by genetic mutation has resulted in a more commercially desirable composition, but potentially less physiologically viable seeds. In an effort to develop a method to improve viability and seed storability in soybean, stem–leaf–pod explants of three low raffinose, low stachyose lines, two of which were also low in phytin, and a check line were fed solutions containing d-chiro-inositol, myo-inositol or d-pinitol, free cyclitols which unload through the seed coat to the developing embryo where they accumulate as fagopyritols, galactinol and galactopinitols, respectively, during seed maturation. Increased galactopinitol and fagopyritol accumulation may substitute for the roles of raffinose and stachyose in low raffinose, stachyose and phytin seeds. Explants of all lines unloaded d-chiro-inositol, myo-inositol and d-pinitol. Fed d-chiro-inositol accumulated in leaf tissues demonstrating uptake into explants. Fed d-chiro-inositol and myo-inositol accumulated in pod wall and seed coat tissues of one or more lines. The results indicate that d-chiro-inositol was unloaded from the seed coat to the embryo in increased amounts after feeding. The potential use of increased maternal d-chiro-inositol for synthesis of fagopyritols in embryos to improve seed performance in low-stachyose and low-phytin soybean seeds is supported. The seed coat cup unloading of fed free cyclitols may provide a model system to test effective unloading of upregulated maternally synthesized cyclitols.

European gorse (Ulex europaeus L.) is a spiny shrub that grows spontaneously in the understorey of forests and heathlands in western Europe. Gorse is a pioneer species and forms large seed banks that can persist for a long time while buried deeply in the soil. Although many studies have been conducted on gorse seed banks in invasive contexts and in scrubland ecosystems, few data are available on forests in a native context. The aim of the present study was thus to report on the variability of seed-bank density in ‘critical’ stages in the forest management of pine stands (five stands) in south-western France. We examined variations in the number of gorse seeds as a function of soil depth but also of the presence and abundance of adult gorse in the understorey. Seed-bank density did not show a clear decrease in seed number with pine stand age, principally because gorse also appears to be able to establish itself in mature pine stands, probably thanks to local disturbances. In the pine stands in our study, the presence and abundance of seeds in the soil appeared to depend mostly on the presence of adult gorse as seeders in the understorey. Finally, we observed that, contrary to what has generally been found in scrubland ecosystems, most gorse seeds were located in the 5–10 cm soil layer rather than in the 0–5 cm soil layer. This depletion of the first 5 cm may be linked to seed germination that was not compensated for by the production of new seeds.

Coffea arabica L. is highly homozygotic and caffeine content in most cultivars is reported to be similar (approximately 1%). The caffeine content was analysed using individual seeds of cultivars Catuaí Vermelho, Mundo Novo and Bourbon Vermelho, and varieties Mokka and Laurina of C. arabica. Seeds were cut transversely in half, a thin slice from each was extracted in a mixture of chloroform and methanol, and caffeine was measured by high performance liquid chromatography. The extraction procedure was validated by comparison with two conventional methods less suited for analysing large numbers of seeds. The results revealed an unexpectedly large variation [Catuaí Vermelho: 3.72–25.9 mg g− 1 (seeds); Mundo Novo: 5.06–18.59 mg g− 1; Bourbon Vermelho: 6.76–16.59 mg g− 1] even in types naturally low in caffeine such as Laurina (1.17–9.97 mg g− 1) and Mokka (2.01–9.64 mg g− 1). The mean values for all cultivars were always in the range of the caffeine content found in the literature. Several studies showed that C. arabica has approximately 10% cross-pollination, but the variation observed in caffeine content in individual seeds was too large to be explained by cross-pollination.

Among 14 species of herbaceous Fabaceae, all six winter annuals show a marked non-deep physiological dormancy of the embryo in addition to physical dormancy. This physiological dormancy is apparent at 23°C, but not at lower temperatures of 10°C and 5°C and disappears after 3 months of dry storage. These results corroborate the hypothesis that combinational dormancy is a double safety mechanism for delaying germination during summer: physical dormancy postpones germination, and even in early softened seeds germination is prevented by physiological dormancy of the embryo. Softened, swollen seeds of Medicago arabica tolerate a subsequent desiccation and remain viable even after five cycles of dehydration and rehydration. The rate of natural softening of M. arabica seeds increases exponentially at higher temperatures, with a Q10 between 3.4 and 5.1, and obeys the Arrhenius equation. This indicates that a chemical reaction might be involved in breakdown of physical dormancy. Winter annuals with hard seeds show similar properties as winter annuals with permeable seeds: the need for afterripening and requirement of lower temperatures delay germination until autumn. Only one species, Vicia sativa, loses physical dormancy during dry storage. Drying during summer might be a supplementary cue for germination in autumn.

Germination and early seedling development are critical for successful stand establishment of plants. Following germination, the cotyledons, which are derived from embryonic tissue, emerge from the seed. Arabidopsis seedlings at post-germinative stages are supported mainly by the supply of nutrition from the cotyledons until vegetative leaves emerge and initiate photosynthesis. The switch to autotrophic growth is a significant transition at the post-germinative stage. Here, we provide evidence that down-regulation of SQUAMOSA PROMOTER-BINDING PROTEIN LIKE13 (SPL13) by microRNA156 (miR156) plays an important role in the regulation of the post-germinative switch from the cotyledon stage to the vegetative-leaf stage. Silent mutations created in the SPL13 sequence in the region that is complementary to the miR156 sequence caused the deregulation of the mutant form of SPL13 (mSPL13) mRNA from miR156. Mutant seedlings over-accumulated miRNA-resistant messages and exhibited a delay in the emergence of vegetative leaves compared to wild-type seedlings. The delay was not observed in control transgenic plants expressing non-mutated SPL13, indicating that the phenotype was caused specifically by the silent mutations and deregulation of SPL13 from miR156. Characterization of the SPL13 promoter indicated that this gene is expressed mainly in the hypocotyl and affects leaf primordium development. These results suggest that the repression of SPL13 by miR156 is essential for normal post-germinative growth in Arabidopsis.

Models that quantify the effects of cropping systems on weed dynamics are useful tools for testing innovative cropping systems. In these models, seed mortality in the soil is a key parameter to account for the cumulated effect of cropping systems over time via the soil seed-bank. Since seed mortality is difficult to measure, our objective was to develop a method to estimate it from easily accessible information. Seeds of 13 weed species were buried 30 cm deep in fields and were recovered regularly for 2 years to measure their viability. Seed mass, dimensions, shape, and protein and lipid contents as well as coat thickness were measured. To estimate seed mortality of species not included in the study, we searched for relationships between mortality rates and seed traits. Seed viability mainly decreased during the second year of burial, with mortality rates ranging from 0.01 to 0.63 seeds·seeds− 1·year− 1, depending on the species. Seed mortality decreased with increasing seed coat thickness. No correlation was found with other measured traits or with seed persistence data in the literature. These results were confirmed when the effects of phylogenetic relatedness with phylogenetically independent contrasts were included. The thickness of the seed coat, which varied between 17 and 231 μm over the range of species studied, can protect the seed from external attacks in the soil and slow down seed decay. This trait can be easily measured via X-ray images and could be used to estimate the seed mortality rate for a wider range of species.

Mature seeds of many legume species are normally characterized by water-impermeable seed coats, a form of physical dormancy. However, observations have suggested that the incidence of mature but permeable (non-dormant) seeds is sometimes substantial. Yet, the ecological processes associated with this non-dormancy have received little attention by plant ecologists. In white clover (Trifolium repens), we therefore studied: (1) the occurrence of initially permeable seeds in wild populations; (2) the relative performance of non-dormant and dormant seeds in plant establishment and reproduction in a field-sown experiment; and (3) the extent to which the trait is affected by humidity and plant genotype in a greenhouse experiment. No less than 35% of all viable seeds from the wild populations proved to be water permeable at maturity. The proportion of permeable seeds within inflorescences ranged from 0 to 100%. In the field-sown experiment, autumn-germinated non-dormant seeds had almost equally good chances of establishing as spring-germinated dormant seeds. Due to a marked head start in growth, the former yielded more flowers (and thus seeds) in the first flowering season. However, the greenhouse experiment proved that variation in the proportion of permeable seed between inflorescences represented a plastic response to humidity conditions during seed ripening, rather than variation among clones (broad-sense heritability ≤ 0.025). Thus the trait is not easily subject to selection.

Seeds of Medicago truncatula, a genomic model species for legumes, exhibit physiological and physical dormancy. Here, the factors influencing the germination behaviour of freshly harvested and stored seeds were investigated using several genotypes. Hardseededness is promoted when mature seeds are equilibrated at relative humidities (RH) below 75%. The release of physical dormancy during imbibition was dependent on the initial water content/RH that the seeds were dried to: the drier the seeds, the longer the imbibition time needed to break physical dormancy. The kinetics of physical dormancy release was slower than that of physiological dormancy, making it possible to uncouple both phenomena. Freshly harvested embryos without seed coverings germinated at the same speed as afterripened seeds. The depth of dormancy varied between different M. truncatula genotypes, from more to less dormant: DZA315.16>A17 (Jemalong)>R108>DZA45.5. This difference was eliminated by removing the endosperm. Collectively, these observations indicate that the endosperm is likely the main factor in the reduced germination of freshly harvested seeds. White light decreased germination speed of dormant seeds whereas it had no effect on non-dormant seeds. Recently harvested seeds were most dormant at temperatures above 17°C, whereas afterripened seeds germinated over a wider range of temperature. Fluridone could efficiently break dormancy, reinforcing the role of abscisic acid (ABA) synthesis. However, dormancy was not affected by gibberellic acid (100 μM GA3) or nitrate. The particular dormancy features unravelled here for M. truncatula, combined with the available genomic resources, make it a new, useful model for genetic and molecular studies which can complement those developed for Arabidopsis.

Cynara cardunculus (L.) seeds require incubation at fluctuating temperatures to terminate dormancy. In this study, we analysed the physiological mechanisms underlying such a requirement, focusing on the role of abscisic acid (ABA) and gibberellin (GA). As a conceptual framework, we considered the possibility that fluctuating temperatures and light trigger a similar set of hormonal processes after stimulus perception. To test this possibility, we (1) carried out hydrotime analysis of germination in seeds exposed to fluctuating temperatures (25/15°C) and constant temperature (20°C) with or without gibberellin (GA3) or red light; (2) determined the responses of seeds incubated at fluctuating or constant temperature to ABA, GA3, fluridone, an inhibitor of ABA biosynthesis, and paclobutrazol, an inhibitor of GA biosynthesis; and (3) determined the ABA content of seeds incubated at fluctuating or constant temperature. Incubation at 25/15°C or 20°C in the presence of GA3 reduced the mean base water potential [ψb(50)] of the population to a similar extent, compared to that observed with seeds incubated at 20°C without GA3. Irradiation with red light also reduced ψb(50) to a lesser extent than incubation in the presence of GA3. At all concentrations tested, exogenously applied GA3 did not promote germination of seeds incubated at 25/15°C. However, paclobutrazol inhibited germination, suggesting that fluctuating temperatures terminate dormancy through de novo GA biosynthesis. Fluctuating temperatures enhanced seed germination in the presence of ABA, but ABA content did not differ between seeds incubated at fluctuating and constant temperatures. This study provides clear evidence for the involvement of hormonal regulation in dormancy termination by fluctuating temperatures.