Prechilling seeds of Sitka spruce (Picea sitchensis [Bong.] Carr.) at 4°C with 30% moisture content for 12–14 weeks (84–98 d) removed conditional dormancy (i.e. they were then able to germinate at 10°C). The non-dormant status was preserved after redrying to 6% moisture content. However, conditional dormancy was gradually reimposed during subsequent air-dry storage at 4°C and 6% seed moisture content in all five seed lots tested. Further investigations with one seed lot showed that reimposition was reversed by a second prechill treatment, but was reimposed again during subsequent air-dry storage. The trend of dormancy reimposition within seed lots over time was quantified by negative exponential relations between ability to germinate at 10°C and duration of air-dry storage. The progress of dormancy reimposition was influenced by seed storage moisture content and was most rapid at 4–10%. At higher moisture contents (15 and 20%) the rate of the reimposition of conditional dormancy was much reduced, while at moisture contents of 25 and 30% further loss in dormancy occurred. Thus it is clear that dormancy reimposition occurred during storage at low water potential rather than solely during desiccation from high to low water potential.

Winter rape has a life cycle which necessitates rapid testing of seed quality for the seed production sector, namely between July after harvest of the previous crop and August when the next crop establishment begins. As this crop is the fourth most important crop in France, and the most important oilseed crop, the French seed testing station GEVES has managed several research programmes to study seed quality in this species. These collaborative programmes have not only associated seed science with seed technology but also informatics and electronic engineering to provide seed testing tools that are already used by the seed sector, such as conductivity, or innovative tools, such as germination time courses monitored by computer imaging. Forthcoming tools have also been looked at in order to control seed storage potential, which is often used nowadays to anticipate the next season. Biochemical markers have been studied, including seed-specific biotinylated proteins or enzymes involved in free radical (reactive oxygen species) scavenging, such as catalase. Both are well correlated with seed ageing and have been tested to provide an ELISA-like assay for seed testing laboratories. Germination monitoring by computer imaging is now used by research and breeding institutes to phenotype varying seed material, including genotype or mutant collections, primed or ageing seed lots, etc. The next step in seed quality testing in oilseed rape is to characterize variation in germination and seed vigour in order to stabilize optimal yield. This aim constitutes a new international project involving high-throughput phenotyping and genotyping of a large collection of genetically diverse rapeseed genotypes (double haploid populations or diverse cultivars).

In Britain, Lobelia urens (L.) (the heath lobelia) occurs in rough grassland, is rare and only found in southern England, where it is at the northern limit of its range. Emergence and survival of L. urens was investigated at six locations in two geographically distinct sites experiencing spring, autumn or no grazing in two consecutive years. Four factors were evaluated qualitatively, as a means of characterizing microhabitats for germination and survival: all permutations of higher plant cover, bryophytes, plant litter and surface depressions. The potential effect of adjacent plants on recruitment was also assessed using the nearest neighbour distance (NND). Grazing created depressions, removed plant litter and increased the proportion of sites with higher plant cover. It also resulted in a more open sward with higher NNDs. None of these changes stimulated recruitment. Instead, small increases in the frequency of some rare or very rare microhabitat types were vital in making grazed rough grassland more suitable for emergence. Reduced litter loads and a greater quantity of moss were two key responses. Overall, survival of L. urens was less than 1% and was particularly favoured by moss and an increase in NNDs. Shading from higher plants, with or without plant litter, decreased emergence, but the precise role of litter was complex and most probably related to its quantity. In an experimental seed bed, only empty depressions favoured emergence. The microhabitat relationships of L. urens were unusually consistent among locations and consecutive years. L. urens requires high soil surface temperatures but also adequate water for large scale recruitment, and such conditions are encouraged by grazing. The particular problems of experimental design and statistical analysis of data from recruitment experiments are also discussed.

Excised Helianthus annuus embryos were capable of precocious germination as early as 7 d after pollination. At this stage they were not capable of withstanding desiccation treatment. The desiccation tolerance was acquired at a stage of development which depended on the rate of water loss to which embryos were subjected. Coincident with the acquisition of tolerance to a rapid desiccation, they gained the capacity to withstand 24 h preincubation in agitated water; this acquisition could be correlated to a sharp decrease of the leaching of electrolytes, amino acids and proteins in the incubation water, suggesting that a change in membrane properties played a key role in this transition. Moreover, the acquisition of tolerance to these stresses was coincident with the increase of endogenous abscisic acid (ABA). ABA applied in vitro to young embryos induced tolerance both to desiccation and to preincubation in water: this in vitro acquisition of tolerance was again correlated with a decrease in the leakage.

It has been proposed that gibberellic acid (GA3) promotes germination by overcoming restrictions in sugar production and utilization in afterripening-responsive dormant caryopses. While their germination rates were similar, germination commenced sooner in afterripened wild oat (Avena fatua L.) caryopses than in dormant caryopses treated with GA3 and dormant excised embryos treated with GA3 plus fructose (Fru). Limited germination occurred in dormant excised embryos cultured with GA3 alone. Carbohydrate concentrations were measured over time in dormant caryopses and excised embryos whose germination was induced with GA3 and GA3 plus Fru. The concentration of sucrose (Suc) in the endosperm declined prior to germination of dormant GA3-treated caryopses. Raffinose (Raf) family oligosaccharides in the embryos of dormant GA-treated caryopses remained relatively constant prior to and shortly after the onset of germination. In contrast, Raf family oligosaccharides in the embryos of afterripened caryopses declined prior to germination. Together this suggests Raf family oligosaccharide utilization is not associated with germination per se. Increased starch levels, which occurred in dormant excised embryos treated with Fru and GA3 plus Fru, were associated with dormancy because similar effects were not apparent in afterripened embryos cultured with Fru. An initial decline in the concentration of Raf family oligosaccharides in dormant embryos cultured with GA3 or GA3 plus Fru seems to be a result of the excision process. GA3 appears to stimulate the germination of dormant embryos by enhancing the uptake or utilization of Fru. It appears that GA3 and afterripening-induced changes in carbohydrate utilization in dormant caryopses are different.

Soil seed banks are important to vegetation recruitment, ecosystem functioning and land management. We evaluated composition of 0–5 cm soil seed banks and relationships of seed banks with forest community types (ranging from low-elevation pinyon–juniper to high-elevation bristlecone pine), vegetation cover and environmental variables within a 40,000-ha relatively undisturbed coniferous forest landscape in Nevada, USA. We collected samples from 36 sites and used the emergence method to assay seed banks. Seed density averaged 479 seeds m− 2 across sites and a total of 39 taxa were detected. Most (79%) of these taxa were perennials and 35 of 39 (90%) were native. Moreover, 62% of seed-bank taxa were in the vegetation of mature forests, an uncommon finding in studies of forest soil seed banks. Seed-bank density, species richness and composition did not display strong relationships with forest community types, vegetation cover or environmental variables. Weak relationships likely arose from the relatively uniform seed-bank density among sites, where 50% of sites had seed densities in the range of 106–282 m− 2. Results suggest that while seed banks on this landscape are not large, they provide recruitment potential for some native perennial species of mature, relatively undisturbed communities.

Previous experiments had shown that microgravity adversely affected seed development in Brassica rapa L. We tested the hypothesis that gravity controls seed development via modulation of gases around the developing seeds, by studying how hypergravity affects the silique microenvironment and seed development. Using an in vitro silique maturation system, we sampled internal silique gases for 16 d late in the seed maturation sequence at 4 g or 1 g. The carbon dioxide level was significantly higher inside the 4-g siliques, and the immature seeds became heavier than those maturing at 1 g. Pollination and early embryo development were also studied by growing whole plants at 2 g or 4 g for 16 d inside chambers mounted on a large-diameter centrifuge. Each day the rotor was briefly stopped to permit manual pollination of flowers, thereby producing cohorts of same-aged siliques for comparison with stationary control material. The loss of starch and soluble carbohydrates during seed development was accelerated in hypergravity, with seeds developing at 4 g more advanced by 2 d than those at 1 g. Seeds produced at 4 g contained more lipid than those at 1 g. Taken together, these results indicate that hypergravity enhances gas availability to the developing embryos. Gravity's role in seed development is of importance to the space programme because of the plan to use plants for food production and habitat regeneration in extraterrestrial settings. These results are significant because they underscore the tight co-regulation of Brassica seed development and the atmosphere maintained inside the siliques.

For endozoochorous species, the quality component of seed dispersal effectiveness depends in part on the treatment seeds receive in the animal's gut. Covering a variety of taxa, diet, digestion system and body size of Prosopis flexuosa seed dispersers, we analysed differences among species in (1) mean retention time of ingested seeds, (2) recovery of viable seeds, (3) seed germination in comparison with seeds collected from trees and (4) germination of seeds after two different periods of retention in the gut. Feeding experiments were conducted with captive individuals of Dolichotis patagonum, Lycalopex gymnocercus, Rhea americana, Chelonoidis chilensis and Lama guanicoe. On the first day, we provided them with fruits containing controlled amounts of seed, and on the subsequent days, we collected faeces in order to recover seeds. We performed germination and viability tests on seeds coming from faeces and collected from trees. The results showed differences among species in the mean retention time of seeds. Chelonoidis chilensis had the longest mean retention time, but its effect on seed recovery and germination was similar to that of the other species, except for L. guanicoe, which showed the lowest seed recovery. When scarification and promotion of seed germination were considered, herbivorous mammals and tortoises (L. guanicoe, D. patagonum and C. chilensis) were the ones increasing germinability, whereas R. americana and L. gymnocercus did not significantly increase final seed germination percentage, which was similar to that for seeds collected from trees. P. flexuosa seeds receive a variety of treatments from endozoochorous dispersers, which might result in an overall fitness benefit for a plant living in unpredictable environments.

An element of storage stability of many orthodox seeds is that the embryo cytoplasm is vitreous at normal storage temperatures. That is, as drying proceeds during seed maturation, intracellular solutions become so concentrated and viscous that diffusional movement is all but eliminated (Williams and Leopold, 1989). In this report the relationship between desiccation tolerance and the ability to form the vitreous state as germination proceeds is examined. As pea seeds (Pisum sativum cv. Alaska) imbibed water up to 18 h, no change in the glass transition temperature was seen, and the embryos remained desiccation-tolerant. Between 18 and 44 h of imbibition, the axes lost the ability to exhibit a distinguishable vitreous transition, and the embryos had lost the ability to survive desiccation. After about 50 h, embryos again showed vitrification but only at markedly lower temperatures, as would be expected to accompany the loss of oligosaccharides (sucrose, raffinose, stachyose and verbascose) and their replacement by monosaccharides during early germination (Koster and Leopold, 1988). Thus, the loss of desiccation tolerance during germination in pea seeds appears to be associated with a loss of the high temperature oligosaccharide: water glass and a subsequent appearance of a new glass transition at a lower temperature resulting from the accumulation of monosacchrides.

Abstract In isolated embryos from dormant barley grains, synergistic effects of fusicoccin (FC) and gibberellic acid (GA3) were observed on the induction of α-amylase mRNA expression. However, no α-amylase mRNA expression could be induced by both agents in embryos from non-dormant grains. Both light- and electron-microscopy studies demonstrated that there were large numbers of starch granules present in mature embryos (mainly in scutellum) from dormant barley grains but none or almost none in embryos from non-dormant grains. Furthermore, the content of reducing sugars in embryos from dormant grains was about half of that from non-dormant grains. In contrast to GA3, FC was able to induce a strong acidification of extracellular pH (pHe). Clamping the pHe to prevent FC-induced acidification, by using 50 mM MES buffer (pH 5.6), caused an inhibition of GA3- or FC-induced α-amylase mRNA expression but did not affect the germination of embryos from dormant grains. In addition, in MES buffer, addition of FC or a combination of FC and GA3increased the germination rate of embryos isolated from dormant grains, though large numbers of starch granules were still present in these embryos. Based on these observations, the presence of starch granules and a low reducing sugar level in embryos from dormant grains is not a factor for control of grain dormancy and germination.

Laboratory characteristics of seeds of 37 species (41 seed types) from the Strandveld Succulent Karoo were used to predict seed bank types according to a modified key of ). Five seed bank strategies were recognized for this vegetation type, i.e. two with transient and three with persistent seed bank strategies. Of the 37 species investigated, 32% (all perennial species) had transient seed bank strategies, while 68% had persistent seed bank strategies. Seed dispersal of these 37 species was mainly anemochorous, although antitelechoric elements such as myxospermy, hygrochasy, heterodiaspory and synaptospermy were found among these species. The seed bank alone will not be sufficient to restore the vegetation of damaged land in the Strandveld Succulent Karoo, since many of the dominant species in the vegetation do not produce persistent seed banks. Many of these species may, however, be dispersed by wind into revegetation areas from surrounding vegetation. Topsoil replacement, seeding and transplanting of selected species will be essential for the successful revegetation of mined areas in this part of Namaqualand.

Germination experiments are becoming increasingly complex and they are now routinely involving several experimental factors. Recently, a two-step approach utilizing meta-analysis methodology has been proposed for the estimation of hierarchical models suitable for describing data from such complex experiments. Step 1 involves fitting models to data from each sub-experiment, whereas Step 2 involves combination estimates from all model fits obtained in Step 1. However, one shortcoming of this approach was that visualization of resulting fitted germination curves was difficult. Here, we describe in detail an improved two-step analysis that allows visualization of cumulated data together with fitted curves and confidence bands. Also, we demonstrate in detail, through two examples, how to carry out the statistical analysis in practice.

Knowledge of processes responsible for seed dormancy can improve our understanding of the evolutionary dynamics of reproductive systems. We examined the influence of the breeding system on primary seed dormancy in Hypericum elodes, an Atlantic–European softwater pools specialist plant that exhibits a mixed mating strategy (the ability to both self- and cross-pollinate) to set seeds. Seeds were obtained through hand pollination treatments performed in a natural population during three consecutive years. Primary dormancy of seeds recovered from each pollination treatment was measured by analysing the seed germination response at dispersal and after various periods of cold stratification. While all collected seeds exhibited physiological dormancy, the degree of primary dormancy was associated with the pollination treatments. Weak and rapid loss of primary dormancy characterized seeds recovered from self-pollinated flowers, while stronger dormancy was found in seeds obtained from cross-pollination. The association between pollination treatments and primary dormancy indicated that the mating system should be considered as a source of variation for dormancy degree, proportional to self- and cross-pollinations (selfing rate) within populations of this species. These results suggest that by shedding seeds with various degrees of dormancy, plants may distribute their offspring across time by means of polymorphism in germination response. We conclude that seed germination alone is not an appropriate fitness measure for inbreeding depression estimates, unless dormancy is removed.

Seed germination requirements of Viburnum lantana were investigated by experiments both in the laboratory and outdoors. Embryo length, radicle emergence and shoot emergence were analysed to determine the level of morphophysiological dormancy (MPD) of seeds. Mean embryo length in fresh seeds was 1.30 mm, and required growth to at least 2.51 mm to germinate. The critical embryo length was 4.1 mm. In the laboratory, the embryo reached 3 mm length after 20 weeks of warm-temperature incubation (20/7 or 25/10°C), which in fact represents a combination of warm + cold stratification. In seeds subjected to cold stratification (1.5 or 5°C) for 24 weeks, embryos hardly grew. Gibberellic acid stimulated embryo growth and germination. In the outdoor phenology test, the embryos grew from 1.30 mm, i.e. fresh seeds sown in September, to 2.98 mm at the end of the following March. In the ‘move-along’ test (laboratory), starting with temperatures of warm stratification [i.e. 25/10°C (4 weeks) →  20/7°C (4 weeks) →  15/4°C (4 weeks) →  5°C (12 weeks) →  15/4°C (4 weeks)], and in the outdoor phenology study on seeds exposed to a similar temperature sequence, radicle emergence percentages reached 73% after 28 and 35 weeks, respectively. V. lantana does not exhibit a delay between root and shoot emergence, dismissing any kind of epicotyl dormancy. Seeds of V. lantana have non-deep simple MPD, a level not detected previously in the genus Viburnum, with the physiological dormancy component overcome by a combination of warm and cold stratification, preferably in that order.

In Arabidopsis, seed germination is a biphasic process involving rupture of the seed coat followed by emergence of the radicle through the micropylar endosperm. Embryo expansion results in seed coat rupture and removal of seed coat imposed dormancy with DELLA proteins blocking embryo expansion in the absence of gibberellins. Exogenous abscisic acid (ABA) treatment does not block seed coat rupture but does block radicle emergence. We used this limited effect of exogenous ABA to further investigate the mechanism by which it blocks the onset of germination marked by seed coat rupture. We show that physical nicking of the seed coat results in exogenous ABA treatment blocking both seed coat and endosperm rupture and this block requires the transcription factors ABI3 and ABI5, but not ABI4. Furthermore, we show that the repression of expression of several EXPANSIN genes (EXPA1, EXPA2, EXPA3, EXPA9 and EXPA20) by exogenous ABA requires ABI5. We conclude that ABI5 plays an important role in the ABA-mediated repression of germination through prevention of seed coat rupture and propose that this involves EXPANSIN related control of cell wall loosening.

The seed coat of soybean (Glycine max (L.) Merrill) must protect the seed but allow water intake. Overprotection, causing impermeability, is assumed to be due to the presence of an impermeable layer in the seed coat, although validation of this assumption has relied on imbibition testing, which tends to be influenced by microfractures in the seed coat. Recent micromorphological analyses using laser-assisted topography microscopy revealed links to the surface roughness (SR) of the seed coat. To verify genetic links between hardseededness and SR, we analysed quantitative trait loci (QTLs) governing SR formation using 148 recombinant inbred lines (RILs) with a genetic linkage map covering 2663.6 cM of all 20 linkage groups of soybean, with 355 DNA markers and 5 phenotype markers. Five QTLs were detected, including previously identified hardseededness QTLs for ratio of seeds absorbing water, namely RAS1 and RAS2, which accounted for 20% of the phenotypic variance, and one near a locus inhibiting seed coat colour (I). These results indicate that the impermeability of soybean seed is genetically related to the reduction of SR.

During soybean seed germination, the expansive growth potential of the embryonic axes is driven by water uptake while cell wall loosening occurs in cells from the elongation zone (EZ). Expansins are regarded as primary promoters of cell wall remodelling in all plant expansion processes, with the expression profiles of the soybean expansins supporting their cell or tissue specificity. Therefore, we used embryonic axes isolated from whole seed and focused on the EZ to study seed germination. Using a suite of degenerate primers, we amplified an abundantly expressed expansin gene at the EZ during soybean embryonic axis germination, which was identified as EXP1 by in silico analyses. Expression studies showed that EXP1 was induced under germination conditions in distilled water and down-regulated by abscisic acid (ABA), which inhibits soybean germination by physiologically restraining expansion. Moreover, we also identified a time window of ABA responsiveness within the first 6 h of incubation in water, after which ABA lost control of both EXP1 expression and embryonic axis germination, thus confirming the early role of EXP1 in the EZ during this process. By contrast, EXP1 levels in the EZ increased even when germination was impaired by osmotically limiting the water availability required to develop the embryonic axes’ growth potential. We propose that these higher EXP1 levels are involved in the fast germination of soybean embryonic axes as soon as water availability is re-established. Taken together, our results show strong EXP1 expression in the EZ and postulate EXP1 as a target candidate for soybean seed germination control.

The micromorphology of a seed is linked to its dispersal and germination, but evolutionary and ecological aspects in Orchidaceae remain unclear. We investigated the seed characters of Paphiopedilum and Cypripedium that might be associated with life form and involved in possible ecological adaptations. A phylogenetic comparative analysis of nine seed micromorphological characters was performed in 24 species from two genera with close phylogenetic relationships but significant differences in their ecological characteristics. Species within Paphiopedilum had larger embryos and a smaller percentage of air space (AS) than those of Cypripedium species. Compared with 16 terrestrial species, two epiphytic Paphiopedilum species had larger embryos and smaller AS. Those larger embryos might ensure more successful seedling establishment while the higher amount of air space in both terrestrial Paphiopedilum and Cypripedium may increase seed buoyancy and enable them to disperse over longer distances. Whereas AS and seed length (SL) are phylogenetically conservative, most other characters examined here had weak signals, indicating clear convergent evolution. Across species, SL was positively correlated with AS, indicating a high degree of seed size–dispersal coordination. These findings may imply a trade-off of seed characters in relation to the possible ecological adaptations required for seedling establishment versus dispersal.