Sunflower (Helianthus annuus L.) seeds show more germination at high temperatures (25–30°C) than at temperatures below 20°C. Osmopriming with polyethylene glycol-6000 for 3–5 days at 15°C strongly increases germination at suboptimal temperatures. This stimulatory effect of priming persists after seed redrying and during subsequent storage at 20°C (55% RH) for at least 14 weeks. However, primed seeds deteriorate faster than untreated seeds during accelerated aging (45°C, 100% RH). The longer the priming treatment, the higher is the amount of germination but at the same time the higher is the sensitivity of seeds to accelerated aging. Priming enhances the respiratory activity of seeds transferred onto water and their ability to convert 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene. These effects remain after drying the seeds and are maintained in part during dry storage, whereas they disappear during accelerated aging. These results suggest that ACC-dependent ethylene production might be a good indicator of seed vigour; it increases with duration of priming and decreases very early during aging, well before significant loss of seed viability. Decrease in ACC conversion to ethylene indicates that aging is probably associated with membrane deterioration since in vivo ACC oxidase activity depends on membrane properties. However, no increase in electrolyte leakage is observed during aging.

Links between germination, DNA replication and β-tubulin accumulation were studied with cabbage (Brassica oleracea L.) seeds, by using flow cytometric analysis of nuclear DNA content and immunodetection of β-tubulin levels. The seeds were incubated in water or 0.1–500 mM hydroxyurea solutions. Radicle tips isolated from dry cabbage seeds revealed most 2C (Go or G1 stage) and some 4C (G2 stage) signals of nuclear DNA contents and a constitutive level of β-tubulin. The onset of DNA replication in the radicle tip was observed between 12 and 24 h of imbibition in water and was preceded by an increase of β-tubulin levels. Incubation of the seeds in 1 mM hydroxyurea retarded DNA replication, whereas an arrest of DNA replication occurred upon incubation in 10 mM hydroxyurea or higher concentrations. The arrest of DNA replication and cell division did not block radicle protrusion and allowed some extension of the radicle. This demonstrated that DNA replication is not a prerequisite for radicle protrusion and initial extension. However, further seedling development, including root growth and root hair development, was dependent on DNA replication. Accumulation of β-tubulin was not affected by hydroxyurea. Thus, it can be deduced that both DNA replication and β-tubulin accumulation are two parallel and independent cell cycle events during seed germination.

Controlled aging experiments were carried out in order to evaluate the changes in seed longevity occurring following the attainment of maximum seed dry weight in foxglove (Digitalis purpurea L.). Seeds harvested in three successive years were stored at ~5% moisture content (fresh weight basis) and 50°C. Seed longevity was greater the later the seeds were harvested. When the data were modelled using the predictive viability model of Ellis and Roberts, these increases were attributable to increases in both the theoretical initial viability (in probits) of the seed-lot, Ki, and in the standard deviation of the normal distribution of seed deaths in time, σ. Furthermore, there was a positive relationship between Ki and σ which was independent of the year of harvest (r = 0.9016, for 11 d.f., P < 0.001). These results do not support the predictive model which relies on the assumption that σ will not differ between seed lots of the same species stored under identical conditions. When seeds were stored at a range of moisture contents (between 5 and 10%) at 50°C, increases in σ during seed development were manifest as changes in the negative logarithmic relationship between σ and moisture content; below an upper limit, the regressions for seed lots at different stages of maturity were significantly different (P < 0.05) but could be constrained to either a common intercept or to a common slope (P < 0.05). It is proposed that it was the inherent variation in individual seed lifespans which increased during seed development. These results raise important concerns regarding the use of predetermined ‘species constants’ to predict the longevity of seed lots during long-term storage in seed banks.

Pretreatment of moist seeds at certain temperatures can reduce seed dormancy, but such observations represent the net effect of pretreatment. Seeds of Sitka spruce (Picea sitchensis [Bong.] Carr.) were raised to 30% moisture content and pretreated at five different temperatures between 5 and 30°C for up to 24 weeks (168 d). Subsequent ability to germinate at 10°C and viability were then determined in order to investigate the effects of pretreatment on seed dormancy and survival. There was a curvilinear, negative semi-logarithmic relationship between seed longevity and pretreatment temperature, such that Q10 for loss in viability increased from 2.6 between 10 and 20°C to 2.8 between 20 and 30°C. Simple multiplicative models combining cumulative normal frequency distributions for each of loss in viability and loss in dormancy were able to describe the changes in ability to germinate at 10°C, after pretreatment at 5, 10 and 30°C. However, in order to quantify the changes in ability to germinate observed at 10°C after pretreatment at 15°C, it was necessary also to invoke a model of dormancy reimposition, while for the results at 20°C it was necessary to postulate both dormancy reimposition and the further loss of this reimposed dormancy. It is concluded that moist seeds of Sitka spruce held at 15 and 20°C cycle between the dormant and non-dormant condition.

cDNAs for the major cysteine endopeptidase (CEpase) of mung bean (Vigna radiata [L.] Wilczek) seedling cotyledons have been cloned using gene-specific primers with the polymerase chain reaction (PCR) in a 3'-RACE system. A cDNA clone for CEpase, pKL042, that is 1221 bp long excluding the poly A tail was isolated. It appears to contain the entire coding sequence for a 362-residue-long polypeptide. The N-terminal sequence for the mature CEpase begins at position 128 of the putative translation product, suggesting removal of an N-terminal hydrophobic signal peptide and additional sequences to produce the mature protease. Northern blot hybridization with CEpase cDNA pKL042 as probe indicates that CEpase transcript is not detectable in the cotyledons or the embryonic axis of dry seeds, but is first detectable in the day 1 cotyledons and in the day 3 axis. The level of CEpase mRNA in both cotyledons and axis increases as growth proceeds. A decline of protease activity, however, is observed after day 3 in the cotyledons, even though the level of protease transcripts continues to increase until day 8. Detachment of the axis from the cotyledons before day 3 results in the prevention of the normal increase in both protease activity and CEpase mRNA.

Biotin is an important vitamin. It is biologically active as a protein prosthetic group, where it functions in enzymatically catalysed carboxylation reactions. It has previously been shown that the ability to synthesize biotin is not necessary for germination of Arabidopsis thaliana seeds, but that this process is required for early seedling growth. This research was conducted to determine if changes in the detection of biotinylated proteins could be observed that reflect changes in the need for biotin-mediated enzyme reactions observed during early soybean seedling growth. A seed specific 75-kDa biotinylated protein present in the embryonic axes and the cotyledons was lost during the first 3 d of germination. Seed specificity, and pattern of expression during germination suggest that this protein is a homologue of the seed specific 65-kDa biotinylated protein previously identified in pea (Pisum sativum). If samples were not treated with 2-mercaptoethanol, three equally spaced proteins at approx. 85 kDa were visible. In the presence of 2-ME these proteins appeared as a single 85-kDa band. This triplet was distinct only in the embryonic axes of dry seeds and not in imbibed seeds or in other plant parts. This demonstrates that imbibitional changes do occur in the pool of biotinylated proteins present in dry soybean seeds, and that 2-ME treatment can inhibit complete identification of the biotinylated proteins present in seed tissues.

The developmental pattern of biotinylated proteins (BP) during embryogenesis and maturation of soybean seed was characterized. Detection of these BP was compared with the development of desiccation tolerance in seeds. Three groups of BP were detected in soybean seeds using a biotin-streptavidin detection method: the first group consisted of a set of three bands with a mean apparent MW of 85 kDa (called BP85), detected in crude extracts of embryonic axes (EA) from non-dehydrated seeds (NDS) and from artificially slow-dehydrated seeds (DS); the second one, BP75, was a single protein with an apparent MW of 75 kDa and was expressed in cotyledons (COT) and EA tissues of NDS and DS; the third group with a mean apparent MW of 35 kDa (BP35), was expressed at high levels only in COT of NDS. BP35 concentration was highest in the early stages of seed development (21 days after flowering — DAF) and decreased as seeds developed, being almost imperceptible after 47 DAF. Conversely, only traces of BP75 and BP85 extracted from EA and COT were detected at early stages of seed development (21–33 DAF). Maximum levels of accumulation of these proteins were expressed at 42–47 DAF and remained constant until harvest maturity. Desiccation-tolerant stage of the seeds was initiated at 47 DAF, which coincided with the stage of maximum accumulation of BP75 and BP85 in the seeds, however, appearance of these proteins could be stimulated by desiccation of immature seeds that had not achieved desiccation tolerance. Therefore changes in biotinylated proteins are coincident with, but not sufficient for, the development of desiccation tolerance.

Protein synthesis in embryonic axes of Norway maple (Acer platanoides) was studied in seeds stratified at 3 or 15°C. At 3°C stratification, germination of seeds started between 6 and 7 weeks; at 15°C stratification germination did not occur. During imbibition of embryonic axes undergoing cold and warm stratification many new proteins became visible. Two of these proteins may be connected with the release from dormancy at 3°C. In seeds stratified at 15°C, these proteins occur in low amounts or are completely absent.

Osmotic priming in a polyethylene glycol solution (300 g/kg water) for 48 h resulted in a partial loss of desiccation tolerance for seeds of Vigna radiata (L.) Wilczek (mung bean). The percentage of germination began to decrease after primed seeds were dried down to water contents less than 0.06 g/g DW. As compared with control seeds, primed mung bean seeds also had poorer storage stability. The decline of storage stability after osmotic priming was correlated with the modifications of seed water sorption properties. Priming significantly increased the amount of water associated with the weak water-binding sites, and reduced the amount of water associated with the strong binding sites and multi-molecular binding sites in seed tissues. The enhancement of molecular mobility in seeds, as a result of such water redistribution, probably accelerates seed deterioration and decreases storage stability.

To colonize newly disturbed areas, weedy species must disperse their seeds in space and/or in time. Cirsium vulgare has poor dispersal in space, but can form a persistent seed bank. Fresh seeds are capable of germinating in either light/dark or constant dark conditions under favourable diurnal temperatures of 25°C: 10°C. Despite this lack of dormancy, a large proportion of seeds do not germinate in the autumn after dispersal. Variation in germination requirements according to seed colour was also examined. Fresh seeds were placed under one of two overwintering treatments at 5°C, alternating light/dark or constant darkness. Some seeds of all colours: white, light, intermediate, dark and black germinated under these conditions. Six months later, ungerminated seeds subjected to the light/dark treatment did not require light for germination when placed under optimal temperatures. However, most seeds that had overwintered in constant darkness required light for germination. In the field, this induced dormancy would prevent seeds from germinating if buried or located in deep shade. Such seeds have the potential of forming a persistent seed bank. The proportion of dormant seeds did not appear to be associated with seed colour. Seeds incubated at 10°C following overwintering under light/dark conditions germinated to a higher percentage than seeds overwintered in darkness, regardless of the light conditions for germination. The results of this study help to explain the contradiction between delayed germination in the field and the lack of seed dormancy.

Soil samples were collected from the original land surface beneath an experimental earthwork, constructed in 1963 at Morden Bog, Dorset, England, and excavated in 1996. Large numbers of seeds of Calluna vulgaris, Erica cinerea and E. tetralix germinated from the samples, but other species present in the original flora, including Ulex minor and several grasses, were absent. The results confirm the previously-documented longevity of Calluna and E. tetralix, and for the first time demonstrate similar longevity in E. cinerea.