Dehydrin and QP47, proteins present in mature pea seeds (Pisum sativum), have been proposed to play protective roles during desiccation. To identify possible relationships between these proteins and desiccation tolerance, their tissue locations and patterns of synthesis and degradation have been examined during germination. Tissue locations were determined by immunocytochemistry using polyclonal antibodies raised against a conserved dehydrin amino acid sequence and against purified QP47. In embryonic axis and cotyledon cells, QP47 and dehydrin were distributed uniformly with no apparent nuclear or organellar specificity. Both proteins were present in 24 h-imbibed axes that had not initiated radicle growth but were completely absent from 24 h-imbibed axes that had begun to grow. The amounts of QP47 and dehydrin in embryonic axes decreased with time after the start of imbibition and were undetectable by 48 h. When germination was prevented by polyethylene glycol (PEG) or abscisic acid (ABA), both proteins remained at their original amounts. Thus, both QP47 and dehydrin disappeared coincidently with the beginning of growth and not simply as a function of the time after imbibition. QP47 persisted in cotyledons until at least 31 days into seedling growth, whereas dehydrin was not detectable in cotyledons after 7 days. Dehydrin, but not QP47, could be re-induced in pea shoots and cotyledons by dehydration. The timing of degradation of both proteins was correlated with the loss of desiccation tolerance during germination of pea axes.

New techniques for the accurate and feasible detection of many seedborne pathogens have been developed over the last 25 years. These include conducive environmental incubation and identification (many seedborne fungi), liquid plating assay (seedborne bacteria), enzyme-linked immunosorbent assay (seedborne viruses) and polymerase chain reaction (PCR) technology. Regardless of the detection methodology, the specificity, sensitivity, reliability, efficiency of the assay and an understanding of pathogen tolerance in a seedlot need to be considered before a technique is an acceptable clinical seed health test. Acceptable seed health tests are tools for disease risk-management and routinely used in seed quality assessment. Seed health issues are increasingly important in international seed trade. With the advent of free trade, many countries are redefining their phytosanitary requirements with the goal of preventing introduction of a devastating pathogen into their country. However, many phytosanitary regulations are implemented without fully understanding the economic threat of a pathogen or a complete scientific analysis of a pathogen's presence in that country. This has led to confusing regulations, unnecessary tests or inspection requirements and unjustified trade barriers. To provide a means to answer scientifically the problems encountered in the worldwide movement of seed, an international movement has emerged to standardize seed health tests and inspection practices for international seed trade. The impact of standardization on the free trade of seed and the goals of the International Seed Health Initiative (ISHI) will be discussed.

Germination responses of nonstratified (conditionally dormant) and of cold-stratified (nondormant) seeds of three Solidago species were tested in darkness at 30/15°C following various lengths of exposure [0 (control), 5 and 30 min, 1, 6 and 14 h] to green (mostly 500–600 nm) or white (400–700 nm) light. Prior to exposures, nonstratified seeds were imbibed in darkness at laboratory conditions for 24 h, and cold-stratified seeds were kept in darkness at 5°C on a moist substrate for 8 wk. Nonstratified seeds of S. altissima, S. nemoralisand S. shortii germinated to 0–46%, 0–25% and 36–99%, respectively, in darkness following 5 min to 14 h of green or white light, whereas cold-stratified seeds germinated to 53–98%, 31–85% and 82–100%, respectively. Thus, sensitivity of seeds to both green and white light increased as they passed from conditional dormancy to nondormancy. Moreover, sensitivity to green and white light was similar to the hierarchy of seed size. Green safe lights must be used with caution in interpreting the results of studies on light/dark responses of seeds.

Regeneration from seed affects species assembly in plant communities, and temperature is the most important environmental factor controlling the germination process. Thermal dependence of seed germination is thus associated with species occurrence in an ecosystem. Hence, we aimed to investigate the role of temperature on seed germination of ten tree species from the western Brazilian Amazon. Seeds were collected in the state of Rondônia, Brazil, and set to germinate under constant temperatures ranging from 10 to 40°C in germination chambers. We calculated germination capacity (G%), germination rate (GR50, reciprocal of germination time), and thermal parameters, such as cardinal temperatures and thermal time requirements. Most species had a large range of temperatures showing G% ≥80%, with optimal temperature varying from 20 to 40°C. Base temperature ranged from 6 to 12°C and ceiling temperatures were mainly >40°C. Astronium lecointei and Parkia nitida showed high germination capacity under temperatures of 35–40°C, while germination of Theobroma cacao dropped from 100% to zero under temperatures between 37 and 40°C. The climax species Cedrela fissilis had the slowest germination time (10 days) and highest thermal time requirement, while seeds of Enterolobium schomburgkii (a late-successional species) germinated within the first day of the experiment. Rapid recruitment of Amazon species could be favoured with treefall disturbance, which increases temperatures in the understory, but sharp limits might be found in the supra-optimal range of temperatures. Such patterns might indicate different regeneration strategies in the tropical rainforest, providing important information regarding seed germination among Amazon species.

The longevity of neem, Azadirachta indica, seeds from African Sahelian (Burkina Faso) and Asian (Sri Lanka) provenances was studied over two years of storage under different conditions of moisture and temperature. After drying to equilibrium moisture content (MC) at different relative humidities at 20°C, seeds were placed in open storage at 20°C or hermetically sealed in packets at temperatures ranging from −20 to +20°C. There was hardly any difference in storage behaviour between seed batches / lots, whatever their provenance. Seeds originating from mature yellow fruits lived longer than seeds from green or brown fruits. In all storage experiments with seeds having MCs ≥ 10%, viability was preserved best at 10−15°C, indicating that neem seed is chilling (and freezing) sensitive. There was no survival longer than 2 years under these conditions. At MCs of 4–8%, seeds were considerably more tolerant of low temperature storage and had 40–60% viability after 2 years at all temperatures tested (−20 to +20°C). However, the seeds were sensitive to imbibitional stress, which could be alleviated by imbibition at temperatures of 25–30°C or above. The difficult storage behaviour of neem seed seems to stem from: (1) the sensitivity to low temperatures at MCs ≥ 10%; (2) the extreme sensitivity to imbibitional stress after storage at ≤ 8% MC; (3) underestimation of the water activity due to the high oil content of the neem seeds, causing unexpected metabolic stress in the higher MC and temperature range.

Seeds show important variation as plant regenerative units among species, but their evolutionary co-variations with other plant characteristics are still poorly understood. Whilst a positive association of seed mass with genome size (GS) and life forms has already been documented, a broad-scale quantification of their evolutionary correlation and adaptive selection has never been conducted. Here, we tested for correlated evolution of seed mass and GS towards distinct selective regimes related to life form in angiosperms. In particular, we tested the hypothesis that the selection toward lighter seeds and smaller genomes is stronger for annual plants, ensuring high regenerative potential. Using multivariate evolutionary models over a dataset containing 3242 species, we showed an overall positive correlated evolution of seed mass and GS deviating from a pure drift process. Instead, evolutionary changes in seed and genome sizes were driven by adaptive selection towards optimal values differing among life forms. Specifically, the evolutionary optima towards which the seed and genome sizes evolve show a covariation toward small values in annuals, intermediate values in perennial herbs and a trade-off between seed mass and GS in woody plants. Moreover, the evolutionary correlation between seed mass and GS is strongest in annuals as an adaption to complete their life cycle in a short time window, when environmental conditions are favourable for regeneration and development to maturity. The asymmetry in the correlated evolution acting on seed and genome sizes due to life form could explain how life-history traits interplay with functional traits and how plants have evolved diverse successful life-history strategies.

In the work reported in this paper, we attempted to elucidate the nature of the different abscisic acid (ABA) sensitivities presented by developing embryos from sorghum [Sorghum bicolor (L.) Moench] lines with contrasting pre-harvest sprouting (PHS) behaviour (Redland B2, susceptible; IS 9530, resistant). We explored two different hypotheses for a possible mechanism: (1) a different functionality of the ABA signalling pathway, and (2) a different rate of ABA degradation/conjugation in the apoplast of embryos from these genotypes. To assess the first possibility, we used an ABA-responsive gene (Rab17) as a reporter of changes in endogenous ABA content, which were artificially induced in embryos from both genotypes by means of fluridone application immediately after anthesis, to reduce ABA content, and embryo incubation in the presence of ABA. A defect in ABA signalling should be seen as a level of Rab17 expression that is independent of endogenous ABA content. For testing the second possibility at two stages of development, embryos from both lines were isolated and incubated in water for different periods. ABA concentrations in embryos and the incubation media were quantified through radioimmunoassay. In contrast to our findings for the resistant IS 9530 line, Rab17 expression did not respond to changes in ABA levels in sensitive Redland B2 embryos. The ABA degradation/conjugation rates in embryos and incubation media did not show clear differences between sorghum lines for any of the developmental stages analysed. These results suggest that a disruption in the ABA signal transduction pathway in Redland B2 underlies the low ABA sensitivity shown by embryos from this line.

The concept of ‘imbibitional damage’ arose when it was observed that considerable leakage of cell contents could occur when dry seed or pollen tissues are plunged directly into water. It is now common practice to imbibe dehydrated tissue slowly, to permit the re-establishment of functional membranes, prior to placing the tissue into liquid water. However, this argument may not hold if the tissue of interest is inherently desiccation-sensitive. Slow drying of desiccation-sensitive (recalcitrant) seeds or excised embryonic axes results in damage at high water contents, because it permits time for aqueous-based deleterious processes to occur. The same argument may apply if partially dried material is re-imbibed slowly, as this technique will also expose the tissue to intermediate water contents for protracted periods. This hypothesis was tested using embryos or axes from seeds of three recalcitrant species (Artocarpus heterophyllus, Podocarpus henkelii and Ekebergia capensis). Excised material was rapidly dried to water contents within the range over which viability is lost during drying, and re-imbibed either rapidly, by plunging directly into water, or slowly, by placing the material on damp filter paper or exposing it to a saturated atmosphere for several hours. Although details of the response differed among species and developmental stage, in all cases direct re-imbibition in water resulted in higher (or similar, but never lower) survival than either of the slow rehydration techniques.

Totalling more than 3000 species, parasitic angiosperms are a most diverse group of organisms. Classified as either shoot or root parasites according to site of attachment, plant parasites are further classified, according to the degree of parasitism, into one of two divisions: holoparasitic or hemiparasitic. Within these divisions, angiosperm parasites are either obligate or facultative.

Obligate parasites cannot survive to maturity without attaching to a host plant, thus the need to germinate in close proximity to a host first characterizes the specialized germination and seed dispersal strategies of these plants. However, seeds of all species of the rootholoparasitic Orobanchaceae and four genera within the Scrophulariaceae have evolved very specialized germination strategies. Once the seeds have undergone an after-ripening period, they require a prolonged imbibition period at temperatures of approx. 30°C before they acquire the potential to germinate. Minute seeds with very little reserves, they must germinate within millimetres of the host root for successful host attachment and continued survival. To this end, germination only occurs in responseto specific chemical germination triggers released from the host root. In Striga hermonthica (Del.) Benth., the perception of the host-derived trigger results in stimulation of endogenous ethylene production and it is this ethylene that initiates germination. It is proposed that the chemical triggers released from the host be consideredas ‘elicitors’ which function primarily as allelochemicals and that the parasite seeds have become adapted in such a way as to have an absolute requirement for external germination triggers.

The proteins produced just prior to maturation desiccation in the developing, orthodox seed, are stored in the desiccated state and recruited as the functional proteome upon imbibition. For the resumption of protein function, these stored proteins must be protected from permanent denaturation while dehydrating, throughout desiccation, and during rehydration. For some forms of damage there is the possibility of repair following imbibition potentially coordinated with de-aggregation into monodispersed polypeptides capable of refolding into a functional configuration. While studying aspects of the natural protection and repair mechanism in seeds, evidence has accrued that those proteins directly involved in translation are particular targets of both protection and protein repair. Such a phenomenon was first described by Rajjou et al. (2008) examining the frequency with which proteins involved in translation were identified as differentially abundant between aged and un-aged Arabidopsis seeds and the translational competence of aged versus un-aged seeds. The inference drawn from these observations was that, of all the stored proteins, it is imperative that those involved in translation endure desiccation, quiescence and rehydration in a functional state if the seed is to survive. Proteins involved in any other process other than translation can be replaced from the stored transcriptome or by de novo transcription but no mRNA is of value without the translational machinery. This has become known as ‘Job's rule’ in honour of the laboratory from which this hypothesis was first put forward (Rajjou et al., 2008). We review in this manuscript the evidence accrued to date on which Job's rule is based.

Seeds of sugar beet (Beta vulgaris L.) were collected at weekly intervals from 3 weeks before to 1 week after commercial harvest time, dried and stored at room temperature (18–22°C). Laboratory germination tests and flow cytometric analyses were performed immediately after harvest (fresh seeds) and five times at weekly intervals during storage (dry seeds). After 6 months of storage, seeds were exposed to a controlled deterioration treatment (CD). The proportion of G2 nuclei in the embryo was constant in the fresh seeds, regardless of their maturity. It decreased, however, after drying and CD, especially in those seeds harvested before maturation drying had commenced. The proportion of endosperm cells in the seed decreased with maturation, and a further decrease was observed after drying and CD. These observations suggest that nuclei with a higher nuclear DNA content were more sensitive to water stress caused by premature desiccation and to deterioration than nuclei with a lower DNA content. Fresh seeds exhibited some germination, but this increased after drying, suggesting that desiccation induced a switch from the developmental to the germination mode. Germination percentages were the highest in dry seeds collected at the commercial harvest time and a week after. This high germinability coincided with the highest proportion of G2 cells in the embryo. It is concluded that flow cytometry provides information about the status of sugarbeet seed maturation, seed quality and storage potential, and can be used for estimation of optimal harvest time.

The enhanced laboratory and field emergence characteristics of osmo- and halo-primed tomato seeds (cv. Pusa Ruby) were related to changes in hydration–dehydration kinetics, modified sorption properties and nuclear magnetic resonance (NMR) relaxation behaviour of humidified and imbibed seeds. Water sorption isotherms were constructed for primed and unprimed seeds by equilibrating to different water activities (aw) at 25°C. Analysis of the isotherms by the D'Arcy–Watt equation revealed that priming reduced the number of strong binding sites and the associated water content, and increased significantly the number of weak binding sites and the associated water content. This redistribution of water, which increased the availability of seed water, may be the reason for the higher speed of germination of primed seeds. The changes in transverse relaxation time (T2) of seed water and its components, measured in vivo using nuclear magnetic resonance spectroscopy, showed interesting differences between primed and unprimed seeds. With an increase in humidification time, the T2 of primed seeds could be resolved into three components with varying mobilities, while the control seeds had only two components until 10 d of humidification. During imbibition, the third component appeared after 2 and 6 h in primed and control seeds, respectively. This component disappeared after the germination process started in all treatments. The third fraction, with very low molecular mobility, which accounted for about 40% of the proton population, was assigned to hydration water of macromolecules. Hence, we propose that better performance of primed seeds may be attributed to the modifications of seed water-binding properties and reorganization of seed water during imbibition, so as to increase the macromolecular hydration water required for various metabolic activities related to the germination process.

Three seedlots of Agathis australis (Lambert) Steud. (kauri pine) were stored hermetically at moisture contents of 1.5–15.5% and at temperatures from −20° to 42°C; and one seedlot of Agathis macrophylla was stored at 7%, 9% or 13% moisture content and at 26°, 6° or −13°C. Seeds of A. australis showed orthodox storage behaviour; except that viability was progressively reduced with drying below about 5% moisture content, and, depending on seedlot, subsequent storage longevities were reduced at all temperatures. Seeds of A. macrophylla again showed essentially orthodox characteristics at 9 and 13% moisture content; but drying to 7% moisture content reduced viability significantly, and of those seeds surviving, a further proportion died as a result of transfer to −13°C, and the remainder survived for 2500 d with little further viability loss. Seeds of neither species conformed fully to orthodox characterisics, nor to those of the intermediate category. The results are discussed in relation to possible causes of this ‘sub-orthodox’ behaviour, especially current seed-harvesting methods, and the limits put upon ex situ conservation of these species in seed banks by their predicted comparatively short storage lives.

Wheat and barley seeds were subjected to accelerated aging conditions at 12% moisture content and 35°C temperature over 28 days of storage, and to heat-shock treatment carried out by 4 h of incubation at 40°C following 16 h of imbibition at 20°C. Heat-treated, aged seeds showed altered germination behaviour and increasing leakage electroconductivity as well as a reduced incorporation of [35S]-methionine into embryo proteins, in comparison with the corresponding untreated seeds. Two-dimensional electrophoresis of labelled proteins from embryos gave evidence of further quantitative and qualitative changes: (a) ‘normal’ germination protein synthesis in wheat and barley was slightly modified by age, but following heat-shock treatment a general reduction of most of the control polypeptides occurred; (b) heat-shock response resulted in the production of several HSPs with different MW and pl, but a uniform general decline in their synthesis was not observed. Highly labelled HSPs (e.g., those with MW 66.7 or 89.3–66.7 kDa in wheat or barley, respectively) did not change over the entire period of aging, while some smaller HSPs with MW >29 kDa either decreased in intensity or disappeared. Conversely, specific low molecular weight HSPs (MW 17–14.2 kDa) were synthesized more in extremely aged embryos of both species. The hypothesis that these polypeptides may be used as biochemical markers in monitoring both vigour loss and heat-shock induced tolerance in aged seeds is discussed.

A cleistogamous species consists of individuals that produce both chasmogamous (open, CH) and cleistogamous (permanently closed, CL) flowers, which facilitates a mixed-mating system. In contrast to what one might expect, CL (obligately selfed) seeds and the plants derived from them can be more fit than CH (potentially outcrossed) seeds and the plants they give rise to. Our aim was to review some theoretical aspects of mixed mating in relation to retention of both CH and CL in cleistogamous species and to determine if data on germination support the notion that CL is advantageous over that of CH. Based on germination (or seedling emergence) of CH vs CL seeds in 29 species in 21 genera and 11 families of monocots and eudicots, CL seeds germinated better in 107 and equally well as in 64 of 252 case studies as CH seeds (67.9%), and the (CH < CL):(CH > CL) ratio was 107/81 (1.32). We conclude that our study lends support to the notion that production of CL seeds by cleistogamous species is advantageous over that of CH seeds. Retention of CH by CL species may be due to the need to prevent complete selfing (s= 1.0) and thus total inbreeding depression (δ), which theory predicts would decrease reproductive success. Some caveats concerning the results of comparative studies on the germination biology of CH vs CL seeds of amphicarpic sensu stricto Fabaceae species and Commelina benghalensis and the aerial cleistogamous grass Danthonia spicata are discussed.

This study investigated the induction and loss of dormancy in oilseed rape (Brassica napus). Twenty genotypes were preliminary screened; from these, two genotypes, RGS003 and Hayola 308, which possess high potential for dormancy induction (HSD) and medium potential to induce secondary dormancy (MSD), were selected. The stratification of seeds at alternating temperatures of 5–30°C (in dark) significantly relieved secondary dormancy, but dormancy was not fully released. The ψb(50) values were −1.05 and −1.06 MPa for the MSD and the HSD before dormancy induction. After inducing dormancy, the ψb(50) values for the MSD and the HSD were increased to −0.59 and −0.01 on day 0 stratification at 20°C. The hydrothermal time (θHT) value was low for one-day stratification for HSD in comparison with other stratification treatments. Water stress can induce dormancy (if the seeds have the genetic potential for secondary dormancy) and warm stratification (in dark) can only reduce the intensity of dormancy. The seeds with a high potential of dormancy induction can overcome dormancy at alternating temperatures and in the presence of light. It can, therefore, be concluded that a portion of seeds can enter the cycle of dormancy ↔ non-dormancy. The secondary dormant seeds of B. napus cannot become non-dormant in darkness, but the level of dormancy may change from maximum (after water stress) to minimum (after warm stratification). It seems that the dormancy imposed by the conditions of deep burial (darkness in combination with water stress and more constant temperatures) might be more important to seed persistence than secondary dormancy induction and release. The dormancy cycle is an important pre-requisite in order to sense the depth of burial and the best time for seed germination.

Water availability regulates timing of germination, especially in those environments where it is a limiting factor. However, the water requirements for germination of most wild Mediterranean species are unknown. In this work we analysed the germination response to water stress of eight Cistaceae species with hard-coated seeds, which are typical for the Mediterranean shrublands. Seeds were exposed to a heat shock in order to overcome physical dormancy and then incubated under five water potentials (0, −0.2, −0.4, −0.6 and −0.8 MPa). Ungerminated seeds following these treatments were transferred to water in order to evaluate the recovery of germination. Additionally, at the end of the experiment, viability of still ungerminated seeds was examined. We analysed final germination percentage, time to reach 50% of the final germination (t 50), recovery of germination and seed viability in relation to water stress treatments. Furthermore, hydrotime analysis was performed; sensitivity to water stress, as characterized by Ψb(50), was checked in relation to seed size of species. Overall, water stress delayed and decreased germination but species showed different germination sensitivities, which were not related to seed size. Recovery of germination was high after disappearance of water stress but seed viability decreased in some species. Consequently, post-fire germination of Cistaceae must be strongly regulated by water availability and their seeds can recover germination capacity when sufficient water is available after long periods of incomplete hydration. However, some seeds die during the wait.

Evidence for effects of saprophytic fungi on buried seed demography is usually obtained from studies involving the simultaneous burial of fungicide-treated seeds and of untreated seeds. However, any potential influence of fungicide treatment on seed dormancy levels is generally ignored in these studies. Also, some studies assume that a combination of several fungicidal compounds provides better protection against a broader range of fungi, ignoring chemical interactions that may potentially occur between different compounds. To investigate these issues, we carried out a 6-month burial experiment using seeds of Anthriscus sylvestris (L.) Hoffm., Centaurea nigra L. and Daucus carota L., and three substrates differing in organic matter content. Three fungicidal compounds, captan, iprodione and mancozeb, were applied alone and in combination, including an untreated control. All fungicidal compounds and combinations thereof provided protection against fungal-induced seed mortality and, except for a low efficacy of iprodione in protecting seeds of Anthriscus, there were no pronounced differences in seed mortality between different fungicide treatments. Captan temporarily inhibited germination in Centaurea, whereas a similar inhibition in Daucus seeds caused by mancozeb was more long lasting, suggesting an induction of secondary dormancy. Organic matter content had only a negligible influence on these results. Our results suggest that the basic conclusions from most seed burial studies are robust with respect to their choice of fungicide. We conclude by discussing further implications of our findings for the design and interpretation of seed burial studies.

Seed salinity tolerance and dormancy play important roles in germination behaviour. The effects of NaCl concentration and different dormancy-breaking treatments on germination were determined for six species growing on the coasts of the Baltic Sea or the Gulf of Riga: Juncus balticus, Triglochin maritima, Triglochin palustre, Anthyllis maritima, Linaria vulgaris and Linaria loeselii. Germination percentages were significantly reduced at NaCl concentrations >100 mM. Germination of J. balticus, T. maritima and T. palustre, but not that of A. maritima and L. vulgaris, recovered after seeds were rinsed with distilled water. Seeds of J. balticus were non-dormant; those of A. maritima had physical dormancy; and those of T. palustre, T. maritima, L. loeselii and L. vulgaris had non-deep to intermediate physiological dormancy, which was broken by cold stratification or gibberellic acid. The seeds of plants growing in habitats periodically exposed to sea water (coastal meadows) were characterized by greater salinity tolerance than seeds of plants growing in habitats less exposed to sea water (dunes).

This is the first report on the SEed Ecological Database (SEED) being developed under the co-ordination of the author. In its first version the database will cover physical characteristics of seeds, such as: slenderness (length/width), flatness (thickness/width) and thousand-seed weight. The list of species encompasses all seed plants (gymnosperms and angiosperms) native or naturalized in Hungary (about 2450 species). Two examples to demonstrate the power of SEED are included. The significantly lower slenderness values of Hungarian C4 grasses (compared with C3 types), shown in the first example, is a new contribution to understanding the ecology of C4 grasses. In the second example heavier seed weights of shade plants are statistically demonstrated, compared to their heliophilous congeners with the same life forms. The 113 congeneric species pairs considered are derived from a wide range of plant families.