Specific chitinases accumulate in seeds of many species as part of their normal developmental programme. Some chitinases can also be induced in developing and germinating seeds in response to microbial attack. All known seed isoforms belong to classes I, II, IV and VII, which are encoded by Chia genes, as well as to the more divergent class III encoded by Chib genes. The study of seed-specific chitinases has contributed significantly to current knowledge of this ubiquitous protein family, including antifungal properties, structure, specificity and catalytic mechanism. Indeed, the first plant chitinase for which a three-dimensional structure was solved had been isolated from barley seeds. Moreover, the finding that a chitinase could rescue a somatic embryo mutant was the first evidence of a non-defensive function. Several lines of evidence have substantiated the biotechnological potential of chitinases to counter plant fungal disease. The recent identification of several seed and fruit chitinases as major panallergens should be taken into account when selecting the genes to be introduced into food crops.

Senna multijuga and Plathymenia reticulata are tropical tree species native to the Brazilian Atlantic Forest and the Brazilian Cerrado, respectively. Seed-coat dormancy variation was evaluated within and among natural populations of these two species. Scarified and non-scarified seeds from different plants within populations were germinated at 28°C, and the percentage of germinated seeds was estimated for both species. Mean germination percentages of non-scarified seeds tended to be higher for P. reticulata populations (40 and 62%) than for S. multijuga populations (9 and 35%). After scarification, germination percentages increased significantly in both species, with all populations showing mean values above 84%. The level of seed dormancy, evaluated through the experiment with non-scarified seeds, differed significantly within and among populations of both species (P < 0.05). The values of the coefficient of genotypic determination were high for populations of both species (b = 0.85). Although this coefficient is an overestimation, since it includes non-genetic maternal effects, its high values suggest that a considerable part of the phenotypic variation in seed dormancy in S. multijuga and P. reticulata is of genetic origin. Variation in seed dormancy can be an important factor for increasing genetic diversity in populations of these species, making them able to respond to environmental changes.

The stimulatory role of 3-methyl-2H-furo[2,3-c]pyran-2-one, a smoke-derived butenolide, on germination and post-germination events is well documented. Previous studies have involved germinating seeds in the continuous presence of the compound. However, commercial growers cannot exploit the potential of the butenolide for large-scale production of crops due to limited availability and environmental constraints. The present study was undertaken to assess the potential of the butenolide as a priming agent of tomato (Solanum esculentum Mill.) seeds. Flow cytometry data revealed that butenolide-primed seeds had a higher percentage of nuclei at the 4C stage than water-primed seeds. Emergence of the radicle was much faster in the primed seeds. After 36 h of imbibition, a higher percentage of the butenolide-primed seeds (22%) exhibited radicle emergence compared to the water-primed seeds (12%). While butenolide-primed seeds initially germinated more rapidly than either water-primed or unprimed seeds in a 48-h period, water-imbibed seeds reached a similar germination level as the butenolide-primed seeds by 60 h of incubation. The butenolide-primed seeds produced significantly (P ≤ 0.05) more vigorous seedlings than water-primed seeds or seeds in the continuous presence of either water or butenolide. A gradual decrease in the seedling vigour index was recorded for both water and butenolide-primed seeds with increased seed storage at room temperature. Nevertheless, the vigour index was significantly greater in the butenolide-primed seeds than the water-primed seeds. Vigour indices were significantly (P ≤ 0.05) higher for the butenolide-primed seeds under various stress conditions (salinity, osmoticum or temperature) compared to control or water-primed seeds. Results of the present study suggest that the butenolide is a good seed-priming agent. Additionally, primed seeds retained the promotive effect for a considerable time. This was also the case for tomato seeds under various simulated field stress conditions.

Most Pinus species are obligate seeders. Thus, knowledge of germination characteristics can help in the understanding, prediction and manipulation of the regeneration and dynamics of pine forests. Seven pine species with contrasting habitat preferences and different genetic pairwise distances are present in the Iberian Peninsula and the Canary Islands: P. halepensis, P. nigra, P. pinaster, P. pinea, P. sylvestris, P. uncinata and P. canariensis. These seven pine species comprise an exceptional experimental set to test some questions related to germination traits, such as: (1) What are the effects of light and temperature on germination, taking into account interpopulation variability? (2) Is there any association of germination traits with habitat (montane versus lowland) preferences? and (3) What is the relationship between germination traits and the genetic distance between pine species? P. nigra, P. sylvestris and P. uncinata seeds showed faster germination rates. Seeds of P. nigra and P. sylvestris reached high total germination percentages in every temperature and light treatment, suggesting an opportunistic germination strategy. Unlike montane pines, lowland pines did show significant effects of temperature on germination response: final germination was higher between 15°C and 20°C than at warmer and alternating temperatures. Relatively low temperatures associated with the winter rainy season would favour germination of most of these species. Nested models showed that population variability was the main source of variation in germination response. Thus, there is no phylogenetic control of the germination response and, surprisingly, germination traits were not related to habitat preferences. As a consequence, we believe that studies of the germination characteristics of a pine species should consider different populations.

Seed dormancy was analysed in Geraniaceae, testing 29,103 seeds of 35 species for seed-coat impermeability to water. Seeds of wild and cultivated species from a number of different climatic/ecological zones (northern Germany, the Canary Islands and South Africa) were investigated. Seeds from cultivated plants and natural populations had similar (±2%) percentages of water-impermeable seeds. The percentages of impermeable seeds at maturity were also mostly constant (±2%) for two different years. The species analysed contained from 0 to 100% impermeable seeds at maturity, and could be classified into three quantitative types of physical dormancy: four species with no impermeable seeds (PY0), 10 species with a maximum of 80% impermeable seeds (PY80), and 21 species with more than 80% impermeable seeds (PY100). Species with different life cycles and from very different habitats, e.g. Geranium pratense or Pelargonium capitatum, belonged to the PY100 group. However, all of the succulent shrubs investigated from the genus Pelargonium were PY0 species. Five years of dry storage at 20 ± 5°C decreased physical dormancy in a species-dependent fashion. Erodium cicutarium, for example, had 95% impermeable seeds at maturity; all of them became permeable during the dry storage and began to germinate immediately after placing them on a moist substrate. Physical dormancy of other species, e.g. Pelargonium zonale, Pelargonium vitifolium or Geranium pratense, was not affected by dry storage. Thus, physical dormancy is a diversely differentiated feature in Geraniaceae with regard to both percentages of impermeable seeds at maturity and maintenance of dormancy under particular conditions.

Restoration of semi-natural grassland communities involves a combination of (1) sward disturbance to create a temporal window for establishment, and (2) target species introduction, the latter usually by seed sowing. With great regularity, particular species establish only poorly. More reliable establishment could improve outcome of restoration projects and increase cost-effectiveness. We investigated the abiotic germination niche of ten poorly establishing calcareous grassland species by simultaneously exploring the effects of moisture and light availability and temperature fluctuation on percentage germination and speed of germination. We also investigated the effects of three different pre-treatments used to enhance seed germination – cold-stratification, osmotic priming and priming in combination with gibberellic acid (GA3) – and how these affected abiotic germination niches. Species varied markedly in width of abiotic germination niche, ranging from Carex flacca with very strict abiotic requirements, to several species reliably germinating across the whole range of abiotic conditions. Our results suggest pronounced differences between species in gap requirements for establishment. Germination was improved in most species by at least one pre-treatment. Evidence for positive effects of adding GA3 to seed priming solutions was limited. In several species, pre-treated seeds germinated under a wider range of abiotic conditions than untreated seeds. Improved knowledge of species-specific germination niches and the effects of seed pre-treatments may help to improve species establishment by sowing, and to identify species for which sowing at a later stage of restoration or introduction as small plants may represent a more viable strategy.

The ability of orthodox seeds to survive long-term dry storage is a key prerequisite for ex situ seed conservation in genebanks. However, only a few credible observations of seed survival for ≥ 200 years have been reported. In this paper, seed survival is shown for three species under suboptimal storage conditions for a documented time of ≥ 203 years and carbon dated at 218–270 years. Two species that germinated are in the Fabaceae (Liparia sp. and Acacia sp.). A water-impermeable seed coat, and hence an inability to equilibrate with ambient relative humidity, may contribute to long-term survival of these species. The third species that germinated (Leucospermum sp.) does not have a water-impermeable seed coat, and long-term survival may be linked to an oxygen-impermeable barrier in the seed. These results for species from the Cape Floristic Region, South Africa, suggest adaptation for extreme longevity in seeds of species of seasonally dry, Mediterranean environments.

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.

Lettuce (Lactuca sativa L.) seeds are inhibited from germinating above an upper temperature limit that is dependent upon cultivar, growing conditions and seed treatments. Thermoinhibition is accompanied by an increasing sensitivity of germination to reduced water potentials (ψ). We have employed a water relations analysis (hydrotime model) of seed germination rates to investigate the basis of thermoinhibition. Germination rates can be characterized by the distribution of base water potentials among seeds in the population (ψb(g)) and a hydrotime constant indicating the total accumulated hydrotime (MPa · h) above the base required for radicle emergence. The hydrotime model adequately described germination time courses across a range of ψ at both high and low temperatures. Increasing temperature caused the ψb(g) distributions to become more positive, accounting for the greater sensitivity to ψ and inhibition of germination. Increases in embryo osmotic potential and in the turgor yield thresholds of both the radicle and the endosperm/pericarp envelope contributed to this change. Seed priming (prehydration and drying) treatments speeded germination by reducing the hydrotime requirement. Priming also resulted in smaller increases in ψb(g) at high temperature, alleviating thermoinhibition by lowering the embryo yield threshold sufficiently to compensate for the increased endosperm resistance. The beneficial effects of priming in lettuce appear to occur primarily in the embryo, rather than in the surrounding envelope tissues.

Scaling of water absorption and water loss by seeds on various soil surfaces was simulated using seed analogues constructed with paper pulp. Three sizes of analogue seeds (large, medium and small) were laid on three types of soil surface (coarse, medium and fine texture). To estimate the amount of water absorbed by a seed during a fixed time interval, the difference in seed weight from the start of the experiment was used. The scaling of water absorption necessary for germination was also studied using actual seeds of 14 species representing a range of seed sizes. Scaling coefficients between the amount of absorbed water by an analogue seed (net water gain) and seed mass were usually lower than 1: small seeds absorbed water more rapidly than large ones. The water loss of analogue seeds was also correlated with seed mass with a scaling coefficient lower than 1, but the amount of water loss itself was far smaller than the absorption. On the other hand, the germination of actual seeds revealed that the amount of water necessary to start germination was proportional to seed mass. Thus, smaller seeds have an advantage over larger seeds in more rapidly attaining the water content necessary for germination. Moreover, small seeds can penetrate through small cracks in the soil surface and thus enjoy a double advantage in a microsite that promotes water absorption and minimizes desiccation.

This study analysed the anatomical structure of the seed coats, identified the location of water uptake and evaluated the effects of alternating temperatures and heat treatment on the breaking of physical dormancy of two species of Fabaceae (Caesalpinioideae), Cassia leptophylla and Senna macranthera, from the Atlantic forest of Brazil. The seed coats of both species consisted of a cuticle (extra-hilar region) or remaining funicle region (hilar region), subcuticular layer, palisade layer with lignin, osteosclereids, sclerified parenchyma and white cells. The palisade layer was formed by elongated macrosclereids with a light line of callose. In thermally scarified seeds of C. leptophylla, water entered through the micropylar canal, and in S. macranthera the water entered through the lens. Alternating temperatures that ranged from 15 to 30°C did not break physical dormancy of either species; however, exposure to 50°C broke seed hardcoatedness, allowing the entrance of water in both species.

To obtain information on possible sources of contamination of the seed harvest of oilseed rape (Brassica napus L., spp. napus) by other varieties (adventitious presence), we investigated the purity of certified seed lots; the abundance and origin of volunteers; and longevity and origin of seeds in the soil seed-bank. This information was acquired through DNA analysis of volunteers collected in the field and seedlings derived from the soil seed-bank. DNA profiles of the volunteers and seedlings were obtained using Inter Simple Sequence Repeat (ISSR) markers, and the profiles were compared with ISSR profiles from an assortment of 14 of the most commonly cultivated oilseed rape varieties from 1985 to 2004. This comparison was performed using the assignment program, AFLPOP. The age of the seed bank germinating to become volunteers was assumed from information on previously cultivated oilseed rape varieties at the site and the year of cultivation. The results showed that plants or seeds belonged to varieties cultivated at the site 4–17 years earlier, suggesting a long persistence of volunteer populations. High volunteer frequencies (6%, 29% and 32%) were found in the three oilseed rape fields analysed and, from soil cores, their soil seed-bank was estimated to be 50–100 seeds m− 2. The ISSR analysis of the 14 reference varieties showed that three of the certified seed lots contained other varieties above the allowed threshold. Our findings are discussed in the context of the coexistence between GM (genetically modified) and non-GM oilseed rape, and future levels of seed purity.

Physical dormancy refers to seeds that are water impermeable. Within the Fabaceae, the structure associated with the breaking of dormancy is usually the lens. This study verified the role of the lens in physical dormancy of seeds of Schizolobium parahyba, a gap species of Fabaceae from the Atlantic Forest of Brazil. The lens in S. parahyba seeds appeared as a subtle depression near the hilum and opposite the micropyle. After treatment of the seeds with hot water, the lens detached from the coat. Blocking water from contacting the lens inhibited water absorption in hot-water-treated seeds. High constant (30°C) and alternating (20/30°C) temperatures promoted the breaking of physical dormancy and germination in non-scarified seeds. Maximum percentage of germination occurred earlier for seeds incubated at 20/30°C than for those incubated at 30°C. Seeds with a blocked lens did not germinate at alternating or high temperatures. This study suggests that alternating temperatures are probably the cause of physical dormancy break of seeds of S. parahyba in gaps in the forest.

Recalcitrant seeds are shed moist from the plant and do not survive desiccation to the low moisture contents required for prolonged storage. It has been widely hypothesised that during desiccation of these seeds a stress induced metabolic imbalance develops that leads to free radical mediated damage and viability loss. We investigated this hypothesis in a comparison of two sympatric species of Acer during late seed development and post-harvest desiccation: A. platanoides (Norway maple) has orthodox seeds and A. pseudoplatanus (sycamore) has recalcitrant seeds. In both species, respiration rates declined to similar levels at shedding, and the extent of defences against free radicals appears no less in sycamore than that in Norway maple. During drying there was no evidence for the accumulation of a stable free radical, increased lipid peroxidation or decline in free radical scavenging enzymes in either species. In addition, there was a very similar, large increase in total tocopherol in both species. This increase in sycamore was largely of alpha-tocopherol, whereas in Norway maple the increase was largely from its precursor, gamma-tocopherol. Arguably this suggests a similar mechanism in both species, but increased oxidative stress in sycamore. In general, the results suggest that, although damage resulting in viability loss was clearly taking place, the limitation to desiccation tolerance did not result from inadequate free radical scavenging. Soluble carbohydrates and dehydrin-like proteins were also measured during late seed development and drying in sycamore and Norway maple. The greater concentrations of sucrose, raffinose and stachyose and amounts of dehydrins in the radicles and cotyledons of Norway maple compared with those in sycamore was consistent with greater desiccation tolerance in the former. Sycamore seeds are dormant and at the tolerant end of the continuum of desiccation sensitivity among recalcitrant species, and this may account for their different response to that of the seeds of other more sensitive recalcitrant species studied.

The five versions of the M.G. Nikolaeva seed dormancy terminology and formulae system are correlated to each other and to the dormancy terminology proposed by Baskin and Baskin [Seed Science Research14, 1–16 (2004)]. The proposal is made that the dormancy formula scheme of Nikolaeva [Botanicheskii Zhurnal86, 1–14 (2001)] be adopted as part of an international seed dormancy classification system, and her logic for derivation of the new formulae for physiological and morphophysiological dormancy is discussed via examples. We conclude that there are several advantages of adopting formulae into a seed dormancy classification system. However, Nikolaeva's scheme, especially the part for morphophysiological dormancy, probably needs a small amount of further refinement.

Mean seed dry mass values were determined for 27 species of trees and shrubs in Amazonian caatinga (a forest-type especially short of nitrogen) and for 11 species in adjacent much taller forest on less poor soil. The tall trees (> 15 m) of caatinga have smaller seeds than the tall trees in adjacent forest on less infertile soil (both overall and in six taxonomically controlled comparisons), and than the tall trees in lowland rainforests elsewhere. The smaller seed size is interpreted in terms of a major advantage of keeping up seed number outweighing the marginal advantages of larger seed size. For trees of caatinga and adjacent forest considered together, there is a significantly greater concentration of P and Mg, and almost significantly greater concentration of N, in the embryo-cum-endosperm fraction of smaller-seeded species, but the content per seed of N, P and Mg is smaller in smaller seeds. The mean contribution of the seed coat (including endocarp for pyrenes) was 17% for dry mass, 3% for content of P, 10% for N and Mg, 15% for K, and 30% for Ca.

Seeds vary widely in the sensitivity of germination to oxygen (O2) partial pressure, depending upon the species, temperature, dormancy state and physiological status of the seeds. Most analyses of the O2 sensitivity of germination have focused on final germination percentages and estimated the O2 percentage in air that is required to reduce germination to a given percentage (usually 50%). In contrast, we have applied a population-based threshold model utilizing time courses of germination to quantify three parameters related to seed germination sensitivity to O2 availability: the median base (or threshold) O2 percentage, the standard deviation of O2 thresholds among seeds in the population, and an oxygen–time constant that relates O2 percentage to germination timing. The model fits germination responses accurately across a wide range of O2 concentrations. The response to O2 was logarithmic in all cases, with the O2 percentage required for 50% germination ranging from 21% to as low as 0.005%, depending upon the species, the temperature and the seed dormancy level. Modelling indicated that some seeds can adapt to low O2 percentages and shift their thresholds to lower values over time. Lower temperatures decreased the minimum O2 threshold, as did after-ripening. Seed priming generally reduced the oxygen–time constant and increased the standard deviation of germination responses, but had relatively little effect on the O2 sensitivity per se. The population-based threshold model can be used to quantify the O2 sensitivity of seed germination and to predict germination rates and percentages when O2 availability is limiting.

When the laboratory methods employed are qualitative, the statistical methodologies used in testing for the adventitious presence (AP) of transgenic material in conventional seed and grain lots are well defined. However, when the response from the method used by the laboratory is quantitative (e.g. percent transgenic DNA), the statistical methodologies developed for qualitative laboratory methods are not fully appropriate. In this paper, we present the details of procedures specific to quantitative laboratory methods. In particular we consider: (1) the assessment of quantitative laboratory method errors using linear modelling; and (2) the process of deciding whether or not a lot meets pre-specified purity standards, including the development of probability calculations needed to develop operating characteristic curves and estimate consumer and producer risks for a given lower quality limit (LQL), acceptable quality limit (AQL) and testing plan. We also describe implementation of this approach in a useful spreadsheet application.

A survey of Pisum genotypes for seed trypsin inhibitors revealed a tenfold range in the extent of inhibition. Approximately 90% of trypsin inhibitory activity was associated with two albumin fractions in selected variant lines. The differences among extreme variants were consistent in three environments, between two sources of trypsin tested and whether expressed on a unit protein or dry weight basis.

A study of the appearance of trypsin inhibitors during seed development in selected highand low-inhibitor lines showed differences in the accumulation pattern of active inhibitors. An endogenous protease was identified in Pisum seed protein preparations, whose in vitro trypsin-like activity was predominant in protein from early stages of seed development, when little or no trypsin inhibitor was present. However, there was no correlation between the amount of this protease and the extent of trypsin inhibitory activity in lines that varied for inhibitor content.

A method, electrical impedance spectroscopy (EIS), is introduced to study seed viability non-destructively. Snap bean (Phaseolus vulgaris L.) seeds were studied by EIS to determine the most sensitive EIS parameter(s) and the optimal range of moisture content (MC) for separation of viable and non-viable seeds. Hydrated seeds exhibited two impedance arcs in the complex plane at the frequency range from 60 Hz to 8 MHz, and impedance spectra of viable and non-viable seeds differed. The hydrated seeds were best-modelled by an equivalent electrical circuit with two distributed circuit elements in series with a resistor (Voigt model). Moisture content and seed viability had strong effects on the EIS parameters. The most sensitive EIS parameters for detecting the differences between viable and non-viable seeds were the capacitance log(C2), the resistance R2, the resistance ratio R2/R1 and the apex ratio, which all represent specific features of the impedance spectrum. The highest differentiation in the EIS parameters between the viable and non-viable seeds occurred in partially imbibed seeds between MC of 40 and 45% (fresh weight basis).