Solute leakage is used as an indicator of membrane damage during desiccation of recalcitrant seeds. We re-examined this phenomenon and its utility by comparing recalcitrant Spartina alterniflora and orthodox Spartina pectinata seeds, as well as drying isolated embryos or embryos isolated after whole-seed drying. During drying, intact seed leakage (electrical conductivity and absorbance at 280 nm) was independent of seed moisture content, dormant/non-dormant state or desiccation tolerance. Embryos from both Spartina species, isolated after drying within the intact seed, exhibited increased leakage, especially below 40% (dry weight basis, DWB) seed moisture, the critical water content for viability loss in Spartina alterniflora. When isolated embryos were dried, the pattern and extent of increased solute leakage were similar in both Spartina species, even though S. pectinata embryos were >95% viable and S. alterniflora embryos were < 20% viable. We conclude that increased solute leakage is an artefact of embryo excision and not an accurate indicator of desiccation damage to recalcitrant Spartina alterniflora seeds.

Glebionis coronaria (L.) Cass. ex Spach is a common Mediterranean weed producing distinctive central and peripheral dormant cypselae with a hard fruit coat, which was previously hypothesized to impose physical dormancy. Analysis of water uptake in cypselae and in naked seeds showed that it preferentially takes place at the basal end of the fruit; however, seeds within an intact pericarp do not fully imbibe when compared with naked seeds. Germination was not significantly different between the two heteromorphs, and afterripening or cold stratification did not increase germination, while warm stratification at 35/20°C, as revealed by logistic regression, resulted in a significant improvement. However, loss of viability was also rapid at these high temperatures. Central and peripheral cypselae generally showed very low germination. In both heteromorphs, faster and higher germination (60–70%) was reached only after extensive scarification of pericarp tissue, and full germination was observed only after complete removal of pericarp tissue. Although the pericarp significantly reduced water uptake, no palisade layer(s) of macrosclereids could be observed. Xylem-vessel elements were found running through the basal end of the pericarp and forming the main point of water entry. We reject the hypothesis that G. coronaria cypselae have physical dormancy. Instead, water uptake and germination are impeded by: (1) directed water uptake, mainly through a pericarp-spanning channel-like structure; and (2) mechanical constraint on embryo growth exerted by the hard pericarp. The channel-like structure forms the principal system for controlling seed germination.

Two possible sources of resistance to pre-harvest sprouting were evaluated in quinoa. They showed dormancy at harvest and significant variations in dormancy level in response to environmental conditions experienced during seed development. The aims of this work were to evaluate the importance of seed coats in the regulation of dormancy in this species, to investigate possible mechanisms of action and to assess association of seed coat properties with changes in dormancy level caused by the environment. Accessions Chadmo and 2-Want were grown under field conditions on different sowing dates during 2 years. Seed coats were manipulated and seed germination was evaluated at different temperatures. Seed coat perforation before incubation led to faster dormancy loss in both accessions. This effect decreased with delayed sowing date, and seeds expressed a level of dormancy not imposed by coats. This suggests the presence of embryo dormancy in the genus Chenopodium. Seeds of the accession 2-Want had a significantly thinner seed coat at later sowing dates, associated with a decreasing coat-imposed dormancy, but this pattern was not detected in Chadmo. The seed coat acts as a barrier to the release of endogenous abscisic acid (ABA) in quinoa, suggested by the increase in germination and a higher amount of ABA leached from perforated seeds. ABA is able to leach from seeds with an intact seed coat, suggesting that differences in seed coat thickness may allow the leakage of different amounts of ABA. This mechanism may contribute to the observed differences in dormancy level, either between sowing dates or between accessions.

The dispersal unit of many species of Brassicaceae is an indehiscent fruit, but relatively few studies have tested the effect of the pericarp on seed germination in this family. Our aim was to determine the effect of the pericarp on seed dormancy in six species of Brassicaceae native to the cold desert of north-west China. Intact dispersal units and isolated seeds of Chorispora sibirica, Euclidium syriacum, Goldbachia laevigata, Spirorrhynchus sabulosus, Sterigmostemum fuhaiense and Tauscheria lasiocarpa were stored dry at ambient laboratory conditions for 0–12 months and tested for germination in light and in dark at 5/2, 15/2 and 30/15°C. The amount of water absorbed by fruits and by seeds within the fruits was determined. For four species, intact fruits, isolated seeds and isolated seeds plus the removed pericarps were used to test for the mechanical versus possible chemical role of the pericarp in seed dormancy. Fresh isolated seeds, which have a fully developed embryo, germinated to lower percentages and rates than afterripened seeds. Thus, seeds have non-deep physiological dormancy. The pericarp significantly reduced germination, but inhibition was not due to lack of water uptake by seeds or to chemical inhibitors. Afterripened seeds of the six species germinated to 0–50% inside the fruits. We conclude that the pericarp plays a dominant role in seed dormancy of the six study species, and it does so by mechanically restricting embryo growth. Thus, the pericarp has the potential to spread germination over an extended period of time.

The genus Conostylis (Haemodoraceae) is endemic to fire-prone south-western Australia. To gain an understanding of the effect of some fire-related germination cues, eight Conostylis taxa were tested in response to water, nitrate, smoke water and karrikinolide (KAR1) under light and dark conditions, when seeds were freshly collected and after a year of burial. The germination of all taxa tested was higher in response to smoke water and KAR1 than in water alone, whereas nitrate did not stimulate germination. Germination was higher in all taxa following 1 year of burial than in fresh seeds. Recently, glyceronitrile has been identified as another chemical in smoke water, apart from KAR1, that can stimulate the germination of certain species. The relative response of eight Conostylis taxa to KAR1, glyceronitrile and smoke water was examined in laboratory-stored seeds. Germination of these taxa was promoted by both smoke water and KAR1, except for C. neocymosa, which had high germination regardless of treatment. Four of the other seven taxa germinated to higher levels in at least one of the glyceronitrile concentrations tested (10, 50 or 100 μM) than in water alone. However, in only two of these taxa, C. aculeata subsp. septentrionora and C. juncea, was germination in glyceronitrile as high as that in smoke water. Thus, the response to glyceronitrile is not uniform across Conostylis taxa. Generally, germination was higher with KAR1 than glyceronitrile, suggesting that although some Conostylis taxa have the capacity to respond to glyceronitrile, KAR1 is the more important germination stimulant for this genus.

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.

Mediterranean characteristics are attributed to the genus Crocus, which is inadequately studied in terms of seed germination. An afterripening requirement is very common in environments with warm and dry periods, and photoinhibition has been detected in many angiosperms inhabiting dry and open areas. The effects of afterripening and light on seed germination were investigated for the first time in 23 native Greek Crocus taxa, collected from various localities with either a Mediterranean or a temperate climate. Germination experiments were conducted in continuous darkness and in light at the optimal temperature for each taxon, with both freshly collected and afterripened seeds; warm stratification (20°C, darkness) was also examined in 22 taxa. A number of selected taxa were additionally investigated with respect to afterripening outdoors, afterripening and warm stratification at higher temperatures (35 and 25°C, respectively), stratification at 20/10°C, dry storage at low temperatures, response to gibberellic acid and phenology of embryo growth. It was postulated that an afterripening requirement is a characteristic of the genus Crocus, and we found that it can be fulfilled in nature during the Mediterranean dry summer. Also, for the vast majority of the taxa, warm stratification and stratification at 20/10°C can both meet the afterripening requirement. Embryos of the taxa studied are underdeveloped and have to grow prior to germination. Intrageneric differences of seed germination were observed only towards light, with photoinhibition being predominant in taxa from drier environments.

Chlorophyll fluorescence (CF) analysis was explored as a potential tool to identify the optimal time to harvest rice seed germplasm for maximum storage longevity. Seeds of 20 diverse genebank accessions were harvested at 24, 31, 38 and 45 d after peak flowering (DAF) and half of each seed lot was sorted by hand, following normal practice at the T.T. Chang Genetic Resources Center. CF analysis was carried out on both non-sorted and sorted seeds, while storage experiments were carried out on sorted seeds. Seed longevity (the time for viability to fall to 50%, p50) was significantly correlated with the skewness, kurtosis, mode and mean of the CF histograms when the data for every accession at all the harvest times were included in the correlation analysis. However, these correlation coefficients were ≤ 0.481. The correlation coefficient between p50 and DAF was similarly low (0.461). For individual accessions, there was wide variation in the correlation coefficients. While for some accessions, there appeared to be a strong relationship between p50 and mean CF that could be used to guide when to harvest seeds in the field, for other accessions, a unique mean CF to inform when to harvest seeds or for use in seed sorting could not be identified; this was also true for DAF over the harvesting schedule used in this study. Given the number and diversity of accessions managed by a genebank, it seems unlikely that CF analysis would be an appropriate tool to help manage the regeneration or processing of seeds intended for storage in the genebank.

Suaeda salsa is an annual herbaceous euhalophyte in the family Chenopodiaceae that produces dimorphic seeds on the same plant under natural conditions. In order to determine the effect of salinity on seed quality traits during seed formation, seeds from plants grown under control conditions and on 200 mM NaCl were used to investigate the effect of NaCl on seed production and seed germination. Results showed that size and weight of both black and brown seeds generated from 200 mM NaCl-treated plants were markedly greater than those from controls. The germination percentage of brown seeds from both control and NaCl-treated plants was higher than that of black seeds. Furthermore, the germination percentage of the black seeds generated from 200 mM NaCl-treated plants was significantly higher than that of the control at different concentrations of NaCl, although germination percentage declined with the increase NaCl concentration. Surprisingly, NaCl did not affect germination of the brown seeds. The germination index and vigour index of both black and brown seeds from the control plants were significantly lower than those of seeds from the different NaCl treatments. Seed starch, soluble sugar, protein and lipid content of both black and brown seeds generated from the 200 mM NaCl-treated plants were significantly higher than those from the control. These results suggest that a certain concentration of NaCl plays a pivotal role in seed vitality of the euhalophyte S. salsa through increasing seed weight and contents of storage compounds such as protein, starch and fatty acids.

Impermeability of the testa hinders efficient penetration of some small chemicals, such as transcriptional inhibitors, through the endosperm and the embryo during seed experiments. In Arabidopsis seeds, 5-bromo-4-chloro-3-indolyl β-d-glucuronic acid, a substrate for β-glucuronidase, did not permeate through the endosperm and embryo efficiently at the stages before testa rupture. The Arabidopsis testa also limited efficient entry of methoxyfenozide, a chemical ligand that was used for inducible gene expression experiments, into seeds. While the detection of a reporter gene at the early imbibitional stages could be replaced by reverse transcription-polymerase chain reaction (RT-PCR), the interference of entry of the chemical ligand into seeds by the testa was still problematic to gene induction experiments. To develop an efficient inducible expression system for gene function analysis in seeds, an inducible expression system with nitrate, which is a testa-permeable ligand, was examined. The vector containing the 2.1-kb upstream sequence of NITRITE REDUCTASE 1 was able to cause expression of a test gene (long non-coding RNA) in imbibed seeds at the stage before testa rupture in a nitrate-dependent manner. This system can be used not only for characterization of genes associated with seed dormancy and germination in basic research, but also for the development of germination recovery or enhancement technologies for agricultural applications.