In central Asia, Agriophyllum squarrosum is the first species to become established during natural succession on sand dunes. However, low germination percentages and thus poor stand establishment greatly inhibit the use of this key species in the stabilization of dunes. The aim of this review is to critically analyse published information on the seed biology of A. squarrosum with particular reference to identifying the factors limiting germination of seeds sown in the field. A conceptual model is used to illustrate the complexities of factors as well as the unknowns we found about the seed/seedling stage of the life cycle of this sand dune annual. A major result of this review is that we now know that high germination percentages can be obtained by storing freshly collected seeds dry at room temperatures for 2 to 3 months to allow dormancy break to occur via afterripening, and then storing them dry at low (e.g. 4–5°C) temperature to prevent them from entering secondary dormancy. Non-dormant seeds should be sown in the field in late spring when wind-blown sand will cover them, thus ensuring that they are in darkness, which promotes germination, at the time summer rains occur.

Induction of nine-cis-epoxycarotenoid dioxygenase 6 (NCED6), an abscisic acid (ABA) biosynthesis gene, alone is sufficient to suspend germination in testa-ruptured seeds, which are at the final step of germination. Molecular consequences of NCED6 induction in imbibed seeds were investigated by RNA sequencing. The analysis identified many unknown and uncharacterized genes that were up-regulated by NCED6 induction, in addition to the major regulators of ABA signalling. Interestingly, other NCEDs were up-regulated by NCED6 induction, suggesting that the major rate-limiting enzymes in the ABA biosynthesis pathway are subject to positive-feedback regulation. ZEAXANTHIN EPOXIDASE and ABSCISIC ALDEHYDE OXIDASE3, which function upstream and downstream of NCED, were also up-regulated in seeds by NCED6 induction, which suggests that the distinct layers of positive feedback loops are coordinately operating in the NCED6-induced seeds. SOMNUS (SOM), which was also up-regulated by NCED6 induction, was the major mediator of enhanced ABA signalling in NCED6-induced seeds. SOM exerted negative effects on GA biosynthesis, which also contributes to a high ABA–GA ratio and reinforces the suppressive state of germination. Besides these coding genes, long intergenic non-coding RNAs (lincRNAs) were also up-regulated upon NCED6 induction (termed N6LINCRs). Conditional expression of N6LINCR1 altered gene expression profiles in seeds. Twenty-six genes were up-regulated and 66 genes were down-regulated by the induction of N6LINCR1. These results suggest that some of N6LINCRs have a regulatory role in gene expression in seeds, which potentially contributes to the regulation of germination by ABA.

In fire-prone ecosystems, many species regenerate after fire from persistent soil seed banks. Species with physically dormant (PY) seeds have dormancy broken by fire-related heat. The magnitude of post-fire recruitment, to predict response to varying fire severity, is commonly estimated by testing dormancy-breaking temperature thresholds of fresh PY seeds. However, seeds spend years in the soil during the inter-fire period, and determining whether dormancy-breaking thresholds change over time is essential to accurately predict population persistence. Germination of four south-eastern Australian PY species from the Fabaceae family (Acacia linifolia, Aotus ericoides, Bossiaea heterophylla and Viminaria juncea) were studied. Dormancy-breaking temperature thresholds vary inter-specifically and the species represented either high or low dormancy-breaking threshold classes. Freshly collected seeds, and seeds that had been buried in the field or stored in dry laboratory conditions for 6 and 18 months were subjected to a fire-related range of heat treatments (40–100°C). Seed ageing increased germination response to heat treatments, effectively lowering the dormancy-breaking thresholds of three species. The fourth species, A. linifolia, initially had a relatively large non-dormant fraction which was lost as seeds aged, with older seeds then displaying PY broadly similar to the other study species. Patterns of threshold decay were species-specific, with the thresholds and viability of low-threshold species declining more rapidly than high-threshold species. The non-dormant fraction did not increase over time for any of our study species. Instead of increasing their non-dormant fraction, as is common in other vegetation types, these fire-prone PY species displayed a change of dormancy-breaking temperature thresholds. This is an important distinction, as maintaining dormancy during the inter-fire period is essential for population persistence. While changes in sensitivity to dormancy-breaking treatments have previously been reported as seeds age, our study provides the first test of changes to temperature thresholds, which increases the range of germination response from the seed bank under varying fire severity.

My purpose is to introduce a special issue of Seed Science Research devoted to papers resulting from material presented at Seed Ecology V held in Caeté, Brazil on 21–25 August 2016. An overview of the field of seed ecology is presented that includes a short summary of what I consider to be the eight basic subcategories of this field, and the five new areas of investigation that have developed as extensions and/or recombinations of basic areas. Seed ecology conferences allow researchers to communicate with each other and build new collaborative relationships. At Seed Ecology V, information was presented that related to each area of seed ecology. The nine papers in this special issue are a small sample of the information presented at the meeting, and each paper is briefly described and placed into one of the subcategories of seed ecology research.

Seed dispersal effectiveness (SDE) is a useful framework to explore the evolutionary and ecological consequences of seed dispersal to plant fitness. However, SDE is poorly studied in tropical open grasslands. Here, we studied both quantitative and qualitative components of SDE in two species of Miconia (Melastomataceae) from ferruginous campo rupestre, a vegetation highly threatened by mining activities. We determined fruit traits and fruit availability and found that fruits of both species are produced in times of resource scarcity at the study site. Based on the number of visits and the number of fruits removed per visit, we calculated the quantitative component of SDE for both species. Finally, we explored the qualitative component of SDE by means of a controlled experiment that simulated the effects of gut passage on seed germination. Bird species differed strongly in the quantitative component of SDE. Gut passage did not affect germination compared with hand-extracted seeds, except for a minor negative effect on germination time in M. pepericarpa. However, seeds within intact fruits showed lower germination percentages compared with hand-extracted seeds. Our data indicate that Miconia species from ferruginous campo rupestre are visited by a diverse assemblage of generalist birds that differ in quantitative, but not qualitative, seed dispersal effectiveness. We argue that planting Miconia species can overcome seed limitation in degraded areas and thus assist ecological restoration after mining abandonment.

Seeds of the winter annual Bromus tectorum lose primary dormancy in summer and are poised to germinate rapidly in the autumn. If rainfall is inadequate, seeds remain ungerminated and may enter secondary dormancy under winter conditions. We quantified conditions under which seeds enter secondary dormancy in the laboratory and field and also examined whether contrasting B. tectorum genotypes responded differently to dormancy induction cues. The study also extends previous hydrothermal time models for primary dormancy loss and germination timing in B. tectorum by using similar models to account for induction and loss of secondary dormancy. Maximum secondary dormancy was achieved in the laboratory after 4 weeks at –1.0 MPa and 5°C. Seeds in the field became increasingly dormant through exposure to temperatures and water potentials in this range, confirming laboratory results. They were released from dormancy through secondary after-ripening the following summer. Different genotypes showed contrasting responses to dormancy induction cues in both laboratory and field. To examine secondary dormancy induction and release in the field in terms of hydrothermal time parameters, we first created a model that allowed mean base water potential (Ψb(50)) to vary while holding other hydrothermal time parameters constant, as in models for primary dormancy loss under dry conditions. The second model allowed all three model parameters to vary through time, to account for changes (e.g. hydrothermal time accumulation) that could occur simultaneously with dormancy induction in imbibed seeds. Shifts in Ψb(50) could explain most changes in dormancy status for seeds retrieved from the field, except during the short period prior to dormancy induction, when hydrothermal time was accumulating. This study illustrates that hydrothermal modelling, and specifically changes in Ψb(50), can be used to characterize secondary dormancy induction and loss in B. tectorum.

Mammal species contribute in different ways to seed dispersal effectiveness, acting as endozoochorous dispersers, scatter-hoarders and seed predators. Seed removal by these functional mammal groups could be affected by anthropogenic drivers, such as grazing management, a common practice in drylands. We evaluated removal of seeds from a native tree species (Prosopis flexuosa) by terrestrial wildlife mammals with different functional roles, on grazed and ungrazed sites and at different times during the fruiting period of Prosopis. We offered Prosopis fruits, each containing 15 seeds, to animals and used camera traps to identify the species removing them. We obtained the number of seeds removed (1 fruit removed = 15 seeds removed) by each animal species and each functional group. Native and domestic mammals removed 65.4% of the total seeds offered; 69.5% of offered seeds were removed from the grazed area and 61% from the ungrazed site. Considering removal times, 64.25% of offered seeds were removed during the beginning of the fruiting period of Prosopis and 67% towards the end of this period. Small mammals acting either as seed predators (Graomys griseoflavus and Akodon dolores) or scatter-hoarders (Microcavia australis) were the functional mammal groups removing the highest amount of seeds. Seed predators removed more seeds from the ungrazed site, whereas the scatter-hoarder did so at the grazed site. In the ungrazed area, it would be important to ensure habitat heterogeneity in order to improve seed removal by functional groups that disperse seeds, such as endozoochorous dispersers and scatter-hoarders.

In the present study a combination of BLAST mediated homology search and 3′ RACE was utilized to isolate the full-length gene of Eleusine coracana alpha prolamin (Ec-α-prolamin) from finger millet. Phylogenetic analysis of Ec-α-prolamin along with related prolamin genes of different cereals and millets shows the clustering of Ec-α-prolamin in a separate group. Secondary structure prediction reveals 59.4% alpha helix structure, a structural hallmark of Ec-α-prolamin. Besides this, the protein also possesses a balanced proportion of all essential amino acids. Expression analysis based on qPCR shows increased accumulation of α-prolamin transcripts in developing grains of finger millet until attainment of seed maturity. Western blotting, using a monospecific anti-α-prolamin antibody, further confirmed the expression of a 22 kDa band in the S3 and S4 stages of developing spikes. The heterologous expression of isolated full-length Ec-α-prolamin could be potentially harnessed for making nutritionally enhanced functional food products and value-added industrial products.

Vitamin E (tocols) is a key metabolite for efficient scavenging of lipid peroxy radicals that cause membrane breakdown during seed ageing. However, in rice, this hypothesis has been tested for very few lines only and without considering intraspecific variation in genomic structure. Here, we present a correlation study between tocols and seed longevity using a diverse rice panel. Seeds of 20 rice accessions held in the International Rice Genebank at the International Rice Research Institute, representing Aus, Basmati/Sadri, Indica, temperate Japonica and tropical Japonica variety groups, were used for tocols analysis (quantification of α-, β-, γ- and δ-tocopherol/tocotrienol by ultra-performance liquid chromatography) and storage experiments at 45°C and 10.9% seed moisture content. To examine the effects of DNA-haplotype on the phenotype, the 700 K high-density single-nucleotide polymorphism marker dataset was utilized. Both seed longevity (time for viability to fall to 50%; p50) and tocols content varied across variety groups related to the heterogeneity in the genetic architecture. Among eight types of tocol homologues, α-tocopherol and γ-tocotrienol were significantly correlated with p50 (negatively and positively, respectively). While temperate Japonica varieties were most abundant in α-tocopherol, Indica varieties recorded 1.3- to 1.7-fold higher γ-tocotrienol than those of other groups. We conclude that the specific ratio of tocol homologues rather than total tocols content plays an important role in the seed longevity mechanism.

In Mediterranean fire-prone ecosystems, annual species specific to post-fire habitats should have a soil seed bank and should be able to germinate after a fire. Therefore, various fire-related cues can be expected to stimulate germination in post-fire annuals. Germination patterns of the rare annual Chaenorhinum rubrifolium (Plantaginaceae) were examined in response to mechanical scarification, heat shock, aqueous smoke, nitrogenous compounds, gibberellic acid, karrikinolide (KAR1), and mandelonitrile (a cyanohydrin analogue, MAN) under dark and photoperiod conditions in the laboratory. Combinations of these treatments were also included in the experiment. Strong physiological dormancy in the seeds of C. rubrifolium was partially broken by several fire-related germination cues, including smoke and nitrate, under light conditions. KAR1 and MAN also stimulated germination, and the highest improvement in germination was achieved in the KAR1 treatment in the presence of light. Heat shock + smoke and KAR1 + MAN combinations had positive synergetic and additive effects on germination under light conditions, respectively. The light played a crucial role in the promotion of germination. The results suggest that multiple fire-related cues operate to stimulate germination in C. rubrifolium, an annual species from the Mediterranean Basin. However, the species may have a broader germination niche than a fire-restricted one.

Adaptive plastic responses in invasive species allow for establishment and persistence despite the lack of genetic matching to new environments. The capacity of annual species to invade habitats to which they are not adapted is likely to be predicated on post-invasion seed trait variation correlated with conditions in the new habitat. To test this, I compared variation in seed traits and germination patterns of Aegilops triuncialis, an invasive annual grass, from 69 sample populations from 24 sites in California (USA) across 13 years. Seed mass, germination fractions, seed viability, and strength of induced dormancy between dimorphic seed pairs were used to investigate plastic variation within and among populations, across two general soil types, and among two widespread maternal genotypes. I found that seed mass variation was constrained although both seed types showed a 3-fold range among populations, 20–25% variation within populations between years, and was positively correlated with longer growing seasons. However, induced seed dormancy in the small seed type was correlated with large seed mass and to late-season precipitation, suggesting that longer growing seasons influence the strength of maternal and sibling chemical signals that induce dormancy in the small seeds. Thus variation in small seed germination fractions varies with the growing conditions experienced by the maternal plant. The greater longevity of the small seed type suggests that plasticity in germination in this species may contribute to seed bank formation, which is atypical of invasive annual grasses in California, and this may contribute to the persistence and spread of A. triuncialis in invaded ranges.

Information from a field perspective on temperature thresholds related to physical dormancy (PY) alleviation and seed resistance to high temperatures of fire is crucial to disentangle fire- and non-fire-related germination cues. We investigated seed germination and survival of four leguminous species from a frequently burned open Neotropical savanna in Central Brazil. Three field experiments were conducted according to seed location in/on the soil: (1) fire effects on exposed seeds; (2) fire effects on buried seeds; and (3) effects of temperature fluctuations on exposed seeds in gaps and shaded microsites in vegetation. After field treatments, seeds were tested for germination in the laboratory, together with the control (non-treated seeds). Fire effects on exposed seeds decreased viability in all species. However, germination of buried Mimosa leiocephala seeds was enhanced by fire in an increased fuel load treatment, in which we doubled the amount of above-ground biomass. Germination of two species (M. leiocephala and Harpalyce brasiliana) was enhanced with temperature fluctuation in gaps, but this condition also decreased seed viability. Our main conclusions are: (1) most seeds died when exposed directly to fire; (2) PY could be alleviated during hotter fires when seeds were buried in the soil; and (3) daily temperature fluctuations in gaps also broke PY of seeds on the soil surface, so many seeds could be recruited or die before being incorporated into the soil seed banks. Thus seed dormancy-break and germination of legumes from Cerrado open savannas seem to be driven by both fire and temperature fluctuations.

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.

Hypseocharis is a genus endemic to the high Andes and sister to all other Geraniaceae genera. Regarding its basal position in Geraniaceae evolution, its germination ecology can provide important insights into the early evolution of physical dormancy. Imbibition tests performed on seeds of two Hypseocharis populations from Bolivia indicate that their seeds indeed have physical dormancy like all other Geraniaceae. These results indicate that physical dormancy in Geraniaceae evolved during the Eocene before the uplift of the Andes mountains and before the events that led to the cross-Atlantic disjunct distribution of Geraniacae.

Post-harvest drying prolongs seed survival in air-dry storage; previous research has shown a benefit of drying moist rice seeds at temperatures greater than recommended for genebanks (5–20°C). The aim of this study was to determine whether there is a temperature limit for safely drying rice seeds, and to explore whether the benefit to longevity is caused by high-temperature stress or continued seed development. Seeds of two rice varieties were harvested at different stages of development and dried initially either over silica gel, or intermittently (8 h day–1) or continuously (24 h day–1) over MgCl2 at temperatures between 15 and 60°C for up to 3 days. Seeds dried more rapidly the warmer the temperature. Subsequent seed longevity in hermetic storage (45°C and 10.9% moisture content) was substantially improved by increase in drying temperature up to 45°C in both cultivars, and also with further increase from 45 to 60°C in cv. ‘Macassane’. The benefit of high-temperature drying to subsequent longevity tended to diminish the later the stage of development at seed harvest. Intermittent or continuous drying at high temperatures provided broadly similar improvements to longevity, but with the greatest improvements detected in a few treatment combinations with continuous drying. Heated-air drying of rice seeds harvested before maturity improved their subsequent storage longevity by more than that which occurred during subsequent development in planta, which may have resulted from the triggering of protection mechanisms in response to high-temperature stress.

Seed longevity is dependent on seed traits and storage conditions. This study evaluated the seed longevity and physical dormancy (PY)-break of two species of Dimorphandra endemic to Brazilian biodiversity hotspots. Longevity was tested in situ, by burying seeds in their natural habitats (12 months), and ex situ, by storage in a cold chamber (5°C; 24 months). Seeds were taken at regular intervals to assess germinability at 30°C (12 h photoperiod) using intact and scarified seeds. Intact seeds (freshly collected, and after 12 months storage) were analysed using scanning electron microscopy. The germinability of freshly collected seeds of both species reached approximately 10% for intact seeds and >85% for scarified seeds. Cold storage maintained seed viability in both species, and broke dormancy for 35% of D. wilsonii seeds. After 12 months, only 55% (for D. exaltata) and 41% (for D. wilsonii) of the buried seeds were recovered; more than 90% of which remained viable in both species. Seeds gradually overcame PY during burial, with a higher germination increase for D. wilsonii (71%) than D. exaltata (32%). Dimorphandra exaltata seeds did not show clear structural changes after cold storage although D. wilsonii seeds evidently experience an increase in the depth of fracture lines. Burial promoted deep seed coat changes in both species, more intense in D. wilsonii, indicating that temperature and humidity variations throughout the year are among the main factors releasing Dimorphandra seeds from PY. The seeds of both studied species overcame PY during burial and are able to form small persistent soil seed banks.

The water uptake by the seed in arid and semi-arid ecosystems may not be continuous, but may occur in hydration and dehydration cycles (HD). In order to investigate the benefits of discontinuous hydration on seed germination of Pilosocereus catingicola subsp. salvadorensis subjected to environmental stresses, an imbibition curve was determined and times X, Y and Z selected, corresponding to 25, 50 and 75% of the first phase of imbibition, respectively. Seeds of two populations (Caatinga and Restinga) were subjected to 0, 1, 2 and 3 HD cycles, with 48 h of dehydration. Water and saline stress responses were tested at five osmotic potentials (0.0, –0.1, –0.3, –0.6 and –0.9 MPa). Discontinuous hydration provided greater tolerance to water and saline stress in the seeds from the Caatinga region. When the seeds were subjected to water stress, the reduction of the mean germination time (MGT) provided by HD cycles decreased with increasing concentration of the solution. In saline stress, the reduction of MGT was greater in the solutions with the highest concentrations. We conclude that the seeds of the same species collected from populations located in different ecosystems have different germination responses after passage through discontinuous hydration and HD cycles. This provides greater tolerance to environmental stresses but with different responses among populations.

Seed germination is vital for persistence of many plant species, and is linked to local environmental conditions. Small increases in temperature during this critical life history transition may threaten species by altering germination timing and success. Such changes in turn may influence population dynamics, community composition and the geographic distributions of species. In this investigation, a bi-directional temperature gradient plate was used to profile thermal constraints for germination in 26 common, threatened and geographically restricted Eucalyptus species (Myrtaceae) from southern Western Australia. These observed data were used to populate models to predict optimum germination responses (mean time to germination, germination timing and success) under current (1950–2000 averages) and future (2070 high greenhouse gas emission climate scenario) mean monthly minimum and maximum temperatures. Many species demonstrated wide physiological tolerance for high germination temperatures and an ability to germinate outside current and forecast future autumn–winter wet season temperatures, suggesting that climatic distribution is a poor proxy for thermal tolerance for Eucalyptus seed germination. Germination for some species is predicted to decline under forecast conditions, but the majority will maintain or improve germination particularly during the cooler winter months of the year. Although thermal tolerance may benefit persistence of many Eucalyptus species in southern Western Australia as warming becomes more severe, large rainfall declines are also forecast which may prove more detrimental to plant survival. Nonetheless, this framework has the potential to identify seed resilience to heat stress in an early life history phase and hence species vulnerability to one characteristic of forecast environmental change.

Obtaining corn hybrid seeds (Zea mays L.) with high vigour depends on the parental lines and the direction of the cross, and this relates to seed desiccation tolerance and composition. This research studied reciprocal crosses between pairs of proprietary, elite parent lines (L1 and L5; L2 and L4) producing hybrid seeds with different qualities attempting to correlate vigour with seed composition, focusing on storage proteins, starch and soluble sugar amounts. Four corn hybrid seed lots produced from reciprocal crosses were compared (HS 15 with HS 51, and HS 24 with HS 42) by assessing germination, vigour, and seedling emergence in the field. Seed composition was assessed in mature, dehydrated seeds. Proteins were extracted, quantified, and analysed by electrophoresis and densitometry. Starch amounts were assessed using a kit and soluble sugars were determined using high performance liquid chromatography with pulsed electrochemical detection. The L1 and L2 lineages, used as female parents, provided seeds with lower vigour; however, the quantification of major protein bands, and sucrose, raffinose and stachyose were similar between seed lot pairs. While both total seed protein and starch varied between reciprocal hybrids for one of the two sets of crosses, the amounts of neither correlated with seed vigour. Interestingly, hybrids with low seed vigour (HS 15, HS 24) accumulated greater amounts of fructose relative to their reciprocal; correlation analysis confirmed these results. We demonstrate different effects on seed vigour dependent on the maternal parent in reciprocal crosses producing hybrid corn seeds. We also show that vigour is negatively correlated with seed reducing sugar contents.

Endozoochorous dispersal of seeds by livestock has long attracted the attention of grassland scientists. However, little is known about seed dispersal after ingestion by Kazakh sheep on dry grasslands in the Tianshan Mountains. The objective of this experiment was to learn more about the recovery and germinability of seeds from 17 plant species after either actual or simulated ingestion (i.e. insertion through a rumen fistula) by Kazakh sheep. The passage time of seeds through the sheep gut ranged from 12 to 96 h. More than 80% of all recovered seeds were defecated 24–48 h after ingestion. The mean retention time of seeds in the gut ranged from 27.3 to 42.2 h. Seed recovery percentage ranged between 12.6 and 17.6% for leguminous species and between 0.8 and 3.2% for gramineous species. Seed recovery percentage was positively correlated with seed mass, but negatively correlated with seed shape. The germination percentages of the gramineous species were greater in the non-ingested treatment (66–98%) than in the simulated ingestion treatment (3–10%). In contrast, for leguminous species, seed germination percentages were greater in the simulated ingestion treatment (23–70%) than in the non-ingested one (5–12%). Seed germination percentage after simulated ingestion was positively correlated with seed mass, but negatively correlated with seed shape. In conclusion, leguminous seeds were more likely than gramineous ones to pass through the gut of Kazakh sheep and then germinate. Free-ranging Kazakh sheep can contribute to the spread of plant species, especially leguminous species, in the Tianshan Mountains.