Time of seedling emergence is an important step in the life cycle of annual plants because it may determine subsequent performance and success. Timing of emergence is especially critical to plant performance in habitats like arable fields which are subject to frequent disturbances. Within-season variation in timing of germination in the range of only a few days is typical for many arable weeds. However, since it is unclear whether such small deviations in germination date translate into fitness differences in the course of the life cycle, the aim of this paper was to quantify the effects of short germination delays on plant performance. We conducted two generalized randomized block experiments in an unheated greenhouse to study the impact of delayed germination (1, 2, 3 and 7 d) with and without competition, respectively, on the fitness of the arable weed species Agrostemma githago (L.). We expected that delayed germination significantly reduces fitness in terms of several life-history traits, and that the decrease of fitness is higher in the presence of competition. Under realistic conditions with competition through barley, Agrostemma plants with delayed germination of 7 d produced 54% fewer shoots, 57% less biomass, 52% fewer flowers, 36% lighter seeds and were 23% shorter as compared to control plants without delayed germination. Without additional stress through competition with barley this pattern was less pronounced. Thus, in the situation of interspecific competition, early emerging seedlings have biologically significant fitness advantages over later emerging seedlings of the same species.

The understanding of the genetic basis of grain dormancy in wheat has rapidly improved in the last few years, and a number of genes have been identified related to that trait. We recently identified the wheat genes TaPM19-A1 and -A2 and we have now taken the first step towards understanding the role of this class of genes in seeds. By investigating the Arabidopsis homologous PM19-Like 1 (PM19L1) we have found that it has a seed-specific expression pattern and, while its expression is higher in dormant than in non-dormant seeds, knock-out mutations produced seeds with increased dormancy. Not only primary dormancy, but also secondary dormancy in response to high temperature was increased by the loss-of-function. We have also examined the function of PM19L1 by localizing the PM19 protein primarily to the cotyledon cells in seeds, possibly in membranes. By investigating the co-expression network of this gene we have found that it is connected to a small group of abscisic acid (ABA)-induced seed maturation and storage-related genes. The function of PM19L1 represents a good opportunity to explore the interactions of key factors that can influence seed dormancy such as ABA, temperature and membrane properties.

The distribution and dynamics of plant populations depend on the recruitment of young individuals, which is influenced by seed production, animal seed dispersal, dispersal distance, and the deposition of seeds in favourable places for seed germination/establishment and seedling survival. In particular, seeds dispersed over long distances will likely encounter new environmental conditions that occur at large spatial scales, with seed and seedling survival influenced by the adaptation of plant populations to soil and climate conditions. In this paper, it is hypothesized that seed germination and seedling survival probabilities depend on seed origin and deposition sites. A reciprocal seed and seedling transplant experiment was carried out with zapote seeds (Manilkara zapota) to determine the effect of origin and deposition sites on seed germination and seedling survival over a year in the Greater Calakmul Region of Mexico. Two origin and two deposition sites were selected that show different soil moisture levels within the habitat of the Baird's tapir, a major seed disperser of M. zapota seeds. The results show that sites of origin and deposition influenced seed germination and seedling survival probabilities. This suggests that the displacement of seeds far from parent trees, while potentially reducing intraspecific competition, does not ensure their survival, and that seeds need to be deposited in microsites within their environmental tolerance for dispersal to be successful. Furthermore, this paper emphasizes the importance of field experiments to provide strong inference about the effects of environmental conditions on recruitment and distribution of plant species.

There is little information about seed dormancy release and germination behaviour in Vitis amurensis, which is one of the most important wild Vitis species. This work aimed to investigate the dormancy release and germination characteristics and their variation in V. amurensis seed. Seed dormancy was released by: (1) stratification at 5, 10, 15, 20, 25 and 15/5°C, respectively; (2) stratification at 25°C followed by stratification at 5°C; (3) 5°C stratification before or after seed drying; and (4) stratification at a series of relative humidities (RHs) at 5 and 25°C. The freshly harvested and dormancy-released seeds were germinated at three fluctuating and four constant temperatures. In V. amurensis: (1) dormancy release could occur at temperature ≤ 25°C, but was maximal at 5°C for all varieties; (2) dormancy release and induction occurred simultaneously during stratification; (3) the rates of dormancy release and induction were dependent on temperature; (4) seed moisture content affected dormancy release with a temperature-dependent pattern; (5) seeds germinated better at fluctuating temperature than at constant temperature. However, the optimum condition for stratification and germination, the rates of dormancy release and induction, and the response of seeds to warm stratification, to drying and to germination temperature varied significantly among the V. amurensis varieties. Our data imply that seed dormancy and germination in V. amurensis not only have intra-species similarity, but also vary within species, which may be due to differences in environmental conditions at the original growth and cultivated sites.

The fruits of Quercus robur are shed containing seeds at high moisture contents which remain desiccation sensitive and exhibit recalcitrant storage behaviour. Little is known of the control of seed development in these and other recalcitrant seeds. In the present work the changing concentrations of four plant growth regulators during seed reserve accumulation was studied over five years on seeds from the same tree. The pattern of reserve accumulation and changing ABA content in seeds differed between years. Although ABA content in the cotyledons increased in line with increasing dry weight to different contents at shedding, similar concentrations existed in cotyledons in each year. Thus ABA did not appear to be influencing dry weight accumulation. However, unlike orthodox seeds the decline in ABA concentration prior to shedding was limited and consistent with a continuing role for ABA in preventing precocious germination. An earlier peak in ABA concentration was associated with greater desiccation tolerance at shedding across years. The concentrations of zeatin and zeatin riboside in cotyledons were similar in each year and declined during reserve accumulation in a similar fashion to that reported for orthodox seeds. By contrast, IAA concentration increased in both the cotyledons and axes in the latter stages of seed development, opposite to that reported for orthodox seeds. It is possible that the increasing IAA concentration in cotyledons and axes and the stable concentration of zeatin and zeatin riboside throughout the latter stages of development in the axes of Q. robur are linked to the maintenance of active metabolism for the rapid initiation of germination upon shedding observed in seeds of this species.

The osmotin gene was introduced into tea [Camellia sinensis (L.) O. Kuntze] somatic embryos at the globular stage of development via microprojectile bombardment, to determine the effects of this stress-tolerance-related gene on the accumulation of storage reserves and acquisition of desiccation tolerance during embryo maturation. Changes in total soluble sugars, starch and proteins in response to osmotin were estimated in the stably transformed embryos, using standard biochemical and histochemical methods, after the globular embryos had matured to the heart stage of development. These changes also were compared with two types of controls, comprising (1) untransformed embryos and (2) embryos that were bombarded without osmotin DNA. The control embryos showed poor reserve accumulation and negligible germination but, in contrast, the embryos bombarded with the osmotin gene showed a several-fold increase in storage reserves, normal maturation and 57–63% germination. When these transformed embryos were desiccated to 15.5% moisture content, only 40% germinated, although a further increase in the accumulation of storage reserves occurred. In contrast, the control embryos became necrotic and failed to germinate. This study demonstrates the use of somatic embryogenesis as a tool for understanding seed recalcitrance in tea, and suggests that transformation with stress-tolerance genes can overcome the developmental problems associated with recalcitrant seeds.

Published information regarding the effect of storage temperature on dormancy alleviation of sunflower achenes is contradictory and ambiguous. In the present study we explored the effect of temperature during dry storage on dormancy release in two sunflower genotypes, including a commercial hybrid and an inbred line. Dry storage at 25°C consistently accelerated dormancy release of achenes compared with 5°C. This response fits the general pattern reported for dry after-ripening in seeds of many other species. Depending on the genotype and the dormancy factor prevailing, higher temperature alleviated embryo dormancy and coat-imposed dormancy. Hormonal pathways involved in these changes were investigated at the physiological level. In both genotypes, sensitivity to abscisic acid (ABA) was reduced by storage at 25°C. Also, but only in one genotype, storage at 25°C reduced ABA levels upon imbibition and increased the response to a gibberellin (GA) synthesis inhibitor and to applied GA3, compared with storage at 5°C; these results support the idea that temperature affects both ABA and GA metabolism and signalling pathways during after-ripening. This information will be useful to define storage conditions for commercial sunflower achenes, and will also help focus future research on the underlying mechanisms of dormancy release during dry after-ripening in sunflower.

Nuclear DNA of wheat embryos was probed with a synthetic oligonucleotide (C3TA3C3TA3) to assess changes in the length of telomeres during storage and germination. Chromosomal DNA of dry embryos, especially those derived from long-term-stored caryopses, was considerably depleted of telomeric repeats, but, shortly after the start of imbibition, the chromosomal DNA became rich in sequences hybridizing to the telomeric probe. Simultaneously, newly synthesized telomere-related sequences appeared in an extrachromosomal form that could be separated from bulk nuclear DNA by velocity and isopycnic centrifugations. It is concluded that, in addition to typical repair events, rapid synthesis of telomeric repeats occurs in wheat embryos at the onset of germination.

Endophytic microbiomes play a beneficial role in the development and protection of plants. However, seed-borne endophytic bacteria have not yet been fully explored. Investigation of parasitic plants, whose existence depends on yet poorly understood and complicated relationships with microorganisms and hosts, is particularly crucial. Endophytic bacteria promote seed conservation and facilitate seed germination in soil. Several root holoparasites from the Orobanchaceae family are the most aggressive broomrape species, often causing serious yield losses in important crops. Parasitic plants are characterized by the production of a large number of some of the smallest seeds in the world's flora, allowing them to stay viable in the soil for several dozen years. This study's aim was to isolate and characterize the seed endophyte and surface bacteria of the most aggressive and widespread broomrape weed, Phelipanche ramosa. We isolated two endophytic bacteria from within the seeds which are closely related to Brevibacterium frigoritolerans and Bacillus simplex described as soil bacteria, highly resistant to environmental conditions, and as plant growth-promoting rhizobacteria. Moreover, we isolated three strains from the surface of non-sterile seeds; all three isolates were related to the Bacillus cereus group.

The desiccation tolerance of Zizania palustris seeds has been debatable, but recently it has emerged that survival to low moisture contents is a function of the temperature during dehydration. Survival of dehydration is greatest at 25–30°C and viability declines as the dehydration temperature is reduced. On the other hand, extended hydrated chilling (stratification) is required to break dormancy. The present contribution examines the ultrastructural condition of embryonic axes of Zizania palustris after dehydration at various temperatures, and following reimbibition and stratification. Axis cells sustained least damage when dehydration was carried out at 25°C and ultrastructural deterioration was more severe with lower temperatures during water loss. Damage sustained as a result of unfavourably low dehydration temperatures was visible when seeds were fixed from the dry state and was generally exacerbated during fully imbibed stratification. However, 25°C represented the optimum dehydration temperature; seeds that had been dried at 30°C also showed considerable ultrastructural disturbance when fixed from the dry state. This was largely reversed during fully imbibed stratification, although signs of the damage that had been sustained still persisted. These observations are in keeping with the germination behaviour of Z. palustris seeds dehydrated at temperatures above and below 25°C. The results are discussed in terms of the predominance of one of two mechanisms of membrane deterioration, depending on the dehydration temperature, which are probably not mutually exclusive. It is hypothesized that, at temperatures above 25°C, damage by free-radical-mediated events may predominate, whereas, at temperatures below the optimum, irreversible lipid-phase transitions may be the major factor resulting in membrane damage.

The viability of Norway maple seeds harvested at mass maturity (57.3% moisture content) was reduced from 100% to only 38% if dried rapidly (at 15% RH and 15°C for 3 days) to 4.7% moisture content. In contrast, slow drying for 32 days (the optimum duration of several investigated) to 29.9% moisture content enabled 93% of the seeds to survive subsequent rapid desiccation to 3.5% moisture content. This is similar to the 95% viability shown by seeds harvested 40 days after mass maturity and then dried rapidly to 4.4% moisture content. However, fruits or seeds harvested at mass maturity and then held moist for 21 days also showed 94 and 91% viability after subsequent rapid desiccation to 3.8 and 3.3% moisture content, respectively. Thus a post-ovule-abscission programme is required before Norway maple seeds are able to tolerate rapid enforced desiccation to low moisture contents, but loss in moisture during this period is not essential to the development of desiccation tolerance.

Embryos taken from late maturation phase seeds of Phaseolus vulgaris cv. Seminole prior to seed desiccation (35–45 DAA) can be induced to germinate in the absence of water by exogenous ethylene. NMR imaging of proton relaxation within the embryo shows changes in water status in putative (and not fully differentiated) vascular tissues of the hypocotyl within 3 h of ethylene administration. Difference imaging revealed that the change was progressive in the hypocotyl towards the radicle tip and was accompanied by changes in water status in the cotyledons. Water within plumular leaves was also affected. Increase in diameter of the hypocotyl–radicle axis (as estimated by pixel counts) was detectable from 3 h. Longitudinal radicle growth was detectable by NMR imaging at 18 h. Visible germination under the conditions used was apparent after 20 h. Changes in water status detected by this technique are an indication of changes in activity (concentration) or motion of water molecules or both. The data are consistent with a mode of action of ethylene in stimulating a redistribution of water within embryo structures from cotyledons to axis via the cotyledonary node and allowing the axis access to water sufficient to support germination. This supports the hypothesis that in vivo, quiescence at this developmental stage is induced and maintained by sequestration of water within the cotyledons.

Astragalus is the largest genus of seed plants with 3000 or more species that occurs naturally on several continents. The genus has some use as a forage and medicine and in industry, and many of the species are rare endemics threatened with extinction. The seeds are reported to be dormant at maturity, and various treatments have been used in an attempt to germinate them. Our primary aim was to determine via a meta-analysis the most effective way(s) to break dormancy in seeds of this species-rich genus. Mechanical and chemical (conc. sulphuric acid) scarification were by far the best of 12 treatments for breaking seed dormancy of the 40 species included in our meta-analysis, whereas prechilling, gibberellin and smoke were ineffective. These results along with those of imbibition tests confirm that seeds of the examined Astragalus species have physical dormancy (PY). Further, PY in these 40 species and (its documented occurrence) in 118 species that could not be included in our meta-analysis transcends climatic and geographic boundaries, edaphic conditions, life cycle/life form types and infrageneric phylogeny. Thus, it seems likely that most species of Astragalus have PY. However, in addition to PY, physiological dormancy (PD), that is, combinational dormancy (PY + PD), has been reported in a few species of Astragalus. This study should be useful to both basic and applied scientists who want to germinate seeds of Astragalus.

Neither ethylene nor 1-aminocyclopropane-1-carboxylic acid (ACC) was able to prevent the induction of secondary dormancy of Amaranthus caudatus at 45°C. Both ethylene (4.5 × 10-9–4.5 × 10-7 M) and ACC (10-3–10-2 M) removed secondary dormancy at 25°C, although ethylene was much more effective. The presence of ethylene for only 10 h was sufficient to remove secondary dormancy in almost all seeds. Incubation of secondary dormant seeds for up to 5 d at 25°C did not change sensitivity to ethylene. The breaking of secondary dormancy by ethylene was prevented by 2,5-norbornadiene (NBD; 1.5 × 10-5–3 × 10-4 M), indicating the physiological action of ethylene. Abscisic acid (ABA; 10-4–10-3 M) increased the requirement for exogenous ethylene. It is suggested that secondary dormancy in A. caudatus seeds might be related to insufficient ethylene production associated with an insufficient amount of ACC.

Assessment of the rehydration procedure has been consistently overlooked in evaluation of factors contributing to successful cryostorage of partially dehydrated, embryonic axes excised from recalcitrant seeds. Conventional rehydration of Trichilia dregeana (Sond.) axes in distilled water resulted in the lack of root pole gravitropism after culture on medium in vitro. In comparison, a strong gravitropic response was observed in axes that had not been dehydrated, and by those rehydrated in a solution containing 1 M CaCl2 and 1 mM MgCl2. However, no marked loss of either cation from the tissue could be detected, whether axes were rehydrated in distilled water or the Ca2+/Mg2+ solution. Starch-packed statoliths differentiated rapidly in both non-dehydrated axes and those rehydrated in the divalent cation solution, but these organelles failed to develop or accumulate much starch following axis rehydration in water, as was the case for the amyloplasts of root cells generally. After rehydration in the Ca2+/Mg2+ solution and 48 h in culture, axis root-cap columella cells accumulated Ca2+, whereas axes rehydrated in water or solutions containing either Ca2+ or Mg2+ alone did not take up Ca2+. Rehydration with the Ca2+/Mg2+ solution also altered distribution of the actin component of the cytoskeleton. When rehydrated in the divalent cation solution, actin was associated with the nucleus and with the statoliths, which were located distally in statocytes of axes. In contrast, actin was largely confined to the perinuclear area in root-cap columella cells of the agravitropic, water-rehydrated axes. The present results indicate a definitive primary role for Ca2+ with Mg2+ in graviperception, via starch metabolism, and in the determination of statolith morphology, which appears to be linked with maintenance of the actin component of the cytoskeleton in root-cap statocytes.

By means of direct manipulation, this study showed that the testa protects the germinating seed and germinant from decay (an advantage in moist soils) and desiccation (an advantage in periodically dry soils) in four species, all of which retain their testas until the cotyledons are lifted above the soil. Furthermore, the testa prevents early spreading of the cotyledons (an advantage when elongating through soil and litter). Almost all germinants of Banksia hookeriana and Leucadendron eucalyptifolium, with their testas removed, died and rotted away after being sown on the soil surface or at a range of depths. Hakea cyclocarpa and H. strumosa showed only minor losses compared with the intact germinants. All naked germinants were smaller than the controls at any depth, except among H. cyclocarpa. Absence of the testa did not differentially affect the length of the hypocotyl in the dark or light, but lateral spread of the cotyledons was accelerated, especially in the dark. Removal of a small fraction of the testa was sufficient to cause general mortality of B. hookeriana germinants but not of H. cyclocarpa or H. strumosa. After imbibition, water content of the naked embryos of B. hookeriana reverted to the air-dry state within 24 h of sowing in air-dry sand, while intact seeds were still moist even after 7 days of drying. Compared with intact seeds, drying periods of 3–7 days decreased the amount of germination and root growth of naked embryos after rewetting, and increased microbial infection of the survivors. Removal of the testa from germinants was sufficient to cause general mortality after only 6 h in air-dry sand. In contrast, most intact germinants with short radicles recovered after drying for 2 days.

Seed–soil contact plays an essential role in the process of germination as seeds absorb water through direct contact with the moist soil aggregates that surround them. Factors influencing seed–soil contact can be considered as those pertaining to soil physical properties (e.g. texture, bulk density, porosity, etc.) and those related to environmental conditions (e.g. temperature, rainfall, frost). Seed–soil contact is furthermore influenced by the specific field management processes that farmers apply, which have developed significantly over the last 30 years. However, the precise effect of cultivation on the actual contact area of the seed with the surrounding soil is based on a series of assumptions and is still largely unknown. This review considers the influence of soil management and its direct impact on seed–soil contact and establishment. We review the state of the art in methodology for measuring seed–soil contact and assess the potential for soil amendments such as plant residues and waste materials to improve seed–soil contact. Engineering the ‘optimal’ seed–soil contact remains a challenge due to the localized variation between the interaction with field management techniques and soil texture, climatic conditions and crop type. The latest imaging approaches show great promise to assess the impact of management on germination. Combining the techniques with the latest network models offers great potential to improve our ability to accurately predict germination, emergence and establishment.

Axes, hypocotyls and radicles excised from dry cotton seeds (Gossypium hirsutum L. cv. M-8, a double haploid) were imbibed for 24 h and compared with axial segments (excised sections of embryos below the cotyledons) of imbibed, intact seeds. Radicles of excised axes had a 7.4-fold increase in length compared with only 5.2- and 5.7-fold increases, respectively, in radicles of intact seeds and in those isolated when dry. Change in hypocotyl length was not as extensive. EM data for hypocotyl and radicle cortical cells from dry and imbibed seeds revealed a major reduction in matrix protein in protein storage vacuoles along with significant organelle development at 24 h from the start of imbibition. This occurred in parallel with a reduction in salt-extracted proteins and an increase in 2% SDS-extractable proteins. SDS-PAGE of protein from low (0.2 M NaCI) and high (1.0 M NaCI) salt extracts showed a reduction in amount of the major storage proteins (53 and 48 kDa), these bands being almost totally absent in gels of protein extracts from imbibed radicles and significantly reduced in hypocotyls, within 24 h from the start of imbibition. These results indicate that initial elongation of hypocotyls and radicles in intact seeds or of excised axes, after 24 h imbibition, involves breakdown of storage proteins in these axial parts to supply nutrients for growth, with very limited contribution from the cotyledons.

Eugenia stipitata occurs along rivers in Western Amazonia and produces berry-type fruits with economic potential. Its large recalcitrant (i.e. desiccation-intolerant) seeds have been proposed as a model to study seed stress response, as no apparent differentiation between the embryonic axis and the fused cotyledons are visible. Here, the longevity of submerged seeds was analysed with a view to understanding adaptive mechanisms to seasonal flooding. Submerged seeds began germinating after 2 months. After 1 year, 87 and 96% total germination was reached when seeds were submerged under a water column of 6 cm (where seedlings could emerge from under the water) and 26 cm (where seedlings could not reach the water surface), respectively. Seedling morphology was altered underwater, with short internodes and rudimentary leaf blades, and when submersion was terminated, seedlings transplanted to nursery conditions recovered a normal phenotype. Furthermore, when seedlings were detached from the seeds, the ‘resown’ seeds produced a second, normal seedling within 9 months. Concentrations of the antioxidant glutathione, which was measured as a stress marker, increased with submersion time in water. Seeds that had developed roots and shoots underwater had higher concentrations of glutathione disulphide than non-germinated seeds, suggesting that the flooding stress was more intense for seedlings than seeds, although more oxidizing cellular redox environments are also consistent with the conditions required for differentiation. Submergence underwater is recommended for storage of the recalcitrant seeds of E. stipitata for up to 1 year.

Efficient seed storage is a shared concern among the growing number of seed banks established for crop improvement or ex situ conservation. Container properties greatly affect seed interactions with the environment and the overall cost and success of seed banking operations. Several material properties contribute to their suitability as seed containers. This paper provides a consolidated list of water vapour permeability properties of thermal plastics commonly used for packaging. Composite packages with layers of film with different properties provide distinct advantages to seed banks. Different seed banks must rank the importance of the various factors depending on their mission and resources. Once the risks, costs and benefits are weighed, an appropriate strategy can be developed that addresses a seed bank's specific needs. Because there are many problems and several solutions, it is likely that strategies will vary among seed banks. This response details variables to consider when selecting seed storage packages, and focuses on water diffusion rates of packages with different compositions. A ‘moisture audit’ will help seed bank operators make informed decisions about packaging.