Capeweed (Arctotheca calendula) seeds were found to be dormant at harvest. Effects of duration and temperature of storage under ‘laboratory’ and ‘natural’ conditions, growth regulators, stratification and age of seeds, were studied on the germination of dormant seeds. Three factors imposed on seeds were found to promote germination of capeweed: (i) allowing seed to age, either in storage, buried in soil or during stratification (germination of 18-month-old seeds was up to 60% higher than that of fresh seeds); (ii) the presence of light (the average germination percentage of seeds exposed to light during storage was 3.2-fold greater than that of seed stored in the dark); and (iii) the application of growth regulators, particularly gibberellic acid, enhanced by scarification (GA3 and ethephon promoted germination by up to 58% when applied to scarified seeds). Other factors, e.g. temperature and depth of storage, influenced germination but were less critical to its success. These results provide a preliminary assessment of the importance of these factors in controlling dormancy in seeds of capeweed.

The structure and function of grassland ecosystems can be altered by a changing climate, including higher temperature and elevated atmospheric CO2 concentration. Previous studies suggest that there is no consistent trend in seed germination and seedling recruitment as affected by these conditions. We collected seeds of two native and two invasive species over 6 years from a field study with elevated CO2 (600 p.p.m.) and temperature (1.5/3.0°C day/night) on the mixed-grass prairie of Wyoming, USA. Seed fill, viability and mass were evaluated and germination tests were conducted under alternating temperatures in growth chambers. Thermal time requirements to reach 50% germination (θ50) and base temperatures (Tb) for germination were determined using thermal time models. Climate change conditions had limited effects on seed fill, viability and mass. The combination of CO2 enrichment and warming increased germination of Bouteloua gracilis. Centaurea diffusa and Linaria dalmatica, two invasive species in this study, had the lowest θ50 and Tb required for germination among all the species studied. Although final germination percentages of these invasive species were not affected by treatments, previous studies reported increased seed production under future climate conditions, indicating that they could be more invasive at the regeneration stage in the future. We conclude that projected future temperature increases will have little effect on seed reproductive traits of native species. In addition, the distribution and abundance of B. gracilis and invasive species may be favoured by global climate change due to enhanced germination or seed production traits caused by elevated parental CO2 and temperature conditions.

The correlation between physiological markers for seed maturity (acquisition of germinability, definitive morphological traits and desiccation tolerance) and timing of the shutdown of nuclear replication activity and β-tubulin synthesis has been investigated in developing pepper seeds. Nuclear replication activity was analysed by flow cytometry. Expression of β-tubulin, a cell-cycle related structural protein, was detected in western blots of total protein extracts. In pepper embryos, DNA synthesis stopped 42–49 days after anthesis (DAA), at which time seeds started to acquire desiccation tolerance. A clear β-tubulin signal was detected up to 63 DAA, only 1 week before the acquisition of complete desiccation tolerance. Both nuclear replication and β-tubulin synthesis can therefore be used as markers for the physiological stage reached by developing pepper seeds. Detection of β-tubulin signals in embryonic tissues can also be helpful in discriminating immature seeds within a commercial seed lot.

Seven ancient seeds, about 1600 years old, were found during an archaeological excavation in Asar Island which is located in south-western Turkey. These seeds were subjected to germination, in vitro callus induction and molecular characterization experiments to test the viability and plant origin of the seeds. Six of the seven seeds had viable seed components (such as cotyledons) and produced callus tissue in Murashige and Skoog medium supplemented with 2 mg l− 1 6-benzylamino purine (BAP), 0.2 mg l− 1 1-naphthaleneacetic acid (NAA), 20 g l− 1 sucrose, 2 mg l− 1 glycine and 7 g l− 1 agar, but the calli from these seeds failed to yield adventitious shoots. DNA samples from callus tissues produced by ancient seeds in vitro were of good quality. The internal transcribed spacer (ITS) region in nuclear DNA (nDNA) of ancient seeds was amplified successfully. The sequences from amplified ITS DNA products of six ancient seeds indicated that their ITS sequences matched those of Anagyris foetida after subjecting them to BLAST searches in international sequence databases (NCBI). A. foetida is a relict species endemic to the Mediterranean region and used as a herbal medicine. We believe that seed characteristics such as the very hard, extremely smooth and shiny testa, toxic anagyrine alkaloid content and their storage in a pot further improved the longevity of these ancient seeds.

Seed germination of the kiwifruit (Actinidia deliciosa A. Chev. C.F. Liang and A.R. Ferguson), a deciduous, perennial vine, is low because of seed dormancy. The main purpose of this study was to characterize kiwifruit seed dormancy and germination in response to environmental factors such as temperature and light. Dormancy of hydrated seeds is alleviated by the perception of a period at low temperatures (stratification) of at least 3 weeks at 2 or 5°C. Alleviation of dormancy is accomplished by incubation at fluctuating temperatures (20/30°C). A red light pulse did not affect germination, whereas a far-red light pulse strongly inhibited germination. This inhibition was readily reverted by a second pulse of red light, indicating that phytochromes are also involved in dormancy alleviation. Although seed germination was inhibited by the fruit pulp, the latter prevented neither perception of low temperature nor changes in the light sensitivity of the seeds. Therefore, it can be advanced that kiwifruit seeds will only germinate after dispersal if the time–temperature requirement for dormancy alleviation and fluctuating temperatures for dormancy termination are fulfilled. Perception of a closed canopy might interfere with dormancy termination.

Seed longevity in situ is a prerequisite for understanding the life histories and community dynamics of species, although long-term longevity under thick tephra has not been documented because of a lack of opportunity and/or awareness. The seed bank for this study was estimated by both germination and flotation tests. Seeds of 17 species have survived with high density, having been buried under thick tephra for 30 years, since the 1977–1978 eruptions on Mount Usu, Hokkaido Island, northern Japan. The total seed density was >1000/m2. Rumex obtusifolius was the most common seed-bank species for 30 years, but decreased in density between 20 and 30 years. More seeds of Hypericum erectum occurred in deeper soil. The total seed density decreased gradually for 30 years, but H. erectum and Juncus effusus did not decline. Native seeds tended to be viable longer than exotic seeds. These results suggest that small, native seeds tend to survive longer with deep burial, while the more numerous weedy, exotic seeds located at the soil surface declined faster. The seed bank provides long-term monitoring of seed survival under natural conditions, and could be used to detect genetic changes.

The Araliaceae is known to have seeds with underdeveloped embryos that must grow prior to radicle emergence, and thus they have morphological (MD) or morphophysiological (MPD) dormancy. Araliaceae is one of about 15 families with woody species in the tropical montane zone, and in Hawaii 15 species occur in the montane. Our purpose was to determine if seeds of the Hawaiian Araliaceae species Cheirodendron trigynum subsp. trigynum have MD or MPD and, if MPD, what level. In a move-along experiment, some seeds were incubated continuously at 15/6, 20/10 or 25/15°C, while others were moved sequentially from low to high or from high to low temperature regimes. Germination percentages and embryo growth were monitored. Also, the effects of cold and warm stratification on dormancy break were determined. Seeds had physiological dormancy (PD) in addition to small embryos that grew prior to germination, and thus MPD. PD was broken slowly ( ≥ 12 weeks), after which embryos grew rapidly, followed by root and shoot emergence. Embryos grew at temperatures suitable for warm stratification; thus, seeds have Type 1 non-deep simple MPD; the dormancy formula is C1bBb. Seeds from Oahu germinated to 94–100% at 15/6, 20/10 and 25/15°C, while those from the Big Island germinated to high percentages only at 15/6 and 20/10°C. Temperature shifts improved germination of seeds from the Big Island, and movement from either low to high or from high to low temperature regimes was effective in promoting germination. This is the first report of non-deep simple MPD in the Araliaceae.

Arrowleaf sida (Sida rhombifolia) is a tropical weed that grows in disturbed areas. Its control by hand pulling and mowing are only partially effective because arrowleaf sida is difficult to pull and quickly sprouts after cutting. We studied the seed-bank dynamics of S. rhombifolia in the region of Campinas, São Paulo State, Brazil for 2 years, following a known number of seeds placed artificially at five burial depths. Predators and pathogens were responsible for the loss of c. 40% of the seeds during the first year of the experiment. The importance of predation declined with depth. Mortality due to fungal attack did not differ among the burial depths, except for seeds placed at the surface (where it was negligible). Seedling emergence only occurred during the first year, after the beginning of the rainy period and when the study area was still free of a dense vegetation cover. Seedlings did not emerge from depths greater than 5 cm. We conclude that seedling emergence and death are significant causes of seed losses from the seed bank of S. rhombifolia but their relative importance is highly dependent on depth of burial.

Interception by plant canopies during wind dispersal can affect the final destination of diaspores. However, how the interaction of wind speed, canopy type and diaspore attributes affects interception of diaspores by the plant canopy has rarely been studied. We investigated canopy interception for 29 species with different diaspore attributes, six canopy types and six wind speeds in controlled experiments in a wind tunnel. Shrub canopy interception of diaspores were controlled by wind speed and diaspore attributes, but the latter had a greater influence on canopy interception than the former. At low wind speed, diaspore wing loading had a large influence on canopy interception, whereas at high wind speed, diaspore projection area had a large influence. The chance of canopy interception at a particular wind speed was additionally affected by the type of canopy. This study increases our knowledge of the dispersal process, corrects the previous understanding of diaspore dispersal potential and improves the theoretical basis for predicting spatial pattern and dynamics of plant populations.

An abscisic acid (ABA)-induced cDNA fragment encoding a putative protein kinase (PK) was obtained by differential screening of a cDNA library from Fagus sylvatica seeds. The full-length clone, named FsPK1, was produced by 5′ rapid amplification of cDNA ends (RACE) extension. This clone contained the 11 catalytic domains present in all protein kinases, but displayed unusual characteristics found only in a few plant PKs. FsPK1 exhibits features of both serine/threonine and tyrosine protein kinases within the catalytic domain, a putative nuclear localization signal within the regulatory domain and the consensus sequence involved in binding of 14-3-3 proteins. The catalytic domain, expressed in Escherichia coli as a fusion protein, showed Ca2+-dependent in vitro kinase activity and dual serine/threonine and tyrosine specificity. Transcription of the FsPK1 gene was reduced by seed stratification at 4°C, and clearly increased when seeds were treated with 0.1 mM ABA, correlating with the inhibition of germination. Interestingly, FsPK1 transcripts were enhanced when ABA (0.1 mM) and calcium (1 mM) were added together, while the addition of EGTA (calcium chelator) and 3,4,5,-trimethoxibenzoic acid 8-(diethylamino) octyl ester (TMB-8, a calcium antagonist) decreased its expression. Furthermore, FsPK1 transcript expression was tissue specific and accumulated only in ABA-treated seeds, but not in any ABA-treated vegetative tissues examined. These results suggest that the expression of the corresponding protein could be related to the inhibition of germination mediated by ABA in a calcium-dependent pathway.

The size and composition of the seed bank of a rare species (Erica ciliaris L.) was analysed. E. ciliaris hybridizes with a common relative in southern England (Erica tetralix). The seed banks of these co-occurring species were measured at a number of sites with a range of vegetation types and different management histories. Additional sets of samples were taken from forestry plantations on former heathland sites, where these species were known to occur. Relatively few hybrid seedlings were found in any of the seed-bank samples, even though their vegetative abundance within the sampling areas was equal to that of the pure plants. However, the abundance in the vegetation of the two pure species was reflected in the seed-bank size on each of the vegetation types, both with and without burning management. The seed banks from the forestry plantations show that the numbers of seeds of both E. tetralix and the hybrid were depleted, but that the seed bank of E. ciliaris was not significantly different from that of open heathland seed banks. The long-lived nature of the seed bank indicates that there are opportunities for habitat restoration on former heathland sites.

Although pear species (Pyrus spp.) are widely cultivated as an important fruit tree in the world, the regulatory mechanism of their seed dormancy is still unclear. In this research, the role of endocarp, seed coat and embryo in sand pear (Pyrus pyrifolia) seed dormancy was analysed by detecting the endogenous abscisic acid (ABA) and analysing germination experiments with different treatments. The results suggest that a combination of testa and embryo impose sand pear seed dormancy. The high concentration of endogenous ABA in the embryo effectively inhibited seed germination. The high concentration of ABA in the endocarp, as well as in the testa, suggests that dissipation and/or degradation of endogenous ABA in imbibed embryos were suppressed to maintain ABA-inhibited germination. The ABA and gibberellic acid (GA) key signalling genes, including ABI3–5, GAI, RGA and RGL2, were cloned and their expression was analysed in ABA-treated embryos and embryos of imbibed nutlets, imbibed true seeds (endocarp removed) and GA-treated true seeds. The results revealed regulation by ABA of the expression of ABA and GA signalling factors controlling the dormancy release of sand pear imbibed seeds.

In the tetraploid Tulipa sylvestris we found larger and heavier seeds that germinated around 28 days earlier than those of the diploid ancestor Tulipa pumila. The faster germination of the tetraploid species is linked to the faster growth of embryos, which reached their final length much earlier than the embryos of the diploid species. In conclusion, we argue a cautionary approach when dealing with comparative studies on ploidy level and germination to avoid misinterpretation of results when set against the natural conditions.

Seeds with epicotyl dormancy reside in soil up to 15 months (or longer), being exposed to a sequence of temperatures, before seedlings completely emerge (i.e. with both roots and shoots). Heretofore, few studies have examined precise temperatures, especially in sequences, for promotion of radicle and cotyledon emergence and how they relate to environmental cues in nature. Viburnum is the best known genus to exhibit epicotyl dormancy and, as such, we investigated the Japanese V.furcatum, hypothesizing a similar kind and level of dormancy. The underdeveloped embryos in mature seeds in October were spatulate shaped, unlike those in other Viburnum species, and they elongated from late June to August of the following year. Radicles emerged after embryo growth until mid-October, followed by cotyledons from mid-April to mid-May. Temperatures required for embryo growth, radicle and cotyledon emergence in the laboratory approximated closely those in the field. Embryo elongation and radicle emergence occurred at warm temperature regimes, and gibberellic acid (GA3) did not substitute for this warm temperature requirements. Following a 120-d cold stratification of seeds with an emerged radicle, shoots emerged from seeds at 10, 15, 15/5, 20/10 and 25/15°C. We identified that seeds of V. furcatum have deep simple epicotyl morphophysiological dormancy like the majority of other Viburnum species. For propagation of the species from seeds, the nearly 2-year period for seedling emergence could be shortened to 8 months: start fresh seeds at 25/15°C (60 d) and then move them through a sequence of 15/5°C (30 d) →  0°C (120 d) →  20/10°C (30 d).

The final stage of development of orthodox seeds is maturation drying, a process often accompanied by a dramatic decline in storage-protein synthesis. When desiccation is imposed prematurely at certain stages prior to the completion of development, a switch in synthetic events is elicited. events unique to development, such as synthesis of storage protein, are terminated, while syntheses associated with germination and growth are initiated. We investigated whether desiccation plays a key role in effecting a decline in expression of genes for storage proteins by acting directly upon the regulatory regions of these developmental genes. The desiccation responsiveness of the 5′ and 3′ regulatory regions of the gene for the pea storage protein vicilin was tested in transgenic tobacco seed. Chimaeric genes were introduced into tobacco, these genes consisted of the coding region of the reporter gene for β-glucuronidase (GUS) and 5′ and 3′ regions from the vicilin gene, or, as controls, the same regions derived from constitutively expressed genes, presumed to be desiccation-insensitive – those from the cauliflower mosaic virus (35S) and nopaline synthase genes. In transgenic seed expressing the gene constructs containing the vicilin 5′ upstream region, GUS activities declined dramatically after imbibition in mature seed and after rehydration of prematurely dried tobacco seed. In contrast, GUS activities increased after seed rehydration when the constitutive viral (35S) promoter replaced the vicilin 5′ upstream region. The 3′ downstream region of the gene construct did not significantly affect the patterns of changes in GUS activities after seed rehydration. In studies of transient gene expression in castor bean cotyledons, premature desiccation led to termination of GUS gene expression only when the gene construct contained the vicilin 5′ upstream region. This region may respond directly to desiccation; alternatively, changes to trans-acting factors important for expression of genes for storage-proteins may occur as a result of drying.

Western blot analysis using antiserum raised against the lysine-rich box common to dehydrins, one class of late-embryogenesis-abundant (LEA) proteins, was used to study the abundance of heat-stable ‘dehydrin-like’ proteins during development and water stress in Ranunculus sceleratus L. achenes (seeds). A 61 kDa dehydrin-like protein was apparently limited to immature seeds (fresh and dried) which had not attained full desiccation tolerance. In contrast, lower-molecular-mass proteins which were induced by desiccation were found only in more mature seeds. The molecular masses of desiccation-induced proteins changed during seed development from 18 kDa in seeds harvested at 13 days post anthesis (DPA) to 31 kDa at harvest maturity, 21 DPA.

Placing seeds at 21 DPA in polyethylene glycol (PEG) at −1.5 MPa reduced seed moisture content and was accompanied by accumulation of 31 kDa protein. This protein was no longer detected when the seeds were transferred to water. In seeds harvested at 13 DPA, PEG induced the synthesis not only of 18 kDa protein (which is associated with dried seeds at this developmental stage), but also of 28 kDa and 31 kDa proteins. These dehydrin-like proteins were also synthesised when seeds at 13 DPA were imbibed in water. These and other data indicate that both quantitative and qualitative changes in dehydrin-like proteins can occur in R. sceleratus, depending on seed maturity and the degree and duration of water stress.

The effectiveness of ATPase in germinated seed may play an important role in the vigour of germination. The activities of tonoplast and plasma membrane ATPases in two maize (Zea mays L.) lines with different vigour of germination were determined. ATP hydrolysis was measured in microsomal fractions from coleoptiles along with the responses to specific inhibitors for the plasma membrane, tonoplast and mitochondrial ATPases as well as for acid phosphatase. Nitrate-sensitive ATPase activity was 1.5–3.0 times lower in the low-vigour line than in the high-vigour line. Kinetic analysis of ATP hydrolysis at different substrate concentrations revealed the existence of two enzymes in the microsomal fractions of the two lines. The Vmax of enzyme 1 in the low-vigour line was a third of that in the high-vigour line. This enzyme was identified as the nitrate-sensitive or tonoplast ATPase on the basis of measurements of ATP hydrolysis in the presence of specific inhibitors at high (8.12mm) and low (0.77mm) ATP concentrations.

During seed germination, metabolism is reactivated, DNA is repaired and cell division is restarted in the meristems. The mechanisms that co-ordinate cell growth and division in maize embryonic axes during germination are not well understood. However, the presence of a factor similar to IGF (insulin-like growth factor) that accelerates germination has been reported. In the present work, the regulation of the cell-cycle restart by bovine insulin [which has been demonstrated to produce similar effects as insulin-like growth factor of maize (ZmIGF) in maize seeds] was studied in germinating embryonic axes. Our results showed that bovine insulin differentially stimulates growth, S6K phosphorylation, S6rp transcript accumulation on the polysomal fraction, as well as de novo DNA synthesis in the radicles and the coleoptiles of the embryonic axis. A stronger and earlier effect was observed in radicles compared to coleoptiles; therefore, the effect of insulin on the cell cycle of the root meristem was studied by flow cytometry. The G1–S transition was stimulated and cell proliferation was induced. Furthermore, it was determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) that bovine insulin increased E2F and PCNA (proliferating cell nuclear antigen) transcription after 15 h of germination and PCNA de novo synthesis at 15 h of germination. These results show that bovine insulin preferentially stimulates growth in the radicles of germinating embryonic axes and suggest that its effect on the G1–S transition and the activation of cell proliferation is mediated by the induction of E2F and PCNA transcription.

Abies cephalonica cone and seed morphometric characteristics as well as seed germination behaviour were investigated during an 8-year-long diachronic study (2007–2015). The research was carried out in Parnitha National Park, in the part of this Greek endemic, silver fir population that was spared from an unprecedented wildfire (2007). A statistically significant interannual and among-tree variation of cone traits has been identified, except for cone diameter. Cone length is correlated to the number of seeds per cone, while the percentage of empty seeds has been consistently high, with extreme values of 29.3 and 81.8% in a masting (2015) and lean crop (2009) year, respectively. There is also a considerable proportion of dead (including infected) and non-germinated seeds, and the eventual germinable fraction is well below 25% with spikes in masting years (39.4 and 60.9% in 2010 and 2015, respectively). Untreated seeds have been tested at a wide range of constant and alternating temperatures and germination is completed within 4–5 weeks at T ≥ 15°C and ca 15 weeks at 5–10°C. A chilling pretreatment of 4–8 weeks (at 2–4°C) concludes germination at 15–20°C within 2 weeks. A significant white light requirement has been observed for untreated seeds at 15–20°C, while germination was light indifferent at lower or higher temperatures. Data obtained in this study confirm the present, field seed germination during springtime; we may also predict an earlier germination (late autumn to mid-winter) in forest gaps, under the predicted, warmer conditions in the future.