A high-molecular-weight aspartyl proteinase complex, sensitive to pepstatin A was purified to near electrophoretic homogeneity from dry seeds ofjack pine (Pinus banksiana, Lamb.). Two partial activities, endo-and exo-proteinolytic, were associated with the complex, judged by the analysis of haemoglobin hydrolysis products. The high activity of this enzyme in dry jack pine seed suggests a possible function in early germination.

Five Euphorbiaceae species (Jatropha macrocarpa, J. hieronymi, J. excisa, Cnidoscolus tubulosus and Manihot guaranitica) were analysed for seed oil content and fatty acid profiles in order to evaluate their potential as sources of biodiesel. The five species are perennial shrubs adapted to arid and semi-arid environments of north-western Argentina. The seed oil content for all species ranged from 27.9 to 38.7% by dry weight. Fatty acid composition consisted mainly of linolenic, myristic, palmitic, stearic, arachidic, oleic and linoleic acids, with linoleic and oleic acid being the most abundant. Among the five species analysed, J. macrocarpa has the best potential to be used as a biodiesel source. The average total seed oil content (35.8%) and a high value of oleic acid (27.3%) together with the lowest values of linoleic acid (55.8%) make this species a promising crop as a source of biodiesel in arid and semi-arid regions.

Sugarbeet (Beta vulgaris) seeds which had undergone an advancement treatment germinated more quickly and showed greater germination in a standard test at 9°C than seeds which had been steeped in water. Both treatments included the fungicide, thiram (tetramethyl thiuram disulphide). Advanced seeds also emerged more quickly and showed higher establishment in the field. The levels of extractable high molecular weight (HMW) RNA and DNA from advanced, thiram-steeped and untreated seeds were compared. Advanced seeds contained significantly (P > 0.05) more HMW RNA and DNA than thiram-steeped or untreated seeds. Extractable DNA levels were higher from thiram-steeped seeds than untreated seeds but not significantly so. Cell-cycle studies on advanced seeds demonstrated an increase from 2% to 24% of root-tip nuclei with DNA contents of 4C and above. This indicates that DNA replication occurred during the advancement treatment. There was no evidence of cell division occurring, indicating that: (1) advanced seeds were arrested post-S-phase and (2) the period following DNA synthesis in sugarbeet seeds is desiccation tolerant.

Mangroves are highly adapted to extreme environmental conditions that occur at the interface of salt and fresh water. Adaptations to the saline environment during germination are a key to mangrove survival, and thereby, its distribution. The main objective of this research was to study the effect of salinity on seed germination of selected mangrove species and the application of a hydrotime model to explain the relationship between water potential of the medium and rate of seed germination. Germination of seeds was examined at 15, 25 and 35°C in light/dark over a NaCl gradient. Germination time courses were prepared, and germination data were used to investigate whether these species behave according to the principles of the hydrotime model. The model was fitted for the germination of Acanthus ilicifolius seeds at 25°C. Final germination percentage was significantly influenced by species, osmotic potential and their interaction at 25°C. Moreover, temperature had a clear effect on seed germination (Sonneratia caseolaris and Pemphis acidula) which interacted with osmotic potential. Only A. ilicifolius seeds behaved according to the hydrotime principles and thus its threshold water potential was –1.8 MPa. Optimum germination rates for seeds of the other species occurred at osmotic potentials other than 0 MPa. The descending order of salinity tolerance of the tested species was Aegiceras corniculatum > Sonneratia caseolaris > Acanthus ilicifolius > Pemphis acidula > Allophylus cobbe, suggesting that the viviparous species (A. corniculatum) is highly salt tolerant compared with the non-viviparous species. The results revealed that seeds of the study species exhibited facultative halophytic behaviour in which they can germinate over a broad range of saline environments. Use of a hydrotime model for mangroves was limited as germination of their seeds did not meet model criteria.

Seeds of neem (Azadirachta indica A. Juss.), a multipurpose tropical tree, have limited desiccation tolerance. Whether their intermediate storage behaviour might be caused by an elevated percentage of 4C nuclei in the embryo at maturity was investigated. Morphological development of neem seeds was monitored on selected trees in Ouagadougou, Burkina Faso. Developing seeds were tested for germinability, and cell cycle events were determined by using flow cytometry and analysing the level of β-tubulin. Germination could occur after 8 weeks of seed development, but normal seedlings resulted only after 10 weeks or more. The change in fruit colour from green to yellow began after approximately 12 weeks of seed development. Immature, 4-week-old embryos about 2 mm in length had 15% of cells in the G2 phase and 60% in the G1 phase of the cell cycle, as indicated by 4C and 2C nuclear DNA levels, respectively. During maturation, the proportion of G2 cells declined to 3% at full maturity and slightly further after drying, and the proportion of G1 cells increased to approximately 90%. A strong β-tubulin signal was observed in tips of young embryonic radicles and cotyledons, but a weak or non-detectable signal was found in 9-week-old ones and in those from green-mature and yellow fruits. Because DNA replication and β-tubulin level were almost negligible at seed maturity, as in orthodox tomato seeds, it is suggested that these factors are not involved in the intermediate storage behaviour of neem seeds.

Hydration memory is a phenomenon in which a seed can tolerate discontinuous hydration periods, displaying enhanced germination after one or multiple hydration–dehydration (HD) cycles; it was described physiologically in a few cactus species around 15 years ago. Although no additional work was done on this subject, it has great biotechnological potential since its analysis would permit predictions about whether a seed can withstand discontinuous hydration; in the long run, the knowledge about its regulation might lead to induction of this resistance, so we aimed to provide an initial approach to the molecular mechanisms that underlie hydration memory. This phenomenon was reproduced successfully in our lab with Ferocactus peninsulae seeds. Using two-dimensional (2D) electrophoresis, we compared expression patterns of proteins involved in seed maturation of seeds and seedlings subjected to an HD cycle treatment. We found differential expression of several proteins possibly involved in primary metabolism, ubiquitination pathway and reserve protein availability regulation in seeds and seedlings subjected to an HD cycle. We also found differential stability of total RNA. These results strongly suggest that the differential expression of proteins is at least partially related to the hydration memory process.

The naturally occurring fungal seed pathogen, Pyrenophora semeniperda, reduces the seed bank of Bromus tectorum but the role of plant litter in this seed–pathogen interaction is unexplored. To investigate the direct and indirect effects of litter on this interaction, we first collected field seed-bank samples from low and high Bromus litter patches. From these data, we explored the relationship between litter depth, seed-bank density and seed mortality from P. semeniperda. Second, we manipulated the fungal stages (conidial spores and mycelium) in/on the litter through sterilization techniques, to measure the direct effect of litter on seed death. Third, for indirect effects, we manipulated litter levels and held seed density and inoculum constant to determine whether Bromus litter could modify the seed zone microsites to favour disease. We found that seed-bank samples from high-litter patches contained higher field-killed seed densities compared with low-litter patches, although the percent difference of disease between litter patch types varied among sites and years (e.g. 80% to 46%). In testing the direct effects of litter on the seed–pathogen interaction, we found that litter can act as a direct inoculum source for the pathogen in the early summer but decreases in disease transmission by the following spring when the litter naturally is in contact with seeds. Investigating indirect effects, we found four times as many pathogen-killed seeds in high-litter treatments as compared with low-litter treatments when inoculum loads and seed densities were held constant. In addition, we found that litter influences the seed–pathogen interaction through density-dependent disease transmission. Our findings demonstrate the ecological importance of litter in semi-arid environments as it influences disease levels of a seed pathogen by direct and indirect means.

Conserving alpine ecosystems and the plant communities they contain using ex situ conservation requires an understanding of seed longevity. Knowledge of seed longevity may determine the effectiveness of ex situ seed banking for alpine plant conservation, and may provide insight into plant recruitment in situ. We sought to determine the influence of elevation and climatic variables, as well as plant and seed traits, on the seed longevity of 57 species inhabiting a unique biome, (sub-)alpine regions of mainland Australia. Seed longevity was estimated using controlled accelerated ageing tests to determine the time taken for seed viability to fall by 50%. We found that, across the study species, like alpine seeds elsewhere in the world, Australian alpine seeds are relatively short-lived and overall shorter-lived than Australian plants in general. Seed mass and elevation explained most of the variation in seed longevity among the Australian alpine species considered. Species with larger seed mass, and collections made at higher elevations, were found to have relatively short-lived seeds. Phylogeny, however, explained very little of the variation in longevity. Our results suggest that viability testing for Australian alpine seeds in ex situ seed banks should be conducted with shorter intervals than for the non-alpine flora. This study highlights how seed longevity in the Australian Alps is not dictated primarily by evolutionary lineage but rather by a complex combination of environmental variables and intrinsic seed characteristics. Potential implications for conservation ex situ and in situ in the context of climate change are discussed.

From previously published studies we have learned that very fast-germinating species have some characteristics in common with cryptoviviparous species: (1) little or no endosperm/perisperm; (2) embryos fully developed and chlorophyllous at seed maturity; and (3) very fast germination, independent of temperature. Thus, we propose that these very fast-germinating seeds are ‘cryptoviviparous-like′.

Various sources of germination heterogeneity of carrot (Daucus carota L.) seeds were investigated: germination conditions (temperature, oxygen tension), size of seeds, position of the umbels on the mother plants, and pollination conditions of male-sterile plants in hybrid seed production. All seeds tested germinated over a large range of temperatures (5–35°C). However, low temperatures (5–10°C) and temperatures above 30°C reduced germination. Seeds were also sensitive to oxygen deprivation, but their sensitivity to hypoxia depended on the cultivar. The germination responses of seeds to temperature and oxygen depended on their size, particularly at sub- and supra-optimal temperatures. The largest seeds (1.8–2.1 mm) of commercial lots usually germinated better at 5°C and were more sensitive to oxygen deprivation than the smallest ones (1.2–1.8 mm). Experiments performed with open pollinated plants showed that seeds produced by umbels of first and second orders germinated faster and at higher percentages than those collected on third-order umbels. Evidence for the involvement of pollination conditions in the germination quality of carrot seeds was given by pollination by honey bees of male-sterile plants cultivated at various distances from the pollen donors. The longer the distance of the mother plants from the pollinating plants, the lower the seed yield, the heavier the mean seed weight, the more difficult the germination of seeds, and the higher their sensitivity to oxygen deprivation.

The flatulence-producing soluble oligosaccharides are an important component of lupin seeds and were assayed to establish the pattern of their accumulation in relation to germinability during seed development and maturation. Maturing yellow lupin cv. Juno seeds were harvested at 5-day intervals from 15 to 45 days after flowering (DAF). Seed fresh mass increased to a maximum at 35 DAF followed by a decrease when axis and cotyledon tissues changed colour from green to yellow. Maximum seed fresh mass corresponded to the maximum seed size. Seed dry mass continuously increased until 40 DAF. About 75% of mature seed dry mass was in cotyledons, 22% in testa, and 3% in axis. Maximum seed germinability occurred at 45 DAF after maximum seed dry mass and desiccation. Maturing yellow lupin seeds were desiccation tolerant. Mature dry seeds contained 10.9% oligosaccharides and 1.5% sucrose. During seed growth stachyose accumulation preceded accumulation of raffinose and verbascose. The highest rate of oligosaccharide accumulation appeared during seed desiccation and correlated with the acquisition of the ability to germinate. The sucrose:oligosaccharide ratio continuously decreased reaching 0.13 in mature seeds. Accumulation of oligosaccharides in maturing yellow lupin seeds is associated with seed germinability and seed desiccation tolerance.

The technique of radioimmunoassay following sample resolution by HPLC was used to assay the amounts of the cytokinins zeatin (Z), zeatin riboside (ZR) and isopentenyladenine (IPA), the combined amounts of gibberellins1+3 (GA1+3), and the amounts of indole acetic acid (IAA) and abscisic acid (ABA) during germination in grains of sorghum. Concentrations of GA1+3 were low throughout germination and did not appear to be related to the time of germination. In the mature, non-germinated grain, the concentration of each of the other plant growth regulators was much higher in the smaller component comprised of the embryonic axis and scutellum than in the much larger endosperm tissue. During the germination period studied (64 h), these concentrations declined, with a peak in the amount of the cytokinin IPA and a small peak in Z+ZR (24 h) in the embryo following the first visible signs of root protrusion and coincident with a large enhancement in amylase activity. The high concentration of ABA in the embryo tissue prior to germination was noteworthy. It is suggested that the interaction of ABA and the cytokinins IPA and Z+ZR may play a significant role in controlling sorghum germination.

Dormancy in rice (Oryza sativa L.) seed is imposed by certain physical and chemical factors associated with its covering structures, i.e.hull and pericarp. The nature of these germination blocks, their mode ofaction, and processes regulating the release of dormancy are not fully understood. Of nine rice cultivars studied, Ching-shi 15, Stejaree 45, PTB10, and Mahsuri are weakly dormant, and Bansphul, Benaful, Kataktara, Dular, and N22 are dormant. Release of seed dormancy in rice by various treatments, oxidative processes and enzymic changes associated with dormancy, and parallelism between natural and artificially imposed dormancy patterns were examined. The influence of the hull in imposing dormancy was stronger and more prolonged than that of the pericarp. Application of GA3 was effective in inducing germination only in weakly dormant cultivars. Dormancy was completely released in all cultivars by subjecting the seeds to moist heat treatment, by removing the hull and pericarp, and by applying GA3 after dehulling. Dormant cultivars had higher O2 uptake rate and peroxidase activity and lower amylase and dehydrogenase activities than the weakly dormant ones. Hull removal substantially decreased peroxidase activity but enhanced amylase and dehydrogenase activities. Nonanoic acid (C90), a short-chain saturated fatty acid (SCSFA), when exogenously applied to non-dormant seeds imposed dormancy. Dry heat treatment or presoaking in 0.01 m KNO3 or 0.1 m H2O2 was very effective in releasing SCSFA-imposed dormancy. Amylase activity was greatly reduced by treatments with nonanoic acid (C90) or ABA. Considering earlier reports and results of the present study, it is proposed that seed dormancy in rice is regulated both by the presence of SCSFAs and ABA in the hull and the pericarp. The relative significance of these substances in cultivars of tropical and temperate origins and its implications in terms of ecogeographic adaptability are discussed.

Recently, a list has been published of angiosperms capable of germinating in less than 24 h (‘very fast germination’). Here, we add three families and 23 species to that list. The main extra families complement ecological groups already recognized – the Cactaceae into the aridity-adapted group and the Tamaricaceae into the floodplain-adapted group. These amended findings on very fast germination (VFG) are integrated into the recent work on the functional ecology of embryo size. They confirm the important connection between germination speed and embryo to seed ratio. Plotting the plant orders containing VFG species on a phylogenetic tree shows that VFG has evolved multiple times throughout angiosperm history, including at least three times within the Caryophyllales. The fact that species with VFG are mainly restricted to advanced clades shows that VFG is a derived trait that evolved as an adaptation to either arid, saline or floodplain habitats.

Freshly harvested oat (Avena sativa L.) seeds are considered to be dormant because they are unable to germinate at relatively high temperatures (above 20°–25°C). This primary dormancy results partly from the structures surrounding or adjacent to the embryo (pericarp, testa and endosperm) and partly lies in the embryo itself. Incubation of imbibed dormant seeds at 30°C induces a thermodormancy which is expressed as germination in a narrower temperature range. Induction of thermodormancy is apparent after 3–8 hours at 30°C, is optimal after about 1–2 days at this temperature, but diminishes with longer treatment times. Although whole grains become more dormant as a result of such treatment, the embryos themselves become less dormant, therefore the tissues covering or adjacent to the embryos must have become more inhibiting or the embryos have become more sensitive to their inhibitory action. The germination and sensitivity to ABA or oxygen of embryos isolated from such seeds are no longer so dependent on temperature. The effect of high temperature on primary dormancy and induction of thermodormancy is discussed with reference to sensitivity to ABA and temperature.

Tomato seeds that have been dried, imbibed and redried (primed) develop internal free space between the embryo and endosperm. Seeds of the ABA-deficient sitiens(sitw) tomato mutant can exhibit internal free space at the completion of seed development even without priming. Both primed and sitwseeds germinate more rapidly than untreated wild-type seeds. To determine whether internal anatomy predicts germination physiology, individual sitwand primed wild-type seeds were sorted into three categories based upon the extent of internal free space observed non-destructively using X-radiography. Category 3 (C3, extensive free space present) sitw seeds completed germination more rapidly than all other seed categories and genotypes in water, in abscisic acid (ABA) or under far-red illumination. The force necessary to puncture the endosperm caps (and testa) of C3 sitw seeds was less, and the percentage of nuclei in C3 sitw radicle tips in the G2 stage of the cell cycle was greater than for all other seed categories. Wild-type seeds exhibited free space following long-term priming, but germination was still prevented by far-red light and ABA, and endosperm cap strength and nuclear DNA contents were not altered. Endo-β-mannanase activity of individual endosperm caps was not consistently related to their resistance to puncture. While internal free space is diagnostic for primed tomato seeds and occurs in a fraction of sitw seeds, it is not predictive of many aspects of germination physiology.

We studied seasonal variation in density and species richness of seeds in the 0–10 cm soil depth layer in primary, secondary and pine forests, and in shrubland and grassland in the Shilin Stone Forest Geographical Park, Yunnan, SW China. Soil samples were collected four times during the year at 3-month intervals. Seeds from 119 species were identified by germination tests in the soil samples. Density and species richness of seeds of herbaceous plants were greater than woody plants at all five sites throughout the year. Sampling time and site differences had significant effects on the mean number of species and on seed-bank density. Mean number of species per sample increased from February, reached the highest value in May, decreased to the lowest value in August and then increased in November. An exception was in the primary forest, where the highest number of species was found in February. Mean seed-bank density peaked in May at all five sites, and no significant differences were found between densities in February, November and August, except for the primary forest. The peak in seed-bank density in May might be due to dispersal of new seeds of spring-fruiting species, combined with persistence of seeds dispersed in previous years. This seasonal variation of individual species was due primarily to differences in species phenology rather than to differences between sites. Four seasonal seed-bank strategies were identified: two transient and two persistent. At all sites, similar numbers of seeds of herbaceous species were found between seasons, but the number of species of trees and shrubs decreased in August.

Variation in dormancy of the wetland sedge, Carex elongata L., was tested using seeds from three wild populations and the garden-grown progeny of one population. Germination experiments, comprising four combinations of temperature and light, were conducted either with fresh-matured or cold-stratified seeds, to assess the relative contribution of populations and mother plants to the total variation. Between-year variation was tested at the population level and, partly, at the individual level, using seeds collected in two consecutive years. Among-population variation accounted for 72%, and among-individual variation for 23%, of the total variance in the first experiment. Significant differences were apparent between one wild population and its garden-grown descendants. Differences in germinability among populations were maintained in the two consecutive years, but a proportion of the variance was due to the contribution of the maternal environment. Weak evidence for genetic control at the individual level was shown by a correlation across years in one population. However, the lack of a main effect at the individual level in the first experiment makes it difficult to assess the relative contribution of the mother plants to the total variation. Our results imply that germination patterns of C. elongata have a genetic basis, but are markedly modified by environmental conditions.

The germination ecology of the winter annual Iberian endemics Iberis pectinata and Ziziphora aragonensis was investigated in order to better understand adaptations of rare species to their natural habitat and to improve ex-situ propagation techniques and management of their habitat. Specifically, we analysed the following aspects: (1) influence of temperature, light conditions and seed age on germination patterns; (2) phenology of germination; (3) germinative response of buried seeds to seasonal temperature changes; and (4) temperature requirements for induction and breaking of secondary dormancy. Germination was substantially lower in darkness than with a photoperiod in both taxa, with this difference being more pronounced in Z. aragonensis. Freshly matured seeds showed conditional physiological dormancy, germinating at low and medium temperatures but not at high temperatures (28/14 and 32/18°C). Germination capability increased with time of dry storage in both species, suggesting the existence of non-deep physiological dormancy. Under greenhouse conditions, germination of both taxa was mostly concentrated in autumn (October–November), while spring percentages were less than 1% of total accumulated germination recorded during the study. I. pectinata and Z. aragonensis seeds buried and exposed to natural seasonal temperature variations in an unheated greenhouse came out of conditional dormancy in summer and re-entered it in winter, thus exhibiting an annual conditional dormancy/non-dormancy cycle. Dormant seeds of both species which were stratified at 28/14 or 32/18°C during an 8-week period, were non-dormant when they were subsequently incubated over a range of temperatures from 5 to 25/10°C. Non-dormant seeds were induced into dormancy when stratified at 5 or 15/4°C for 8 weeks, showing a particularly low germination response at high temperatures. Recommendations for wild-population reinforcement programmes and for the management of the natural habitat of both endemics are discussed.