Seeds of Aconitum napellus subsp. castellanum were physiologically dormant at maturity in early autumn, with underdeveloped embryos. Thus they have morphophysiological dormancy (MPD). Embryos in fresh seeds were on average 1.01 mm long, and they had to grow to 3.60 mm before radicle emergence. Cold stratification at 5°C for 5 months with light enhanced the mean embryo length to 2.73 mm (SE = 0.13) and seed germination to 20%. However, with higher temperatures (15/4, 20/7, 25/10, 28/14 and 32/18°C) embryo growth was small, with no seeds germinating. Optimal germination was achieved after 4 months of cold stratification at 5°C followed by incubation at 20/7°C for 1 month with light, when germination ranged between 70 and 79%, depending on seed age, locality and year of collection. Cold stratification could be substituted by the application of GA3 solution, since mean embryo length in seeds incubated at 25/10°C for 1 month with light was 3.52 mm and the germination was 80%. Since cold stratification was the only requirement for the loss of MPD, the longest embryo growth occurred during this treatment, and GA3 promoted MPD loss, we concluded that A. napellus seeds have intermediate complex MPD. Germination was higher in 4-month stored than in freshly matured seeds. A pronounced variability in germinative patterns at inter-annual and inter-population level was recorded.

Proteomic approaches based on two-dimensional gel electrophoresis, mass spectrometry and database search are widely used to address questions about the development, physiology and quality of seeds. Identification of proteins is of great importance in proteomic analyses. For seed crops without full genome information, cross-species protein identification by mass spectrometry-driven sequence similarity search can be used. However, this approach is risky due to protein polymorphism between different species. Species-specific expressed sequence tag (EST) databases are an invaluable resource, which complements mass spectrometry data analysis for protein identification. Here, we illustrate a modified method of protein identification and characterization using species-specific EST databases and peptide mass fingerprinting with an example of protein identification. This method is reliable, supplements the existing methods, and improves the efficiency and accuracy of protein identification for seed crops for which complete genome information is not available.

On the basis of previous research, we predict that Mediterranean grasslands should show larger-seeded annuals in: (1) more-arid grasslands; (2) more-fertile soils; (3) less-grazed grasslands; and (4) grasslands with lower intensities of seed predation by ants. To test these predictions, we set 29 sampling units of 50 m × 50 m in a 1000 km2 grassland area in Central Spain, and characterized them according to the former factors. We then recorded annual vegetation using ten quadrats of 20 cm × 20 cm in each sampling unit. Seed size at the community level was described using six variables: (1) mean seed mass; (2) standard deviation of seed mass; (3) weighted mean seed mass (by species frequencies); (4) proportion of small-seeded annuals; (5) proportion of medium-seeded annuals; and (6) proportion of large-seeded annuals. Most climate variables (mean annual temperature, length of the summer drought, water balance and mean annual precipitation) correlated with seed-size descriptors, showing that large-seeded annuals increase in warmer and more-arid communities. Mean seed size was modelled as a function of mean annual temperature and grazing pressure. According to this model, warmer and less-grazed communities tend to show a smaller mean seed size. These results confirm the importance of seed-size descriptors at the community level in Mediterranean grasslands, and the role of climate and grazing as major drivers in these communities. Conversely, hypotheses about soil fertility and seed predation by ants were not supported by our results.

Antioxidants protect seeds from oxidative damage during storage, supporting the maintenance of seed viability and the ability to germinate post-storage. No data on antioxidants during long-term storage are available for the seeds of the halophyte Suaeda maritima. Therefore, changes in lipid-soluble antioxidants in the tocopherol family (α-, γ-, δ-tocopherol), were investigated in seeds stored for up to 16 years at 4°C at a seed moisture content of 10–13%, as well as changes in the water-soluble antioxidant glutathione (GSH) and electrolyte leakage. Seed oil content was also measured and determined to be 22%. During the first 3 years of storage, seed viability remained high and the concentration of total tocopherol was stable. Thereafter, both seed viability and α-tocopherol concentration rapidly decreased and electrolyte leakage increased, while γ-tocopherol and δ-tocopherol concentrations did not correlate with seed viability. Although the concentrations of neither GSH nor glutathione disulphide (GSSG) alone were correlated with seed viability, the glutathione half-cell reduction potential (EGSSG/2GSH) was strongly correlated with viability throughout storage and increased before the onset of viability loss. Hence, in this species EGSSG/2GSH appeared to be an ‘early warning’ system preceding viability loss while α-tocopherol concentration changed concomitantly with viability.

Detecting the presence of genetically modified plants (adventitious presence of unwanted transgenic plants, AP) from outcrossing species such as maize requires a method that lowers laboratory costs without losing precision. Group testing is a procedure in which groups that contain several units (plants) are analysed without having to inspect individual plants, with the purpose of estimating the prevalence of AP in a population at a low cost without losing precision. When pool (group) testing is used to estimate the prevalence of AP (p), there are sampling procedures for calculating a confidence interval (CI); however, they usually do not ensure precision in the estimation of p. This research proposes a method to determine the number of pools (g), given a pool size (k), that ensures precision in the estimated proportion of AP (that is, it ensures a narrow CI). In addition, the study computes the maximum likelihood estimator of p under pool testing and its exact CI, considering the detection limit of the laboratory, d, and the concentration of AP per unit (c). The proposed sample procedure involves two steps: (1) obtain a sample size that guarantees that the mean width of the CI () is narrower than the desired width (ω); and (2) iteratively increase the sample size until is smaller than the desired width (ω) with a specified degree of certainty (γ). Simulated data were created and tables are presented showing the different possible scenarios that a researcher may encounter. An R program is given and explained that will reproduce the results and make it easy for the researcher to create other scenarios.

The dynamics of many plant populations essentially depend upon seed and seedling stages, and a persistent seed bank may give species an opportunity to disperse through time. Seed burial is a decisive prelude to persistence and may strongly influence seed-bank dynamics. The fate of buried seeds depends on species-specific traits, environmental conditions and possibly also burial mode. We tested seed germination, seedling emergence and growth of the co-occurring herbaceous flood-meadow species Arabis nemorensis, Galium wirtgenii, Inula salicina, Sanguisorba officinalis and Selinum carvifolia in response to the experimental manipulation of burial depth (0, 1, 2, 4, 8, 12 cm) and substrate type (sand, clay). Increasing burial depth led to decreased germination, emergence and growth in all species studied, and seedling growth differed significantly between substrate types. The responses of species differed on an individual basis, but also showed a higher-ranking pattern based on seed size. Larger-seeded species were able to emerge from greater depths and experienced less depth-mediated growth inhibition than smaller-seeded species, which, in turn, had higher survival rates during burial and were less likely to experience fatal germination. Based on these results, we suggest that herbaceous flood-meadow species have developed two different seed-size based strategies for coping with the extreme recruitment conditions prevailing in flood meadows, the balance of which seems to be maintained by disturbance events.

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.