In nature, fruit and seed production in many plants have been shown to be pollen limited. Pollen limitation is demonstrated when open-pollinated plants that are hand-supplemented (Ps) with outcross pollen produce more fruits and/or seeds than open-pollinated controls that are not hand-pollinated (Po). There are three categories of results in such studies: Ps > Po, Ps = Po and Ps < Po, in which case pollen limitation indices are positive, zero and negative, respectively. In an index widely used to calculate pollen limitation, 1 – (Po/Ps), the bounds for Ps ≥ Po are 0 to + 1, whereas the bounds for Ps < Po are 0 to –∞. The first aim of this review was to show how the pollen limitation index can be modified so that the bounds of Ps < Po are 0 and –1, whereupon the index gives equal weight to the best performer (Ps or Po) and worst performer (Ps or Po). In addition to seed quantity, pollen supplementation can affect seed quality, including germinability. Thus, our second aim was to summarize the results of studies that have also tested the effect of pollen limitation on seed germination. In short, the 30 case studies in 15 families, 16 genera and 18 species that we identified show that seed germination percentage increased, was not affected or decreased by pollen supplementation in 12, 11 and seven cases, respectively. The effect of pollen limitation on seed germination, which can be quite large, has not been considered in developing population growth models to determine the effect of pollen limitation on λ.

Seeds often survive in the soil in a dormant state, but their persistence is endangered by micro-organisms that could feed on them. Seed–microbe interactions in the soil are, however, poorly understood. We used dormant caryopses of red rice to study the defence response induced by pronase, a mixture of proteases secreted by Streptomyces griseus, a non-pathogenic bacterium. Pronase was shown to activate the plant immune reaction, indicating that its activity was recognized as a potential microbial attack. The defence reaction included extracellular alkalinization and superoxide production, and the former was necessary to activate the latter, since buffering at pH 6 inhibited the oxidative burst. Alkalinization was sufficient to trigger the oxidative burst, as superoxide production increased when caryopses were incubated in buffered solutions of increasing pH without pronase. Release of proanthocyanidins was observed, with or without pronase. These diverse mechanisms are hypothesized to cooperate in reinforcing seed protection. Finally, time profiles of superoxide production by dormant and non-dormant red rice caryopses during imbibition did not support a relationship between extracellular superoxide and dormancy breaking or germination. Thus, the role of this reactive oxygen species in red rice imbibed caryopses appears to be essentially aimed at defence against attacks by challenging micro-organisms.

The objective of this study was to evaluate the effect of day length after flowering on pollen tube elongation, embryo formation and seed development. The quinoa varieties used in this study were Amarilla de Marangani (valley type) and NL-6 (sea-level type). After sowing, the quinoa plants were cultivated in growth cabinets. From sowing to flowering, plants were exposed to a 15 h day length regime. After flowering, the plants were grown under either a 15 h or 11 h day length regime. The elongation of the pollen tube and the formation of the early embryo were not inhibited in either Amarilla de Marangani or NL-6 under the 11 or 15 h day length regimes. Although growth of the embryo in NL-6 was not inhibited by the 15 h day length regime after flowering, the same was not observed in the case for Amarilla de Marangani. In Amarilla de Marangani, seed diameter at 8 and 14 days after flowering under the 11 h day length regime was larger than that of seeds grown under the 15 h day length regime. Thus, the decrease in the number of seeds in Amarilla de Marangani grown under the 15 h day length regime may be caused by the suspension of embryo growth after fertilization.

The genus Aegilops belongs to the secondary gene pool of wheat and has great importance for wheat cultivar improvement. As a genus with only annual species, regeneration from seeds in Aegilops is crucial. In several species in Aegilops, spikes produce different seed morphs, both in size and germination patterns. However, little is known about the ecology of seed germination, nor about the seed longevity in this genus. Here we investigated the germination phenology of Ae. neglecta under laboratory and field conditions and assessed longevity of different seed morphs of five additional Aegilops species using controlled ageing tests. Large seeds were short-lived and germinated faster than small seeds in most of the species. Field experiments with Ae. neglecta showed that large seeds of the dimorphic pair germinated 3 months after dispersal in contrast to 14 months for smaller seeds. Differences in longevity were detected not only in dimorphic seed pairs, but also among seeds from different positions on the spike. Our results indicate that different longevities in seed morphs of Aegilops may reflect a different soil seed bank persistence, with smaller seeds able to maintain a higher viability after dispersal than larger ones, thereby spreading seedling emergence over two years. Differences of seed germination and longevities between seed morphs in Aegilops may have important implications for ex situ seed conservation and reinforce the hypothesis of a bet-hedging strategy in the germination ecology of this genus.

Morphophysiological dormancy (MPD) is predominantly found in seeds of temperate regions and is uncommon in arid biomes. MPD has been reported in a number of Hibbertia (Dilleniaceae) species of temperate Australia, and in a single species of the arid zone, H. glaberrima. This study aimed to examine the dormancy and germination ecology of seeds of H. glaberrima. Seeds were subjected to temperature stratification treatments designed to mimic summer and autumn conditions in the Pilbara region of Western Australia. Seed germination and embryo growth were measured. We also tested the interaction between temperature stratification and cycles of drying and wetting designed to mimic sporadic rainfall events. All temperature and moisture treatments were tested in combination (+/–) with the smoke-derived chemical karrikinolide (KAR1). Exposing dormant seeds to temperatures suitable for warm stratification (35°C) for ≥ 8 weeks, followed by incubation at 25°C, resulted in significantly higher germination compared with non-stratified seeds. Exposing seeds to dry/wet cycling in conjunction with temperature stratification did not significantly increase germination. Exposure to KAR1 increased germination under most conditions. Once seeds are shed during October to December, they are exposed to hot and sporadically wet conditions over summer, allowing MPD to be overcome in a proportion of the seed population. Seeds may germinate in autumn (March to April), in conjunction with cooler temperatures. More deeply dormant individuals may require more than one summer to overcome dormancy. Similar to other species occurring in fire-prone ecosystems, fire also plays a crucial role in the germination ecology of H. glaberrima.

Large animals tend to disperse seeds over long distances via ingestion and defecation due to their large home range and capacity to move among different habitats for feeding. The aim of this study was to investigate the potential of endozoochorous seed dispersal by five herbivores: Ovis vignei, Capra aegagrus, Gazella subgutturosa, Cervus elaphus, Capreolus capreolus and two omnivores: Sus Scrofa and Ursus arctos in the Golestan National Park, northeast of Iran, by a greenhouse germination experiment. A total of 3107 seedlings belonging to 154 different plant taxa were germinated from 655 dung samples collected in three different habitats. Plant families that most frequently germinated were Poaceae and Brassicaceae. Urtica dioica was the most abundant germinating seed, accounting for 20% of all the seedlings recorded in our dung samples, whereas the most frequently observed species was Portulaca oleracea, which occurred in 24% of our samples. We showed that 54% of the seeds germinating were dispersed by only one of the mammals studied. Herbs and graminoids were the most frequently dispersed growth forms by the herbivores and the wild boar, whereas brown bears mostly dispersed shrubs. The seedling composition in the dung samples was strongly correlated with the local flora especially for non-selective feeders, like red deer. The differences observed in the number of plants and frequency of different growth forms dispersed among the studied mammals reflect their body size, digestive physiology, and dietary and habitat preferences. Our findings highlight the different and complementary roles of large herbivores and omnivores as long-distance seed dispersal vectors.

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.

We assessed inter-seasonal dynamics of seed banks, dormancy and seed germination in three endemic Chihuahuan Desert succulent species, under simulated soil warming conditions. Hexagonal open top-chambers (OTCs) were used to increase soil temperature. Seeds of Echinocactus platyacanthus (Cactaceae), Yucca filifera and Agave striata (Asparagaceae) were collected and buried within and outside OTCs. During the course of one year, at the end of each season, seed batches were exhumed to test viability and germination. Soil temperature in OTCs was higher than in control plots. Yucca filifera seeds always had high germination independently of warming treatment and season. Agave striata seeds from OTCs had higher germination than those from control plots. Agave striata exhibited low germination in fresh seeds, but high germination in spring. Seeds from this species lost viability throughout the experimental timeframe, and had no viable seeds remaining in the soil. Echinocactus platyacanthus showed high germination in fresh seeds and displayed dormancy cycling, leading to high germination in spring, low germination in summer and autumn, and high germination in winter. Germination of this species was also higher in seeds from OTCs than those from control plots. Echinocactus platyacanthus formed soil seed banks and its cycle of inter-seasonal dormancy/germination could be an efficient physiological mechanism in a climate change scenario. Under global warming projections, our results suggest that future temperatures may still fall within the three studied species’ thermal germination range. However, higher germination for A. striata and E. platyacanthus at warmer temperatures may reduce the number of seeds retained in the seed bank, and this could be interpreted as limiting their ability to spread risk over time. This is the first experimental study projecting an increase in soil temperature to assess population traits of succulent plants under a climate change scenario for American deserts.

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.

Drying reduces seed moisture content, which improves subsequent seed survival periods. Diverse maximum temperatures have been recommended to limit or avoid damage to seeds, but some high-temperature drying regimes may improve subsequent seed quality. Seeds from 20 different accessions of five rice (Oryza sativa L.) variety groups (aromatic, Aus, Indica, temperate Japonica, tropical Japonica) were harvested over several seasons at different stages of maturation and either dried throughout at 15°C/15% relative humidity (RH) or for different initial periods (continuous or intermittent) in different drying regimes at 45°C before final equilibrium drying at 15°C/15% RH. Subsequent seed longevity in hermetic storage at 45°C with 10.9% moisture content was determined. In no case did initial drying at 45°C provide poorer longevity than drying at 15°C/15% RH throughout. There was a split-line relation, which did not differ amongst investigations, between longevity after initial drying at 45°C relative to that at 15°C/15% RH throughout and harvest moisture content, with a break point at 16.5% (a seed moisture status of about –14 MPa). Below 16.5%, relative longevity did not differ with harvest moisture content with little or no advantage to longevity from drying at 45°C. Above 16.5%, relative longevity showed a positive relation with harvest moisture content, with substantial benefit from drying at 45°C to subsequent longevity of seeds harvested whilst still moist. Hence, there are temporal (immediately ex planta cf. subsequent air-dried storage) and water status discontinuities (above cf. below 16.5%) in the effect of temperature on subsequent air-dried seed longevity.

Seed longevity is influenced by many factors, a widely discussed one of which is the seed lipid content and fatty acid composition. Here, linear and non-linear regressions based on machine learning were applied to analyse germinability and seed composition of a set of 42 oilseed rape (Brassica napus L.) accessions grown under the same single environment and at the same time following a period of up to 31 years storage at 7°C. Mean viability was halved after 27.0 years of storage, but this figure concealed a major influence of genotype. There was also wide variation with respect to fatty acid composition, particularly with respect to oleic, α-linolenic, eicosenoic and erucic acid. Linear regression (rL) revealed significant correlation coefficients between normal seedling appearance and the content of α-linolenic acid (+0.52) and total oil (+0.59). Multivariate regression using artificial neural networks including a radial basis function (RBF), a multilayer perceptron (MLP) and a partial least square (PLS) recognized underlying structures and revealed high significant correlation coefficients (rM) for oil content (+0.87), eicosenoic acid (+0.75), stearic acid (+0.73) and lignoceric acid (+0.97). Oil content or a combination of oleic, α-linolenic, arachidic, eicosenoic and eicosadienoic acids and glucosinolates resulted in highest model fitting parameters R2 of 0.90 and 0.88, respectively. In addition, the glucosinolate content, predominantly in the Brassicaceae family and ranging from 4.6 to 79.5 µM, was negatively correlated with viability (rL = ‒0.43). Summarizing, oil content, some fatty acids and glucosinolates contribute to variations in average half-life (15.2 to 50.7 years) of oilseed rape seeds. In contrast to linear regression, multivariate regression using artificial neural networks revealed high associations for combinations of parameters including underestimated minor fatty acids such as arachidic, stearic and eicosadienoic acids. This indicates that genetic and seed composition factors contribute to seed longevity. In addition, multivariate regressions might be a successful approach to predict seed viability based on fatty acids and seed oil content.

There are a number of studies describing the gross range of morpho-anatomical variability in epiphytic Tillandsia species, but the interspecific variation in seed traits remain largely unexplored, although these play an important role in determining dispersal and establishment success. In order to evaluate interspecific variation in seed morphology, anatomy and germination, we sampled six Tillandsia species from the Yucatan peninsula, Mexico, distributed along a precipitation gradient. We studied morpho-anatomical traits (seed length, seed mass, ratio of coma to seed, ratio of embryo to endosperm), seed terminal velocity in still air, and performed histochemical analyses and germination trials under controlled conditions. Tillandsia recurvata differs from the other five species in the structure of the plumose coma; it was the only species lacking an endosperm and showed distinct seedling development. Among the species, bigger seeds were related to longer comas, and had higher germinability. Overall, seed terminal velocity was invariably slow, compared with reports of other anemochorous species, suggesting a high dispersal potential. Taxonomical and ecological implications of our results are discussed.

Seeds are notoriously high in polyphenols and polysaccharides, which reduce RNA quality and yield, and interfere with downstream applications. We present simple modifications to a rapid RNA extraction protocol for use with seeds. The inclusion of polyethylene glycol in place of polyvinylpyrrolidone reduced polyphenol and polysaccharide contamination. In addition, replacing NaCl with KCl improved the RNA yield from Arabidopsis seeds still bound by mucilage. On extraction of Arabidopsis seed recovered from field soils clean RNA pellets with no accompanying gelatinous matrix (polysaccharide) were seen, with A260/230 ratios greater than 1.8 confirming the lack of polysaccharide carry-over. When Brassica oleracea and Sinapis arvensis seeds were extracted, 260/230 ratios greater than 1.8 were seen. RNA yields in excess of 10 µg per 100 mg seed suitable for RT-QPCR were obtained.