Seed persistence, desiccation tolerance, and dormancy play a crucial role in plant population and community dynamics. However, these life-history traits remain largely understudied in perennial herbaceous species, particularly in tropical ecosystems. We evaluated the seed storage behaviour, potential longevity, soil seed bank, seed dormancy alleviation in the field and the effects of after-ripening temperature and time on seed dormancy alleviation in Carajasia cangae – an endangered perennial forb endemic to the ironstone outcrops of the Eastern Amazon. We performed germination experiments to examine the effect of storage conditions (−20, 5 and 28°C, as well as field storage) and time on seed viability, mean germination time and percentage. Our results suggested that C. cangae seeds form a transient soil seed bank and show orthodox storage behaviour. The seeds' longevity was favoured in all controlled storage conditions in relation to soil-stored seeds (field). However, the marked loss of seed viability in less than 1 year, regardless of storage condition, indicates a low potential for long-term germplasm conservation through seed banking. Seed dormancy was fully alleviated after 3 months of field storage during the dry season. Moreover, seeds stored for 6 months at 28°C had their dormancy partially alleviated, indicating that environmental conditions found throughout the dry season in the species habitat are required to alleviate its seed dormancy. A transient seed bank type is favoured by predictable seasonal variations in climate in the region, species iteroparity and seed dormancy alleviation during the dry season, which delays germination until the onset of the next rainy season.

The previous study indicated that ubiquitination is involved in the freezing tolerance of hydrated seeds. Parthenolide (PN), inducing the ubiquitination of MDM2, an E3 ring-finger ubiquitin ligase, adversely affects the freezing tolerance of hydrated lettuce seeds. Therefore, a proteomics analysis was conducted to identify PN's targets in hydrated seeds exposed to cooling conditions. Several pathways, including oxidative phosphorylation (KEGG00190), amino sugar and nucleotide sugar metabolism (KEGG00520), and biosynthesis of nucleotide sugars (KEGG01250), were enriched in the PN treatment under slow-cooling conditions (3°C h−1, P < 0.05). Among the proteins in oxidative phosphorylation, the expression of NADH dehydrogenases and ATP synthases (ATPsyn) decreased in PN treatment. In contrast, uncoupling proteins increased after PN treatment, which led to the dissociation of the electron transport chain from ATP synthesis. Treatments with rotenone, dicoumarol, and oligomycin (i.e., oxidative phosphorylation inhibitors) decreased the survival rate of hydrated seeds under freezing conditions, which indicated that energy metabolism was related to the freezing tolerance of hydrated seeds. The predicted interactions between PN and MDM2-like proteins of Lactuca indicated that LsMDM2-5 forms two potential hydrogen bonds with PN. Furthermore, based on AlphaFold predictions and yeast 2-hybrid results, MDM2-5 might interact directly with NADH2. The knockdown of MDM2-5 by RNAi caused a higher level of NADH2 and ATPsyn and a higher freezing tolerance of hydrated seeds. This indicated that MDM2 played negative roles in regulating ATP synthesis and freezing tolerance of hydrated seeds.

Endozoochory, the dispersal of seeds through the animal gut passage, plays a significant role in vegetation dynamics. The success of endozoochorous seed dispersal depends on each stage of the process: ingestion by animals, gut passage, and post-dispersal events after defecation. After the deposition of seeds through feces, the effects of feces on the initial stages of seedling establishment, including seed germination and seedling growth, can significantly impact overall survival. The pattern of fecal effects on plant species depends on the animal species. In this study, we investigated the effects of feces presence on seed germination and early seedling growth using feces of the Korean water deer (Hydropotes inermis argyropus). We conducted a germination experiment on 12 plant species belonging to 10 plant families, which are known to germinate in the feces of Korean water deer. The study compared the seed germination rate and seedling length after germination between seeds sown with and without feces of the Korean water deer. In general, we found that the presence of deer feces per se had no significant effects on seed germination and early growth stages. However, additional research on post-dispersal events such as long-term growth, fecal type, and germination conditions is needed to fully understand the costs and benefits of endozoochory.

Seeds are crucial for plant reproduction, dispersal and agriculture. Seed quality and vigour greatly impact crop production by enabling rapid and uniform germination under various environmental conditions. This leads to healthy seedlings that can withstand both biotic and abiotic stresses, which are particularly important in the context of the accentuation of global climate change. Upon imbibition during germination sensu stricto, seeds release exudates, complex mixtures of organic and inorganic molecules, into the microenvironment surrounding them, known as the spermosphere. These exudates play a pivotal role in seedling development and overall plant fitness by influencing microbial selection, growth and interactions in the spermosphere, ultimately shaping the plant's microbiome. Proteins such as enzymes with protection properties have previously been demonstrated to be released by the seeds in their exudates. However, limited information is available pertaining to peptides in seed exudates. Here, we developed an experimental protocol to extract and identify peptides in the spermosphere of germinating common bean seeds. We showed that our methodology was successful in identifying a broad spectrum of peptides and that extraction solvent choice impacts peptide identification both qualitatively and quantitatively. We also show the possibility of using online prediction tools to predict the properties of identified peptides based on their amino acid sequence. We propose that this approach may be used to identify potential molecules that could be used as candidates for developing strategies to enhance seed quality and improve crop productivity.

Many plants developed unique adaptations such as protective seed coatings or eatable fruits/seeds that aid in seed dispersal by animals. Birds are among the animals that perform endozoochoric diaspore dispersal. In this study, we used mechanically demucilaginated seeds and seeds covered with artificial mucilage with the aim of determining the protective potential of the mucilage envelope against seed digestion during endozoochory by birds. We removed the mucilage from the seeds of Plantago lanceolata, Plantago ovata, Plantago psyllium, and Linum usitatissimum and fed the seeds to pigeons. Using naturally non-mucilaginous seeds of Amaranthus albus, Brassica napus and Nigella sativa, we tested the survival of diaspores covered with artificial mucilage (P. ovata husk) in the digestive systems of pigeons. Among the demucilaginated diaspores (2700 per taxon), no seeds of L. usitatissimum, 1 seed of P. lanceolata, 1 seed of P. ovata and 7 seeds of P. psyllium passed through the bird gut intact. This indicated that after removing the mucilage envelope, few diaspores can pass through the digestive system. Among diaspores with artificial mucilage, 7 seeds (from 8100 total) passed through the bird gut intact. This indicated that the artificial mucilage did not support the survival of the seeds as they passed through the pigeon digestive system. Furthermore, germination experiments revealed a decrease in the germination rate of demucilaginated seeds. They moulded rapidly, and only a few of the seeds germinated. The results of this study demonstrate the significant role of the mucilage in preventing digestion and supporting diaspore germination during endozoochory.

Seeds rely on temperature to adjust their germination timing by modulating primary and secondary dormancy. The knowledge regarding an intraspecific variation in the germination responses to supra-optimal temperatures during imbibition within the Solanum lycopersicon species and its relation with pre- and post-harvest environments is limited. Here, we studied the impact of imbibition at 35°C in 17 genotypes selected from a multiparent advanced generation intercross (MAGIC) population. We discovered a high genetic variability in the germination responses to heat, leading to thermotolerance, thermoinhibition or thermodormancy with different depths. While thermodormancy appeared more profound than primary dormancy, there was no correlation between the deepness of primary and thermodormancy. Post-harvest treatments influenced considerably germination at supra-optimal temperatures. Dry storage beyond the apparent loss of primary dormancy led to an increased proportion of thermotolerant or thermoinhibited seeds at the expense of thermodormancy in a genotype-dependent manner, thereby revealing cryptic genetic variation. Prolonged cold imbibition also led to increased thermodormancy in genotypes that produced thermotolerant and thermoinhibited seeds. The thermal history before and after flowering influenced primary dormancy and the germination response to heat during imbibition in a genotype-dependent manner, with high temperatures leading to increased thermotolerance or thermoinhibition at the expense of thermodormancy, suggesting transgenerational plasticity despite the domestication of the species. The high potential of the MAGIC population for quantitative trait loci mapping and causal polymorphism identification will be helpful in deciphering the regulatory mechanisms that lead to the plasticity of thermoinhibition or thermodormancy, as well as their connection to the parental environment.

For many decades, seed germination data have been modelled by probit analysis. In particular, it is the basis of the seed viability equation used, in the first instance, to describe the decline in germination of seeds in storage, but then also the rate of the decline, depending on seed moisture content and the temperature of storage. The underlying assumption of a probit model is that the response follows a normal distribution, in this case, loss of the ability to germinate over time. Probit analysis also takes into account the binomial error associated with germination data. Many statistical packages have probit analysis as an option within the generalized linear modelling framework; here, we present code for applying probit analysis in the free software, R. Codes are provided for fitting a single survival curve, for a single seed lot stored in a constant storage environment; for fitting multiple survival curves and evaluating the effect of constraining parameters for the different seed lots; and lastly, to model the moisture relations of seed longevity. The code bases provided could also be used in pollen and fern/bryophyte spore longevity modelling.

Although the seeds of various Orchidaceae members can be readily germinated on sterile nutrient media, numerous species exhibit complete asymbiotic germination failure. Also, while seed morphology in orchids has been previously linked to dispersal, associations with germinability have not been widely explored. We compiled seed morphometric and germination data for 203 orchid species globally, drawing from international literature and our lab's unpublished findings. Based on (a) final germination percentage and (b) pre-treatment duration, two distinct groups of asymbiotic germination behaviour emerged – termed compliant and defiant, respectively. Additionally, a statistically significant relationship between germinability and the embryo-to-seed (E:S) length ratio was observed. E:S values tended to be lower in species with defiant germination and higher in those with compliant germination. We further correlated seed morphometric and germination data with phylogenetically and ecologically significant parameters (subfamily, growth form, mycoheterotrophy level, climatic zone and habitat shadiness). Notably, lower E:S values and defiant germination predominate in orchids of shaded habitats, while compliant germination is more prevalent in open habitats. Lastly, considering the mycoheterotrophy continuum, we propose that the reduction in the E:S ratio is linked to the adaptation of various orchid lineages to shaded habitats through both an increased dispersal ability and their greater reliance on fungal symbiosis.

We have reviewed seed dormancy and germination in the Rubiaceae, the fourth-largest angiosperm family (in terms of species richness), in relation to ecology, life form, biogeography and phylogeny (subfamily/tribe). Life forms include trees, shrubs, vines and herbs, and tropical rainforest trees have the greatest number of tribes and species. The family has five kinds of embryos: investing, linear-full, linear-underdeveloped, spatulate and spatulate-underdeveloped, and seeds are non-dormant (ND) or have morphological (MD), morphophysiological (MPD) or physiological (PD) dormancy. Except for the occurrence of the investing embryo only in dry fruits of Dialypetalanthoideae, each kind of embryo is found in dry and fleshy fruits of Dialypetalanthodies and of Rubioideae. In tropical and temperate regions, there are species with ND seeds and others whose seeds have MD, MPD or PD. A complete seed dormancy profile (i.e. some species with ND seeds and others whose seeds have MD, MPD or PD) was found for tropical rainforest trees and shrubs and semi-evergreen rainforest shrubs. Dormancy-break occurs during cold or warm stratification or dry-afterripening, depending on the species. Some tropical species have long periods of dormancy-break/germination extending for 4–5 to 30–40 weeks. Soil seed banks are found in 5 and 15 tribes of Rubiaceae in tropical and temperate regions, respectively. With increased distance from the Equator, diversity of life forms and seed dormancy decreases, resulting in only herbs with PD at high latitudes. We conclude that the low species richness of Rubiaceae in temperate regions is not related to low diversity of seed dormancy/germination.

Although seed trait variations and their relationship to the ecological niche have been studied extensively at the species level, they do not necessarily reflect variations at the population level. In this study, we explored the intra-specific variation in relative embryo length, seed mass and germination speed in 40 populations of Daucus carota distributed across Europe and North America. By including information on local climate conditions, we aimed to examine the impact of the geographical origin on various seed functional traits and to detect potential local adaptation. No significant difference was observed in final seed germination for European and North American seeds incubated at 20°C, nor in seed viability. In European populations, relative embryo length significantly increased with increasing seed mass, but no such relation was found in North American populations. Larger relative embryo length at dispersal resulted in increased germination speed in both European and North American populations. Populations in drier areas typically had seeds with larger relative embryo lengths. Precipitation-related climate variables showed a negative relationship with relative embryo length, indicating a reduction in relative embryo length with increased precipitation. No clear relationship between climate and seed mass was observed. We can conclude that seed functional traits of D. carota are adapted to local climate conditions, as a clear gradient was observed in the relative embryo length of D. carota, which was associated with germination speed and climate. This gradient was less pronounced in North America, which can be explained by its relatively recent introduction to the continent.

The endemic tree Neltuma caldenia Burk. and the shrub Neltuma flexuosa var depressa F.A. Roig (Fabaceae; subfam: Mimosoideae) are two promising species from the central region of Argentina, with high potential for use in the restoration of disturbed environments, for extensive livestock grazing and apiculture. Both species have seeds with physical dormancy. Ecological study of native species is important from the point of view of rehabilitation of degraded areas by natural regeneration or via seed-based programmes. The objective of this study was to evaluate soil seed bank persistence and seedling traits to understand variation among different populations of each of the study species growing along an ecological gradient and to identify potential components driving this variation. Home environments influenced seed bank persistence, which was higher in populations originating from more arid and unpredictable environments where it could act as a bet-hedging strategy between years and seasons. We also observed differences associated with seedling growth traits between the species and populations. Populations with higher seed persistence were associated with greater seedling growth in N. caldenia. The rapid elongation rate of N. caldenia seedlings growing in large populations and unstable environments could help their ability to escape drought by accessing deeper soil moisture and would confer a high relative competitive ability. In contrast, N. f. var depressa displayed higher seed persistence, which was associated with more arid and unstable conditions and correlated with lower seedling growth, possibly due to an inbreeding depression effect, resulting from the presence of mother plants in low-densities or being isolated from other individuals. There was no effect of population size on seed persistence. To better understand seed persistence and associated seedling trait variation, future studies need to consider the genotype, environment and landscape conditions.

Seed longevity influences the success of ex situ storage and preservation of plant genetic diversity and is thus a critical factor in conservation efforts. Rapid seed ageing experiments at high temperature and high humidity have been widely used to classify seed longevity for hundreds of plant species, with potential implications for longevity in ex situ conservation. In this approach, radicle emergence (R) is normally used as a measure of the viability of the seeds. However, R could overestimate the level of normal seedling development and, consequently, the perceived longevity of seeds. Here, seed lifespan for 33 alpine species was compared to assess whether germination criteria could affect seed longevity parameters. Seeds were exposed to controlled ageing [45°C, 60% relative humidity (RH)] and regularly sampled for germination assessment as both radicle emergence (R) and radicle plus cotyledon emergence (R + C). The time taken in storage for viability to fall to 50% (p50) was determined using probit analysis, including either R or R + C data. A coefficient of overestimation of seed longevity (OESL, %) was determined. The results highlight significant differences in seed longevity estimates both across species and the germination criteria. For 17 species, seed longevity estimated by R was significantly higher than that estimated using R + C, resulting in large variation in OESL (0.54–9.01 d). The introduction of OESL facilitates effective screening for seed longevity and recovery, enhancing the overall efficiency of conservation strategies for diverse species.

Nypa fruticans Wurmb is both a relic plant and a true mangrove. In China, wild populations are distributed only on Hainan Island and face significant challenges in regeneration from seedlings. This study explored the underlying causes of recruitment limitation by examining seed morphological traits from three distinct populations (Haikou, Wenchang and Wanning) and analysing seed germination and seedling growth characteristics under varying conditions. The key findings are as follows: fruiting and seed-setting rates for N. fruticans were notably low, standing at only 21 and 40%, respectively. The Wanning population exhibited significantly higher rates compared to the other two populations. Under natural conditions, the germination and seedling emergence rates were also modest, at 36.58 and 22.99%, respectively. The germination and emergence rates of the Wanning population were significantly greater than those of the Haikou and Wenchang populations. Meanwhile, seeds from a single population did not differ in germination rates among three in situ N. fruticans habitats, but seedling emergence rates differed significantly. Optimal conditions for seed germination involved a light intensity of 60%, a salinity of 5‰ and a flooding time of 8 h/day. In natural settings, these three environmental factors fall short of the ideal conditions. The study underscores that light, salinity and flooding are primary factors contributing to the limitations in N. fruticans seedling recruitment. In addition to advocating increased investment in scientific research and technology to address seed source issues, we recommend heightened efforts in habitat restoration, in situ conservation and the optimization of relocation and field return strategies to bolster N. fruticans populations.

The performance of plants in any one generation can be influenced not just by the prevailing biotic and abiotic factors, but also by those factors experienced by the parental generation. These maternal effects have been recorded in an array of plant species, but most studies tend to focus on abiotic factors over two generations. Here we show that maternal effects in the annual forb Senecio vulgaris may be influenced by beneficial arbuscular mycorrhizal fungi and insect herbivory over four successive generations. These effects were very much determined by seed provisioning, wherein C:N:P ratios were altered by both fungi and aphids. There was little evidence of epigenetic changes induced by the fungi or insects, instead the driving forces seemed to be allocation of N and P to the seeds. However, changes in seed chemistry were not cumulative over generations, often decreases in seed nutrient content were followed by recovery in subsequent generations. The changes in seed stoichiometry can have important consequences for viability, germination and subsequent seedling growth rates. We conclude that studies of maternal effects need to be conducted over multiple generations, and also need to be multifactorial, involving variation in abiotic factors such as water and nutrients, combined with biotic factors.

Seed genebanks must maintain collections of healthy seeds and regenerate accessions before seed viability declines. Seed shelf life is often characterized at the species level; however, large, unexplained variation among genetic lines within a species can and does occur. This variation contributes to unreliable predictions of seed quality decline with storage time. To assess variation of seed longevity and aid in timing regeneration, ten varieties of pea (Pisum sativum L.), chickpea (Cicer arietinum L.) and lentil (Lens culinaris Medikus subsp. culinaris) from the Australian Grains Genebank were stored at moderate temperature (20°C) and moisture (7–11% water, relative humidity [RH] ~30%) and deterioration was assessed by yearly germination tests for 20 years. Decline in germination was fit to a sigmoidal model and the time corresponding to 50% germination (P50) was used to express seed longevity for each genetic line. The feasibility of using RNA fragmentation to assess changed seed health was measured using RNA integrity number (RIN) from RNA extracted from seeds that were stored for 13 and 20 years. Seed lots of legume grains that maintained high survival throughout the 20 years (i.e. they aged slower than other lines) had higher RIN than samples that degraded faster. RIN was lower in embryonic axes compared with cotyledons in the more deteriorated samples, perhaps indicating that axes exhibit symptoms of ageing sooner than cotyledons. Overall, RIN appears to be associated with longevity indicators of germination for these legumes and indicating that RIN decline can be used to assess ageing rate, which is needed to optimize viability monitoring.

Seeds of many Amazonian floodplain forest trees are dispersed during high-water periods and spend weeks or months underwater until the flooding retreats. To assess whether prolonged seed submersion affects germination and early seedling development, an experiment was carried out in a greenhouse with seeds of Campsiandra laurifolia, Cassia leiandra, Crataeva tapia, Ilex inundata, Macrolobium acaciifolium, Nectandra amazonum, Pouteria glomerata, Psidium acutangulum, Sorocea duckei, and Vitex cymosa. They are common in this type of forest, differ in fruit type, number of seeds per fruit, fruit dimensions, and fresh mass and have fruits or seeds that can float. Seeds were collected in a Central Amazonian floodplain forest (flooded approximately 6 months year−1; water column of 5 m) and germinated in (1) irrigated soil or (2) underwater (water column of 5–7 cm) for 6 months. Seeds that germinated underwater were transferred to drained soil. Seeds of all species germinated underwater and developed seedlings when transferred to soil. However, submersion reduced the germination percentage of Psidium acutangulum, N. amazonum, P. glomerata and V. cymosa. Six species delayed germination in water. Ca. leiandra, I. inundata and P. glomerata did not differ in mean germination time in drained soil and underwater, whereas S. duckei seeds germinated faster underwater. Seed submersion negatively affected seedling growth (shoot length) of three species but did not affect seedling biomass. Timing of fruit dispersal, fruit buoyancy and high number of seeds per fruit can be critical for species with seeds that are not as able to cope with long-term submersion.

The environment experienced by a plant before and after reproduction can have a profound effect on the behaviour of the progeny after shedding. Maternal environmental effects on seeds are important for phenology and fitness in plants, especially for bet-hedging reproductive strategies. Maternal tissues that disperse with seeds are important for dormancy in many species, particularly those with coat-imposed dormancy and those that disperse in indehiscent fruits. Maternal nitrogen status, temperature and photoperiod modify maternal tissues and also influence the developing zygote. During seed development on the mother, the progeny may acquire environmental information directly, but there is also evidence for maternal–filial signalling and the epigenetic inheritance of environmental information through the germline.

Several experimental tools allow researchers to manipulate environmental variables to simulate future climate change scenarios during in situ seed ecology studies. The most common ones are designed to modify a single environmental variable. For example, open-top chambers (OTCs) increase temperature or rain-out shelters decrease precipitation. However, changes in environmental variables in the future are expected to happen simultaneously, and at present, an understanding of their combined effects in natural environments is limited. Here, we present a passive novel OTC design that simultaneously increases the soil temperature and decreases soil moisture. We assessed the performance of the design during 1 year in a high-mountain environment and reported its effects on the organic and topsoil layers. The modified OTC reduced the soil volumetric water content throughout the study period. Overall, chambers increased the mean day air temperature by 3.3 °C (at 10 cm above the soil surface), the mean day soil surface temperature by 1.35 °C and the mean day below the soil surface temperature by 1.30 °C (at −5 cm) and 1.25 °C (at −10 cm). Remarkably, surface and soil temperatures remained warmer at night (+0.65 at soil surface, +0.41 at −5 cm and +0.24 at −10 cm). We detail the design plans, tools and materials needed for its construction. Furthermore, we recommend on how to use it during seed ecology studies. This tool can help increase our understanding of the potential responses of seeds and seedlings to the combined effects of warming temperatures and a decrease in precipitation.