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.

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.

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.

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.

Seed germination is a pivotal period of plant growth and development. This process can be divided into four major stages, swelling absorption, seed coat dehiscence, radicle emergence and radicle elongation. Cupressus gigantea, a tree native to Tibet, China, is characterized by its resistance to stresses such as cold, and drought, and has a high economic and ecological value. Nevertheless, given its unique geographic location, its seeds are difficult to germinate. Therefore, it is crucial to explore the mechanisms involved in seed germination in this species to improve the germination efficiency of its seeds, thereby protecting this high-quality resource. Here, our findings indicate that seed germination was enhanced when exposed to a 6-h/8-h light/dark photoperiod, coupled with a temperature of 20°C. Furthermore, the application of exogenous GA3 (1 mg/ml, about 2.9 mM) stimulated the germination of C. gigantea seeds. Subsequently, proteomics was used to detect changes in protein expression during the four stages of seed germination. We identified 34 differentially expressed proteins (DEPs), including 13 at the radicle pre-emergence stage, and 17 at the radicle elongation stage. These DEPs were classified into eight functional groups, cytoskeletal proteins, energy metabolism, membrane transport, stress response, molecular chaperones, amino acid metabolism, antioxidant system and ABA signalling pathway. Most of them were found to be closely associated with amino acid metabolism. Combined, these findings indicate that, along with temperature and light, exogenous GA3 can increase the germination efficiency of C. gigantea seeds. Our study also offers insights into the changes in protein expression patterns in C. gigantea seeds during germination.

Seed dormancy is an important trait associated with pre-sprouting and malting quality in barley. Genome-wide association studies (GWASs) have been used to detect quantitative trait loci (QTLs) underlying complex traits in major crops. In the present study, we collected 295 barley (Hordeum vulgare L.) accessions from Australia, Europe, Canada and China. A total of 25,179 single nucleotide polymorphism (SNP)/diversity arrays technology sequence markers were used for population structure, linkage disequilibrium and GWAS analysis. Candidate genes within QTL regions were investigated, and their expression levels were analysed using RNAseq data. Five QTLs for seed dormancy were identified. One QTL was mapped on chromosome 1H, and one QTL was mapped on chromosome 4H, while three QTLs were located on chromosome 5H. This is the first report of a QTL on the short arm of chromosome 5H in barley. Molecular markers linked to the QTL can be used for marker-assisted selection in barley breeding programmes.

Alpine plant species are particularly vulnerable to climate change. Temperature fluctuations are projected to be most severe at high elevations. Even small shifts in temperature have major consequences on phenology, reproduction, and community composition. Early life stages are arguably the most important processes in the fitness of an individual plant and the dynamics and persistence of plant populations. These initial developmental stages are expected to be more vulnerable to changes in climate than adult life stages. To understand how early life stages of alpine plant species will respond to warming temperatures, seeds and seedlings of two species were exposed to three different temperature regimes. Temperatures were based on current and projected conditions under low and high emission scenarios. Two rare alpine species performed better under warmer temperatures at both the germination and seedling stages. The results show that early life stages of alpine plants may not be at high risk from warming temperatures; however, there are many other shifting climatic factors to consider, resulting from climate change beyond temperature alone.

The assessment of seed quality and physiological potential is essential in seed production and crop breeding. In the process of rapid detection of seed viability using tetrazolium (TZ) staining, it is necessary to spend a lot of labour and material resources to explore the pretreatment and staining methods of hard and solid seeds with physical barriers. This study explores the TZ staining methods of six hard seeds (Tilia miqueliana, Tilia henryana, Sassafras tzumu, Prunus subhirtella, Prunus sibirica, and Juglans mandshurica) and summarizes the TZ staining conditions required for hard seeds by combining the difference in fat content between seeds and the kinship between species, thus providing a rapid viability test method for the protection of germplasm resources of endangered plants and the optimization of seed bank construction. The TZ staining of six species of hard seeds requires a staining temperature above 35 °C and a TZ solution concentration higher than 1%. Endospermic seeds require shorter staining times than exalbuminous seeds. The higher the fat content of the seeds, the lower the required incubation temperature and TZ concentration for staining, and the longer the staining time. And the closer the relationship between the two species, the more similar their staining conditions become. The TZ staining method of similar species can be predicted according to the genetic distance between the phylogenetic trees, and the viability of new species can be detected quickly.

Climate variability is expected to increase due to climate change, with projected increases in temperature and erratic rainfall patterns. These changes will alter the environmental cues sensed by seeds, and therefore will impact plant recruitment. This study investigated the effects of seed functional traits (germinability, germination time, synchrony and seed mass) on germination responses of several sub-tropical native Australian plant species under different environmental factors (water stress, salinity and pH). The effect of a hot water pre-treatment was also tested on Fabaceae seeds with known physical dormancy. Seed traits, environmental factors and seed pre-treatments had significant effects on final germination percentage and germination time. Seed mass and time to 50% germination (t50) were also positively correlated. In contrast, pH did not affect germination and there was no interaction between pH and any of the measured seed functional traits. Some species showed a high thermal tolerance to germination and germination was indifferent to light conditions for all species. Results showed that certain seed functional traits interact with environmental factors to influence germination percentage and time. These findings highlight the importance of considering seed functional traits when determining a species germination response under a changing climate. In addition, the findings provide important knowledge to better guide seed-based land restoration programmes.

Desiccation tolerance (DT) of seeds, one of the plant's environmental adaptation mechanisms, allows them to survive as seeds in a quiescent state under extremely water-deficient conditions during the plant's life cycle, followed by seed germination and seedling establishment under favourable water conditions. The seed-DT is lost after radicle emergence; however, there is a developmental period called the ‘DT window’ during which the germinating seeds can re-induce DT following a cue from their ambient low water potential (i.e. mild osmotic stress). The DT re-inducibility within the DT window has been used as a model biosystem for understanding molecular mechanisms that activate/supress DT in a number of plant species. However, the characteristics of the DT window for species particularly important to the agroindustry are still largely fragmented. Here, physiological analyses were performed, aiming to elucidate the properties of the DT window in tomato, a model species for Solanaceae, holding a key strategic position for the seed industry and commercial use around the world. We revealed that (i) the DT window of tomato seeds is closed when the developing radicle reaches about 4 mm after germination, (ii) the most effective ambient water potential to re-induce DT into seeds is about −1.5 MPa and (iii) there is organ specificity of DT re-induction with hypocotyls, showing a longer DT window than cotyledons and roots in post-germination seeds.

Statistical significance, or lack thereof, is often erroneously interpreted as a measure of the magnitude of effects, correlations between variables or practical relevance of research results. However, calculated P-values do not provide any information of this sort. Alternatively, effect sizes as measured by effect size indices provide complementary information to results of statistical hypothesis testing that is crucial and necessary to fully interpret data and then draw meaningful conclusions. Effect size indices have been used extensively for decades in the medical, psychological and social sciences but have received scant attention in the plant sciences. This Technical Update focuses on (1) raising awareness of these important statistical tools for seed science research, (2) providing additional resources useful for incorporating effect sizes into research programmes and (3) encouraging further applications of these tools in our discipline.