The Genebank Standards for Plant Genetic Resources recommend that genebanks periodically monitor the change in viability of their stored germplasm to ensure timely regeneration. The Australian Grains Genebank (AGG) has been recording germination data on orthodox seeds from tropical grain crops, stored under long-term storage (−20°C), for 40 years. Real-time viability data, collated from germination tests carried out on seedlots which had been in storage for a minimum of 20 years, from six agriculturally important grain crops (Sorghum bicolor, Phaseolus vulgaris, Glycine max, Vigna radiata, Cajanus cajan and Vigna angularis) was analysed by probit analysis. For each species independently, a common loss in viability was observed for all seedlots showing a consistent decline in viability during storage; with observed longevity estimates (σ; time for viability to fall by 1 NED/probit) of 17.4, 30.7, 33.2 49.6, 48.8, 63.5 and years for Vigna angularis, Cajanus cajan, Phaseolus vulgaris, Glycine max, Sorghum bicolor, and Vigna radiata, respectively. Common values of σ were subsequently used to determine species-specific viability monitoring intervals, based on the results of the last germination test. Dynamic monitoring intervals are a cost-efficient strategy that will avoid the over-use of seed through too frequent viability monitoring whilst still ensuring the timely regeneration of material. With funding shortfalls often reported as the main contributing factor to regeneration and viability testing backlogs, the ability for genebanks to maximise cost-efficiencies, where possible, is paramount to secure the genetic integrity of stored germplasm; particularly as collections continue to grow.

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.

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.

The plant species Sesbania tomentosa (‘ōhai; Fabaceae) is endemic to the Hawaiian Islands, federally listed as endangered in the USA and has been proposed for categorization as Vulnerable on the IUCN Red List. In 2021, c. 12,000 seeds from 12 seed lots collected during 1990–1992 from across the Hawaiian Islands were discovered in ambient herbarium conditions (55% relative humidity (RH) at 20 °C). International gene bank standards suggest drying seeds in equilibrium with 15% RH and stored at −18 °C. To investigate seed viability, we mechanically scarified then sowed 15 seeds from each accession at daily alternating regimes of 12 h light and 12 h dark at temperatures of 25/15 °C, respectively. Germination was observed after 7 days and ended after 34 days. Mean final germination was 88.9 ± SD 0.1% (range 73–100%). Each seed lot was accessioned into the National Tropical Botanical Garden's Seed Bank and Laboratory. In seeds with a water-impermeable seed coat (i.e. physical dormancy), such as S. tomentosa, seeds can desorb but not absorb water. Therefore, if the seeds were initially dried, although exposed to high RH for up to 32 years, seed equilibrium RH may have remained low, which may in part explain the observed high germinability. This study holds significance for managers who are working to conserve this endangered Hawaiian species and suggests that even suboptimal conditions may still yield highly viable seeds several decades into the future.

Cumulative oxidation of cellular macromolecules during storage reduces seed longevity. This study was undertaken to unravel the physiological and biochemical changes in rice seeds that contribute to deterioration during storage. Rice seeds maintained at three different seed moisture contents (SMC; 10, 12 and 14%) were stored in airtight glass jars. Half of the jars were flushed with nitrogen gas to provide modified oxygen conditions, while the other half were sealed with natural air (21% O2). Seed quality in terms of germination and antioxidant defence mechanisms was monitored after 3 and 6 months of storage at 25°C. The results showed that seeds performed better when stored at low SMC (10 and 12%), whereas the deterioration process accelerated in seeds stored at higher SMC (14%). Coupling high SMC with the availability of oxygen in the storage environment produced a negative effect on seed quality and longevity. Results from the antioxidant analysis showed more activity in seeds stored with oxygen at high SMC (14%) compared to lower SMC stored in modified oxygen conditions. Therefore, it is recommended that storage with low moisture levels (12%) or below (10%) is the best to preserve rice seed quality. However, at higher moisture levels (14%), the availability of oxygen in storage is more harmful to seed lifespan and quality.

Sexual propagation of Agave plants is an incipient cultivation method, these plants withstand drought and adverse growing conditions; therefore, research on Agave’s diversity, seed processing, and storage could support its cultivation on marginal lands. The aim of this work was to evaluate seed morphology, germination, and seedling genetic diversity of six seed origins (species × provenance) of Agave plants collected in five provenances from Mexico. Seed longevity was evaluated in two seed origins after a 10-year storage period. Seed morphology and seedling genetic variation results demonstrated intra- and interspecific variation within Agave salmiana and with the other seed origins, respectively. After a 10-year storage period seed germination of two A. salmiana seed origins remained relatively stable, storage conditions, and seed variables of this work can serve as reference parameters for future analyses. To the best authors’ knowledge, this is the first report of Agave’s seed longevity evaluation after a 10-year storage period.

The J. Derek Bewley Career Lectures presented at the triennial meetings of the International Society of Seed Science support early-career seed scientists by providing retrospective views, from those late in their careers, of lessons learned and future implications. Ambition, ability, inspiration, foresight, hard work and opportunity are obvious career requirements. The importance of mentoring and teamwork combined with the clear communication of results, understanding and ideas are emphasized. The role of illustration in research, and its dissemination, is outlined: illustration can support hypothesis development, testing and communication. Climate change may perturb the production of high-quality seed affecting conservation as well as agriculture, horticulture and forestry. An illustrative synthesis of the current understanding of temporal aspects of the effects of seed production environment on seed quality (assessed by subsequent seed storage longevity) is provided for wheat (Triticum aestivum L.) and rice (Oryza sativa L.). Seed science research can contribute to complex global challenges such as future food supplies from seed-propagated crops in our changing climate whilst conserving biological diversity (through seed ecology and technologies such as ex situ plant genetic resources conservation by long-term seed storage in genebanks), but only if that research can be – and then is – applied.

Serotiny is a strategy in which the retention of mature seeds in parent structures allows plants to cope with environmental variability like heat, drought or fire. Although this phenomenon might be common in Cactaceae, and particularly in Melocactus, it has generally been scarcely addressed. The main goal of our work is to investigate if there are seeds hidden in the cephalium of Melocactus matanzanus and if there are, determine whether or not these seeds maintain their viability. We also discuss some advantages the cephalium may offer as diaspore after the death of individuals. Cephalia collected from dead individuals were divided into four slices and their seeds counted; we also assessed the viability and photoblastic response of the seeds by using growth chambers at 25/30°C, and by a cut test on the seeds that did not germinate. Our results showed retention of viable seeds of different ages in all slices of the cephalium. Seeds were photoblastic positive with germination between 11–22% and viability above 50% in the portion of the lots that did not germinate.

UK trees require increased conservation efforts due to sparse and fragmented populations. Ex situ conservation, including seed banking, can be used to better manage these issues. We conducted accelerated ageing tests on seeds of 22 UK native woody species, in order to assess their likely longevity and optimize their conservation in a seed bank. Germination at four ageing time points was determined to construct survival curves, and it was concluded that multiple samples within a species showed comparable responses for most species tested, except for Fraxinus excelsior. Of all species studied, one could be classified as very short-lived, four as short-lived and 17 as medium, with none exceeding the medium category. The most important finding of this manuscript is that although some taxonomic trends were observed, the results indicate the need for caution when making broad conclusions on potential seed storage life at a species, genus or family level. Longevity predictions were compared to actual performance of older collections held in long-term storage at the Millennium Seed Bank, Kew. Although most collections remain high in viability in storage after more than 20 years, for short-lived species at least, there is some indication that accelerated ageing predicts longevity in seed bank conditions. For species with reduced potential longevity, such as Fagus sylvatica and Ulmus glabra, additional storage options are recommended for long-term gene banking.

Vitamin E is known to scavenge lipid peroxy radicals and has a purported role in preventing seed deterioration during storage. In our previous studies using 20 rice varieties from different variety groups, the specific ratio of vitamin E homologues rather than total vitamin E content was associated with seed longevity. To validate this result, we extended the experiment to a rice panel composed of 185 Aus (semi-wild rice) varieties. Seed longevity values were determined through storage experiments at 45°C and 10.9% seed moisture content (MC). Eight types of vitamin E homologues (α-, β-, γ- and δ-tocopherol/tocotrienol) were quantified by ultra-performance liquid chromatography. The theoretical initial viability in NED, Ki, was positively correlated with γ- and δ-tocopherols and negatively correlated with α-tocotrienol. The time for viability to fall to 50% during storage at elevated temperature and relative humidity, p50, was positively correlated with δ-tocopherol. The harvest MC was negatively correlated with all seed longevity traits. Taking this factor into account in a genome-wide association (GWA) analysis, we were able to correct false positives. A consistent major peak on chromosome 4 associated with −σ−1 was detected with a mixed linear analysis. Based on rice genome annotation and gene network ontology databases, we suggest that RNA modification, oxidation–reduction, protein–protein interactions and abscisic acid signal transduction play roles in seed longevity extension of Aus rice. Although major GWA regions were not overlapped across traits, three genetic markers, on chromosomes 1, 3 and 4, were associated with both δ-tocopherol and Ki and two markers on chromosome 1 and 8 were associated with both δ-tocopherol and p50.

Drought and high temperature each damage rice (Oryza sativa L.) crops. Their effect during seed development and maturation on subsequent seed quality development was investigated in Japonica (cv. Gleva) and Indica (cv. Aeron 1) plants grown in controlled environments subjected to drought (irrigation ended) and/or brief high temperature (HT; 3 days at 40/30°C). Ending irrigation early in cv. Gleva (7 or 14 days after anthesis, DAA) resulted in earlier plant senescence, more rapid decline in seed moisture content, more rapid seed quality development initially, but substantial decline later in planta in the ability of seeds to germinate normally. Subsequent seed storage longevity amongst later harvests was greatest with no drought because with drought it declined from 16 or 22 DAA onwards in planta, 9 or 8 days after irrigation ended, respectively. Later drought (14 or 28 DAA) also reduced seed longevity at harvest maturity (42 DAA). Well-irrigated plants provided poorer longevity the earlier during seed development they were exposed to HT (greatest at anthesis and histodifferentiation; no effect during seed maturation). Combining drought and HT damaged seed quality more than each stress alone, and more so in the Japonica cv. Gleva than the Indica cv. Aeron 1. Overall, the earlier plant drought occurred the greater the damage to subsequent seed quality; seed quality was most vulnerable to damage from plant drought and HT at anthesis and histodifferentiation; and seed quality of the Indica rice was more resilient to damage from these stresses than the Japonica.

The long-standing hypothesis that seed quality improves during seed filling, is greatest at the end of seed filling, and declines thereafter (because seed deterioration was assumed to begin then), provided a template for research in seed quality development. It was rejected by investigations where seed quality was shown to improve throughout both seed development and maturation until harvest maturity, before seed deterioration was first observed. Several other temporal patterns of seed quality development and decline have also been reported. These are portrayed and compared. The assessment suggests that the original hypothesis was too simple, because it combined several component hypotheses: (a) the seed improvement (only) phase ends before seed deterioration (only) commences; (b) there is only a brief single point in time during seed development and maturation when, in all circumstances, seed quality is maximal; (c) the seed quality improvement phase coincides perfectly with seed filling, with deterioration only post-seed filling. It is concluded that the search for the single point of maximum seed quality was a false quest because (a) seed improvement and deterioration may cycle (sequentially if not simultaneously) during seed development and maturation; (b) the relative sensitivity of the rates of improvement and deterioration to environment may differ; (c) the period of maximum quality may be brief or extended. Hence, when maximum quality is first attained, and for how long it is maintained, during seed development and maturation varies with genotype and environment. This is pertinent to quality seed production in current and future climates as it will be affected by climate change and a likelihood of more frequent coincidence of brief periods of extreme temperatures with highly sensitive phases of seed development and maturation. This is a possible tipping point for food security and for ecological diversity.

Seeds of 15 diverse rice accessions, representing aus, indica, temperate japonica and tropical japonica subpopulations, were produced under temperate climate conditions in Korea and used for vitamin E analysis and seed storage experiments at 45°C and 10.9% seed moisture content. High γ-tocotrienol was significantly positively correlated with seed longevity. In addition, a high β-tocopherol proportion relative to δ-tocopherol was significantly negatively correlated with seed longevity. Using high-density single-nucleotide polymorphism marker data, DNA haplotype analysis showed clear allelic variations in the region of two S-adenosylmethionine synthetase genes: LOC_Os04g42095 and LOC_Os11g15410, which regulate the conversion of δ-tocopherol into β-tocopherol. Four indica accessions with rare and subpopulation-specific alleles showed a 2.3-fold lower β-/δ-tocopherol ratio compared with accessions from other subpopulations.

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.

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.

Knowledge of the effects of burial depth and burial duration on seed viability and, consequently, seedbank persistence of Palmer amaranth (Amaranthus palmeri S. Watson) and waterhemp [Amaranthus tuberculatus (Moq.) J. D. Sauer] ecotypes can be used for the development of efficient weed management programs. This is of particular interest, given the great fecundity of both species and, consequently, their high seedbank replenishment potential. Seeds of both species collected from five different locations across the United States were investigated in seven states (sites) with different soil and climatic conditions. Seeds were placed at two depths (0 and 15 cm) for 3 yr. Each year, seeds were retrieved, and seed damage (shrunken, malformed, or broken) plus losses (deteriorated and futile germination) and viability were evaluated. Greater seed damage plus loss averaged across seed origin, burial depth, and year was recorded for lots tested at Illinois (51.3% and 51.8%) followed by Tennessee (40.5% and 45.1%) and Missouri (39.2% and 42%) for A. palmeri and A. tuberculatus, respectively. The site differences for seed persistence were probably due to higher volumetric water content at these sites. Rates of seed demise were directly proportional to burial depth (α=0.001), whereas the percentage of viable seeds recovered after 36 mo on the soil surface ranged from 4.1% to 4.3% compared with 5% to 5.3% at the 15-cm depth for A. palmeri and A. tuberculatus, respectively. Seed viability loss was greater in the seeds placed on the soil surface compared with the buried seeds. The greatest influences on seed viability were burial conditions and time and site-specific soil conditions, more so than geographical location. Thus, management of these weed species should focus on reducing seed shattering, enhancing seed removal from the soil surface, or adjusting tillage systems.

Interchangeable effects of temperature, moisture content and oxygen on seed longevity have been mostly examined to estimate seed viability during long-term dry storage, whereas few experiments have studied seed viability under near-natural conditions to evaluate seed persistence in the soil. To this end, we artificially aged seeds of Ranunculus baudotii, a hydrophyte widely distributed in temporary ponds constituting an abundant soil seed bank. Seeds were exposed to controlled ageing at three different relative humidities (RH) under both aerobic and anoxic conditions. Their viability, water content, membrane damage, oxidative stress and anti-oxidant enzymatic defence activity were evaluated. Seed survival was longer at higher relative humidity (97% RH), and lowest at a relative humidity (90% RH) simulating moist but not waterlogged soils. Anoxic conditions showed a protective role on viability at lower moisture contents (70% RH). Seed viability was negatively associated with hydrogen peroxide content and correlated with anti-oxidant enzyme activities, but not with membrane damage. Altogether, these results suggest negative roles for moist soils and anoxia in determining seed persistence in the field, but at higher moisture contents the negative effects of anaerobia diminished. The anti-oxidant system activation, even under unfavourable conditions, might recover seeds once all protective processes can operate, pointing out the plasticity of mechanisms involved in seed loss viability.

Post-harvest drying prolongs seed survival in air-dry storage; previous research has shown a benefit of drying moist rice seeds at temperatures greater than recommended for genebanks (5–20°C). The aim of this study was to determine whether there is a temperature limit for safely drying rice seeds, and to explore whether the benefit to longevity is caused by high-temperature stress or continued seed development. Seeds of two rice varieties were harvested at different stages of development and dried initially either over silica gel, or intermittently (8 h day–1) or continuously (24 h day–1) over MgCl2 at temperatures between 15 and 60°C for up to 3 days. Seeds dried more rapidly the warmer the temperature. Subsequent seed longevity in hermetic storage (45°C and 10.9% moisture content) was substantially improved by increase in drying temperature up to 45°C in both cultivars, and also with further increase from 45 to 60°C in cv. ‘Macassane’. The benefit of high-temperature drying to subsequent longevity tended to diminish the later the stage of development at seed harvest. Intermittent or continuous drying at high temperatures provided broadly similar improvements to longevity, but with the greatest improvements detected in a few treatment combinations with continuous drying. Heated-air drying of rice seeds harvested before maturity improved their subsequent storage longevity by more than that which occurred during subsequent development in planta, which may have resulted from the triggering of protection mechanisms in response to high-temperature stress.

Vitamin E (tocols) is a key metabolite for efficient scavenging of lipid peroxy radicals that cause membrane breakdown during seed ageing. However, in rice, this hypothesis has been tested for very few lines only and without considering intraspecific variation in genomic structure. Here, we present a correlation study between tocols and seed longevity using a diverse rice panel. Seeds of 20 rice accessions held in the International Rice Genebank at the International Rice Research Institute, representing Aus, Basmati/Sadri, Indica, temperate Japonica and tropical Japonica variety groups, were used for tocols analysis (quantification of α-, β-, γ- and δ-tocopherol/tocotrienol by ultra-performance liquid chromatography) and storage experiments at 45°C and 10.9% seed moisture content. To examine the effects of DNA-haplotype on the phenotype, the 700 K high-density single-nucleotide polymorphism marker dataset was utilized. Both seed longevity (time for viability to fall to 50%; p 50) and tocols content varied across variety groups related to the heterogeneity in the genetic architecture. Among eight types of tocol homologues, α-tocopherol and γ-tocotrienol were significantly correlated with p 50 (negatively and positively, respectively). While temperate Japonica varieties were most abundant in α-tocopherol, Indica varieties recorded 1.3- to 1.7-fold higher γ-tocotrienol than those of other groups. We conclude that the specific ratio of tocol homologues rather than total tocols content plays an important role in the seed longevity mechanism.

A study under glasshouse conditions determined the effect of germination stimulants, potassium nitrate and ethephon, on emergence of several pure lines of wild oat from soil. Effect of the two chemicals on stimulating emergence was variable and no synergism noted when used together. Examination of exhumed seed showed that these treatments reduced seed viability. By counting the number of dead and adding to those that emerged, total seed removed from the soil was obtained. All treatments removed seed from the soil seedbank, especially when seed was old, near the soil surface, was primary seed, or from low or intermediate dormancy lines. However, because of the insensitivity of a large proportion of the seed and the high rates required it is unlikely that either potassium nitrate or ethephon has practical use for field application.