Uncaria rhynchophylla (Rubiaceae), a woody liana with significant medicinal value, has been used as a traditional Japanese and Chinese medicine. While effective seed production is required for breeding and efficient seedling production, the physiology of sexual reproduction remains largely unknown in this species. Therefore, we first observed the flowering behaviour, and next attempted artificial pollination using flowering individuals in a greenhouse. In this study it became clear that this species sets seeds by allogamy, but not by autogamy. The obtained seeds showed about a 90% germination rate. We also examined seed desiccation tolerance and storage conditions which are important to preserve the genetic resources. Seeds of this species were found to have a characteristic of the orthodox type, having high desiccation tolerance. Seeds after 6 months of storage at + 22, −20 and −160°C showed comparable germination rates to the seeds before storage.

Understanding seed moisture desorption and adsorption isotherms is important for seed quality maintenance and better predicting seed storage lifespan. Freshly harvested oilseed rape and barley seeds were dried and then rehydrated twice. Seed equilibrium relative humidity (eRH) and moisture content (MC) were determined at different humidity levels so that two cycles of desorption and adsorption could be constructed. In addition, seeds were dried to 30% RH and then rehydrated to 50% RH for five cycles to determine whether they shift to the adsorption isotherm. Monolayer MC was determined using the Gugenheim-Anderson-de Boer model. Storage experiments were conducted for seeds equilibrated at 30, 40, 50, 60 and 70% RH for two cycles of desorption and adsorption at 45°C. Isotherm curves’ shapes were similar for oilseed rape and barley, although spanning a greater MC range in barley. The hysteresis effect was observed for oilseed rape and barley seeds when dried over silica gel at <10% RH. However, this effect was only observed for barley seeds when dried to 30% RH, but not for oilseed rape seeds. Longevity was greater for adsorbing seeds than desorbing seeds at a given eRH, however, there was no significant difference in σ (the standard deviation of the normal distribution of seed deaths over time)–MC log–log relationship. The relationship shifted for seeds on the second cycle. In conclusion, if seed lots are stored at a specific RH, reaching equilibrium by desorption or adsorption can strongly influence their longevity. Also, when seeds of different species are dried to low RH, they will respond differently to a subsequent increase in RH, which could profoundly affect their longevity.

Understanding the relative longevity of different seed lots, perhaps of different species or genotypes, but also following production under different environments or using different cultivation methods, or following different post-harvest treatments, is relevant to anyone concerned with the retention of seed lot viability and vigour during storage. However, different scientists over the years have used different conditions to assess seed lot longevity, as well as different variables as the measure of ‘longevity.’ Here, we give some of the backgrounds to how two standard protocols, with an open and closed system respectively, were derived, and explain why we consider p50, defined as the time during storage when seed lot viability, as measured through a germination test, has declined to 50%, is a suitable longevity trait parameter.

Seeds kept in long-term storage are essential for maintaining genetic resources of crops and other plant materials in seed banks and national germplasm systems. But seeds undergo chemical changes over time as part of the ageing process that result in reduced germination rates and seedling normalcy. For example, oilseed crops such as peanuts are particularly vulnerable to oxidation and rancidification due to their high oil content. To test the effect of storage time on seed oil in peanuts, we grew different accessions and harvested fresh seeds to compare the oil composition of new seeds with seeds aged in storage for varying lengths of time. Out of the nine fatty acids detected and measured by gas chromatography, five including oleic, gadoleic, behenic, lignoceric and cerotic acids differed significantly between new and old seeds. Additionally, old seeds accumulated up to four oxidation products together averaging about 1% of the total oil composition. The concentration of these oxidation products was positively correlated with the age of the seeds, increasing linearly up to more than 6% of the total in the oldest seeds. The presence and concentration of oxidation products measured using simple chromatography techniques can be used as an initial indication of quality and viability in older seed inventories stored in germplasm repositories.

Cultivated northern wild rice (NWR; Zizania palustris L.) has been bred and studied since the 1950s. One challenge facing researchers is the lack of storage options, due to the seed's unorthodox behaviour. This study evaluated varying storage temperature and moisture conditions for the maintenance of seed viability and dormancy breaking in Minnesota-grown NWR. First, seeds were placed in non-submerged, freezing storage (NSFS) for 12–26 weeks, then in submerged, cold storage (SCS) for 2 weeks. The addition of SCS increased germination (%G) relative to NSFS alone (<0.1% NSFS, 15% NSFS and SCS), indicating that NSFS does not kill seeds but also does not break seed dormancy. Next, the required length of SCS was evaluated by placing seeds in NSFS for 12 weeks and then in SCS for 2–14 weeks. A longer SCS period increased %G from 3 to 79%, at 2 and 14 weeks of SCS, respectively. Lastly, seeds were placed in NSFS, followed by SCS, at varying intervals over a 29-week period. Across lines, germination increased from 20 to 76% between 4 and 7 weeks of SCS, respectively, then plateaued. The results of this study indicate that NSFS could be used to store NWR seeds, but at least 7 weeks in SCS is required to overcome dormancy. Additionally, while NSFS did not break seed dormancy, physiological changes related to stratification processes were occurring in non-submerged, freezing conditions. Results also suggest that the genotypic variation in NWR could be utilized for selection to improve germination and storage viability.

Seed longevity is a complex process of key ecological and agronomic importance. DNA damage is a significant factor affecting seed ageing. Likewise, epigenetic changes can control gene expression and, therefore, seed response to ageing. The aim of the present work was to investigate the effect of ageing on nucleic acid stability and to identify reliable molecular markers that might help to monitor epigenetic changes within plant genetic resources during conservation. DNA profiles, evaluated by RAPD (random amplified polymorphic DNA), and methylation patterns, obtained by MSAP (methylation-sensitive amplification polymorphism), were compared in non-aged and aged Mentha aquatica seeds and plants produced by them. Germination decreased to 50% by storing seeds at 35°C and 12% wc for 28 days. RAPD profiles were 99% similar in these aged seeds compared to non-aged seeds. However, seedlings produced from the aged seeds showed a 13% dissimilarity compared to seedlings produced from the non-aged seeds. About 8% difference in the MSAP epigenetic profile was detected in seeds after storage and 16% difference was detected in the seedlings produced from them. This indicates that stress from high temperature and humidity during storage induced changes on the methylation state of seeds, and that changes were also detectable in the regenerated plants. Our results suggest that DNA integrity was compromised in seeds during ageing, and on seedlings produced by aged seeds. Genotype screening techniques such as RAPD and MSAP have the potential as markers of nucleic acid stability during seed ageing.

Seeds are increasingly being stored in seed banks for ex situ conservation, food security, and restoration reserves. Although it is hypothesized that seeds of many species can survive the dry, freezer conditions (−20°C) of seed banks for many years, shelf life is unknown for many species. In particular, changes to dormancy and germination requirements following freezer storage have not yet been studied extensively. To improve our understanding of how seed dormancy and germination respond to seed banking conditions, we evaluated seed of six restoration-priority species in the southwestern United States, which is a region of increasing restoration need. Germination tendencies of seeds that were and were not exposed to a −20°C treatment for 4–6 months were evaluated under 22 cold-moist stratification and incubation treatments to broadly assess changes to dormancy and germination requirements and speed. Direct or indirect (via interactions with stratification and/or incubation treatments) differences in dormancy breaking and germination were observed in seeds for four of the six species studied. Specifically, storage temperature accounted for differences in the final germination proportion, germination during stratification, and/or rate of germination in five of the six species. Notably, seeds of the one species that exhibit a combination of physiological and physical dormancy showed significant differences in all germination measures. However, while significant differences were found between seeds exposed to short-term seed bank storage and those that were not, these differences were small and may not be biologically meaningful, indicating that seed banking these species should not change how they are used for restoration or reintroduction purposes.

We studied the variability of germination, dormancy and viability loss of Hirschfeldia incana seeds in relation to seed size. Seeds were stored at 35°C under humid [75% relative humidity (RH)] or dry (33% RH) conditions. Seed germination and electrolyte leakage were evaluated periodically. Small seeds had lower longevity at humid or dry storage conditions (5 or 407 days, respectively) than large or intermediate seeds (7–9 or 536–727 days, respectively). Moreover, H. incana shows variability in seed dormancy related to seed size within a population, with small seeds having lower dormancy (13%) than intermediate (50%) or large seeds (72%). Dormancy was partially released after a short storage at 35°C and humid conditions. Under dry storage conditions, endogenous dormancy cycles were observed for over a year, and longer times of storage had a dormancy-breaking effect through dry after-ripening. Results suggest a dual strategy producing non-dormant seeds with low longevity that will germinate immediately after dispersal, and seeds with greater longevity that will delay germination. Membrane permeability increased linearly with ageing at both humid and dry storage (R2 = 0.60). Small seeds showed greater conductivity than intermediate or large seeds (0.7, 0.4 or 0.3 mS g–1 dry weight, respectively, at the 80% germination). The conductivity test could be used to evaluate the quality of H. incana seeds and would allow us to identify dormant (non-germinating) seed lots as viable. However, the influence of storage conditions and variability within a seed population on seed longevity should be taken into account when evaluating seed quality.

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.

Witchweed (Striga asiatica Lour. ♯4 STRLU) seed remained viable for 6 yr under open-shelf laboratory conditions and for 14 yr when deep buried under field conditions. Seed kept in normal atmosphere of the laboratory for < 1 through 6 yr had the following germination percentages: 2, 95, 95, 94, 83, 38, and 7. Newly harvested seed will not germinate because of postmaturity ripening requirements. There were no viable seed after 7 yr of shelf storage in the laboratory. Seed buried in the field gave 60 to 90% germination the first season after production. Annual recovery and testing showed a gradual reduction in viability to 10% at year 14 for the deepest buried seed; shallower buried seed expired sooner. No germination occurred thereafter.

The germination behavior of giant foxtail and shattercane seed was investigated to determine whether environment in which maternal plants grow and progeny seed develop affects seed dormancy. Giant foxtail and shattercane seed was collected in 1985, 1991, and 1992. Plants from this seed were grown to maturity in the field and the greenhouse in 1993 and 1994. Experiments were conducted using mature progeny seed collected from these plants to determine the influence of environment of maternal growth on seed germination. Greenhouse-raised giant foxtail seed was more dormant than field-raised seed. Progeny of the 1991 giant foxtail collection had the most dormant seed, whereas progeny of the 1992 collection had the least dormant seed. Field-raised shattercane seed was more dormant than greenhouse-raised seed, but this dormancy was easily overcome by placing them in fluctuating temperatures. Moist stratification of giant foxtail seed increased its overall germination and reduced the differences in germination between individual collections and between field- and greenhouse-raised seed lots. Moist stratification of shattercane seed resulted in increased variability in germination within and among seed lots. Subjecting shattercane seed to freezing and thawing was most detrimental and caused an 89% reduction in germination due to seed death. These data indicate that before overwintering, giant foxtail seed differed in germination percentage, but overwintering reduced these differences. Shattercane seed produced in different environments had similar germination percentage. This suggests that successful overwintering depends on its ability to survive freezing and thawing conditions.

Laboratory and growth room experiments were conducted to determine effects of CGA-133205, oxabetrinil, and flurazole on grain sorghum hybrids ‘DeKalb 46’ and ‘DeKalb 48’ after the seed were treated with the herbicide safeners and stored for up to 24 wk at various relative humidity levels. Each safener treatment had an adverse effect on grain sorghum seed as reflected by fewer normal seedlings at 4 d when averaged over storage time and relative humidity levels. Flurazole and oxabetrinil increased the number of ungerminated seed of both hybrids, and 0.4 and 0.8 g ai kg–1 CGA-133205 increased ungerminated seed of DeKalb 46. At ≥ 8 wk after storage, a reduction in germination was observed for seed treated with flurazole or oxabetrinil compared to seed stored without safener. This reduction also occurred with 0.4 and 0.8 g kg–1 CGA-133205, but only at 24 wk after storage. Each safener treatment minimized reduction in seedling shoot fresh weight by herbicides compared to the no-safener check through 24 wk after storage.

Germination was investigated in 1-yr and 8-yr-old seed lots of barnyardgrass. Each seed lot was divided into a portion stored dry at 25 C (afterripening) and a portion buried in the field (stratification) in central Europe (latitude 50°N, longitude 14°E). The afterripened seed lost dormancy within 2 yr after dispersal, whereas buried seed passed through annual dormancy/nondormancy cycles. The seed was germinated at five constant temperatures between 17 and 35 C with an 18/6-h light/dark photoperiod. Germination was affected by both seed age and storage conditions. Germination percentage and rate in 8-yr-old afterripened and each of the stratified seed lots varied with temperature. Optimum temperatures for germination were between 27 and 31 C, and the range of adequate temperatures increased with seed age. A common base temperature for germination was 11.7 C. In 1-yr-old afterripened material, the proportion of germinating seed (< 5%) and germination rate were similar at all temperatures. Barnyardgrass thus revealed a plastic strategy of germination. Stratification during the first winter removed dormancy and allowed germination at a narrow range of temperatures. This constraint on optimum germination temperature decreased with increased seed age. Seedling emergence, thus, may vary according to whether seed population originates from the previous year or from old soil seed banks.

In fire-prone ecosystems, many species regenerate after fire from persistent soil seed banks. Species with physically dormant (PY) seeds have dormancy broken by fire-related heat. The magnitude of post-fire recruitment, to predict response to varying fire severity, is commonly estimated by testing dormancy-breaking temperature thresholds of fresh PY seeds. However, seeds spend years in the soil during the inter-fire period, and determining whether dormancy-breaking thresholds change over time is essential to accurately predict population persistence. Germination of four south-eastern Australian PY species from the Fabaceae family (Acacia linifolia, Aotus ericoides, Bossiaea heterophylla and Viminaria juncea) were studied. Dormancy-breaking temperature thresholds vary inter-specifically and the species represented either high or low dormancy-breaking threshold classes. Freshly collected seeds, and seeds that had been buried in the field or stored in dry laboratory conditions for 6 and 18 months were subjected to a fire-related range of heat treatments (40–100°C). Seed ageing increased germination response to heat treatments, effectively lowering the dormancy-breaking thresholds of three species. The fourth species, A. linifolia, initially had a relatively large non-dormant fraction which was lost as seeds aged, with older seeds then displaying PY broadly similar to the other study species. Patterns of threshold decay were species-specific, with the thresholds and viability of low-threshold species declining more rapidly than high-threshold species. The non-dormant fraction did not increase over time for any of our study species. Instead of increasing their non-dormant fraction, as is common in other vegetation types, these fire-prone PY species displayed a change of dormancy-breaking temperature thresholds. This is an important distinction, as maintaining dormancy during the inter-fire period is essential for population persistence. While changes in sensitivity to dormancy-breaking treatments have previously been reported as seeds age, our study provides the first test of changes to temperature thresholds, which increases the range of germination response from the seed bank under varying fire severity.

In each of two experiments, freshly harvested seeds of two cultivars of rice were dried and then rehydrated to different moisture levels. In experiment 1, seed equilibrium relative humidity (eRH) and moisture content (MC) were determined at each moisture level so that moisture desorption and adsorption isotherms could be constructed. Seed storage experiments were also carried out for seeds equilibrated at each moisture level, in sealed aluminium foil packets at 45°C. In experiment 2, storage experiments at 45°C were carried out on seeds dried to 12% MC and seeds dried to lower moisture levels and then rehydrated to 12% MC.

The moisture adsorption isotherm was shifted to lower MC at a given eRH compared with the desorption isotherm. This hysteresis effect was seen both when seeds were dried to <6% MC and then allowed to adsorb moisture to different levels, and when seeds were dried to different levels (10, 8, 6 or 4% MC) and then allowed to adsorb moisture up to 12% MC. The log-log relationship between seed longevity, σ [the standard deviation of the normal distribution of seed deaths over time, as defined in the Ellis and Roberts (1980) seed viability equations] and seed storage MC did not vary depending on whether the seeds were desorbing or adsorbing moisture. The relationship between σ and eRH was better described by a log-log model than a log-linear model and did vary depending on whether seeds were desorbing or adsorbing moisture: at a given eRH, the longevity of adsorbing seeds was greater than that of desorbing seeds. The implications for seed storage are discussed.

Miconia chartacea is a widely distributed tree in Brazil, occurring at altitudes ranging from 300 m to 1900 m in the Caatinga, Cerrado and Atlantic Forest biomes. In this work we attempted to classify M. chartacea seeds regarding their behaviour during storage and their germination syndrome and to determine, from a storage test in Cerrado soil and laboratory conditions in situ and ex situ, the longevity of seeds, as well as the capacity of the species to form a soil seed bank. The results suggested that M. chartacea seeds form a transient soil seed bank in the Cerrado and can be classified as orthodox in terms of storage behaviour, although the seeds are dispersed with a relatively high water content. The life span of seeds was favoured in soil-stored seeds in comparison with dry storage at 25°C, whereas storage at low temperatures prevented a decrease of the seed's germinability with storage time (330 d). M. chartacea seeds are dispersed during the dry season and germinate during the next rainy season, which can be classified as an intermediate–dry germination syndrome. Seeds of this species are dispersed in the Cerrado when temperatures and soil moisture are relatively low, which favours the formation of a soil seed bank, considering that the seeds tolerate desiccation and their longevity is favoured by low temperatures. A transient seed bank type is favoured by the loss of viability in storage at warm temperatures linked to the rainy season, and the predictable seasonal variations in climate in the region, with germination being restricted to the beginning of the rainy season.

Plant genetic resources are conserved by genebanks mainly in the form of seeds. In most of the cases, the dried seeds can be stored for a considerable period of time, but eventually seed deterioration results in the inability to generate healthy seedlings. Prolonging seed longevity during storage reduces the frequency of regeneration, which is beneficial from a genetic as well as a management point of view. To reduce the rate of deterioration, cool and dry storage conditions are usually practised for long-term seed storage. In spite of the growing body of evidence that seed deterioration is predominantly caused by oxidative processes, the importance of seed storage under anoxic conditions has received little attention from the genebank community. Herein, we report on the effects of anoxia on seed viability, the oxygen uptake by dry seeds in closed containers and the permeability for oxygen of various seed storage containers. Our results confirm that the ageing of dry seeds is accelerated by the presence of oxygen in the storage environment. Therefore, we recommend that genebanks store dry seeds under anoxic conditions to prolong their longevity during ex situ conservation. To reduce the initial rate of viability loss, we further recommend that the period of temporary storage after seed harvest be minimized and also that the seeds are kept during this period under controlled conditions, including anoxia.

Dehydrins are late embryogenesis abundant (LEA) proteins that accumulate during seed maturation and in response to abiotic stresses in vegetative tissues. They are thought to protect cellular components from dehydration stress. However, whether they play a role in survival in the dry state is not clear. In this study, an RNA interference (RNAi)-construct against the seed-expressed dehydrin of Arabidopsis thaliana, LEA14 (At2g21490), was introduced to wild-type plants, which led to a strong reduction in transcript abundance of the target gene as well as that of two other seed-expressed dehydrin homologues, XERO1 (At3g50980) and RAB18 (responsive to abscisic acid 18, At5g66400) in the transformants. Mature, dry seeds from the RNAi plants germinated to at least 95% after rehydration, indicating that seed desiccation tolerance was not affected, while they exhibited a twofold reduction in longevity. When stored at 75% relative humidity and 35°C, the seeds of two independent RNAi lines lost 50% of their viability in 10 d and 5 d, respectively, while it took 17 d for wild-type seeds to lose 50% viability. In addition, when seeds were imbibed in the presence of 100 mM NaCl, the seeds of RNAi plants exhibited reduced germination compared to wild-type seeds, suggesting that at least one of the three seed-specific dehydrins plays a role both against deterioration during storage at low moisture content and when imbibed tissues are submitted to salt stress at high moisture.

Introduction. Rambutan is a tropical fruit species with recalcitrant seeds. Despite the expansion of exotic fruit cultivation in Brazil, lots of which fruit species, including rambutan, need basic information, especially in relation to propagation and storage of seeds, which are important for genetic improvement studies, maintenance of genetic sources and seedling production. Materials and methods. A completely randomized design was adopted with treatments distributed in a factorial arrangement, 3 × 4, referring to three seed storage conditions [room temperature conditions; a dry chamber with (18 ± 2) °C and 60% relative humidity; and a cold chamber with (10 ± 2) °C and 70% relative humidity] and four storage times (0, 7, 14 and 21 d). Each treatment of 10 seeds was replicated five times. Data on seedling emergence, emergence rate, plant height, number of leaves and length of main root were submitted to variance analysis and means were separated using Tukey’s test. Correlation analysis between seed moisture and seedling emergence was performed. Results and discussion. Our results indicated that dry chamber conditions promoted the statistically significantly highest seedling emergence after 7 d of storage. Cold chamber conditions promoted an extremely low seedling emergence independently of time. Conclusion. Rambutan seeds can be stored in a dry chamber for 7 d without losing viability; after 14 d of storage the loss of emergence is 60%.