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.

Lettuce (Lactuca sativa L.) and onion (Allium cepa L.) seeds have relatively short longevity during storage and their germination is sensitive to environmental stress. Seed priming (controlled hydration followed by drying) can improve seed germination under stressful conditions, inducing faster and more uniform germination over broader temperature ranges, but it can also reduce seed longevity in storage. Controlled deterioration (CD) tests are often employed to study longevity by ageing seeds rapidly at elevated temperature and moisture content, and primed seeds are particularly sensitive to CD conditions. As reactive oxygen (O2) species are thought to be involved in seed deterioration, we tested whether storage under reduced O2 atmospheres (0 and 2% O2) would extend the longevity of primed and non-primed seeds under low relative humidity (RH) (33% RH+37°C) and CD (75% RH+50°C) storage conditions. The longevity of both non-primed and primed lettuce seeds in low RH storage was extended by anaerobic environments, but the effect of O2 was much less under CD conditions. In onion, only primed seeds exhibited a beneficial effect of low O2 atmospheres under both types of ageing conditions. In both species, storage under anaerobic conditions was beneficial for extending the longevity of primed seeds, but was not able to ameliorate fully the negative effect of priming on storage life.

Several of the procedures necessary for cryopreservation of excised zygotic embryonic axes are known to be accompanied by emission of damaging levels of reactive oxygen species (ROS). These have been shown to be associated with shoot apical meristem necrosis, curtailing subsequent axis development to root production only, particularly for tropical/subtropical recalcitrant-seeded species. Here we report on the application of the principles of electrochemistry in the generation of strongly reducing, high pH cathodic water, by electrolysis of a solution containing calcium and magnesium chloride. The cathodic water in which Strychnos gerrardii axes were immersed for 30 min following dehydration, and importantly, after dehydration followed by cryopreservation, was shown to have strongly antioxidative properties in counteracting the damaging effects of ROS bursts and promoting shoot development. In a parallel experiment, axes of Boophane disticha exhibited enhanced total antioxidant activity when exposed to cathodic water both immediately following excision, and after flash-drying. For both species, the efficacious effects of cathodic treatment were manifest after the axes had been in culture for 4 h, suggesting that ROS were not quenched at source, but probably counteracted by enhancement of activity of endogenous antioxidants. Cathodic water therefore affords a non-toxic means of amelioration of oxidative, stress-related damage, which, coupled with the strongly fungicidal activity of the acidic, anionic water fraction, offers significant, and apparently non-injurious, advances towards successful cryopreservation of germplasm – and probably generally improved success of in-vitro-based procedures for plant tissues.

Legumes are crops that develop in cropping systems with relatively low inputs and are suitable to a more sustainable agriculture. Successful crop establishment, which is crucial for reliable plant production, depends on seed quality, environmental factors and genotypes. We studied pea and bean germination and seedling growth at various temperatures (5–40°C) and water potentials ( − 0.2 to − 1.5 MPa) using winter and spring pea and two common bean seeds produced in different conditions. The germination base temperature was − 1.1°C for pea seeds, and seeds of the winter genotype germinated more rapidly than those of the spring genotype. The base temperature for bean seed germination was 5.1–9.6°C, depending on the seed lot. The germination base water potential was about − 2 MPa for both species. The base temperatures for shoot elongation were higher (3–6°C) than those for germination. A review of the literature on other legumes confirmed that the differences in the responses of the legume seeds and seedlings to different temperatures were associated with their geographic origin. These results help understanding of pea and bean crop establishment, provide crop model parameter values and contribute to the search for genetic variability.

Although mature seeds of the monogeneric conifer family Cephalotaxaceae sensu stricto have underdeveloped embryos, no definitive studies have been done to classify dormancy in this family. Our primary purpose was to determine the kind of dormancy in seeds of Cephalotaxus wilsoniana and to put the results into a broad phylogenetic context for gymnosperms. The species is of horticultural and medicinal value, and information is needed on how to propagate it efficiently from seeds. Embryo growth and germination were monitored for seeds at warm, cold and warm plus cold temperatures, and germination was monitored for seeds subjected to: (1) cold →  warm →  cold →  warm; and (2) warm →  cold →  warm →  cold →  warm temperature sequences. The effects of gibberellic acids GA3 and GA4 were tested on radicle emergence in ungerminated seeds and on shoot emergence in root-emerged seeds. Germination was promoted by ≥ 36 weeks of warm stratification followed by ≥ 8 weeks of cold stratification, but only if seeds were returned to high temperatures. The underdeveloped embryo must increase in length by >120% before the radicle emerges. Neither GA3 nor GA4 was effective in promoting radicle emergence; however, both plant growth regulators increased rate (but not percentage) of shoot emergence in root-emerged seeds. We conclude that seeds of C. wilsoniana have the deep simple level of morphophysiological dormancy (MPD), C1b-C3-B1b; thus, warm stratification followed by cold stratification and then warm-temperature incubation are required for germination. In gymnosperms, MPD is known in cycads, Ginkgo and now in three families of conifers.

Fleshy fruits fall on to the ground together with cleaned seeds previously ingested by primary dispersers, offering a wide range of fruits and seeds to the ground foragers. Although nutritional properties strongly differ between fruits and seeds, this different seed presentation (cleaned seeds versus seeds within the pulp) has not been addressed in seed removal studies. This study reports on the removal of fruits versus their seeds in five fleshy-fruited species in a temperate forest. We found that rodents removed most of the seeds and partially consumed most of the fruits, preferring seeds to fruits. Rodents bit the fruits to extract the seeds, leaving most of the pulp. We found a preference ranking for the seeds (Sorbus aucuparia>Ilex aquifolium>Sorbus aria>Rosa canina>Crataegus monogyna) but no preferences were found for the fruits, probably due to their similarities in pulp constituents. Seed and fruit choice were affected by chemical and physical properties and not by their size. The presence of alternative and preferred seeds (nuts) delayed the encounter of the fruits and seeds and diminished their removal rates. We found that higher rodent abundance is not necessarily associated with higher removal rates of fleshy fruits. Rodent abundance, fruit size and seed size are minor factors in the removal of fleshy fruits and their seeds. This study underlines that scatter-hoarding rodents are important removers of fleshy fruits and their seeds, producing a differential seed removal depending on the seed presentation (with or without pulp), the nutritional properties of the seeds (but not of the fruits) and the presence of alternative food.

Group testing is a procedure in which groups that contain several units (plants) are analysed without having to inspect individual plants, with the purpose of estimating the prevalence of genetically modified plants (adventitious presence of unwanted transgenic plants, AP) in a population at a low cost, without losing precision. When pool (group) testing is used to estimate the proportion of AP (p), there are several procedures that can be used for computing the confidence interval (CI); however, they usually do not ensure precision in the estimation of p. This research proposes a formula for determining the required number of pools (g), given a pool size (k), for estimating the proportion of AP plants using the Dorfman model. The proposed formula ensures precision in the estimated proportion of AP because it guarantees that the width (W) of the CI will be equal to, or narrower than, the desired width (ω), with a probability of γ. This probability accounts for the stochastic nature of the sample variance of p. We give examples to show how to use the proposed sample-size formula. Simulated data were created and tables are presented showing the different scenarios that a researcher may encounter. The Monte Carlo method was used to study the coverage and the level of assurance achieved by the proposed sample sizes. An R program that reproduces the results in the tables and makes it easy for the researcher to create other scenarios is given in the Appendix.