Abscisic acid (ABA) is a plant hormone that regulates seed dormancy and germination. Seeds undergo changes in both ABA content and sensitivity during seed development and germination in response to internal and external cues. Recent advances in functional genomics have revealed the integral components involved in ABA metabolism (biosynthesis and catabolism) and perception, the core signalling pathway, as well as the factors that trigger ABA-mediated transcription. These allow for comparative studies to be conducted on seeds under different environmental conditions and from different genetic backgrounds. This review summarizes our understanding of the control of ABA content and the responsiveness of seeds to afterripening, light, high temperature and nitrate, with a focus on which tissues are involved in its metabolism and signalling. Also described are the regulators of ABA metabolism and signalling, which potentially act as the node for hormone crosstalk. Integration of such knowledge into the complex and diverse events occurring during seed germination will be the next challenge, which will allow for a clearer understanding of the role of ABA.

Dormant barley grains cannot germinate at 30°C and this inability to germinate is imposed mostly by the glumellae which have been suggested to limit oxygen supply to the embryo. Hypoxia imposed either artificially or by the glumellae to embryos from dormant grains, increases embryo sensitivity to abscisic acid (ABA) and promotes the accumulation of ABA during the first hours after imbibition. Expression of candidate genes involved in ABA synthesis (HvNCED), catabolism (HvABA8OH1) and signalling (HvABI5, HvVP1 and HvPKABA) was analysed in embryos isolated from dormant whole or de-hulled grains incubated in air or in hypoxia (5% oxygen). The presence of the glumellae enhanced the expression of genes involved in ABA metabolism and signalling with respect to that observed in de-hulled grains incubated in air. These results suggest that at least part of the observed physiological responses to the presence of the glumellae are regulated at the level of gene expression. However, hypoxia imposed on dormant de-hulled grains did not mimic the presence of the glumellae in terms of expression of candidate genes. Hypoxia mimics the presence of the glumellae in terms of dormancy maintenance and ABA accumulation and sensitivity, but its effects appear to operate through different mechanisms.

MicroRNAs (miRNAs) are involved in developmental programmes of plants, including seed germination and post-germination. Here, we provide evidence that two different miRNA pathways, miR156 and miR172, interact during the post-germination stages in Arabidopsis. Mutant seedlings expressing miR156-resistant SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE13 (mSPL13), which has silent mutations in the miR156 complementary sequence, over-accumulated SPL13 mRNA and exhibited a delay in seedling development. Microarray analysis indicated that SCHNARCHZAPFEN (SNZ), an AP2-like gene targeted by miR172, was down-regulated in these mutants. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and miRNA gel blot analyses showed that the MIR172 genes were up-regulated in mSPL13 mutants. These results suggest that the miRNA regulation cascades (miR156⊣SPL13 → miR172⊣SNZ) play a critical role during the post-germination developmental stages in Arabidopsis.

Germination and early seedling development are critical for successful stand establishment of plants. Following germination, the cotyledons, which are derived from embryonic tissue, emerge from the seed. Arabidopsis seedlings at post-germinative stages are supported mainly by the supply of nutrition from the cotyledons until vegetative leaves emerge and initiate photosynthesis. The switch to autotrophic growth is a significant transition at the post-germinative stage. Here, we provide evidence that down-regulation of SQUAMOSA PROMOTER-BINDING PROTEIN LIKE13 (SPL13) by microRNA156 (miR156) plays an important role in the regulation of the post-germinative switch from the cotyledon stage to the vegetative-leaf stage. Silent mutations created in the SPL13 sequence in the region that is complementary to the miR156 sequence caused the deregulation of the mutant form of SPL13 (mSPL13) mRNA from miR156. Mutant seedlings over-accumulated miRNA-resistant messages and exhibited a delay in the emergence of vegetative leaves compared to wild-type seedlings. The delay was not observed in control transgenic plants expressing non-mutated SPL13, indicating that the phenotype was caused specifically by the silent mutations and deregulation of SPL13 from miR156. Characterization of the SPL13 promoter indicated that this gene is expressed mainly in the hypocotyl and affects leaf primordium development. These results suggest that the repression of SPL13 by miR156 is essential for normal post-germinative growth in Arabidopsis.

Seeds of Medicago truncatula, a genomic model species for legumes, exhibit physiological and physical dormancy. Here, the factors influencing the germination behaviour of freshly harvested and stored seeds were investigated using several genotypes. Hardseededness is promoted when mature seeds are equilibrated at relative humidities (RH) below 75%. The release of physical dormancy during imbibition was dependent on the initial water content/RH that the seeds were dried to: the drier the seeds, the longer the imbibition time needed to break physical dormancy. The kinetics of physical dormancy release was slower than that of physiological dormancy, making it possible to uncouple both phenomena. Freshly harvested embryos without seed coverings germinated at the same speed as afterripened seeds. The depth of dormancy varied between different M. truncatula genotypes, from more to less dormant: DZA315.16>A17 (Jemalong)>R108>DZA45.5. This difference was eliminated by removing the endosperm. Collectively, these observations indicate that the endosperm is likely the main factor in the reduced germination of freshly harvested seeds. White light decreased germination speed of dormant seeds whereas it had no effect on non-dormant seeds. Recently harvested seeds were most dormant at temperatures above 17°C, whereas afterripened seeds germinated over a wider range of temperature. Fluridone could efficiently break dormancy, reinforcing the role of abscisic acid (ABA) synthesis. However, dormancy was not affected by gibberellic acid (100 μM GA3) or nitrate. The particular dormancy features unravelled here for M. truncatula, combined with the available genomic resources, make it a new, useful model for genetic and molecular studies which can complement those developed for Arabidopsis.

Seeds of Aconitum napellus subsp. castellanum were physiologically dormant at maturity in early autumn, with underdeveloped embryos. Thus they have morphophysiological dormancy (MPD). Embryos in fresh seeds were on average 1.01 mm long, and they had to grow to 3.60 mm before radicle emergence. Cold stratification at 5°C for 5 months with light enhanced the mean embryo length to 2.73 mm (SE = 0.13) and seed germination to 20%. However, with higher temperatures (15/4, 20/7, 25/10, 28/14 and 32/18°C) embryo growth was small, with no seeds germinating. Optimal germination was achieved after 4 months of cold stratification at 5°C followed by incubation at 20/7°C for 1 month with light, when germination ranged between 70 and 79%, depending on seed age, locality and year of collection. Cold stratification could be substituted by the application of GA3 solution, since mean embryo length in seeds incubated at 25/10°C for 1 month with light was 3.52 mm and the germination was 80%. Since cold stratification was the only requirement for the loss of MPD, the longest embryo growth occurred during this treatment, and GA3 promoted MPD loss, we concluded that A. napellus seeds have intermediate complex MPD. Germination was higher in 4-month stored than in freshly matured seeds. A pronounced variability in germinative patterns at inter-annual and inter-population level was recorded.

Detecting the presence of genetically modified plants (adventitious presence of unwanted transgenic plants, AP) from outcrossing species such as maize requires a method that lowers laboratory costs without losing precision. 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 AP in a population at a low cost without losing precision. When pool (group) testing is used to estimate the prevalence of AP (p), there are sampling procedures for calculating a confidence interval (CI); however, they usually do not ensure precision in the estimation of p. This research proposes a method to determine the number of pools (g), given a pool size (k), that ensures precision in the estimated proportion of AP (that is, it ensures a narrow CI). In addition, the study computes the maximum likelihood estimator of p under pool testing and its exact CI, considering the detection limit of the laboratory, d, and the concentration of AP per unit (c). The proposed sample procedure involves two steps: (1) obtain a sample size that guarantees that the mean width of the CI () is narrower than the desired width (ω); and (2) iteratively increase the sample size until is smaller than the desired width (ω) with a specified degree of certainty (γ). Simulated data were created and tables are presented showing the different possible scenarios that a researcher may encounter. An R program is given and explained that will reproduce the results and make it easy for the researcher to create other scenarios.