Dormancy-breaking requirements and level of morphophysiological dormancy (MPD) were determined for seeds of Ilex formosana and I. uraiensis from the subtropical region, and seeds of I. rotunda from both the subtropical and tropical regions of Taiwan. We hypothesized that some Ilex species would have deep simple MPD broken by warm stratification. Germination of seeds and embryo growth was monitored at 30/20, 25/15, 20/10, 15/5°C and at 25°C. Seeds were cold-stratified and then incubated at 25/15°C, and seeds treated with GA3 and GA4 were incubated at 25/15°C. Fresh seeds reached 50% germination after 11–45 weeks of warm stratification. Pre-treatment with GA increased germination percentages of I. formosana and I. rotunda (tropical) but not I. uraiensis and I. rotunda (subtropical), while cold stratification did not promote germination of either species but increased the germination rate of I. rotunda (tropical). Embryo length in seeds of all species increased ≥710% prior to root emergence, and growth occurred during warm stratification. The positive response to GA and relatively short time for beginning of germination and to reach 50% germination indicate non-deep simple MPD in seeds of I. formosana and I. rotunda (tropical). The negative response to GA and long time for beginning of germination and to reach 50% germination indicate deep simple MPD in seeds of I. uraiensis and I. rotunda (subtropical). Thus, in both the subtropical and tropical regions of Taiwan, the seeds of Ilex species have non-deep simple and deep simple MPD that are broken by warm stratification. Furthermore, GA treatment increases the germination rate and percentage of Ilex seeds with non-deep simple MPD, and cold stratification promotes the seed germination rate of Ilex species with non-deep simple MPD in tropical region.

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.

DNA synthesis in organelles seems to be an early event during maize germination. Mitochondria and plastids from maize embryo axes have been separated using differential and gradient centrifugations and DNA polymerase activity has been studied in extracts from each organelle. It has been found that the enzymes from both organelles show some differences from other plant organellar polymerases in terms of response to specific inhibitors, DNA templates and cations. A comparison of different plant organellar DNA polymerases is presented.

Induction of nine-cis-epoxycarotenoid dioxygenase 6 (NCED6), an abscisic acid (ABA) biosynthesis gene, alone is sufficient to suspend germination in testa-ruptured seeds, which are at the final step of germination. Molecular consequences of NCED6 induction in imbibed seeds were investigated by RNA sequencing. The analysis identified many unknown and uncharacterized genes that were up-regulated by NCED6 induction, in addition to the major regulators of ABA signalling. Interestingly, other NCEDs were up-regulated by NCED6 induction, suggesting that the major rate-limiting enzymes in the ABA biosynthesis pathway are subject to positive-feedback regulation. ZEAXANTHIN EPOXIDASE and ABSCISIC ALDEHYDE OXIDASE3, which function upstream and downstream of NCED, were also up-regulated in seeds by NCED6 induction, which suggests that the distinct layers of positive feedback loops are coordinately operating in the NCED6-induced seeds. SOMNUS (SOM), which was also up-regulated by NCED6 induction, was the major mediator of enhanced ABA signalling in NCED6-induced seeds. SOM exerted negative effects on GA biosynthesis, which also contributes to a high ABA–GA ratio and reinforces the suppressive state of germination. Besides these coding genes, long intergenic non-coding RNAs (lincRNAs) were also up-regulated upon NCED6 induction (termed N6LINCRs). Conditional expression of N6LINCR1 altered gene expression profiles in seeds. Twenty-six genes were up-regulated and 66 genes were down-regulated by the induction of N6LINCR1. These results suggest that some of N6LINCRs have a regulatory role in gene expression in seeds, which potentially contributes to the regulation of germination by ABA.

Southern European peninsulas have repeatedly played notable roles as refuges in the natural history of flora during periods of glaciation. Euonymus latifolius (Celastraceae) is a relict species from the last Würmian glaciation in the Iberian Peninsula. It still lives with isolated populations in favourable, cool mountainous microhabitats, being an extremely rare, critically endangered species. These Iberian populations are often high-priority targets for conservation due to their long-term persistence and unique evolutionary trajectory. Previously, it has not been feasible to promote significant programmes for reintroduction and/or population reinforcement of this singular plant species due to the great difficulties of conventional propagation. In this study, we analysed the effects of temperature, light and gibberellic acid (GA3) on the germination responses of E. latifolius to develop an effective protocol for seed germination as a main outcome. The results are coherent with the climatic temperature conditions recorded broadly in the Iberian Peninsula in the past and in the current refuge locations for the taxon. The germination responses of E. latifolius are compatible with those of seeds with intermediate physiological dormancy. In particular, the seeds required a 10-week warm period (20/7°C + 15/4°C) followed by 20 weeks of cold period (5 + 1.5°C) to break dormancy and achieve germination values over 90%. GA3 also promoted germination (80%). Therefore, we developed the first effective protocol for promoting E. latifolius seed germination and, thus, sexual propagation, to facilitate urgent ex situ actions in the current climate change context.

Soil tillage operations stimulate germination of buried seeds in cultivated lands, allowing them to perceive light as a germination-promoting factor. The time of burial and the effect of changing environmental factors affect the physiological state of the seeds, which may lead to an extreme light-sensitivity and very low fluence response (VLFR) through phytochrome A. This paper describes the influence of the progressive process of dormancy breakage, which is accompanied by the acquisition of extreme light-sensitivity, on processes associated with endosperm weakening and embryo growth potential in the VLFR-mediated promotion of Datura ferox seed germination. Our results show that endosperm weakening is mainly limited by β-mannosidase enzyme activity after far-red light stimulation, which is highly dependent on the dormancy level of the seeds. In addition, stimulation of the embryo growth potential by far-red irradiation did not require an extreme light-sensitivity to very low fluence of photons to reach its maximum response, and it was not completely correlated with expansin gene expression in the embryo. Our work indicates that responses of endosperm weakening and embryo growth potential to far-red irradiation, dependent on dormancy level, have different requirements for stimulation by the signalling network initiated by phytochrome A during the course of the very low fluence response in Datura ferox seeds.

Testing for the unintended or adventitious presence (AP) of genetically modified (GM) events in seed lots is a common practice to comply with regulatory requirements and good stewardship practices. A subsample of a seed lot is typically tested for AP levels, and then statistical methods are used to estimate the upper level of AP in the remainder of the lot with a given level of confidence. For large seed lots, a binomial distribution is typically assumed, but for seed lots where the tested sample is a substantial proportion of the overall seed lot, a hypergeometric distribution is typically assumed. Due to the destructive nature of AP seed testing, we suggest that this latter method may overestimate confidence of low AP in the remaining seed.

Protein storage vacuoles (PSVs) from radicles and cotyledons of dry cotton seeds were isolated by differential centrifugation following homogenization in glycerol. Protein complement analysis of isolated PSVs with one dimensional SDS-PAGE gels revealed similar major storage proteins, viz. 53 and 48 kDa, with differences in lower molecular mass proteins. Radicle PSVs have apparently more 35-kDa and less 22-kDa storage protein than do cotyledon PSVs. The mineral composition of whole radicles, cotyledons and isolated PSVs from radicles and cotyledons was determined by atomic absorption spectroscopy and colorimetric elemental analyses. The concentration of calcium (Ca), magnesium (Mg), potassium (K) and phosphate (P) was lower in isolated PSVs from radicles than from cotyledons, resulting in a marked difference in the Mg/Ca and (Mg+Ca)/K ratios in PSVs from these two sources. Analysis of radicle and cotyledon tissue from dry seeds for mineral distribution with EDX and scanning electron microscopy revealed major concentrations of Mg, K and P in PSVs. These observations indicate that PSVs in radicles are similar in protein and mineral composition to PSVs in cotyledons. PSVs in radicles have the potential function as storage organelles to provide minerals and nutrients for radicle growth during imbibition and germination.

‘Seed heteromorphism’ is a broadly- and loosely-defined term used to describe differences in size/mass, morphology, position on mother plants and ecological function (e.g. dispersal, dormancy/germination) of two or more seeds or other diaspores produced by an individual plant. The primary aim of this review paper was to characterize via an in-depth classification scheme the physical structural design (‘architecture’) of diaspore monomorphism and diaspore heteromorphism in angiosperms. The diaspore classification schemes of Mandák and Barker were expanded/modified, and in doing so some of the terminology that Zohary, Ellner and Shmida, and van der Pijl used for describing diaspore dispersal were incorporated into our system. Based on their (relative) size, morphology and position on the mother plant, diaspores of angiosperms were divided into two divisions and each of these into several successively lower hierarchical layers. Thus, our classification scheme, an earlier version of which was published in the second edition of ‘Seeds’ by Baskin and Baskin, includes not only heteromorphic but also monomorphic diaspores, the Division to which the diaspores of the vast majority of angiosperms belong. The scheme will be useful in describing the ecology, biogeography and evolution of seed heteromorphism in flowering plants.