Global DNA methylation and cellular 5-methylcytosine and H4 acetylated patterns in primary and secondary dormant seeds of Capsella bursa-pastoris (L.) Medik. (shepherd’s purse)

Despite the importance of dormancy and dormancy cycling for plants’ fitness and life cycle phenology, a comprehensive characterization of the global and cellular epigenetic patterns across space and time in different seed dormancy states is lacking. Using Capsella bursa-pastoris (L.) Medik. (shepherd’s purse) seeds with primary and secondary dormancy, we investigated the dynamics of global genomic DNA methylation and explored the spatio-temporal distribution of 5-methylcytosine (5-mC) and histone H4 acetylated (H4Ac) epigenetic marks. Seeds were imbibed at 30 °C in a light regime to maintain primary dormancy, or in darkness to induce secondary dormancy. An ELISA-based method was used to quantify DNA methylation, in relation to total genomic cytosines. Immunolocalization of 5-mC and H4Ac within whole seeds (i.e., including testa) was assessed with reference to embryo anatomy. Global DNA methylation levels were highest in prolonged (14 days) imbibed primary dormant seeds, with more 5-mC marked nuclei present only in specific parts of the seed (e.g., SAM and cotyledons). In secondary dormant seeds, global methylation levels and 5-mC signal where higher at 3 and 7 days than 1 or 14 days. With respect to acetylation, seeds had fewer H4Ac marked nuclei (e.g., SAM) in deeper dormant states, for both types of dormancy. However, the RAM still showed signal after 14 days of imbibition under dormancy-inducing conditions, suggesting a central role for the radicle/RAM in the response to perceived ambient changes and the adjustment of the seed dormancy state. Thus, we show that seed dormancy involves extensive cellular remodeling of DNA methylation and H4 acetylation.

Critical role of air and soil temperature in the development of primary and secondary physical dormancy in Albizia julibrissin (Fabaceae)

Physical dormancy (PY) is typically induced by seed coat impermeability that develops once the moisture content of seeds drops below a species-specific threshold. Considering this, we utilized Albizia julibrissin (Fabaceae) to ask (i) whether seeds that mature on the outer branches of trees (directly exposed to sunlight) are more likely to be impermeable than seeds matured under canopy cover; (ii) whether this difference might be explained by the maternal environment in which the seeds mature; and (iii) which conditions impose secondary dormancy following dispersal? Temperature was tracked in both shaded and sun-exposed seed pods throughout the growing season using data-loggers. Temperatures remained lower in pods under canopy cover than those exposed to direct sunlight. Consequently, the moisture content of seeds collected from sun-exposed branches were significantly lower than seeds matured under canopy cover, thereby producing a higher percentage of impermeable seeds. A dispersal-mimicking experiment revealed that seeds matured in sun-exposed branches and subsequently dispersed to an open site for 4 months were more likely to develop impermeability (i.e. secondary dormancy). The opposite was found to be true for seeds matured in shaded branches and subsequently dispersed to a canopy-covered site. We conclude that the microclimate of both the maternal environment in which seeds mature, and the site to which they disperse, determines the development of primary and secondary dormancy, respectively.

Temperature Regulation of Primary and Secondary Seed Dormancy in Rosa canina L.: Findings from Proteomic Analysis

Temperature is a key environmental factor restricting seed germination. Rose (Rosa canina L.) seeds are characterized by physical/physiological dormancy, which is broken during warm, followed by cold stratification. Exposing pretreated seeds to 20 °C resulted in the induction of secondary dormancy. The aim of this study was to identify and functionally characterize the proteins associated with dormancy control of rose seeds. Proteins from primary dormant, after warm and cold stratification (nondormant), and secondary dormant seeds were analyzed using 2-D electrophoresis. Proteins that varied in abundance were identified by mass spectrometry. Results showed that cold stratifications affected the variability of the highest number of spots, and there were more common spots with secondary dormancy than with warm stratification. The increase of mitochondrial proteins and actin during dormancy breaking suggests changes in cell functioning and seed preparation to germination. Secondary dormant seeds were characterized by low levels of legumin, metabolic enzymes, and actin, suggesting the consumption of storage materials, a decrease in metabolic activity, and cell elongation. Breaking the dormancy of rose seeds increased the abundance of cellular and metabolic proteins that promote germination. Induction of secondary dormancy caused a decrease in these proteins and germination arrest.

Effect of storage temperature on occurrence of secondary dormancy in buckwheat seeds

Buckwheat (Fagopyrum spp.) is an important crop in many parts of the world, and many varieties have been developed. One of the important breeding issues to be addressed is the low resistance to preharvest-sprouting which causes loss in yields and quality of buckwheat grain. The occurrence of dormancy in cultivated buckwheat, including presence of secondary dormancy has yet to be clarified. In this study, we investigated the effects of storage temperature and periods using nine common buckwheat (Fagopyrum esculentum), seven Tartary buckwheat (Fagopyrum tataricum) and one wild buckwheat (Fagopyrum homotropicum). In some Tartary buckwheats stored at 5°C, the germination of seeds stored 10 days harvest was significantly less than the germination of seeds collected and sown at harvest maturity; the germination of other germplasm, including common buckwheat, was not affected by storage. This result indicates that Tartary buckwheat can acquire secondary dormancy. In addition, the wild buckwheat did not germinate at any storage temperature up to 30 days after harvest maturity; therefore, this buckwheat is considered to be promising breeding material for increasing resistance to preharvest-sprouting resistance.

A Perspective on Secondary Seed Dormancy in Arabidopsis thaliana

Primary seed dormancy is the phenomenon whereby seeds newly shed by the mother plant are unable to germinate under otherwise favorable conditions for germination. Primary dormancy is released during dry seed storage (after-ripening), and the seeds acquire the capacity to germinate upon imbibition under favorable conditions, i.e., they become non-dormant. Primary dormancy can also be released from the seed by various treatments, for example, by cold imbibition (stratification). Non-dormant seeds can temporarily block their germination if exposed to unfavorable conditions upon seed imbibition until favorable conditions are available. Nevertheless, prolonged unfavorable conditions will re-induce dormancy, i.e., germination will be blocked upon exposure to favorable conditions. This phenomenon is referred to as secondary dormancy. Relative to primary dormancy, the mechanisms underlying secondary dormancy remain understudied in Arabidopsis thaliana and largely unknown. This is partly due to the experimental difficulty in observing secondary dormancy in the laboratory and the absence of established experimental protocols. Here, an overview is provided of the current knowledge on secondary dormancy focusing on A. thaliana, and a working model describing secondary dormancy is proposed, focusing on the interaction of primary and secondary dormancy.

The role of soil temperature and seed dormancy in the creation and maintenance of persistent seed banks of Nassella trichotoma (serrated tussock) on the Northern Tablelands of New South Wales

A large and persistent soil seed bank characterises many important grass weeds, including Nassella trichotoma (Nees) Hack. ex Arechav. (serrated tussock), a major weed in Australia and other countries. In the present study we examined the effects of constant and alternating temperatures in regulating primary and secondary dormancy and the creation and maintenance of its soil seed bank in northern NSW, Australia. One-month-old seeds were stored at 4, 25°C, 40/10°C and 40°C, in a laboratory, and germination tests were conducted every two weeks. Few seeds germinated following storage at 4°C, compared with seeds stored at 25°C, 40/10°C and 40°C. Nylon bags containing freshly harvested seeds were buried among N. trichotoma stands in early summer, and germination tests conducted following exhumation after each season over the next 12 months. Seeds buried over summer and summer plus autumn had higher germination than seeds buried over summer plus autumn plus winter, but germination increased again in the subsequent spring. Seeds stored for zero, three, six and 12 months at laboratory temperatures were placed on a thermogradient plate with 81 temperature combinations, followed by incubation at constant 25°C of un-germinated seeds. Constant high or low temperatures prolonged primary dormancy or induced secondary dormancy whereas alternating temperatures tended to break dormancy. Few temperature combinations resulted in more than 80% germination.