The MKK3 MAPK cascade integrates temperature and after-ripening signals to modulate seed germination.

The timing of seed germination is controlled by the combination of internal dormancy and external factors. Temperature is a major environmental factor for seed germination. The permissive temperature range for germination is narrow in dormant seeds and expands during after-ripening (AR) (dormancy release). Quantitative trait loci analyses of preharvest sprouting in cereals have revealed that MKK3, a mitogen-activated protein kinase (MAPK) cascade protein, is a negative regulator of grain dormancy. Here, we show that the MAPKKK19/20-MKK3-MPK1/2/7/14 cascade modulates the germination temperature range in Arabidopsis seeds by elevating the germinability of the seeds at sub- and supraoptimal temperatures. The expression of MAPKKK19 and MAPKKK20 is induced around optimal temperature for germination in after-ripened seeds but repressed in dormant seeds. MPK7 activation depends on the expression levels of MAPKKK19/20, with expression occurring under conditions permissive for germination. Abscisic acid (ABA) and gibberellin (GA) are two major phytohormones which are involved in germination control. Activation of the MKK3 cascade represses ABA biosynthesis enzyme gene expression and induces expression of ABA catabolic enzyme and GA biosynthesis enzyme genes, resulting in expansion of the germinable temperature range. Our data demonstrate that the MKK3 cascade integrates temperature and AR signals to phytohormone metabolism and seed germination.

Seed production determines the entrance to dormancy of the inflorescence meristem of Pisum sativum and the end of the flowering period.

Flowering plants adjust their reproductive period to maximize the success of the offspring. Monocarpic plants, those with a single reproductive cycle that precedes plant senescence and death, tightly regulate both flowering initiation and flowering cessation. The end of the flowering period involves the arrest of the inflorescence meristem activity, known as proliferative arrest, in what has been interpreted as an evolutionary adaptation to maximize the allocation of resources to seed production and the viability of the progeny. Factors influencing proliferative arrest were described for several monocarpic plant species many decades ago, but only in the last few years studies performed in Arabidopsis have allowed to approach proliferative arrest regulation in a comprehensive manner by studying the physiology, hormone dynamics, and genetic factors involved in its regulation. However, these studies remain restricted to Arabidopsis and there is a need to expand our knowledge to other monocarpic species to propose general mechanisms controlling the process. In this work, we have characterized proliferative arrest in Pisum sativum, trying to parallel available studies in Arabidopsis to maximize this comparative framework. We have assessed quantitatively the role of fruits/seeds in the process, the influence of the positional effect of these fruits/seeds in the behavior of the inflorescence meristem, and the transcriptomic changes in the inflorescence associated with the arrested state of the meristem. Our results support a high conservation of the factors triggering arrest in pea and Arabidopsis, but also reveal differences reinforcing the need to perform similar studies in other species.

Soil salinity regulates spatial-temporal heterogeneity of seed germination and seedbank persistence of an annual diaspore-trimorphic halophyte in northern China.

BACKGROUND AND AIMS: Seed heteromorphism is a plant strategy that an individual plant produces two or more distinct types of diaspores, which have diverse morphology, dispersal ability, ecological functions and different effects on plant life history traits. The aim of this study was to test the effects of seasonal soil salinity and burial depth on the dynamics of dormancy/germination and persistence/depletion of buried trimorphic diaspores of a desert annual halophyte Atriplex centralasiatica. METHODS: We investigated the effects of salinity and seasonal fluctuations of temperature on germination, recovery of germination and mortality of types A, B, C diaspores of A. centralasiatica in the laboratory and buried diaspores in situ at four soil salinities and three depths. Diaspores were collected monthly from the seedbank from December 2016 to November 2018, and the number of viable diaspores remaining (not depleted) and their germinability were determined. RESULTS: Non-dormant type A diaspores were depleted in the low salinity "window" in the first year. Dormant diaspore types B and C germinated to high percentages at 0.3 and 0.1 mol L-1 soil salinity, respectively. High salinity and shallow burial delayed depletion of diaspore types B and C. High salinity delayed depletion time of the three diaspore types and delayed dormancy release of types B and C diaspores from autumn to spring. Soil salinity modified the response of diaspores in the seedbank by delaying seed dormancy release in autum and winter and by providing a low-salt concentration window for germination of non-dormant diaspores in spring and early summer. CONCLUSIONS: Buried trimorphic diaspores of annual desert halophyte A. centralasiatica exhibited diverse dormancy/germination behavior in respond to seasonal soil salinity fluctuation. Prolonging persistence of the seedbank and delaying depletion of diaspores under salt stress in situ primarily is due to inhibition of dormancy-break. The differences in dormancy/germination and seed persistence in the soil seedbank may be a bet-hadging strategy adapted to stressful temporal and spatial heterogeneity, and allows A. centralasiatica to persist in the unpredictable cold desert enevironment.

Multilayer regulatory landscape and new regulators identification for bud dormancy release and bud break in Populus.

For trees originating from boreal and temperate regions, the dormancy-to-active transition, also known as bud dormancy release and bud break, are crucial processes that allow trees to reactive growth in the spring. The molecular mechanisms underlying these two processes remain poorly understood. Here, through integrative multiomics analysis of the transcriptome, DNA methylome, and proteome, we gained insights into the reprogrammed cellular processes associated with bud dormancy release and bud break. Our findings revealed multilayer regulatory landscapes governing bud dormancy release and bud break regulation, providing a valuable reference framework for future functional studies. Based on the multiomics analysis, we have determined a novel long intergenic noncoding RNA named Phenology Responsive Intergenic lncRNA 1 (PRIR1) plays a role in the activation of bud break. that the molecular mechanism of PRIR1 has been preliminary explored, and it may partially promote bud break by activating its neighbouring gene, EXORDIUM LIKE 5 (PtEXL5), which has also been genetically confirmed as an activator for bud break. This study has revealed a lncRNA-mediated regulatory mechanism for the control of bud break in Populus, operating independently of known regulatory pathways.

Seed dormancy, climate changes, desertification and soil use transformation threaten the Mediterranean endemic monospecific plant Petagnaea gussonei.

This study investigated the germination capacity (endogenous factor) of Petagnaea gussonei (Spreng.) Rauschert, an endemic monospecific plant considered as a relict species of the ancient Mediterranean Tertiary flora. This investigation focused also on the temporal trends of soil-use, climate and desertification (exogenous factors) across the natural range of P. gussonei. The final germination percentage showed low values between 14 and 32%, the latter obtained with GA3 and agar at 10 °C. The rising temperatures in the study area will further increase the dormancy of P. gussonei, whose germination capacity was lower and slower at temperatures higher than 10 °C. A further limiting factor of P. gussonei is its dormancy, which seems to be morpho-physiological. Regarding climate trends, in the period 1931-2020, the average temperature increased by 0.5 °C, from 15.4 to 15.9 °C, in line with the projected climate changes throughout the twenty-first century across the Mediterranean region. The average annual rainfall showed a relatively constant value of c. 900 mm, but extreme events grew considerably in the period 1991-2020. Similarly, the land affected by desertification expanded in an alarming way, by increasing from 21.2% in 2000 to 47.3% in 2020. Soil-use changes created also a complex impacting mosaic where c. 40% are agricultural areas. The effective conservation of P. gussonei should be multilateral by relying on germplasm banks, improving landscape connectivity and vegetation cover, and promoting climate policies.

Hormonal profiles in dormant turions of 22 aquatic plant species: do they reflect functional or taxonomic traits?

BACKGROUND AND AIMS: Turions are vegetative, dormant overwintering organs formed in aquatic plants in response to unfavourable ecological conditions. Contents of cytokinin (CK), auxin metabolites and abscisic acid (ABA) as main growth and development regulators were compared in innately dormant autumnal turions of 22 aquatic plant species of different functional ecological or taxonomic groups with those in non-dormant winter apices in three aquatic species and with those in spring turions of four species after their overwintering. METHODS: The hormones were analysed in miniature turion samples using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry. KEY RESULTS: In innately dormant turions, the total contents of each of the four main CK types, biologically active forms and total CKs differed by two to three orders of magnitude across 22 species; the proportion of active CK forms was 0.18-67 %. Similarly, the content of four auxin forms was extremely variable and the IAA proportion as the active form was 0.014-99 %. The ABA content varied from almost zero to 54 µmol kg-1 dry weight and after overwintering it usually significantly decreased. Of all functional traits studied, hormone profiles depended most on the place of turion sprouting (surface vs bottom) and we suggest that this trait is crucial for turion ecophysiology. CONCLUSIONS: The key role of ABA in regulating turion dormancy was confirmed. However, the highly variable pattern of the ABA content in innately dormant and in overwintered turions indicates that the hormonal mechanism regulating the innate dormancy and its breaking in turions is not uniform within aquatic plants.

The dimorphic diaspore model Aethionema arabicum (Brassicaceae): Distinct molecular and morphological control of responses to parental and germination temperatures.

Plants in habitats with unpredictable conditions often have diversified bet-hedging strategies that ensure fitness over a wider range of variable environmental factors. A striking example is the diaspore (seed and fruit) heteromorphism that evolved to maximize species survival in Aethionema arabicum (Brassicaceae) in which external and endogenous triggers allow the production of two distinct diaspores on the same plant. Using this dimorphic diaspore model, we identified contrasting molecular, biophysical, and ecophysiological mechanisms in the germination responses to different temperatures of the mucilaginous seeds (M+ seed morphs), the dispersed indehiscent fruits (IND fruit morphs), and the bare non-mucilaginous M- seeds obtained by pericarp (fruit coat) removal from IND fruits. Large-scale comparative transcriptome and hormone analyses of M+ seeds, IND fruits, and M- seeds provided comprehensive datasets for their distinct thermal responses. Morph-specific differences in co-expressed gene modules in seeds, as well as in seed and pericarp hormone contents, identified a role of the IND pericarp in imposing coat dormancy by generating hypoxia affecting abscisic acid (ABA) sensitivity. This involved expression of morph-specific transcription factors, hypoxia response, and cell wall remodeling genes, as well as altered ABA metabolism, transport, and signaling. Parental temperature affected ABA contents and ABA-related gene expression and altered IND pericarp biomechanical properties. Elucidating the molecular framework underlying the diaspore heteromorphism can provide insight into developmental responses to globally changing temperatures.

Viability and dormancy of the Clematis vitalba aerial seed bank.

Old man's beard (Clematis vitalba L.) is a liana species that has become invasive in many areas of its introduced range. Seeds are produced in abundance and are both physiologically and morphologically dormant upon maturity. To understand the importance of seeds to its invasiveness, changes in viability and dormancy of the aerial seed bank were tracked throughout the after-ripening period and during storage. Seeds collected every second month for 2 years were subjected to germination tests. Other seeds stored in outdoor ambient conditions or in a dry, chilled state were dissected before, during, and after imbibition, as well as during incubation, to measure embryo size. Less than 72% of seeds on the mother plant were viable. Viable seeds remained completely morpho-physiologically dormant throughout autumn, even when treated with nitrate. Physiological dormancy declined in response to seasonal changes, yet morphological dormancy did not change until seeds had been exposed to appropriate germination conditions for several days. Fully dormant autumn seeds decayed at higher rates during incubation than partially or fully after-ripened seeds, which were also more germinable and less dormant. Furthermore, seeds incubated in complete darkness were more likely to decay or remain dormant than those exposed to light. This study demonstrates that fewer than three-quarters of seeds produced are viable and further decay occurs after dispersal, yet total fertility is still very high, with enormous propagule pressure from seeds alone. Viable seeds are protected with two forms of dormancy; morphological dormancy requires additional germination cues in order to break after seasonal changes break physiological dormancy.

Exploring fine tuning between phytohormones and ROS signaling cascade in regulation of seed dormancy, germination and seedling development.

In higher plants, seed is a propagule which ensures dissemination and survival of species. Developmental phases of a seed comprise embryogenesis, maturation and germination paving a way to its final fate i.e. seedling establishment. The final stage of seed maturation is marked by dehydration, acquisition of dessication tolerance and induction of dormancy. A precise Abscisic acid (ABA) to Gibberellins (GA) ratio, accumulation of miRNA 156, low level of reactive oxygen species (ROS) and enzyme inactivity govern seed dormancy. This also prevent pre harvest sprouting of the seeds. Overtime, stored seed mRNAs and proteins are degraded through oxidation of specific nucleotides in response to ROS accumulation. This degradation alleviates seed dormancy and transforms a dormant seed into a germinating seed. At this stage, ABA catabolism and degradation accompanied by GA synthesis contribute to low ABA to GA ratio. GA as well as ROS acts downstream, to mobilize reserve food materials, rupture testa, enhance imbibition and protrude radicle. All these events mark seed germination. Further, seedling is established under the governance of auxin and light. ABA and GA are master regulators while auxin, cytokinins, ethylene, jasmonic acid, brassinosteroids act through interdependent pathways to tightly regulate seed dormancy, germination and seedling establishment. In this review, the role of phytohormones and ROS in accordance with environmental factors in governing seed dormancy, promoting seed germination and thus, establishing a seedling is discussed in detail.

Fruit dehiscence mechanism and release of dimorphic seeds with different germination properties in Commelina erecta.

Commelina erecta is a successful weed species. The aims of this study were to analyse the morpho-anatomy of the fruit and dimorphic seeds of the weed C. erecta, the dynamics and type of dormancy, and water entry. Flowers and fruits at different development stages were processed using standard anatomical techniques. Besides, experiments of imbibition, germinability and water entry were performed on both seed types. In the fruit of C. erecta, free and coated seeds are developed within dehiscent and indehiscent carpels, respectively. Dehiscent carpels open through a region of mechanical weakness in the dorsal vascular bundle. This region does not form in the indehiscent carpel. The main anatomical differences between the two seed types were observed in the testa and in the number of covering layers. Imbibition experiments showed that the covering of both seed types is water permeable, so these seeds lack physical dormancy and may exhibit physiological dormancy. Germinability experiments showed that the dormancy in free seeds is variable throughout the reproductive season, whereas, in coated seeds, it is high throughout the reproductive season. The embryotega is an area where the hardness of the seed coat is interrupted and facilitates water entry. Differences in the morpho-anatomy of carpels result in the formation of dimorphic seeds with different covering layers and different germination properties. These different properties allow some seeds germinate immediately after falling from the mother plant, and others to be incorporated into the seed bank. These results are useful for designing weed management strategies in agroecosystems.

Dormancy release of seeds of Podophyllum hexandrum Royle accompanied by changes in phytochemicals and inorganic elements.

Podophyllum hexandrum Royle is an alpine medicinal plant of considerable importance, and its seed dormancy severely inhibits population renewal. Although cold stratification can break dormancy to a certain extent, the migration and accumulation of phytochemicals and inorganic elements in the seeds during dormancy release and their functions remain unclear. Changes in phytochemicals and inorganic elements in different seed parts were analyzed during dormancy. The key differential phytochemicals and inorganic elements were screened and their association with dormancy release and their roles in dormancy release were explored. The results showed that dormancy release may have occurred following the decrease in palmitic acid and linoleic acid content in the seeds and the increase in 2,3-dihydro-3,5-dihydro-6-methyl-4 (h)-pyran-4-one content in the endosperm. Meanwhile, 6-propyltridecane and hexadecane in the seed coat may enhance the water permeability of seeds to speed up germination. Mg may migrate from the seed coat to the endosperm and seed embryos, whereas Co may migrate from the seed embryo to the seed coat. Ca, Mn, Mg, and Co are involved in various physiological metabolic processes, which may facilitate the dormancy release of P. hexandrum seeds. These findings have enhanced our understanding of the mechanisms of dormancy release in P. hexandrum seeds and can serve as a reference for the development of more effective dormancy-breaking techniques for the conservation of this endangered medicinal plant.

Dormancy-release and germination improvement of Korean bellflower (Campanula takesimana Nakai), a rare and endemic plant native to the Korean peninsula.

Korean bellflower (Campanula takesimana Nakai) is a rare and perennial herb with medicinal and ornamental values, is endemic to the Ulleung Island of Korea. In this study, we investigated the dormancy-release and germination characteristics of C. takesimana (Campanulaceae) seeds by subjecting them to varying temperatures (5, 10, 15, 20, and 25°C and diurnal/nocturnal temperatures of 15/6, 20/10, and 25/15°C), cold stratification periods (0, 4, 8, or 12 weeks at 5°C), and gibberellic acid (GA3) concentrations (0, 10, 100, or 1,000 mg·L-1 at 15/6°C and 25/15°C) to identify the ideal seed propagation conditions. The seeds were stimulated to germinate (at 25°C, 12-h photoperiod with fluorescent lamps at 40 ± 10 µmol∙m-2∙s-1) after cold stratification. To examine the germination characteristics, the seeds were tested for water imbibition and found to readily absorb water. The seeds exhibited underdeveloped embryos during dispersal, showed final germination of 37.00% ± 4.43 at 25°C and were not influenced by temperature. The seeds subjected to 0, 4, 8, or 12 weeks of cold stratification germinated at a success rate of 22.00% ± 4.76, 87.00% ± 6.80, 79.00% ± 2.52, and 77.00% ± 1.91, respectively. Additionally, the germination characteristics, which were based on final germination, mean germination time, and germination velocity (Timson index), were significantly greater in the seeds pretreated with 1,000 mg·L-1 GA3 at 25/15°C than in seeds pretreated with 0 mg·L-1 GA3. Overall, the seeds broke dormancy with GA3 and short-term cold stratification. Therefore, we concluded that C. takesimana seeds have non-deep, simple, morphophysiological dormancy, and pretreatment with cold stratification and GA3 is required for effective seed propagation.

Global seed dormancy patterns are driven by macroclimate but not fire regime.

Seed dormancy maximizes plant recruitment in habitats with variation in environmental suitability for seedling establishment. Yet, we still lack a comprehensive synthesis of the macroecological drivers of nondormancy and the different classes of seed dormancy: physiological dormancy, morphophysiological dormancy and physical dormancy. We examined current geographic patterns and environmental correlates of global seed dormancy variation. Combining the most updated data set on seed dormancy classes for > 10 000 species with > 4 million georeferenced species occurrences covering all of the world's biomes, we test how this distribution is driven by climate and fire regime. Seed dormancy is prevalent in seasonally cold and dry climates. Physiological dormancy occurs in relatively dry climates with high temperature seasonality (e.g. temperate grasslands). Morphophysiological dormancy is more common in forest-dominated, cold biomes with comparatively high and evenly distributed precipitation. Physical dormancy is associated with dry climates with strong seasonal temperature and precipitation fluctuations (e.g. deserts and savannas). Nondormancy is associated with stable, warm and wetter climates (e.g. tropical rain forest). Pyroclimate had no significant effect on the distribution of seed dormancy. The environmental drivers considered in this study had a comparatively low predictive power, suggesting that macroclimate is just one of several global drivers of seed dormancy.

Intermediate complex morphophysiological dormancy in seeds of Aconitum barbatum (Ranunculaceae).

BACKGROUND: Seed dormancy and germination are key components of plant regeneration strategies. Aconitum barbatum is a plant commonly found in northeast China. Although it has potential for use in gardening and landscaping, its seed dormancy and regeneration strategy, which adapt to its natural habitat, are not well understood. Our aim was to identify conditions for breaking A. barbatum seed dormancy and determine its dormancy type. Embryo growth and germination were determined by collecting seeds over time in the field. Laboratory experiments that control light, temperature, and stratification period were conducted to assess dormancy breaking and germination, and GA3 was used to identify dormancy type. RESULTS: Seeds of A. barbatum have undeveloped embryos with physiological dormancy at maturity in autumn. The embryo-to-seed length ratio increases from 0.33 to 0.78 before the emergence of the radical. Under natural environmental conditions, embryo development begins in early winter. Laboratory experiments have shown that long-term incubation under 4 °C (cold stratification) promotes embryo development and seed dormancy break. With an extension of cold stratification, an increase in germination percentages was observed when seeds were transferred from 4 °C to warmer temperatures. Seeds exposed to light during incubation show a higher germination percentage than those kept in the dark. Seed germination can also be enhanced by a 100 mg/L GA3 concentration. CONCLUSIONS: Seeds of A. barbatum display intermediate complex morphophysiological dormancy at maturity. In addition to the underdeveloped embryo, there are also physiological barriers that prevent the embryo from germinating. Dormancy breaking of A. barbatum seeds can be achieved by natural winter cold stratification, allowing seeds to germinate and sprout seedlings at the beginning of the following growing season. Our findings provide valuable insights into the seed dormancy and regeneration strategy of A. barbatum, which could facilitate its effective utilization in gardening and landscaping.

Methodological and physiological study of seed dormancy release in Tilia henryana.

Tilia henryana is a rare tree of the Tilia family, found exclusively in China. Its seeds have severe dormancy features that limit its normal conditions of reproduction and renewal. Its seeds have severe dormant characteristics that limit its normal conditions of reproduction and renewal. The Dormancy in T. henryana seeds is a comprehensive dormancy (PY + PD) caused by mechanical and permeability barriers of seed coat and the presence of germination inhibitor in endosperm. L9 (34) orthogonal test was used to determine the best procedure for releasing the dormancy of T. henryana seeds, that is, first treating the seeds with H2SO4 for 15 min, followed by the application of 1 g L-1 GA3, stratification at 5 °C for 45 days, and finally germination at 20 °C, which can achieve a 98% seed germination rate. Large amounts of fat are consumed throughout the dormancy release process. As quantities of protein and starch marginally increase, soluble sugars are continuously decreased. Acid phosphatase and amylase activities increased rapidly, and the combined enzyme activities of G-6-PDH and 6-PGDH related to the PPP were also significantly increased. The levels of GA and ZR continued to increase, while the levels of ABA and IAA gradually decreased, among which GA and ABA changed most rapidly. The total amino acids content continued to decrease. Asp, Cys, Leu, Phe, His, Lys and Arg decreased with dormancy release, while Ser, Glu, Ala, Ile, Pro and Gaba showed an upward trend. The physical dormancy of T. henryana seeds is broken with H2SO4 in order to make the seed coat more permeable, which is a prerequisite for germination. As a result, the seeds can absorb water and engage in physiological metabolic activities, particularly the hydrolysis and metabolism of fat, which supply a significant amount of energy for dormancy release. In addition, rapid variations in the levels of different endogenous hormones and free amino acids, induced by cold stratification and GA3 application, are another important factor promoting the quick physiological activation of seeds and breaking the endosperm barrier.

Multiomics strategies for decoding seed dormancy breakdown in Paris polyphylla.

BACKGROUND: The disruption of seed dormancy is a complicated process and is controlled by various factors. Among these factors, membrane lipids and plant hormones are two of the most important ones. Paris polyphylla is an important Chinese herbaceous species, and the dormancy trait of its seed limits the cultivation of this herb. RESULTS: In this study, we investigate the global metabolic and transcriptomic profiles of Paris polyphylla during seed dormancy breaking. Widely targeted metabolomics revealed that lysophospholipids (lysoPLs) increased during P. polyphylla seed dormancy breaking. The expression of phospholipase A2 (PLA2), genes correlated to the production of lysoPLs, up-regulated significantly during this process. Abscisic acid (ABA) decreased dramatically during seed dormancy breaking of P. polyphylla. Changes of different GAs varied during P. polyphylla seeds dormancy breaking, 13-OH GAs, such as GA53 were not detected, and GA3 decreased significantly, whereas 13-H GAs, such as GA15, GA24 and GA4 increased. The expression of CYP707As was not synchronous with the change of ABA content, and the expression of most UGTs, GA20ox and GA3ox up-regulated during seed dormancy breaking. CONCLUSIONS: These results suggest that PLA2 mediated production of lysoPLs may correlate to the seed dormancy breaking of P. polyphylla. The conversion of ABA to ABA-GE catalysed by UGTs may be the main cause of ABA degradation. Through inhibition the expression of genes related to the synthesis of 13-OH GAs and up-regulation genes related to the synthesis of 13-H GAs, P. polyphylla synthesized more bioactive 13-H GA (GA4) to break its seed dormancy.

Non-deep physiological dormancy and germination characteristics of Primula florindae (Primulaceae), a rare alpine plant in the Hengduan Mountains of southwest China.

Timing of seed germination is directly related to the survival probability of seedlings. For alpine plants, autumn-dispersal seeds should not germinate immediately because the cold temperature is not conducive to the survival of seedlings. Seed dormancy is a characteristic of the seed that prevents it from germinating after dispersal. Primula florindae is an alpine perennial forb endemic to eastern Tibet, SW China. We hypothesized that primary dormancy and environmental factors prevent seeds of P. florindae to germinate in autumn and allow them to germinate at the first opportunity in spring. We determined how GA3, light, temperature, dry after-ripening (DAR) and cold-wet stratification (CS) treatments affect seed germination by conducting a series of laboratory experiments. Firstly, the effects of gibberellic acid (GA3; 0, 20, and 200 mg L-1) on germination of freshly shed seeds at alternating temperatures (15/5 and 25/15 °C) were immediately investigated to characterize seed with a physiological dormancy component. Then, the fresh seeds treated with 0, 3, and 6 months of after-ripening (DAR) and cold-wet stratification (CS) were incubated at seven constant (1, 5, 10, 15, 20, 25, and 30 °C) and two alternating temperatures (5/1, 15/5, and 25/15 °C) at light and dark conditions. Fresh seeds were dormant, which only germinated well (>60%) at 20, 25, and 25/15 °C in light but not at ≤15 °C and to higher percentages in light than in dark. GA3 increased germination percentage of fresh seeds, and DAR or CS treatments increased final germination percentage, germination rate (speed), and widened the temperature range for germination from high to low. Moreover, CS treatments reduced the light requirement for germination. Thus, after dormancy release, seeds germinated over a wide range of constant and alternating temperatures, regardless of light conditions. Our results demonstrated that P. florindae seeds have type 2 non-deep physiological dormancy. Timing of germination should be restricted to early spring, ensuring a sufficient length of the growing season for seedling recruitment. These dormancy/germination characteristics prevent seeds from germinating in autumn when temperatures are low but allow them to germinate after snowmelt in spring.

Seed germination variability: why do genetically identical seeds not germinate at the same time?

For survival in the natural environment, plants have evolved a 'bet-hedging' strategy where individual variation is high and a range of phenotypes is produced. When faced with unpredictable environmental conditions, fluctuation in seed behaviour is a beneficial trait that allows plant species to survive, particularly if seedlings from early-germinated seeds die. However, this is not a desired trait from an agricultural perspective, where a set of uniformly growing seedlings is required. Whilst variability in seed behaviour is unavoidable, over the centuries humans have attempted to select seeds with minimum variability for agricultural use. In the model plant Arabidopsis, even non-stratified seeds in the same silique germinate variably, and it remains elusive how this variability is manifested from genes to a physiological outcome and what molecular mechanisms of bet-hedging facilitate this diversity. Will the re-introduction of valuable wild alleles into domesticated crops contribute to this variability between individual seeds by promoting bet-hedging? Recent advances have shed light on possible molecular pathways of germination that are affected at the level of single seeds and single cells. Here, we review the hormonal, molecular, and cellular mechanisms that might affect the germination outcome of individual genetically identical seeds.

Morphological and anatomical changes during dormancy break of the seeds of Fritillaria taipaiensis.

Fritillaria taipaiensis P. Y. Li is the most suitable species planted at low altitudes among other species used as Tendrilleaf Fritillary Bulb, whose seeds embracing the morphological and physiological dormancy need to experience a long-dormant time from sowing to germination. In this study, the developmental changes of F. taipaiensis seeds during dormancy period were observed by morphological and anatomical observation, and the cause of long-term dormancy of seeds was discussed from the perspective of embryonic development. The process of embryonic organogenesis was revealed during the dormancy stage by the paraffin section. The effects of testa, endosperm and temperature on dormant seeds were discussed. Furthermore, we found that the mainly dormant reason was caused by the morphological dormancy, which accounted for 86% of seed development time. The differentiation time of the globular or pear-shaped embryo into a short-rod embryo was longer, which was one of the chief reasons for the morphological dormancy and played an important role in embryonic formation. Testa and endosperm with mechanical constraint and inhibitors involved in the dormancy of F. taipaiensis seeds. The seeds of F. taipaiensis, the average ambient temperature of 6-12°C for morphological dormancy and 11-22°C for physiological dormancy, were unsuitable for seed growth. Therefore, we suggested that the dormancy time of F. taipaiensis seeds could be shortened by shortening the development time of the proembryo stage and stratification for the different stages of dormancy.

The organ-building process of the rod embryo was revealed during dormancy.Morphological after-ripening is one of the main reasons for the seed dormancy of Fritillaria taipaiensis.The Morphological long dormancy is mainly caused by the long period of proembryo development.