The Role of Chilling in the Germination of some Scottish Montane Species

Summary

Seeds of 32 montane species were collected throughout northern Scotland. Germination responses, particularly the effects of chilling on breaking dormancy, were examined using a serial test incorporating single- and double-chilling treatments, alternating 20°C/10°C incubation temperatures, gibberellic acid and, for some species, nicking of the seed coat. Where practicable, any ungerminated seeds were ultimately dissected to assess their viability. Only three species had an absolute requirement for pre-chilling before seeds germinated. A further eight species required chilling to break the dormancy of 15% or more seeds; otherwise chilling generally increased the extent and rate of germination. Unusually, chilling induced dormancy in the seeds of four species whereas warm conditions induced dormancy which was not broken by subsequent chilling amongst seeds of Draba incana. Germinability and germination rates for nine species were regressed on the altitude, latitude and oceanicity of the plants' origin to assess the relative effects of chilling. These three environmental factors accounted for up to 98% of the variability in the germination parameters but few regressions attained statistical significance. Broad patterns suggest that chilling had a decreasing effect on both the extent and, rate of germination as the altitude of the seed source increased. A similar pattern, but only for germinability, was seen with respect to latitude.     

Seed dormancy/germination traits of seven Persicaria species and their implication in soil seed-bank strategy

In order to predict species-specific potential to form persistent soil seed-banks and to characterize the dynamics of their seed-banks, the seed dormancy/germination traits of seven Persicaria (Polygonum s.lat.) species sharing lakeshore habitats in central Japan were examined. Strict light requirements for seed germination were not observed in any of the species examined. Although all species required moist chilling (0–6 weeks) to break seed dormancy and were sensitive to temperature fluctuation, the degree of both responses varied between species. Seed germination of Persicaria hydropiper (L.) Spach, Persicaria lapathifolia (L.) S.F. Gray, and Persicaria longiseta (De Bruyn) Kitag. was more accelerated by temperature fluctuation and required shorter chilling periods compared with Persicaria japonica (Meisn.) H. Gross, Persicaria maackiana (Regel) Nakai, Persicaria thunbergii (Sieb. et Zucc.) H. Gross, and Persicaria sieboldi (Maisn.) Onki. Secondary dormancy was induced in all species at higher temperatures (24 and 30°C). A persistent seed-bank strategy suggested by the dormancy/germination traits of the studied species was also demonstrated by seedling emergence from surface soils collected from the natural habitat immediately before seed dispersal, as well as by viable seed persistence for 13 months in the field in a seed burial experiment. In the natural habitat, the species with longer chilling requirements occurred in various microhabitats, including the interior of moist tall grasslands, whereas the species having higher sensitivity to temperature fluctuation were most frequently found in sparsely vegetated microhabitats.     

Seed germination traits of Trapella sinensis (Trapellaceae), an endangered aquatic plant in Japan: Conservation implications

We investigated the effects of cold stratification, temperature, light, and oxygen conditions on seed germination of Trapella sinensis Oliver, an endangered aquatic plant in Japan. Seeds had physiological dormancy, and final germination rate increased with an increasing period of cold stratification. Seeds of T. sinensis had an almost absolute requirement for aerobic conditions to germinate. Also, alternating temperatures significantly promoted germination regardless of light conditions, although final germination percentage was twice to four times higher in light under constant temperature conditions. Suitable sites for germination of T. sinensis appear to be shallow water with alternating temperatures and sufficient oxygen and sediment anoxia, caused by eutrophication of water bodies, may have prevented recruitment of T. sinensis from seed.     

Effects of temperature, light, nitrate and pre chilling on seed germination of grassland plants

The effects of factorial combinations of alternating (20/10 ^oC) and constant temperature (20 ^oC), of light (intermittent) and dark, and of distilled water and nitrate on the germination of a range of species of indigenous grassland were investigated in 1979, and in 1980 when the effects of pre-chilling were also studied. Species differed greatly in their response to the eight sets of environmental conditions in 1979. With the exception of Lolium perenne ssp. perenne and Cynosurus cristatus, only a small percentage of seeds of most species was able to germinate in constant temperature in the dark. However, when light was supplied there was moderate germination of Anthoxanthum odoratum, Cerastium fontanum ssp. glabrescens, Festuca rubra, Holcus lanatus and Poa trivialis. Alternating temperature greatly increased the germination of most species and nitrate further increased germination of Agrostis capillaris, Deschampsia caespitosa, H. lanatus, Poa annua and P. trivialis. In alternating temperatures, light increased germination of these species even more than did nitrate. A. capillaris, D. caespitosa and P. annua required all three factors for maximum germination, and another 7% of seeds of A. capillaris also required gibberellic acid. Germination responses following sequential application of factors often differed from those resulting from simultaneous treatment: in particular, the germination of Ranunculus species was greatly enhanced. Although seeds of the species tested in 1980 were more germinable than those of the same species in 1979, they responded similarly to the different factors. Light increased the germination of both Taraxacum officinale and Plantago lanceolata in constant but not in alternating temperature, while nitrate was much more stimulatory to the latter species. About half the seeds of P. lanceolata were dormant. Pre-chilling at 4^oC for 7 days increased subsequent germination of all species when followed by constant temperature, except of A. capillaris in the dark and C. cristatus (already maximal) in the light. When followed by alternating temperature in the dark, pre-chilling greatly decreased germination of A. capillaris. In the light, where germination of most species was maximal, there was little effect of pre-chilling. Longer durations (21 and 42 days) of chilling of older seed gave similar results to the 7 day pre-chilling.     

Physiological and molecular mechanisms underlying the integration of light and temperature cues in Arabidopsis thaliana seeds

The relief of dormancy and the promotion of seed germination are of extreme importance for a successful seedling establishment. Although alternating temperatures and light are signals promoting the relief of seed dormancy, the underlying mechanisms of their interaction in seeds are scarcely known. By exposing imbibed Arabidopsis thaliana dormant seeds to two‐day temperature cycles previous of a red light pulse, we demonstrate that the germination mediated by phytochrome B requires the presence of functional PSEUDO‐RESPONSE REGULATOR 7 (PRR7) and TIMING OF CAB EXPRESSION 1 (TOC1) alleles. In addition, daily cycles of alternating temperatures in darkness reduce the protein levels of DELAY OF GERMINATION 1 (DOG1), allowing the expression of TOC1 to induce seed germination. Our results suggest a functional role for some components of the circadian clock related with the action of DOG1 for the integration of alternating temperatures and light signals in the relief of seed dormancy. The synchronization of germination by the synergic action of light and temperature through the activity of circadian clock might have ecological and adaptive consequences.     

The Mechanism of Low Temperature Dormancy in Mature Seeds of Syringa Species

The mechanism of seed dormancy at low temperatures (15-9°C) was investigated in the seeds of Syringa josikaea, S. reflexa and S. vulgaris. Low temperature dormancy in Syringa species was mainly imposed by endosperm embedding the radicle. Different degrees of embryo dormancy may occur in S. reflexa seeds. In most cases the low temperature dormancy was broken completely by removing the endosperm around the radicle. The endosperm did not seem to contain significant quantities of germination inhibitors, and the results indicate that it prevents germination mainly due to its mechanical resistance. The mechanical resistance of endosperm did not change during chilling or during induction of dormancy by high temperature incubation. The strength of the endosperm decreased rapidly in non-dormant seeds before visible germination. Similar changes were not observed in dormant seeds. Generally, the strength of the endosperm was lower in the non- (or less) dormant species S. josikaea and S. vulgaris than in the more dormant S. reflexa seeds. The growth potential of the embryos, measured as their ability to germinate in osmotic solutions (mannitol or polyethylen glycol 4000), was increased by chilling and by GA₃-treatment. The growth potential of untreated S. josikaea and S. vulgaris embryos was generally higher than that of S. reflexa embryos. Acid ethyl acetate fractions of methanol extracts from embryos of all three species contained substances with GA³-like activity in the lettuce hypocotyl test. The activity was found at Rf 0.9–1.0 on paper chromatograms run in distilled water. No significant changes in the activity were detected during chilling or prior to visible germination.     

Effects of Chilling, Light and Nitrogen-containing Compounds on Germination, Rate of Germination and Seed Imbibition of Clematis vitalba L.

Effects of chilling (5 °C) period, light and applied nitrogen(N) on germination (%), rate of germination (d to 50% of totalgermination; T_50%) and seed imbibition were examined inClematisvitalba L. In the absence of chilling, light and N, germinationwas minimal (3%). When applied alone, both chilling and N increasedgermination. Chilling for 12 weeks increased germination to64%, and 2.5 mM NO^-₃or NH⁺₄increased germination to 10–12%.Light did not increase germination when applied alone, but didwhen applied in combination with chilling and/or N. Half theseed germinated when light was combined with 2.5 mM NO^-₃or NH⁺₄.The influence of chilling, light and/or N on germination wasgreater when combined, than when either factor was applied alone.Both oxidized (NO^-₃) and reduced (NH⁺₄) forms of N increasedgermination, but non-N-containing compounds did not, suggestingthe response was due to N and not ionic or osmotic effects. Without additional N, T_50%decreased from 16–20 d at zerochilling, to around 5 d at 8 and 12 weeks chilling. AlthoughT_50%was not influenced by an increase in NO^-₃or NH⁺₄from 0.5to 5.0 mM , it did increase with additional applied N thereafter.However, the magnitude of the N effect was small compared tothat of chilling. Like germination, seed imbibition increasedwith a longer chilling period, but in contrast imbibition decreasedslightly with increased applied NO^-₃or NH⁺₄. It is argued thatincreased imbibition is not directly related to an increasein total germination, but that it may be related to the rateof germination. Possible mechanisms involved in the reductionin dormancy ofC. vitalba seed are discussed. Clematis vitalba L.; germination; dormancy; imbibition; rate of germination; chilling; light; nitrate; ammonium; nitrogen; phytochrome     

Nitrate Reductase Independent Stimulation of Seed Germination in Sisymbrium officinale L. (Hedge Mustard) by Light and Nitrate

After a 72 h preincubation in darkness at 15 °C seed germinationof Sisymbrium officinale L. (hedge mustard) at 24 °C wasstimulated by a combination of red light and nitrate. In thepresence of nitrate the seeds escaped from the inhibiting effectof far-red irradiation with an escape time of approx. 8 h. Afterred light, the exposure of seeds to nitrate could be delayedfor 3 h without affecting maximal germination. Prolonged delayresulted in a decrease of the germination response. The possibilitythat nitrate reduction was involved in the stimulation of germinationwas studied by pre-incubating seeds for 72 h in nitrate andsubsequently transferring them to water and irradiating withred light. During the first 8 h period after the red irradiationin which induction of germination occurred, total nitrate levels(endogenous + leachate) remained constant, indicating an absenceof nitrate reductase activity. During the next 8 h visible germinationstarted and total nitrate levels declined, suggesting inductionof nitrate reduction. It is concluded that nitrate reductiondocs not play a role in the induction of germination. The conclusionwas supported by the lack of inhibition of seed germinationby sodium chlorate and sodium tungstate in spite of an inhibitionof nitrate reduction of 80 and 100%, respectively. The contrastsbetween our results and hypotheses concerning the mechanismof action of nitrate in seed germination are discussed Sisymbrium officinale L., hedge mustard, germination, light, nitrate, nitrate reductase     

Effects of seed moisture content,warm, chilling,and exogenous hormone treatments and germination temperature on the germination of blackthorn seeds

Blackthorn (Prunus spinosa L.) germination is often low, so new methods need to be developed with a view to improving nursery yields and to inform decision-making on natural regeneration. To this end, the effects of seed moisture content (MC) levels in combination with warm and chilling treatments on blackthorn seed dormancy release were investigated. In another experiment, the effect on seed germination of warm and chilling treatments in combination with exogenous hormones was investigated. Following treatment, the seeds were allowed to germinate at a constant 15°C with 8 h lighting per day or 20 (dark)/30°C (light). Seed lot effects were evident, but were consistent across treatments. Seeds adjusted to the lower target MC level (TMC) maintained high germination potential over a longer period of treatment than in those held in the fully imbibed (FI) state. The highest germination was achieved in the TMC seeds that were given six weeks warm treatment followed by 32 weeks chilling. Hormone treatments significantly reduced the amount of chilling needed to release dormancy in TMC seeds, but not in the FI seeds. Overall, germination response was better at 15°C test temperature than at 20/30°C.     

Echinochloa crus‐galli seed physiological dormancy and germination responses to hypoxic floodwaters

• Hypoxic floodwaters can seriously damage seedlings. Seed dormancy could be an effective trait to avoid lethal underwater germination. This research aimed to discover novel adaptive dormancy responses to hypoxic floodwaters in seeds of Echinochloa crus‐galli, a noxious weed from rice fields and lowland croplands.
• Echinochloa crus‐galli dormant seeds were subjected to a series of sequential treatments. Seeds were: (i) submerged under hypoxic floodwater (simulated with hypoxic flasks) at different temperatures for 15 or 30 days, and germination tested under drained conditions while exposing seeds to dormancy‐breaking signals (alternating temperatures, nitrate (KNO₃), light); or (ii) exposed to dormancy‐breaking signals during hypoxic submergence, and germination monitored during incubation and after transfer to drained conditions.
• Echinochloa crus‐galli seed primary dormancy was attenuated under hypoxic submergence but to a lesser extent than under drained conditions. Hypoxic floodwater did not reinforced dormancy but hindered secondary dormancy induction in warm temperatures. Seeds did not germinate under hypoxic submergence even when subjected to dormancy‐breaking signals; however, these signals broke dormancy in seeds submerged under normoxic water. Seeds submerged in hypoxic water could sense light through phytochrome signals and germinated when normoxic conditions were regained.
• Hypoxic floodwaters interfere with E. crus‐galli seed seasonal dormancy changes. Dormancy‐breaking signals are overridden during hypoxic floods, drastically decreasing underwater germination. In addition, results indicate that a fraction of E. crus‐galli seeds perceive dormancy‐breaking signals under hypoxic water and germinate immediately after aerobic conditions are regained, a hazardous yet less competitive environment for establishment.

     

ENVIRONMENTAL CONTROL OF SEED GERMINATION IN THLASPI ARVENSE (CRUCIFERAE)

The effect of environmental conditions during storage and imbibition on germination was investigated in field pennycress (Thlaspi arvense L.), a weed species that can behave as a winter or a summer annual. Freshly harvested seeds of an inbred line with a cold requirement for flowering exhibited primary dormancy that was rapidly lost following 1 month of afterripening in a dry state. Nondormant seeds were positively photoblastic. The light effect was mediated through phytochrome since germination was promoted by red light and inhibited by far red light. Seedling emergence was also inhibited by light filtered through a canopy of wheat leaves. Germination of field pennycress seeds was considerably more sensitive to moisture stress than two sympatric species, wild oat (Avena fatua L.) and wheat (Triticum aestivum L., cv. ERA). Seeds of the latter two species were chosen in order to compare the effect of water potential on germination in field pennycress with that in sympatric species. It was concluded that the major environmental factor limiting nondormant field pennycress seeds on the soil surface was water availability. Imbibition of fully afterripened seeds at low temperatures (6 C) induced a deep secondary dormancy. In contrast to primary dormancy, cold-induced dormancy was not alleviated by red light, alternating temperatures (21/5 C), or 2 months of dry storage at 6, 15, or 35 C. However, exogenous gibberellin A₃ or 24 weeks of dry storage resulted in germination in cold-induced dormant seeds. Secondary dormancy was not observed in fully afterripened seeds that were preincubated at 21 C for 1 or 2 days prior to the cold treatment. These results may explain the failure in field experiments to observe the cold-induced secondary dormancy that limits spring emergence in other winter annuals (J. Baskin, C. Baskin, Weed Res. 1979 19: 285–292).     

Rate of Change in Dormancy Level and Light Requirement in Weed Seeds During Stratification

The rate of change in dormancy level and light requirement,induced during stratification at 3.2 °C, was investigatedin seeds ofStellaria media, Cerastium fontanum, Veronica agrestisandTaraxacumofficinale.Two stratification environments, soil and wet filterpaper in petri-dishes, were used. On nine occasions during a6 week stratification period, germination was tested under threelight conditions at 3.5/18.5 °C: (1) darkness; (2) light;and (3) short-term light exposure followed by darkness. Priorto stratification, germination in all species was 89% in lightand 53% in darkness. Within 2 weeks of stratification, germinabilityinV. agrestisandS. mediaseeds decreased in all treatments. InC.fontanumandT. officinaleseeds, germinability also decreasedafter 2 weeks but only in the dark treatment, indicating inductionof a light requirement. After two more weeks of stratification,the induced dormancy inS. mediaseeds stratified in soil becameweaker and the light requirement inT. officinaleseeds was lost.Differences between the two stratification environments and/orinteractions between light conditions, stratification environmentsand time were found for all species. These results suggest that:(1) the dormancy level and the light requirement of seeds maychange dramatically over relatively short-time periods duringstratification; and (2) germinability depends on the stratificationenvironment experienced by the seeds. Predicting the dormancylevel or light requirement in a seed batch is difficult andrequires a thorough knowledge of the effect of the stratificationconditions used.Copyright 1997 Annals of Botany Company Burial; Cerastium fontanumBaumg.; chickweed; cold stratification; common mouse-ear; dandelion; dormancy; light requirement; seed; Stellaria media(L.) Vill.; speedwell; Taraxacum officinalegroup; Veronica agrestisL.; weed     

Effect of Gibberellic Acid on Dark and Light Germination at Different Temperatures of Calluna, Ledum andRhododendron Seeds

The aim of the experiments was to study the effects of gibberellic acid (GAs) on the germination of Calluna vulgaris L., Ledum palustre L. and Rhododendron lapponicum (L.) Wahlenb. seeds under different environmental conditions. Under continuous light from white fluorescense tubes (3000 lux), untreated seeds of Calluna were partly dormant at all temperatures studied (9, 15, 21, 27, 27/9, 8/16 hours). Percentage of dormant seeds increased, however, with decreasing temperature, and it varied also from seed lot to seed lot. Untreated seeds of Ledum were dormant in light at 9° and 15°C but not at higher temperatures. Untreated seeds of Rhododendron were completely dormant in light at temperatures from 13° to 24°C. Seeds of all species were completely dormant in darkness both at 15° and 27°C. GA₃ stimulated greatly the germination of all species under all studied environmental conditions. The used concentrations (0.2–3.2 mM) gave nearly 100% germination in most cases. At 9°C the dormancy in some seed lots of Calluna and Ledum was only partly broken by the used concentrations of GA₃.     

Some effects of nitrate-treated soil upon the sensitivity of buried redroot pigweed (Amaranthus retroflexus L.) seeds to ethylene, temperature, light and carbon dioxide

Abstract Fresh dormant redroot pigweed (Amaranthus retroflexus L.) seeds were buried 5 cm deep in the field at Stoneville, MS in November 1981. Potassium nitrate (200 kg ha ¹) or nothing was applied to the soil in the fall of 1981 and the late winter of 1982. Seeds were recovered at intervals under darkness during the following 2 years and tested for responses to ethylene, temperature, light and carbon dioxide. During the first overwintering, nitrate enhanced loss of primary dormancy and increases seed sensitivity to temperature, light and ethylene. The loss of dormancy reached a maximum at 25 to 30 weeks (early summer) after burial. Examination of the recovered seeds indicated that about 80% of the non-treated seeds and 98% of the nitrate-treated seeds germinated in situ during the period of maximum loss of dormancy. Thus, after one overwintering period, about 20% of the original buried seed population remained dormant in nontreted soil and 2% remained dormant in the nitratetreated soil. After the second overwintering, the percentages of dormant seeds remaining in nontreated or treated soil were both only 1–2%. Nitrate reduced dormancy and enhanced germination in early summer following the first overwintering. Regardless of treatment, the remaining 1 2% of seeds in soil after the second year were of low sensitivity to the germination stimuli (ethylene, temperature, light) and constituted the long-lived portion of the original seed population.     

Germination and Dormancy of Reed Canary-Grass Seeds (Phalaris arundinacea)

Effects of various chemical and physical factors on the germination of several seed lots of reed canary-grass (Phalaris arundinacea L.) have been studied. Germination at the optimum constant temperatures of 24 to 27°C was significantly stimulated by the following treatments: moist chilling in light, red light given during the first 3 days of imbibition, three 2-h periods at 12°C given during the second day of imbibition, ethylene, increased oxygen tension and soaking in aerated water for 4 days. Dry storage at 20–30°C had no effect on the germination ability of the seeds. No significant quantities of germination inhibitors were found either in water or methanol extracts of seed dispersal units. By comparing three cultivars with various degrees of seed dormancy, respiration measurements showed that there was a significant positive correlation between oxygen uptake prior to visible germination and germination capacity. Similarly, germination-stimulating treatment significantly enhanced oxygen uptake prior to visible germination.     

Dose-Response Analysis of Factors Involved in Germination and Secondary Dormancy of Seeds of Sisymbrium officinale: II. Nitrate

The role of nitrate as a promoter of germination of Sisymbrium officinale seeds was examined in optimal light conditions. It was shown that the requirement for nitrate was absolute. This was true for all seed lots used. The probit of germination in water was log-linearly related to the level of endogenous nitrate. Preincubation at 15°C resulted in an immediate decrease in germination, whereas in 25 millimolar KNO₃ the decrease was delayed. The decline of germination in water was strongly correlated with the rate at which nitrate leached from the seeds. The germination response to a range of KNO₃ concentrations was followed during preincubation at 24-hour intervals. During the entire 264-hour preincubation period increasingly higher nitrate concentrations were required to maintain a response. This resulted in a right-hand shift of the dose-response curve parallel to the x axis. After 120 hours the high maximum germination level started to decline. The dose-response curves could be simulated by an equation from the receptor-occupancy theory. It is proposed that induction of secondary dormancy is a result of a decrease of the number of nitrate receptors. After 24 and 48 hours of preincubation, the nitrate-response curves were biphasic. The biphasic character could be related to the level of endogenous nitrate and to a differential requirement for nitrate of two fractions of the seed population. Similarities with the behavior of fluence-response curves after prolonged dark incubation led to the hypothesis that phytochrome and nitrate share the same site of action.     

Are Seasonal Dormancy Patterns in Arabidopsis thaliana Regulated by Changes in Seed Sensitivity to Light, Nitrate and Gibberellin?

Imbibed seeds of Arabidopsis thaliana (L.) Heynh., passed annuallythrough a pattern of changes in dormancy. Dormancy was brokenin summer and re-induced in autumn-winter. A second small germinationflush occurred in early spring. The role of sensitivity to light,nitrate and gibberellins (GAs) in regulating annual dormancypatterns and germination was studied with the use of GA-deficient(gal-2) and wild-type seeds. Dark-incubated seeds were exposedto a natural temperature regime for periods up to 18 monthsand at regular intervals germination capacity of portions ofseeds was tested at laboratory conditions. Germination datafitted as logistic dose response curves showed that sensitivityto light varied with the seasons in both genotypes. From interpretationof curve parameters, it is proposed that the observed sensitivitychanges involve alterations in the number of receptors, in thebinding characteristics of the receptors and/or in the responsechain initiated by ligand-receptor interaction. In this responsechain GA biosynthesis is stimulated (wild type) and sensitivityto GAs is enhanced (wild type, gal -2). GA sensitivity is alsodirectly influenced by temperature, thus without the interferenceof light. However, the significance of direct regulation ofGA requirement seemed to diminish with prolonged incubationoutdoors, whereas reversible changes in light sensitivity remainedclear. Therefore, we propose that seasonal dormancy patternsare mainly regulated by changes in sensitivity to light. GAsensitivity contributes to this pattern but is not primarilycontrolling dormancy. The GA requirement for germination isobvious as gal-2 seeds did not germinate at any time of theyear when deprived of applied GAs. However, GA biosynthesisis not required for dormancy control, as a dormancy patternwas also observed in the absence of the capacity to synthesizeGAs. Nitrate or sensitivity to nitrate did not contribute tothe regulation of dormancy and germination of this species.Copyright1994, 1999 Academic Press Arabidopsis thaliana (L.) Heynh., curve fitting, dormancy, fluence response curve, germination, gibberellin, gibberellin dose response curve, hormone mutant, light, mouse-ear-cress, nitrate, phytochrome, receptor, seasonal dormancy pattern, sensitivity     

Seedling emergence patterns and dormancy/germination physiology of <Emphasis Type="Italic">Primula modesta</Emphasis> in a subalpine region

On the basis of the germination/dormancy responses of seeds to temperature and light, and local seed rain, we attempted to interpret the seedling emergence patterns of Primula modesta Bisset et Moore (Primulaceae) in two different types of habitats in a subalpine zone of Mt. Asama: an oligotrophic flat moor and a grassland with relatively dense herbaceous vegetation cover. The seasonal pattern of seedling emergence was well explained by the dormancy/germination physiology revealed in laboratory germination tests. The seeds were demonstrated to have a strict light requirement even after experiencing moist chilling, which might facilitate the incorporation of the seeds into the soil seed bank. Despite sufficient seed production, the seedlings emerging were far less at the grassy site than the moor site, but the number of seedlings was significantly dependent on the seed rain within previous season and on the litter cover of the microsite in both sites. Therefore, the spatiotemporal patterns of seedling emergence in the habitats could be well explained by the spatiotemporal patterns of seed rain and safe-sites for germination.