Non-dormant seeds of Orobanche aegyptiaca were incubated at water potentials of 0 to –1.33 MPa and at constant temperatures from 5 to 29°C. Effects of water potential and temperature on final germination were modelled. In general, germination increased with increased temperature from 5 to 20°C and decreased above 26°C. Maximum germination occurred at 20–26°C and 0 MPa. Germination was reduced as the water potential decreased. Water potential also affected the temperature range over which high germination was observed; at 0 MPa high germination occurred over 9° (17–26°C) compared with 3° at -1.25 MPa (17–20°C). The optimum germination temperature also tended to decrease with a decrease in water potential. Final germination could be accounted for by seed-to-seed variation in the population assuming that each seed had a minimum temperature for germination and a maximum temperature above which it would not germinate. Seed-to-seed variation in these characteristics was assumed to be normally distributed, and it was further assumed that the two characteristics were independent. Effects of water potential on these temperature requirements were quantified, and the resulting empirical model accounted for final germination with reasonable accuracy (R2= 0.96).

A survey of Pisum genotypes for seed trypsin inhibitors revealed a tenfold range in the extent of inhibition. Approximately 90% of trypsin inhibitory activity was associated with two albumin fractions in selected variant lines. The differences among extreme variants were consistent in three environments, between two sources of trypsin tested and whether expressed on a unit protein or dry weight basis.

A study of the appearance of trypsin inhibitors during seed development in selected highand low-inhibitor lines showed differences in the accumulation pattern of active inhibitors. An endogenous protease was identified in Pisum seed protein preparations, whose in vitro trypsin-like activity was predominant in protein from early stages of seed development, when little or no trypsin inhibitor was present. However, there was no correlation between the amount of this protease and the extent of trypsin inhibitory activity in lines that varied for inhibitor content.

Requirements for dormancy break and embryo growth were determined for seeds of the western North American species, Osmorhiza depauperata. Seeds were collected in August 2001 from Sandia Crest (3200 m elevation) and Las Huertas (2300 m), New Mexico (USA). Embryos in fresh seeds were c. 0.6 mm long, and they had to grow to c. 9–10 mm before the radicle emerged from the mericarp. Embryo growth occurred at low temperatures (1 and 5°C), and seeds germinated to high percentages at 1°C during 32 weeks of incubation in the light. No seeds germinated at 5, 15/6, 20/10, 25/15 or 30/15°C during 32 weeks of incubation. Although a 4–18 week warm-temperature (25/15°C) pretreatment increased germination rates at 1°C, it was unnecessary for a high percentage of seeds to germinate. Gibberellic acid (GA3, 10–1000 mg l–1) did not substitute for cold stratification. Seeds from the low-elevation population contained larger embryos and required less time to germinate than those from the high-elevation population. O. depauperata seeds have deep complex morphophysiological dormancy (MPD), which is similar to two other western North American congeners and an Asian congener, but different from two eastern North American congeners. Results from this study suggest that: (1) phylogenetic niche conservatism has played a role in the persistence of deep complex MPD in the three western North American species of Osmorhiza; and (2) the stimulatory effect from a warm pretreatment in species needing only cold stratification for dormancy break is a preadaptation that initiated the development of an absolute warm requirement in species needing both warm and cold stratification.

The desiccation sensitivity of seeds of Inga vera Willd. subsp. affinis, a recalcitrant-seeded tree from Brazil, was analysed, focusing on water relations and cell-cycle aspects, including DNA content and the microtubular cytoskeleton. Seeds were collected at four developmental stages, dried to different moisture contents (MCs), assessed regarding water activity and set to germinate. Samples of fresh (non-dried) developing and mature seeds were used for assessment of DNA content by flow cytometry. Immunohistochemical detection of microtubules (MTs) was done in mature seeds at different MCs. Slight desiccation of immature seeds increased germination, but further drying resulted in a quick decline of germinability. During seed development the desiccation sensitivity decreased slightly, but DNA content of the embryonic axis cells remained constant, suggesting no relation between those two parameters. Embryonic axis cells of mature seeds showed abundant cortical microtubule arrays, which were not affected by mild desiccation, but broken down by further drying. It appeared that, upon rehydration, damaged cells were not able to reconstitute the microtubular cytoskeleton. The failure of germination of Inga vera seeds after drying could not be attributed to cellular damage to DNA synthesis and mitosis, since the radicle protruded by means of cell elongation, without a need for cell division. However, the breakdown of MTs during desiccation, and their subsequent inability to reassemble upon rehydration, may be related to the decreased germination, since MTs are required for cell elongation.

Using flow cytometry, we determined relative DNA amounts expressed as C-values, of large numbers of nuclei isolated from embryo root tips in three seed lots of pepper (Capsicum annuum) and tomato (Lycopersicon esculentum). The nuclei in embryos of fully matured dry pepper and tomato seeds predominantly revealed 2C signals, which indicates that the cells of maturing seeds have arrested at G1 phase of the cell cycle. Priming of pepper and tomato seeds in −1.1, −1.3 or −1.5 MPa polyethylene glycol (PEG) for 14 days reduced the mean time to germination. In all seed lots an induction of 4C signals was found after priming, indicating that during priming the cells of the embryonic root tip had replicated their DNA and arrested at the G2 phase of the cell cycle. For both tomato and pepper, the frequency of 4C signals was highest at the lowest PEG concentration (osmotic potential of −1.1 MPa). A significant inverse correlation was found between the frequency of root tip cells expressing 4C DNA signals and the mean time to germination for each individual pepper and tomato seed lot. However, the effect of a specific osmotic treatment differed between seed lots of the same cultivar, with regard to both the decrease in the mean time to germination and to the percentage of 4C cells, suggesting specific differences among seed lots in the response to priming. The present results indicate that flow cytometry can be used to follow the progress in nuclear replication activities during priming. The advancement in DNA synthesis can thus be used to measure the efficiency of a priming treatment with regard to enhancement of seed lot germination rate.

The assumption that seeds imbibe most of the water required for germination as liquid through seed-to-soil contact has been a dominant theme in germination research and seeding technology. Under most conditions, seeds are also exposed to water vapour during imbibition, but the relative contributions of liquid and vapour are difficult to assess. In water uptake models that include vapour, procedures used to estimate potential vapour imbibition have underappreciated the effect of distance on diffusion rate. At the same time, the amount of seed-to-soil contact and the liquid bridge from soil water to the seed tend to be greatly overestimated, considering the soil water contents often found in the field. Most researchers have recorded an approximately equal time to germination at soil water contents ranging from field capacity to nearly permanent wilting point, and little response to bulk density, soil type or seed–soil contact. While hydraulic conductivity decreases by several orders of magnitude as soil water content, bulk density and seed-contact decrease, relative humidity remains near 100%. There are several experiments demonstrating timely germination in water vapour alone. The combined evidence contradicts the assumption that seed–soil contact is important for imbibition of water by seeds. Water vapour should be considered the primary source of water for seeds in unsaturated soils.

We assessed inter-seasonal dynamics of seed banks, dormancy and seed germination in three endemic Chihuahuan Desert succulent species, under simulated soil warming conditions. Hexagonal open top-chambers (OTCs) were used to increase soil temperature. Seeds of Echinocactus platyacanthus (Cactaceae), Yucca filifera and Agave striata (Asparagaceae) were collected and buried within and outside OTCs. During the course of one year, at the end of each season, seed batches were exhumed to test viability and germination. Soil temperature in OTCs was higher than in control plots. Yucca filifera seeds always had high germination independently of warming treatment and season. Agave striata seeds from OTCs had higher germination than those from control plots. Agave striata exhibited low germination in fresh seeds, but high germination in spring. Seeds from this species lost viability throughout the experimental timeframe, and had no viable seeds remaining in the soil. Echinocactus platyacanthus showed high germination in fresh seeds and displayed dormancy cycling, leading to high germination in spring, low germination in summer and autumn, and high germination in winter. Germination of this species was also higher in seeds from OTCs than those from control plots. Echinocactus platyacanthus formed soil seed banks and its cycle of inter-seasonal dormancy/germination could be an efficient physiological mechanism in a climate change scenario. Under global warming projections, our results suggest that future temperatures may still fall within the three studied species’ thermal germination range. However, higher germination for A. striata and E. platyacanthus at warmer temperatures may reduce the number of seeds retained in the seed bank, and this could be interpreted as limiting their ability to spread risk over time. This is the first experimental study projecting an increase in soil temperature to assess population traits of succulent plants under a climate change scenario for American deserts.

Floodplain meadow ecosystems are characterized by high water level fluctuations and highly variable soil water potentials. Additionally, climate change scenarios indicate an increasing risk for summer drought along the northern Upper Rhine and the Middle Elbe River, Germany. While adult plants often persist even after strong changes in water availability, early life phases, such as seed germination and seedling establishment, might be more vulnerable. Therefore we tested whether reduced soil water potentials will affect the germination of meadow species and whether the response varies between (1) forbs indicative of wet and dry habitats and (2) seeds originating from sites along the rivers Elbe and Rhine. We exposed seeds of 20 floodplain meadow species with different moisture requirements from five plant families to a water potential gradient ranging from 0 to − 1.5 MPa. While across species germination percentage and synchrony decreased, germination time increased at reduced water potentials. Germination of the species indicative of dry habitats decreased more strongly, was slower and less synchronous at reduced water potentials than that of species indicative of wet habitats. Seeds from sites along the rivers Elbe and Rhine did not differ in their germination characteristics. We propose that species of wet sites follow an all-or-nothing-strategy with fast and synchronous germination to maximize competitive advantages, betting on a high probability of moist conditions for establishment (optimists). In contrast, species from dry sites appear to follow a bet-hedging strategy with a moisture-sensing mechanism for unsuitable conditions (pessimists), resulting in a slower and less synchronous germination.

Early plant establishment through seed germination and seedling emergence is a crucial process that determines seedling number, emergence time distribution and the early growth of seedlings, all of which are affected by soil climate and soil structure. In the current context of climate change, in which increasing the diversity of cultivated species is considered desirable, and new tillage practices are considerably modifying top-soil surface characteristics, we need to improve our ability to model the effects of the environment on plant establishment. Using a trait-based and model-based framework, we aimed to identify general relationships between seed and seedling traits (e.g. seed mass and lipid content, seedling diameter, base temperature) and germination and emergence model parameters (e.g. time to mid-germination, shoot elongation rate) measured for 18 genotypes belonging to 14 species. Relationships were also investigated among model parameters or traits. Germination rates were faster for species with a high base temperature and for species with seed reserves located principally in the embryo (rather than the endosperm or perisperm). During heterotrophic growth, maximal shoot length and elongation rate increased with seed dry mass. The sensitivity of seedlings to soil obstacles was negatively related to shoot diameter. Thus apart from the known effects of seed mass on seedling establishment, we found that seed reserve location, seedling shoot diameter and shape affected germination rate and emergence success. Such generic rules linking plant traits to germination and emergence parameters enhance our understanding of the determinants of environmental effects on plant establishment success.

Germination over 3 years was followed in 25 soil samples of 31 × 31 × 5 cm each from primary and secondary stands in Białowieża forest, Poland. The number of seedlings emerging and the number of species declined over the 3 years. In year 1 the mean densities of emerging seedlings for the primary and secondary stands were similar (2142 m−2 and 2488 m−2 respectively); in year 3 the difference was greater (248 vs 498 m−2) but not statistically significant. The trend reflected the greater longevity of the seed bank species characteristic of the secondary forest. Two functional groups of herb and shrub species were distinguished in the seed bank based on (a) frequency as adults in the present vegetation, and (b) relative abundance in the seed banks of primary and secondary forest. Group A species need large-scale gaps in the tree canopy; most have very small seeds (<0.15 mg mean dry mass) and are very persistent in the soil. Group B species benefit from single-treefall gaps in the tree canopy, but appear to be suited primarily to gaps in the canopy of the herb layer and in the root layer made mainly by animals; their seeds are not as small (mostly >0.15 mg) and not as persistent in the soil. Group B species made up 86% of the seedlings emerging over 3 years in soil from the primary stand, and 56% of those from the secondary stand. In the laboratory all five Group B species tested showed some inhibition of germination in darkness, but only one showed unequivocal inhibition by far-red-enrichment of low irradiance light.

Community composition and ecosystem processes during succession may be driven partly by traits of plant species that attain dominance. Here, we addressed the hypothesis that Stipa pulcherrima, the dominant grass of abandoned continental grasslands, controls seedling recruitment of co-occurring species through chemical effects of its litter. Eight species with successful and unsuccessful recruitment under field conditions were selected (four familial pairs) to study experimentally the effects of leaf leachate under four temperature regimes. Since fungi developed in leachate-treated Petri dishes, in another experiment seeds were surface sterilized to remove confounding effects of fungi on recruitment. Leachate affected various stages of seedling recruitment: it significantly reduced seed germination (by 33–94%) and radicle elongation, and it delayed germination of seedlings of all species. In two families, species with unsuccessful field recruitment were more negatively affected than the successful ones. In a third family, the species with successful recruitment was more negatively affected, and in the fourth there were no differences. Similar germination responses after exclusion of fungi through seed-surface sterilization suggested that leachate was responsible for the observed effects on recruitment. Besides other traits and physical/microclimatic effects of accumulating litter, S. pulcherrima influences plant community dynamics and may potentially affect ecosystem processes through its secondary compounds.

In the range of water contents studied (0.1–0.4 g H2O g dw−1), Coffea arabica seeds were less sensitive to desiccation than C. costatifructa, C. racemosa and C. sessiliflora seeds. At 0.20 g H2O g dw−1, 53% of C. arabica seeds germinated after direct immersion in LN (rapid cooling, 200°C min−1), but none of them developed into normal seedlings. By contrast, in C. costatifructa, C. racemosa and C. sessiliflora, when seeds were dehydrated to the optimal water content (0.19, 0.28 and 0.31 g H2O g dw−1, respectively), the percentages of seeds which developed into normal seedlings after LN exposure were 26, 78 and 31% of the desiccation control, respectively. Normal seedlings could be recovered from cryopreserved C. arabica seeds only if they were desiccated to 0.20 g H2O g dw−1 and precooled slowly to −50°C prior to immersion in LN. Precooling seeds at 2°C min−1 allowed 25% of seeds to develop into normal seedlings. The thawing rate had no effect on the survival of cryopreserved C. arabica seeds. In all cryopreservation experiments, the total germination did not reflect the percentage of seeds which developed into normal seedlings. Examination of excised embryos indicated a partial explanation of this difference since only the shoot apex was destroyed in abnormal embryos, whereas the hypocotyl and radicle were normal.

Although many studies have been conducted on seeds with a water-impermeable seed or fruit coat (physical dormancy), the primary site of water entry into these seeds after dormancy-breaking treatments is still controversial. Thus, the role of lens, hilum, micropyle and extrahilar regions in water uptake of seeds treated to break physical dormancy was examined in Vigna oblongifolia and Sesbania sesban (Fabaceae) following pretreatment with sulphuric acid and hot water. Morphology of seed surfaces in treated versus non-treated seeds of both species was examined with scanning electron microscopy. Most seeds of V. oblongifolia first cracked in the hilum when pretreated with sulphuric acid, but they cracked in both the hilum and extrahilar regions when pretreated with hot water. However, in S. sesban seeds, a crack formed only in the lens following either acid scarification or hot-water treatments, and the seeds imbibed water only through the lens. These results indicate that the primary site of water entry into seeds following physical dormancy break can vary with species and treatments. Slow, early imbibition via the hilum, and subsequent rapid imbibition via the lens, may not be detected unless seeds are monitored for several days. Time allowed for imbibition studies may, at least in part, explain various interpretations about the role of the lens in physical dormancy reported in the literature.

The effects of desiccation and temperature on the germination capability of Carica papaya L. were investigated for seeds extracted from three commercial fruit batches. More than 50% of freshly isolated, cleaned (sarcotesta removed) but undried seeds germinated at 26°C. However, desiccation to approx. 20%seed RH reduced germination at this temperature to < 10%. A substantial increase in germination at alternating temperatures (33/19°C) indicated that desiccation induced seed dormancy rather than viability loss. Dormancy could be removed in a large proportion of the population by the application of a single heat shock to rehydrated seeds for 4 h at 36°C, with subsequent return to 26°C for germination. Longer (days) and shorter (minutes) periods of heat shock were less effective for releasing dormancy. Heat shock was generally applied 5 or 14 d after rehydration had started, but the treatment was equally effective after imbibition for only 1 d. Light was always applied during both imbibition and the post-heat shock treatment, but was not essential during the actual heat shock treatment. Rehydration and post-heat shock temperature treatments in the range of 16°C to 36°C revealed the same optima of 26°C. Dormancy was re-imposed in heat-shocked seeds when they were subsequently dried to seed relative humidities of 25 to 75%(5 to 11% moisture content [fresh weight basis]), but this state could be removed by a further heat shock. The longer heat-shocked seeds were held on agar-water at 26°C prior to re-drying, the greater the level of desiccation intolerance.

This review focuses on the possible evolutionary status and functional significance of recalcitrance and certain associated ovule/seed characters, within the framework of modern systems of angiosperm classification. The presence of recalcitrant seed viability in 45 dicotyledonous families is significantly associated with bitegmic and crassinucellate ovules and with nuclear endosperm development, all considered ancestral (plesiomorphic) character states of the ovule; as well as with greater seed size, woody habit and tropical habitat, also regarded as ancestral character states in the dicotyledons. In many species with recalcitrant seeds, the predominant storage reserve is carbohydrate. Recalcitrance is significantly associated with the exalbuminous type of reserve storage. It is proposed that in large recalcitrant seeds, the transfer of the main storage function from endosperm to embryo was probably an early development. In many species with recalcitrance, the ovules/seeds are characterized by extensive vascularization of the integument(s)/seed coat or by a pachychalaza. Pachychalazy is proposed to be a significant functional adaptation for a more efficient transfer of nutrients to the embryo/seed. Recalcitrance and some of the other character states proposed to be plesiomorphic in dicotyledons are also present in some gymnosperms, including presumed sister groups of the dicotyledons. In relatively advanced dicotyledonous families, mostly with orthodox seeds, recalcitrance probably persisted only in isolated relict members. Determination of character polarity is particularly problematic at lower taxonomic levels, because there is always the possibility that, in some taxa, a character state such as recalcitrance may have arisen secondarily as a reversal. Available evidence supports our view that seed recalcitrance can be regarded as a relatively ancestral character state in dicotyledons.

Raffinose family oligosaccharides (RFO) accumulate in many developing seeds and are degraded during seed germination. However, acidic α-galactosidase (AGAL) activity and subcellular location do not correlate with raffinose depletion; alkaline α-galactosidases (AGA) may be responsible for RFO hydrolysis in germinating seeds. Three cDNA clones for AGA/SEED IMBIBITION PROTEIN were obtained from the Pioneer Hi-Bred maize expressed sequence database. Two of the clones were expressed in Escherichia coli, and the recombinant proteins, when incubated with naturally occurring galactosides or p-nitrophenyl α-d-galactose, exhibited AGA activity with maximum catalysis at pH 7.5 (ZmAGA1) or pH 8.5 (ZmAGA3). No raffinose biosynthetic capacity was observed with either enzyme. Maximal α-galactosidase activity in mature dehydrated, germinating and germinated maize (Zea mays) seeds occurred at pH 7.5. ZmAGA1 was the sole family member detected in seeds and maize Hi-II, embryo-derived, callus cells. Its transcript accumulated when seed germination was interrupted by heat, cold or dehydration stress, but not in response to NaCl. Tissue prints localized transcripts to the scutellum or the embryo axis, depending on the stress applied. In maize Hi-II callus cells, transcripts accumulated when callus was subjected to heat stress (42 °C), during which ZmAGA1 transcript accumulation was further induced by sucrose. Galactosides in a variety of forms, including raffinose, partially repressed the sucrose-induced accumulation of transcript in heat-stressed callus.

Late embryogenesis abundant (LEA) proteins accumulate in developing seeds prior to maturation drying and are presumed to help protect embryos from desiccation stress. The unusual solubility properties of these proteins, such as resistance to heat coagulation, have led to suggestions that they alter the hydration properties of cellular constituents. Hydration characteristics and water potential range at which wheat heat-soluble LEA proteins were expressed have been determined. Levels of heatsoluble proteins decline in germinating seeds but can be induced by dehydration (ψ ≤ −0.5 MPa) in crown meristematic tissue that is desiccation tolerant. The heatsoluble extract from mature wheat embryos contained proteins and sugars at about a 1:1 (w/w) ratio. Only about half of the sugars could be removed by exhaustive dialysis; the rest appeared to be tightly associated with the proteins. The water sorption characteristics of undialysed and dialysed heat-soluble protein/sugar fractions were compared with other water-soluble proteins (bovine serum albumin, lysozyme or gluten) and with sucrose. At relative humidities greater than 50%, the amount of water absorbed by protein-sugar mixes was a function of the sugar content. For the same sugar content, the heat-soluble protein preparation absorbed 2–3 times more water than a lysozyme/sucrose preparation. The rate at which heat-soluble protein fractions dried was also different to desorption rates of lysozyme/sucrose mixes. While lysozyme/sucrose mixtures dried either very rapidly (within 20 min) or very slowly (about 2 months) depending on the sugar content, desorption rates of the heat soluble protein-sugar preparations were intermediate (2–10 days) and modulated by sugar concentration. Based on the presumption that the hydrophilic properties of LEA heat-soluble proteins are important to their function, it is suggested that these proteins function to control drying so that cells stay at critical water potentials for the proper time. In this respect, the heat-soluble LEA proteins do not prevent desiccation, but serve as hydration buffers.

Links between germination, DNA replication and β-tubulin accumulation were studied with cabbage (Brassica oleracea L.) seeds, by using flow cytometric analysis of nuclear DNA content and immunodetection of β-tubulin levels. The seeds were incubated in water or 0.1–500 mM hydroxyurea solutions. Radicle tips isolated from dry cabbage seeds revealed most 2C (Go or G1 stage) and some 4C (G2 stage) signals of nuclear DNA contents and a constitutive level of β-tubulin. The onset of DNA replication in the radicle tip was observed between 12 and 24 h of imbibition in water and was preceded by an increase of β-tubulin levels. Incubation of the seeds in 1 mM hydroxyurea retarded DNA replication, whereas an arrest of DNA replication occurred upon incubation in 10 mM hydroxyurea or higher concentrations. The arrest of DNA replication and cell division did not block radicle protrusion and allowed some extension of the radicle. This demonstrated that DNA replication is not a prerequisite for radicle protrusion and initial extension. However, further seedling development, including root growth and root hair development, was dependent on DNA replication. Accumulation of β-tubulin was not affected by hydroxyurea. Thus, it can be deduced that both DNA replication and β-tubulin accumulation are two parallel and independent cell cycle events during seed germination.

The effects of temperature on dormancy loss, germination and viability were investigated in seeds of Aesculus hippocastanum L. harvested over a 4-year period. Release from embryo dormancy in freshly harvested seeds was manifest in two phases of morphological growth: initially, when the seed lot was only partially released, axis emergence resulted primarily from cotyledonary petiole extension without radicle extension; subsequently, when the seed lot was totally released, axis emergence of all seeds was followed immediately by extension to >1 cm through growth of the radicle. Germination (axis emergence and radicle extension) at 16°C was a function of pre-treatment period at 2–11°C. The rate of dormancy loss (probit germination d−1) increased linearly below a ceiling temperature for the chilling response; this temperature was estimated to vary from 13°C to 16°C for two seed lots harvested in separate years. Dormancy periods for individual seeds within both seed lot populations can be described by cumulative normal distributions; the predicted standard deviation of chilling units below the ceiling temperature (i.e. thermal time) was 186°C d. Visible germination occurred during the process of stratification at 2°C, starting after 21–25 weeks. By contrast, three years of hy-drated seed storage at 16°C, which was a non-permissive temperature for dormancy loss, resulted in little pre-emergence of the axis during stratification; approximately one third of the seeds remained germinable. The implications of these quantitative analyses of the physiological processes in recalcitrant seeds for the development of improved storage methods are discussed.

There are two ideas regarding changes in the physiological and ecological tolerances and requirements within plant lineages through geological time. One is that these attributes have changed very little, or not at all (trait stasis), and the other is that they have undergone considerable change (adaptation), as plants shifted to new climatic and vegetation zones. We tested these ideas for seed dormancy-breaking and germination requirements of four species in a subclade of Aristolochia subgenus Siphisia: the three temperate species, A. macrophylla and A. tomentosa (the basal species in the subclade) of eastern USA and A. manshuriensis of East Asia, and the Mediterranean-climate species A. californica endemic to California, USA. A long period at cold-stratifying temperatures was required for growth of the underdeveloped embryo and seed germination in A. californica, whereas embryos grew and seeds germinated in the other three species at warm temperatures, either before or after they were cold stratified. Thus, seeds of A. californica have either intermediate or deep complex morphophysiological dormancy (MPD), whereas those of the three temperate species have either morphological dormancy or non-deep simple MPD. Further, there were quantitative differences in temperature requirements for dormancy-break and germination between the Appalachian A. macrophylla, which did not differ from its sister species A. manshuriensis, and the lowland A. tomentosa. Thus, within this lineage there has been both trait stasis and divergence (adaptation) in the physiology and ecology of seed dormancy and germination.