Seeds of Sisymbrium officinale display physiological dormancy and require nitrate to germinate. Rupture of the testa precedes radicle protrusion through the endosperm (germination sensu stricto). While both endosperm rupture and testa rupture (TR) required nitrate, endosperm rupture was fully inhibited by abscisic acid (ABA) but TR was not inhibited. The gibberellic acid (GA)-synthesis inhibitor paclobutrazol prevented TR, which was reverted by exogenous GA4 but not by nitrate. The orientation of TR was transverse, which prompted the question whether seeds elongate prior to radicle protrusion, concurrent with an increase in water content. Between 9 h and 1 d no increase in length or water content was observed. During incubation in ABA the length of imbibed seeds without TR did not increase between 1 and 5 d, whereas nitrate added to ABA induced TR and a 94% increase in length. At the same time the water content of seeds without TR increased by 18%, while the water content of seeds with TR increased by 38%. Length and water content were correlated in a single-seed analysis for seeds with TR, but not for seeds without TR. Increased length was also observed in Arabidopsis seeds with nitrate-induced TR. These results indicate that prior to endosperm rupture dormancy release by nitrate is accompanied by TR, seed elongation and an increase in water content. A new multiphasic model is proposed for the imbibition curve, where the second phase of the classical triphasic curve is split into three sub-phases, of which phases IIB and IIC are associated with TR.

Rudimentary, broad and small linear embryos occur among members of the most primitive (basal) extant angiosperms, collectively called the ANITA grade (i.e. Amborella, Nymphaeales and Austrobaileyales). Amborella (rudimentary) and Austrobaileyales (rudimentary in Austrobaileyaceae, Illiciaceae and Schisandraceae and small linear in Trimeniaceae) have kinds of embryos that are known to be underdeveloped; consequently, they must grow inside the seed prior to radicle emergence (germination). On the other hand, it is not known if broad embryos need to grow before radicles can emerge, and whether they are underdeveloped or fully developed. Thus, we addressed the question: ‘Is the broad embryo of Nymphaeales also underdeveloped?’. Although the embryo length : seed length ratios in Nymphaea Albert Greenburg, N. capensis var. zanzibariensis and N. immutabilis were 0.311, 0.349 and 0.234, respectively, embryos did not grow prior to radicle emergence. Thus, they are fully developed at seed maturity. If Amborella and Nymphaeales are equally the most basal angiosperms, as some molecular phylogenetic studies indicate, then we must conclude that the broad and rudimentary embryos are equally primitive.

Imbibitional stress was imposed on neem (Azadirachta indica) seeds by letting them soak for 1 h in water at unfavourable, low temperatures before further incubation at 30°C. Sensitivity to low imbibition temperatures increased with a decrease in seed moisture content (MC). To investigate a possible involvement of seed age in the extent of imbibitional damage, initially high-quality seed lots that differed in storage history (10 weeks versus 10 months) were examined at 4 and 7% MC (fresh weight basis). After 10 months of storage, the 7% MC seeds had become sensitive to imbibitional stress. Further drying (1 week) to 4% MC affected aged seeds more than non-aged seeds. Barrier properties of cellular membranes in axes excised after 1 d of rehydration were estimated using a spin-probe technique. The proportion of cells with intact membranes increased with increasing imbibition temperature. For each temperature tested, there were more cells with leaky membranes after 10 months than after 10 weeks of dry storage. Localization of embryo cells displaying loss of turgor and abnormal cellular structure was accomplished using cryo-planing, followed by cryo-scanning electron microscopy. Inspection of the cryo-planed surfaces confirmed that imbibitional damage was temperature dependent, occurring at the periphery. Ageing increased the number of imbibitionally damaged, peripheral cell layers. Germination was estimated to fail when less than 70% of axis cells were alive. We conclude that ageing increases the sensitivity to imbibitional stress. Both the fast ageing and the sensitivity to imbibitional stress might explain the apparent controversies about neem seed desiccation tolerance and storage behaviour.

Drought and high temperature each damage rice (Oryza sativa L.) crops. Their effect during seed development and maturation on subsequent seed quality development was investigated in Japonica (cv. Gleva) and Indica (cv. Aeron 1) plants grown in controlled environments subjected to drought (irrigation ended) and/or brief high temperature (HT; 3 days at 40/30°C). Ending irrigation early in cv. Gleva (7 or 14 days after anthesis, DAA) resulted in earlier plant senescence, more rapid decline in seed moisture content, more rapid seed quality development initially, but substantial decline later in planta in the ability of seeds to germinate normally. Subsequent seed storage longevity amongst later harvests was greatest with no drought because with drought it declined from 16 or 22 DAA onwards in planta, 9 or 8 days after irrigation ended, respectively. Later drought (14 or 28 DAA) also reduced seed longevity at harvest maturity (42 DAA). Well-irrigated plants provided poorer longevity the earlier during seed development they were exposed to HT (greatest at anthesis and histodifferentiation; no effect during seed maturation). Combining drought and HT damaged seed quality more than each stress alone, and more so in the Japonica cv. Gleva than the Indica cv. Aeron 1. Overall, the earlier plant drought occurred the greater the damage to subsequent seed quality; seed quality was most vulnerable to damage from plant drought and HT at anthesis and histodifferentiation; and seed quality of the Indica rice was more resilient to damage from these stresses than the Japonica.

Both phenotypic and genetic variation can affect the establishment success of exotic species. Using the weed Senecio vernalis Waldst. & Kit. with a continuous westward expansion of the native geographic range, we asked to what degree seed traits and germination patterns differ among different habitat disturbance types in the introduced region. We hypothesized that seed traits and germination pattern differ between different disturbance types, with lighter seeds and enhanced germination speed in highly disturbed habitats. Since S. vernalis colonizes successfully various disturbed habitats, we expected to find a high phenotypic genetic variation. Individuals from 19 populations were sampled from one region in Eastern Germany to study seed trait variation and germination success by means of a germination experiment and to assess genetic patterns using amplified fragment length polymorphism (AFLP) markers. Contrary to our expectation, habitat disturbance types did not differ in seed traits, germination characteristics or genetic variation. Instead, we found highest phenotypic and genetic variation within populations. Maximum percentage of germination was positively affected by seed mass. In addition, there was a weak but significant association between AFLP band patterns and population size and seed width, indicating that some variation in seed traits has a genetic component. In conclusion, the absence of a population differentiation at this geographic scale makes it unlikely that adaptation to different disturbance regimes is involved. Instead, a high variation in seed traits, overall high germination and genetic variation within populations under a wide range of habitat disturbance conditions seems to enable the offspring to colonize various habitat disturbance types.

The effects of maternal air temperature and soil moisture upon seed physiological dormancy (PD) alleviation of an Australian native Asteraceae were investigated. From the onset of flowering, Actinobole uliginosum plants growing ex situ were subjected to either a warm (mean 26°C) or cool (mean 17°C) temperature regime, with adequate or limited water availability. In the warm environment, the reproductive phase was accelerated, and plants yielded fewer seeds over a shorter, earlier harvest period, when compared to those in the cool environment. Initial germination of all seeds was low ( < 20% at 15°C) due to PD, which was gradually alleviated by a dry after-ripening (DAR) treatment (34/20°C, 40% relative humidity, in darkness). Seeds from plants grown in the warm environment were more responsive to DAR than seeds from the cool environment, but maternal plant water availability had little effect on dormancy status. Germination was higher at 15°C than at 25/15°C, reaching a plateau of c. 80% germination after 20 weeks DAR. Before DAR, application of GA3 had little impact on seeds, which would consequently be classified as having deep PD if tested at the time of dispersal. However, DAR caused seeds to become increasingly responsive to GA3, reaching 97% germination at 15°C following just 4 weeks of DAR, which would indicate non-deep PD if seeds were tested following a period of warm, dry storage. Maternal air temperature regulates PD status of A. uliginosum, such that seeds collected from a warmer environment are likely to be more responsive to DAR. Post-harvest storage in an environment suitable for DAR affects seed response to GA3, which has implications for germination stimulation and dormancy classification.

LEAFY (LFY)-like proteins are plant-specific transcription factors that play essential roles in plant growth and development. In this study, a LEAFY homologue in soybean [Glycine max (L.) Merr.], designated as GmLFY, was cloned from inflorescences by the rapid amplification of cDNA ends (RACE) method. Sequence analysis showed that GmLFY cDNA contained a 1221 bp open reading frame, encoding 407 amino acid residues with typical characteristics of transcription factors. Subcellular localization in onion epidermal cells indicated that GmLFY protein was located in the nucleus. Analysis of the GmLFY genomic structure showed that the GmLFY gene has two introns with similar spliced sites as LFY-like genes in other plants. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that GmLFY was predominantly expressed in reproductive organs, such as inflorescences, pods and developing seeds. Further analysis showed that the expression level of GmLFY was higher in the middle stage than in the early or late stages of seed development, suggesting an essential regulatory role of GmLFY in soybean seed development.

The plant allelochemical coumarin effectively inhibits the germination of Brassica parachinensis (B. parachinensis) seeds. Quantification of endogenous phytohormones showed that contents of abscisic acid (ABA), ABA glucose ester, gibberellin A20 (GA20), GA3, GA15, GA24, GA9 and GA4 were higher in germinating seeds than in seedlings. Moreover, the presence of coumarin significantly reduced the content of bioactive GA4 which is thought to positively regulate seed germination. Histochemical staining and spectrophotometry of reactive oxygen species (ROS) revealed that exogenous GA3 and GA4+7 could effectively promote the production of endogenous ROS during germination and that the GA synthesis inhibitor paclobutrazol could effectively inhibit production of ROS. Coumarin significantly inhibited the accumulation of ROS, especially superoxide anion radical (${\rm O}_2^{{\cdot}{-}} $). This inhibitory effect could be restored by the addition of exogenous GA3 and GA4+7. Coumarin also inhibited the activity of the ROS-degrading enzymes such as superoxide dismutase, catalase and peroxidase as well as β-amylase in seeds and seedlings. Taken together, we propose a model for the regulation of seed germination in B. parachinensis by coumarin, Gas and ROS, in which coumarin may delay seed germination by reducing endogenous GA4, thus decreasing the accumulation of ROS.

Mangosteen (Garcinia mangostana L.), rambai (Baccaurea motleyana Muell.- Arg.) and jelentik (Baccaurea polyneura Hook. f.) are tropical fruit species believed to have recalcitrant seeds. The seeds showed no dormancy; they germinated easily. At harvest, the mean moisture contents (fresh weight basis) were 53.54, 51.20 and 44.90% for G. mangostana, B. motleyana and B. polyneura, respectively. G. mangostana seeds lost viability when their moisture content fell to about 24% while B. motleyana seeds lost viability below 35.5% moisture content. However, for B. polyneura, the seeds could be dried to low moisture content with high survival. The viability was still high when the moisture content was reduced to 13.46%. At this moisture content, the percentage germination was 91.76% and it was found that the seeds survived cyopreservation with 8.3% viability. For B. motleyana axes, the loss of viability occurred when their moisture content fell to about 36% (15% viability with predominantly callus formation) while for B. polyneura axes, viability was reduced to 33-67% when the moisture was 27–30%. At various moisture contents, the seeds of the fruit species studied were exposed for 48 h to 7°C and −4°C. G. mangostana seeds did not survive either temperature. Baccaurea seeds survived 7°C but failed to survive −4°C. No axes from B. motleyana seeds at various moisture contents survived cryopreservation. However, some viability (20–30%) was observed in B. polyneura axes cryopreserved at a moisture content of about 27%. At this moisture content no normal growth was obtained; callus formation was observed. It appears that the seeds vary in the degree of desiccation sensitivity. They also vary in size. Seeds of G. mangostana are larger than Baccaurea seeds and thus, more sensitive to desiccation. Seed and embryonic axis structure may also play a role in desiccation sensitivity.

Six pasture legumes, common in Syrian grasslands, were investigated. Medicago orbicularis and M. rotata had the largest number of hard seeds, with >90% still hard 5 months after shedding; Trifolium stellatum had the least with <30% of hard seeds; T. campestre, M. rigidula and T. tomentosum were intermediate. Hard-seededness in M. orbicularis and M. rotata varied between years. Scarification of hard seeds resulted in almost complete germination in all species. Alternating temperatures (10/20°C) reduced the rate of germination of scarified T. campestre seeds, but had no effect on germination of scarified or unscarified seeds of T. stellatum. The breakdown of hard-seededness in the field in Syria did not begin until 3 months after shedding and greatly increased when seeds overwintered. Breakdown of hard-seededness was slower when seeds were buried at 5 and 10 cm than when seeds remained on the soil surface. Seedling emergence after the rains started in November was highly correlated with the number of soft seeds observed in October. Similarly, a high correlation was found between the percentage of hard seeds in October and carry-over in the seed bank. In small-seeded species (Trifolium spp.), 39% of the initial seed bank was lost in 2 years by causes other than emergence as seedlings. Such losses were only 7% in larger-seeded species (Medicago spp.). The implications of hard-seededness on the utilization and survival of legumes in grasslands are discussed.

Seeds can synchronize their germination and dormancy cycles to regular seasonal environmental changes. The present work sought to evaluate the in-situ longevity of the buried seeds of two species of Syngonanthus and two species of Comanthera (Eriocaulaceae) in the region of natural occurrence (Serra do Cipó Range, south-eastern Brazil), and to relate their germination responses to seasonal climatic changes. Samples were exhumed bimonthly and germination was tested under a 12-h photoperiod (30 μmol m− 2s− 1) at optimal germination temperature. The seeds of Comanthera showed high deterioration after the first year of burial, especially C. elegans, which reflects their incapacity to form viable soil seed banks. The buried seeds of C. bisulcata, S. anthemidiflorus and S. verticillatus acquired secondary dormancy during the rainy season (spring/summer, higher temperatures) which was alleviated during the subsequent dry season (autumn/winter, lower temperatures). It is concluded that C. bisulcata, S. anthemidiflorus, and S. verticillatus form seed banks of the persistent type and demonstrate consecutive cycles of germination/dormancy, accompanying annual seasonal changes for at least 2 years.

Although Asteraceae species are important in the cold deserts of Central Asia, little is known about their seed dormancy and germination. We determined dormancy breaking and germination requirements of three annual Asteraceae, Echinops gmelinii, Epilasia acrolasia and Koelpinia linearis. Achenes (seeds) were tested for germination in light and in darkness over a range of alternating temperatures after various periods of burial outdoors and of dry storage. Germination phenology was monitored for seeds sown in irrigated and non-irrigated sand, and temperature requirements for dormancy break were determined under wet and dry conditions. Effects of pericarp and phyllaries on germination of E. acrolasia and E. gmelinii, respectively, were determined. Low percentages of 20-day-old seeds of E. acrolasia and K. linearis were non-dormant and germinated to low percentages over the range of temperatures, whereas all seeds of E. gmelinii were dormant. As seeds of the three species afterripened, they germinated over the range of temperatures. Whether seeds germinated in autumn or spring depended on the amount of sand moisture. Mechanical resistance of the pericarp and phyllaries reduced germination of E. acrolasia and E. gmelinii, respectively. Temperature requirements for germination as seeds come out of dormancy do not correspond to any of the known five types of non-deep physiological dormancy (PD). Thus, a sixth type is recognized in which germination occurs over the same range of temperatures in the early and late stages of dormancy break. Type 6 allows seeds to germinate at high or at low temperatures, whenever sand moisture is non-limiting.

Glebionis coronaria (L.) Cass. ex Spach is a common Mediterranean weed producing distinctive central and peripheral dormant cypselae with a hard fruit coat, which was previously hypothesized to impose physical dormancy. Analysis of water uptake in cypselae and in naked seeds showed that it preferentially takes place at the basal end of the fruit; however, seeds within an intact pericarp do not fully imbibe when compared with naked seeds. Germination was not significantly different between the two heteromorphs, and afterripening or cold stratification did not increase germination, while warm stratification at 35/20°C, as revealed by logistic regression, resulted in a significant improvement. However, loss of viability was also rapid at these high temperatures. Central and peripheral cypselae generally showed very low germination. In both heteromorphs, faster and higher germination (60–70%) was reached only after extensive scarification of pericarp tissue, and full germination was observed only after complete removal of pericarp tissue. Although the pericarp significantly reduced water uptake, no palisade layer(s) of macrosclereids could be observed. Xylem-vessel elements were found running through the basal end of the pericarp and forming the main point of water entry. We reject the hypothesis that G. coronaria cypselae have physical dormancy. Instead, water uptake and germination are impeded by: (1) directed water uptake, mainly through a pericarp-spanning channel-like structure; and (2) mechanical constraint on embryo growth exerted by the hard pericarp. The channel-like structure forms the principal system for controlling seed germination.

Physical seed dormancy is a common attribute among plants, and a wide range of dormancy-release mechanisms have been described, but their ecological significance is rarely tested through comparative study. This study tests whether dormancy-release responses to wet heat in four legume species with physical dormancy are correlated with habitat: two wetland species (Mimosa pigra and Parkinsonia aculeata, both dispersed primarily by water) and two terrestrial species (Acacia nilotica and Prosopis pallida, both dispersed primarily through vertebrate herbivores). Dormancy release was compared at three moisture levels (80% relative humidity, saturated and submerged) at constant (20–45°C) and diurnally fluctuating (20/40°C) temperatures for 14 d. Seed viability was tested by germinating at 25°C. The functional relationship between temperature and dormancy release after 14 d differed between species: submerged seeds of the two wetland species showed a quadratic response, with low rates of imbibition below 20–25°C and complete imbibition at around 40°C; P. pallida seeds showed a linear positive relationship, whereas there was no temperature response for A. nilotica seeds below 45°C. Surprisingly, dormancy release after 14 d was relatively insensitive to moisture levels, although rate of dormancy release was generally slower under drier conditions. Dormancy release was not influenced by fluctuating temperatures. Seed viability was largely unaffected by temperature or moisture regime, although it did differ with species and was lower for non-dormant seeds. Our results suggest that a functional dormancy-release response to wet heat provides important fitness benefits for wetland species, but not for species dispersed through vertebrate herbivores, for which it may be maladaptive.

We have studied by means of cytology and autoradiography the effect of benzyladenine (BA, a synthetic cytokinin) on DNA metabolism during early maize germination.The data indicate that BA stimulates nuclear DNA replication. The doubling of the amount of nuclear DNA in BA-treated axes occurs earlier than in nontreated axes, and there is a three-fold increase in the mitotic index at 24 h of germination. These results provide further corroboration for the suggestion that the stimulation of DNA synthesis observed relates to a nuclear replicative type of synthesis. Possible mechanisms of action of BA are discussed.

The conditions of osmopriming of maize seeds have been established. At an osmotic potential of–1.7 MPa, seed germination is inhibited and seeds osmoprimed for up to 3 weeks show an improved germinability when the osmotic agent is removed. This improvement also applies to seeds that lost vigour because of improper storage conditions. At the biochemical level, embryo axes from osmoprimed seeds can incorporate precursors into DNA, RNA and proteins although at a low level; after removing the osmotic agent, all 3 types of macromolecules are synthesized at much higher levels. No evidence of DNA replication or cell division was found during osmopriming; however, mitotic figures appear several hours earlier in germinated, osmoprimed root tissues compared with the time at which they appear in non-osmoprimed tissues. The behaviour of osmoprimed maize is compared with that of osmoprimed seeds from other plant species.

Seed germination of the hemiparasitic angiosperm Striga hermonthica is elicited by compounds present in the root exudates of the host plant. A variety of chemicals can substitute for the host-derived signal although the exact mechanism through which these act is unknown. In the present study, thidiazuron (TDZ), a cytokinin-active urea derivative was found to stimulate germination. This thidiazuron-induced germination was shown to be mediated by endogenous ethylene, by a similar mechanism to germination induced by host-root exudate and the synthetic stimulant GR-24. Comparing the effects of TDZ, host-root exudate, GR-24 and the ethylene precursor 1-aminocyclopropane-1-car-boxylic (ACC) suggests that ethylene production increases as a result of increased ACC synthesis and is consistent with a model for Striga seed germination in which host-derived signals and other stimulants act by eliciting the synthesis of ethylene via a stimulation of ACC synthase activity. All germination stimulants tested trigger a rapid increase in O2 uptake by conditioned seeds. The patterns of O2 uptake following TDZ or GR-24 treatments show similarities and three distinct phases of respiration are apparent. In contrast, stimulation of O2 uptake by ethylene results in a different pattern, with no distinct phases obvious following the initial burst of activity.

On the basis of embryo and seed colour, morphology, accumulation of fresh and dry weight, seed development of Chinese cabbage (Brassica rapa subsp. pekinensis) can be divided into 10 discrete but contiguous stages. Precocious germination (PG) occurs in the seeds of a Chinese cabbage mutant on the parent plant mainly during the maturation period (from stage 5 to 8), with either the radicle or the cotyledon protruding out of the testa. All plants of the mutant line produce some PG seeds, but among the seeds, only 18% (maximum) germinate precociously. The developing mutant seeds have higher water contents and lower dry weights than those of the wild-type and are less tolerant of desiccation. The testa structure of Chinese cabbage seed is similar to that of other Brassica species. In developing seeds, it consists of the epidermis, subepidermis, palisade layer and pigmented layers, while in the mature dry seeds, all the cell layers except the palisade layer are crushed into non-cell structures and are stacked on the outer and inner faces of the palisade layer; the aleurone layer is fused to the testa. An alteration in testa structure of the mutant seeds (both non-germinated and precociously-germinated seeds) is probably related to PG. The secondary cell wall materials are less, or not deposited on the radial and basal walls of the palisade cells on the adaxial side of the testa, which could result in a weaker mechanical restraint, thus leading to PG.

Untreated (control) tomato seeds germinated at temperatures ranging from 15 to 35°C. There was a positive linear relationship between the rate of germination (expressed as the reciprocal of time to obtain 50% germination) and temperature up to the thermal optimum. At higher temperatures, the negative relationship was also linear. The thermal optimum for seed germination was 27–28°C and the lower and maximal temperatures were around 9 and 40°C, respectively. Germination was strongly reduced in atmospheres containing less than 10% oxygen. After pretreatment at 15°C in a polyethylene glycol-8000 solution at –1.0 MPa, seeds germinated faster in a wider range of temperatures, and this stimulatory effect remained after drying back the seeds. Such a pretreatment did not change the optimal and maximal temperatures of germination, but decreased the lower temperature to about 6°C. The beneficial effect of osmoconditioning increased as the duration of the treatment increased and was maximal after 5–7 days. Osmotic pretreatment also reduced the sensitivity of seeds to oxygen deprivation; 50% of the seeds germinated within 7 days in 5% oxygen. The range of temperatures and the concentrations of oxygen which were effective in osmoconditioning were very similar to those which allowed the germination of untreated seeds. In particular, the optimal temperature (27–28°C) and oxygen concentrations (more than 10%) for osmoconditioning and germination were the same. These results confirm that the beneficial effect of osmotic pretreatment (priming) corresponds to the realization of germination sensu stricto (phase II of the germination process).

The rice (Oryza sativa L.) cv. Taichung 65, a japonica subspecies, was used to characterize the isoform, protein and gene expression patterns of endo-β-mannanase during and after seed germination. Activity assays and isoform analyses of whole grains or seed parts (scutellum, aleurone layer and starchy endosperm) revealed that seeds began to express endo-β-mannanase activity at 48 h from the start of imbibition at 25°C, after the completion of germination of most seeds. Three isoforms of endo-β-mannanase (pI 8.86, pI 8.92 and pI 8.98) were detected in the aleurone layer and starchy endosperm, but only two (pI 8.86 and pI 8.92) were present in the scutellum. The endo-β-mannanase in the starchy endosperm was mainly from the aleurone layer. Western blot analysis, using a tomato anti-endo-β-mannanase antibody, indicated that an endo-β-mannanase protein was present in an inactive form in dry grains. The amount of this protein decreased in the scutellum, but increased in the aleurone layer during and after germination. Thus, the increase in endo-β-mannanase activity in rice grains may be due to the activation of extant proteins and/or the de novo synthesis of the enzyme. Northern blot analysis showed that four putative rice endo-β-mannanase genes (OsMAN1, OsMAN2, OsMAN6 and OsMANP) were expressed in germinating and germinated rice grains. However, OsMANP was not expressed in the scutellum. The amount of OsMAN6 mRNA decreased after the completion of germination and paralleled the decline in endo-β-mannanase protein. In the aleurone layer, the increase of OsMAN2, OsMAN6 and OsMANP mRNA was prior to the increase of endo-β-mannanase protein.