The germination dynamics of Ilex aquifolium L. were studied in central Spain using a 3-year field sowing experiment. Ten triads of bags (n = 30 seeds per bag) were buried randomly in four different microhabitats: inside the holly woodland, along the forest edge, in open grassland and under isolated shrubs. Bags were harvested annually, and seeds were analysed for seed viability, embryo development changes and germination. Holly seeds did not germinate until the second year. Embryo development and seed germination were faster at the forest edge, with a total germination proportion of 63% of sown seeds. Germination in the holly woodland (48%), under the shrubs (46%) and in grassland (46%) were very similar. Thus, recruitment of Ilex populations in this region is not limited by requirements for germination. After the third year, the seed bank was reduced by 80–90% of the initial seeds, so it can be considered as a short-persistent seed bank. Holly seed behaviour can enhance recruitment in semi-open areas, such as forest edges. Linked to the short persistence of its seed bank, this species can be considered an intermediate strategist between mature forest specialists and early successional species. This work contributes to a deeper knowledge of the germination ecology of I. aquifolium, in an effort to determine the relative importance of germination/ dormancy processes within holly population dynamics.

Epizoochory is widely recognized as an effective long-distance seed dispersal mechanism. Nevertheless, few studies have focused on the investigation of its influencing factors. One of the key aspects of epizoochory is the adhesive interaction between seeds and furs. We describe a new method to quantify experimentally and standardize the adhesivity of seeds to animal fur, as a measure of epizoochorous dispersal potential. The method excludes the impact of animal behaviour and environmental factors, and allows the ranking of species according to their adhesivity score. We measured adhesivity scores for 66 species on the furs of seven mammals. Deep furs with long, rough, undulated hairs implanted at a large angle were most suited for seed adhesion, while seeds adhered less well to shallow furs with short, smooth, straight hairs implanted at small angles. Seeds with specialized adhesive appendages had higher adhesivity scores than seeds with unspecialized appendages and seeds without appendages. However, an interaction effect between certain seed and fur types exists. Although seed morphology is a good predictor for seed adhesivity on fur, less well-adhering seed types often still have relatively high adhesivity scores. Therefore, it is likely that nearly all species are, to some extent, able to disperse epizoochorously.

Expansins are a multi-gene family of proteins involved in changes in cell wall properties. In seeds where the embryo is completely surrounded by the endosperm, dormancy breakage requires weakening of the micropylar endosperm, increased embryo growth potential or both. Cell wall alterations are fundamental components of these processes, and expansins are thought to participate in them. Here, we explore the possible involvement of expansins in the control of germination by phytochrome in Datura ferox L. seeds. Based on the conserved sequences of known expansin genes, corresponding primers were designed to investigate the expression of expansin mRNAs by reverse transcriptase polymerase chain reaction (RT-PCR). One expansin mRNA was detected in micropylar endosperm of D. ferox, while two mRNAs were present in the embryo prior to radicle emergence. Expansin transcript content is promoted by red (R) light, both in the micropylar endosperm and the embryo; these effects of R are far-red light (FR) reversible, displaying a typical low-fluence response (LFR) in a way fully consistent with the photocontrol of germination. On the other hand, when the promotion of germination by the LFR is antagonized by exposing the seeds to continuous FR through a high-irradiance response (HIR) or by ABA, the inhibitory treatments do not affect expansin mRNA abundance. The results support the participation of expansins in the promotion of germination by LFR of phytochromes, and suggest that inhibition of germination by HIR or ABA does not include a reduction in the transcription of expansin genes.

The physical and chemical seed characteristics of 11 dipterocarp species (four Dipterocarpus, two Dryobalanops and five Shorea species) were quantified to examine variations among species. We measured dry weight, pericarp thickness and concentrations of nitrogen, total phenol, condensed tannin, lignin, lipid, starch and total sugar. Although there was low intraspecific variation among parent trees, each species had unique characteristics. We found significant correlations between chemical defensive compounds (i.e. lignin and total phenol) and nitrogen concentration. However, these relationships were no longer significant when an outlier was removed. Fruiting frequency was not significantly correlated with any chemical compound. Seeds of Shorea species had a higher concentration of lipid than seeds of Dipterocarpus. The implications of variation in the physical and chemical characteristics of dipterocarp seeds are discussed.

Both the California chaparral species, Emmenanthe penduliflora Benth. (Hydrophyllaceae), and a tobacco native to the Great Basin Desert of south-western Utah, Nicotiana attenuata Torr. ex Wats. (Solanaceae), germinate in response to component(s) of wood smoke. Nitrogen oxides (NO and NO2), in amounts produced by a fire, have been proposed to be germination signals for E. penduliflora. We examined the germination response of dormant seeds of E. penduliflora and N. attenuata to aqueous solutions of smoke adjusted to different pHs, and two NO donors [sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP)]. The smoke solutions, at pH 4 or 5, induced the maximum germination response. Aqueous solutions of SNP and SNAP, releasing NOx as high as 42 μM, had no effect on germination. Additionally, NO2– could not be detected in aqueous smoke extracts derived from combusted cellulose or wood. Therefore, unidentified cellulose combustion factors, rather than NOx, are likely to be the ecologically relevant germination signals.

The effects of temperature on embryo growth, radicle emergence and cotyledon emergence of Gagea lutea (Liliaceae), a perennial herb widely distributed in Europe, eastern Siberia, the Kurile Islands, Sakhalin and the Far East, were monitored outdoors and in laboratory tests. In Japan, this species inhabits open secondary grasslands and deciduous forests. Seeds with an underdeveloped embryo are dispersed in late May/early June in Hokkaido. The embryo elongates in autumn, and the radicle emerges from the seed in mid-October to mid-November, at temperatures of about 15/4°C. However, cotyledons do not emerge until April, after seeds with an emerged radicle are covered with snow (near 0°C) for about 4 months. In laboratory experiments, temperatures of 25/15°C or 20/10°C followed by 5–10°C were required for embryo growth and radicle emergence. Rate and percentage of cotyledon emergence were promoted by keeping seeds with an emerged radicle under snow. The optimum temperature for cotyledon emergence after 81 d under snow was 15/5°C. Thus, G. lutea has deep simple epicotyl morphophysiological dormancy, and this is the first report of epicotyl dormancy in the genus.

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.

Dormancy in seeds of Dodonaea viscosa is due to a water-impermeable seed coat (physical dormancy, PY). Thus, mechanically scarified seeds imbibed water (c. 95% increase in mass) and germinated to high percentages over a wide range of temperature regimes in both white light and darkness, whereas non-scarified seeds did not take up water. Dry heat at 80–160°C and dipping in boiling water for 1–60 s also broke dormancy in a high percentage of the seeds, and continuous far-red light was not inhibitory to germination. However, dry storage in the laboratory for >1 year did not overcome dormancy. Seeds made water-permeable by boiling imbibed water, and thus germinated, at a much slower rate than those made water-permeable by mechanical scarification. We suggest that boiling opened the ‘water gap’ in the seed coat (not yet described in Sapindaceae but present in other taxa with PY) and that water entered the seed only through this small opening, thereby accounting for the slow rate of imbibition and subsequent germination. Physical dormancy has now been shown to occur in seeds of this polymorphic, worldwide species from Australia, Brazil, Hawaii, Mexico and New Zealand. The low level of dormancy reported for seed lots of D. viscosa in China, India and Pakistan is probably due to collection of seeds before they dried to the critical moisture content for development of water-impermeability of the seed coat. Germination of non-dormant seeds over a wide range of temperatures and in white light, far-red (leaf-canopy shade) light and darkness are part of the germination strategy of D. viscosa and of other taxa whose seeds have PY at maturity.

Seed mass distribution in grassland communities of slopes of contrasting aspect was analysed in dolomite regions of the Pannonian Basin. Species frequencies were obtained for four pairs of data sets, which originated from corresponding south- and north-facing dolomite grasslands, thus forming four independent case studies. The data sets comprised 5–15 sample plots and 51–114 (average 85) species. The species were classified using an eight-class system reflecting their mean seed mass (MSM) records (class 1 being the lowest, MSM ≤ 0.2 mg; class 8 being the highest, MSM > 50 mg). Seed mass class distributions derived from slopes of contrasting aspect showed strong significant differences in chi-square tests for trend for all the four case studies. Small-seeded species (classes 1 and 2) showed a positive balance for the south-facing slopes, whereas large-seeded species (classes 5, 6, 7 and 8) were more frequent on north-facing slopes. Species with intermediate seed mass (classes 3 and 4) were not distinctive between the slopes. These results represent strong evidence of increased seed mass in the vegetation of north-facing grasslands, when compared to their south-facing counterparts. Among the phenomena potentially responsible for the new findings, we discuss the roles of microclimatic effects (especially drought stress and light regime), grass litter, interspecific competition and seed predation.

The concept of ‘imbibitional damage’ arose when it was observed that considerable leakage of cell contents could occur when dry seed or pollen tissues are plunged directly into water. It is now common practice to imbibe dehydrated tissue slowly, to permit the re-establishment of functional membranes, prior to placing the tissue into liquid water. However, this argument may not hold if the tissue of interest is inherently desiccation-sensitive. Slow drying of desiccation-sensitive (recalcitrant) seeds or excised embryonic axes results in damage at high water contents, because it permits time for aqueous-based deleterious processes to occur. The same argument may apply if partially dried material is re-imbibed slowly, as this technique will also expose the tissue to intermediate water contents for protracted periods. This hypothesis was tested using embryos or axes from seeds of three recalcitrant species (Artocarpus heterophyllus, Podocarpus henkelii and Ekebergia capensis). Excised material was rapidly dried to water contents within the range over which viability is lost during drying, and re-imbibed either rapidly, by plunging directly into water, or slowly, by placing the material on damp filter paper or exposing it to a saturated atmosphere for several hours. Although details of the response differed among species and developmental stage, in all cases direct re-imbibition in water resulted in higher (or similar, but never lower) survival than either of the slow rehydration techniques.

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.

For recalcitrant seeds, mortality curves of germination versus water content typically imply a wide range of desiccation sensitivities within a seed population. However, seed to seed differences in water content, during desiccation, may confound our interpretation of these mortality plots. Here, we illustrate this problem for two batches of Vitellaria paradoxa (Sapotaceae) seeds collected in 1996 and 2002. Whole seeds were desiccated to various target water contents (TWCs) using silica gel. During desiccation, smaller seeds in the population dried most rapidly. Consequently, there was a significant linear relationship between whole-seed water content and seed mass during the drying process. In addition, following desiccation to low TWCs, only the largest seeds in the population retained viability. Taken together, this suggests that the larger seeds survived, not as a consequence of great relative desiccation tolerance, but as a result of taking longer to desiccate. Subsequently, the critical water content (CWC) for viability loss was calculated, based on the assumptions that in the seed population whole-seed water content during desiccation was normally distributed and the smallest, and hence driest, seeds were killed first. Using this approach, the driest seeds in the population that were killed, at each TWC, were always below a single CWC (c. 20% and 26% in 1996 and 2002, respectively). In subsequent experiments the effect of seed size variation on the response to desiccation was confirmed by conducting desiccation screens on seeds sorted into two discrete size classes, i.e. the seed-lot heterogeneity in mass was reduced. Using this approach, the mortality curves had a steeper slope. Furthermore, data for 24 tropical tree species from the Database of Tropical Tree Seed Research (DABATTS) revealed that seed lots with less variability in mass had steeper mortality curves. Thus, taken together, the data suggest that, at least for whole seeds, the wide range of desiccation sensitivities typically inferred is an artefact of seed to seed variation in mass, and hence water contents, during drying.

Despite the importance of the palm family, Arecaceae, little has been systematically documented about the seed behaviour of the many species. The post-harvest seed behaviour of Phoenix reclinata, the highly utilized wild date palm species distributed along the eastern seaboard of Africa, is investigated in the present study. While both embryo and endosperm water concentration declined as the seeds of Phoenix reclinata matured, they remained relatively high: this is a characteristic of (but not confined to) non-orthodox seeds. The ultrastructure of embryo cells, and the finding that negligible water uptake was required for the initiation of germination, were in keeping with the possible non-orthodox nature of the seeds. A developmental study revealed that between the acquisition of full germinability and complete pre-shedding maturity, germination performance appeared to be constrained, suggesting the presence of an inhibitor. Pre-treatment by soaking, mechanical or acid scarification had no significant promotory effect on either rate or totality of germination of mature P. reclinata seeds, while use of water transiently at 100°C was highly deleterious. However, germination of partially dehydrated seeds was initiated sooner if they had been soaked or scarified. Mature P. reclinata seeds tolerated dehydration to a mean embryo water concentration of 0.40 g g–1 (dry mass basis; dmb), but at 0.14 g g–1, both rate and totality of germination were adversely affected. However, viability of seeds dehydrated to the mean embryo water concentration 0.40 g g–1 declined during storage for 16 weeks. It is concluded that P. reclinata seeds are non-orthodox, and are best categorized as showing intermediate post-harvest behaviour.

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.

Assessment of the rehydration procedure has been consistently overlooked in evaluation of factors contributing to successful cryostorage of partially dehydrated, embryonic axes excised from recalcitrant seeds. Conventional rehydration of Trichilia dregeana (Sond.) axes in distilled water resulted in the lack of root pole gravitropism after culture on medium in vitro. In comparison, a strong gravitropic response was observed in axes that had not been dehydrated, and by those rehydrated in a solution containing 1 M CaCl2 and 1 mM MgCl2. However, no marked loss of either cation from the tissue could be detected, whether axes were rehydrated in distilled water or the Ca2+/Mg2+ solution. Starch-packed statoliths differentiated rapidly in both non-dehydrated axes and those rehydrated in the divalent cation solution, but these organelles failed to develop or accumulate much starch following axis rehydration in water, as was the case for the amyloplasts of root cells generally. After rehydration in the Ca2+/Mg2+ solution and 48 h in culture, axis root-cap columella cells accumulated Ca2+, whereas axes rehydrated in water or solutions containing either Ca2+ or Mg2+ alone did not take up Ca2+. Rehydration with the Ca2+/Mg2+ solution also altered distribution of the actin component of the cytoskeleton. When rehydrated in the divalent cation solution, actin was associated with the nucleus and with the statoliths, which were located distally in statocytes of axes. In contrast, actin was largely confined to the perinuclear area in root-cap columella cells of the agravitropic, water-rehydrated axes. The present results indicate a definitive primary role for Ca2+ with Mg2+ in graviperception, via starch metabolism, and in the determination of statolith morphology, which appears to be linked with maintenance of the actin component of the cytoskeleton in root-cap statocytes.

Leymus Racemosus, The Mammoth Wild Rye, Is A Rhizomatous Perennial Grass, Mainly Distributed In The Moving Or Semi-Stabilized Sand Dunes In Deserts Of The Junggar Basin In Xinjiang, China. The Revival Ability Of The Young Seedling After Periods Of Desiccation Can Be Influenced By Several Factors: (1) The Stage Of Seedling Development – The Later The Stage At Dehydration, The Longer The Root Length And The Lower Is The Percentage Of Seedlings That Survive; (2) The Length Of The Period Of Desiccation – The Longer The Period That The Seedlings Are Under Desiccation, The Lower Is The Percentage Of Seedlings That Survive; (3) Endosperm Size – The Smaller The Proportion Of Endosperm That Remains In The Caryopses, The Lower Is The Percentage Of Seedlings That Revive, Determined By (A) The Stage Of Seedling Development, And (B) The Proportion Of The Endosperm That Is Removed By Cutting; And (4) Caryopsis Size – The Larger The Polymorphic Caryopses, The Higher Is The Percentage Of Young Seedlings That Revive From Periods Of Desiccation. The Physiological And Ecological Implications Of L. Racemosus Seedling Desiccation Tolerance Are That Under Extreme Desert And Unpredictable Environmental Conditions, The Chances Of Seedling Establishment Are Increased.

As a consequence of previous lack of success in cryostoring axes excised from newly shed seeds of Trichilia dregeana, the effects of the mode of axis excision on seedling production were investigated. Although vigorous root production occurred, no shoots were produced when the cotyledons were severed as closely as possible to the axis surface (explant-type 0). In contrast, shoot production was increasingly facilitated when small to larger segments of cotyledonary tissue were left attached to the axes (explant-types 1, 2 and 4), which did not compromise axis drying rate. However, root growth of explant-types 1, 2 and 4 was negatively affected, probably by leakage into the medium of an inhibitory or toxic substance(s) from the cut surfaces of the cotyledonary tissue. Microscopical examination revealed that the cotyledons were sessile, and their insertions were contiguous with the shoot apex in axes from newly shed seeds, leading to the suggestion that failure of shoot production by type 0 explants in vitro was the direct consequence of the proximity of the wound sites to the apical meristem. When seeds were stored hydrated for 6 months, the shoot apex had elongated, positioning the apical meristem some distance from the top of the cotyledonary insertions. In contrast to axes excised as type 0 explants from newly shed seeds, the equivalent explants from the stored seeds rapidly formed shoots and leaves in vitro. This indicates that the developmental status of axes, when excised, dictates failure or success in their further development in vitro, and that this aspect needs to be resolved before any further manipulations for cryostorage are attempted.