Dormancy release during hydrated storage in Lolium rigidum seeds is dependent on temperature, light quality, and hydration status

The influence of temperature, light environment, and seed hydration on the rate of dormancy release in Lolium rigidum (annual ryegrass) seeds during hydrated storage (stratification) was investigated. In a series of experiments, seeds were subjected to a range of temperatures (nine between 5 degrees C and 37 degrees C), light (white, red, far-red, and dark), and hydration (4-70 g H(2)O 100 g(-1) FW) during stratification for up to 80 d. Samples were germinated periodically at 25/15 degrees C or constant 15, 20, or 25 degrees C with a 12 h photoperiod to determine dormancy status. Dark-stratification was an alternative, but not equivalent dormancy release mechanism to dry after-ripening in annual ryegrass seeds. Dormancy release during dark-stratification caused a gradual increase in sensitivity to light, but germination in darkness remained negligible. Germination, but not dormancy release, was greater under fluctuating diurnal temperatures than the respective mean temperatures delivered constantly. Dormancy release rate was a positive linear function of dark-stratification temperature above a base temperature for dormancy release of 6.9 degrees C. Dormancy release at temperatures up to 30 degrees C could be described in terms of thermal dark-stratification time, but the rate of dormancy release was slower at < or =15 degrees C (244 degrees Cd/probit increase in germination) than > or =20 degrees C (208 degrees Cd/probit). Stratification in red or white, but not far-red light, inhibited dormancy release, as did insufficient (<40 g H(2)O 100 g(-1) FW) seed hydration. The influence of dark-stratification on dormancy status in annual ryegrass seeds is discussed in terms of a hypothetical increase in available membrane-bound phytochrome receptors.     

ROS production and protein oxidation as a novel mechanism for seed dormancy alleviation

At harvest, sunflower (Helianthus annuus L.) seeds are dormant and unable to germinate at temperatures below 15 degrees C. Seed storage in the dry state, known as after-ripening, is associated with an alleviation of embryonic dormancy allowing subsequent germination at suboptimal temperatures. To identify the process by which dormancy is broken during after-ripening, we focused on the role of reactive oxygen species (ROS) in this phenomenon. After-ripening entailed a progressive accumulation of ROS, namely superoxide anions and hydrogen peroxide, in cells of embryonic axes. This accumulation, which was investigated at the cellular level by electron microscopy, occurred concomitantly with lipid peroxidation and oxidation (carbonylation) of specific embryo proteins. Incubation of dormant seeds for 3 h in the presence of hydrogen cyanide (a compound that breaks dormancy) or methylviologen (a ROS-generating compound) also released dormancy and caused the oxidation of a specific set of embryo proteins. From these observations, we propose a novel mechanism for seed dormancy alleviation. This mechanism involves ROS production and targeted changes in protein carbonylation patterns.     

Dormancy release and seed ageing in the endangered species <Emphasis Type="Italic">Silene diclinis</Emphasis>

The influence of seed testa color, temperature and seed water content on dormancy release and seed viability loss in the endangered, endemic species Silene diclinis (Lag.) M. Laínz was evaluated. Dormant heterogeneous seeds (black, red and grey colored) were exposed to three different temperatures (5, 20, and 35°C) and two relative humidities (33 and 60%) in order to assay their dormancy release. Longevity behavior was studied for the three colored seeds, storing samples at nine different combinations of temperature (5, 20 and 35°C) and relative humidities (33, 60 and 90%). According to our findings, seed heteromorphism was not related to neither break of dormancy nor seed storage behavior. Silene diclinis seeds present dormancy after collection, and need an after-ripening period to germinate. Temperature and relative humidity are positively correlated with dormancy release and seed ageing. Therefore, both factors must be carefully controlled during seed manipulation in the laboratory for long term seed conservation purposes. When seeds are stored immediately after collection (dormant), if the temperature of storage is above the base temperature for dormancy release found in this work (between 2.7 and 1.6°C), seeds may eventually overcome dormancy. On the other hand if seeds are stored after an after-ripening period, storage at low temperature does not induce secondary dormancy.     

THE EFFECT OF THE INTERACTION OF TEMPERATURE WITH AFTER-RIPENING REQUIREMENT AND COMPENSATING TEMPERATURE ON GERMINATION OF SEED OF FIVE SPECIES OF ROSA

Seeds of 5 rose species, Rosa multiflora Thunb. ‘Cathayensis,’ R. × reversa Waldst. & Kit., R. setigera Michx. ‘Beltsville,’ R. setigera Michx. ‘Serena,’ and R. wichuraiana Crepin, varied in after-ripening requirement from 30 days at 4.4 C for R. multiflora to 90 days for R. setigera ‘Serena.’ The compensating temperature varied from near 12.8 C for R. × reversa to a value near 29.4 C for R. setigera ‘Beltsville.’ In this report compensating temperature is used to describe that temperature at which mature, moist seed does not germinate, after-ripening does not take place, and dormancy does not change. Seed germination was reduced by interruption of the after-ripening period with intervals at temperatures above the compensating temperature. The interruptions were more effective in reducing germination when more frequent and when the temperature during the interval was higher. Species differed in their sensitivity to high-temperature reduction of germination. Those having the longest after-ripening requirement were most sensitive. Germination of seeds which had the minimum after-ripening treatment was repressed more by high temperature than germination of those seeds which had an excess of after-ripening. The decrease in germination resulted from imposition of a secondary dormancy of the embryo, and probably also from reversal of the after-ripening effect upon the primary dormancy imposed by the seed coat.     

The changing window of conditions that promotes germination of two fire ephemerals, Actinotus leucocephalus (Apiaceae) and Tersonia cyathiflora (Gyrostemonaceae)

BACKGROUND AND AIMS: Following a period of burial, more Actinotus leucocephalus (Apiaceae) and Tersonia cyathiflora (Gyrostemonaceae) seeds germinate in smoke water. The main aim of this study was to determine whether these fire-ephemeral seeds exhibit annual dormancy cycling during burial. This study also aimed to determine the effect of dormancy alleviation on the range of light and temperature conditions at which seeds germinate, and the possible factors driving changes in seed dormancy during burial. METHODS: Seeds were collected in summer, buried in soil in mesh bags in autumn and exhumed every 6 months for 24 months. Germination of exhumed and laboratory-stored (15 degrees C) seeds was assessed at 20 degrees C in water or smoke water. Germination response to light or dark conditions, incubation temperature (10, 15, 20, 25 and 30 degrees C), nitrate and gibberellic acid were also examined following burial or laboratory storage for 24 months. In the laboratory seeds were also stored at various temperatures (5, 15, 37 and 20/50 degrees C) for 1, 2 and 3 months followed by germination testing in water or smoke water. KEY RESULTS: The two species exhibited dormancy cycling during soil burial, producing low levels of germination in response to smoke water when exhumed in spring and high levels of germination in autumn. In autumn, seeds germinated in both light and dark and at a broader range of temperatures than did laboratory-stored seeds, and some Actinotus leucocephalus seeds also germinated in water alone. Dormancy release of Actinotus leucocephalus was slow during dry storage at 15 degrees C and more rapid at higher temperatures (37 and 20/50 degrees C); weekly wet/dry cycles further accelerated the rate of dormancy release. Cold stratification (5 degrees C) induced secondary dormancy. By contrast, no Tersonia cyathiflora seeds germinated following any of the laboratory storage treatments. CONCLUSIONS: Temperature and moisture influence dormancy cycling in Actinotus leucocephalus seeds. These factors alone did not simulate dormancy cycling of Tersonia cyathiflora seeds under the conditions tested.     

Interchangeable effects of gibberellic acid and temperature on embryo growth, seed germination and epicotyl emergence in Ribes multiflorum ssp. sandalioticum (Grossulariaceae)

Morphophysiological dormancy was investigated in seeds of Ribes multiflorum Kit ex Roem et Schult. ssp. sandalioticum Arrigoni, a rare mountain species endemic to Sardinia (Italy). There were no differences in imbibition rates between intact and scarified seeds, suggesting a lack of physical dormancy, while methylene blue solution (0.5%) highlighted a preferential pathway for solution entrance through the raphe. Embryos were small at seed dispersal, with an initial embryo:seed ratio (E:S) of ca. 0.2 (embryo length, ca. 0.5 mm), whereas the critical E:S ratio for germination was three times longer (ca. 0.6). Gibberellic acid (GA(3), 250 mg · l(-1)) and warm stratification (25 °C for 3 months) followed by low temperature (<15 °C) enhanced embryo growth rate (maximum of ca. 0.04 mm · day(-1) at 10 °C) and subsequent seed germination (radicle emergence; ca. 80% at 10 °C). Low germination occurred at warmer temperatures, and cold stratification (5 °C for 3 months) induced secondary dormancy. After radicle emergence, epicotyl emergence was delayed for ca. 2 months for seeds from three different populations. Mean time of epicotyl emergence was affected by GA(3) . Seeds of this species showed non-deep simple (root) - non-deep simple (epicotyl) morphophysiological dormancy, highlighting a high synchronisation with Mediterranean seasonality in all the investigated populations.     

Morphophysiological dormancy in seeds of two North American and one Eurasian species of Sambucus (Caprifoliaceae) with underdeveloped spatulate embryos

In contrast to previous reports, the endocarps ("seed coats") of Sambucus species are not impermeable to water; thus, the seeds do not have physical dormancy. Seeds of the North American species Sambucus canadensis and S. pubens and of the European species S. racemosa have spatulate shaped embryos that are ～60% fully developed (elongated) at seed maturity. The embryo has to extend to the full length of the seed to germinate. Embryos in freshly matured seeds of S. canadensis and in those of S. pubens grew better at 25°/15°C than at 5°C, whereas the rate of embryo growth in S. racemosa was higher at 5°C than at 25°/15°C. Seeds of all three species germinated to significantly higher percentages in light (14-h photoperiod) than in darkness. Fresh seeds of neither species germinated during 2 wk of incubation over a range of thermoperiods. Warm followed by cold stratification broke dormancy in seeds of S. canadensis and in those of S. pubens. Thus, seeds of these two North American species have deep simple morphophysiological dormancy (MPD). In comparison, seeds of the European species S. racemosa required a cold stratification period only for dormancy break, and thus they have intermediate complex MPD. GA(3) was much more effective in breaking dormancy in seeds of S. racemosa than it was in those of S. canadensis or S. pubens.     

Secondary dormancy in Avena fatua: Effect of temperature and after-ripening

To evaluate the effect of after-ripening on secondary dormancy induction in pure genetic lines of Avena fatua L., seed samples were periodically removed from standard conditions of storage and the caryopses then subjected to anoxia. Anoxia did not induce secondary dormancy in SH430, a line characterized by no primary dormancy at harvest maturity; secondary dormancy was induced in caryopses of other lines that had been after-ripened to over-come primary dormancy ranging in duration from a few days (CS40, CS166) to several months (AN51, AN127). Germination response to low GA₃ concentrations indicated that secondary dormancy in CS40 and CS166 was less intense than in AN51 and AN127. The longer the period of dry after-ripening prior to anoxia treatment, the lower the intensity of secondary dormancy induced. After a period of dry after-ripening, which was characteristic for each line, anoxia became an ineffective dormancy-inducing treatment. Caryopses selected for their response to dormancy induction by anoxia were subjected to temperatures from 5 to 35°C to investigate the effect of low (5 to 18°C) and high (20 to 35°C) temperatures on both thermo- and secondary dormancy induction. SH430 was not responsive to any treatment, while CS40, CS166 and AN51 were induced into a thermo-dormancy at temperatures above 20°C and CS166 and AN51 were induced into secondary dormancy by anoxia at temperatures from 5 to 35°C. The effect of anoxia on secondary dormancy induction in a range of pure genetic lines is discussed with reference to primary dormancy, after-ripening and temperature.     

Discovering the type of seed dormancy and temperature requirements for seed germination of Gentiana lutea L. subsp. lutea (Gentianaceae)

Aims There are a number of mechanisms that regulate germination; among these, seed dormancy, one of the most important, is an adaptative mechanism in plants to promote survival by dispersing germination in space and time until environmental conditions are favourable for germination. The main goals of this study were to determine the temperature requirements for seed dormancy release and germination of Gentiana lutea subsp. lutea, to identify the class and level of seed dormancy and to suggest an optimal germination protocol.Methods Seeds belonging to two different localities were subjected to various pre-treatments, including cold stratification (0 and 5°C), warm stratification (25/10°C) and different combinations of these, and then incubated at a range of constant temperatures (5–25°C) and 25/10°C. Embryo growth during pre-treatments and incubation conditions were assessed at different times by measuring the embryo to seed length ratio (E:S ratio). The final germination percentage (FGP) and the germination rate (t 50) were calculated.Important findings Fleshy mature seeds of G. lutea subsp. lutea have linear underdeveloped embryos. Cold stratification at 0°C was effective in overcoming the physiological dormancy (PD) and promoted embryo growth and subsequent germination. After cold stratification at 0°C, both the root and the shoot emerged readily under a wide range of temperatures. G. lutea subsp. lutea seeds showed an intermediate complex morphophysiological dormancy (MPD). As regards the optimal germination protocol for this taxon, we suggest a period of cold stratification at ca. 0°C followed by seed incubation at 10–20°C. The optimal germination temperatures found for seeds of this taxon, as well as its pre-chilling requirement at 0°C, suggest that it is well adapted to a temperate climate; this behavior highlights an increasing threat from global warming for G. lutea, which could reduce the level of natural emergence in the field, prejudicing also the long-term persistence of the natural populations in Sardinia.     

Underdeveloped embryos and germination in Aristolochia galeata seeds

Aristolochiaceae have been described as having seeds with underdeveloped embryos and morphological or morphophysiological dormancy. Aristolochia galeata is a native climber found in the Cerrado biome, associated with road and gallery forest edges. The aims of this study were to investigate: embryo growth rate, morphology and seed germination parameters under different treatments. Embryos were excised to obtain embryo length at four stages: initial, seeds after coat rupture, radicle tip protrusion and cotyledon emergence from the seed coat. Germination tests were conducted at 30 °C under three nitrate concentrations (1, 10 and 20 mM), fluctuating temperature (27/20 °C) and light and dark conditions. We found that seeds have underdeveloped embryos, which take about 301 ± 178 h (±SD) to achieve seed coat rupture, another 205 ± 126 h to reach radicle protrusion and 176 ± 76 h more to the final stage of cotyledon emergence. Germinability was above 52% in all treatments, except in the dark (15%). For all treatments, average germination time was above 290 ± 123 h. Potassium nitrate increased germinability to >87%. No particular treatment was required for embryo development, but seeds in the population that continued to germinate after 1 month were probably in various states of non-deep, simple morphophysiological dormancy. Increased germinability in nitrate treatments and light requirement for germination could prevent germination under unsuitable environmental conditions and be a strategy to increase seedling establishment in the cerrado.     

濒危植物巴东木莲种子休眠与萌发特性的研究

巴东木莲(Manglietia patungensis)为我国特有种, 属国家重点保护植物。为找出其生殖环节中的致危因素, 作者对巴东木莲种子休眠与后熟过程中的形态和萌发特性进行了研究。结果表明, 巴东木莲种胚发育不完全可能是种子休眠的主要原因, 在其后熟过程中胚不断分化、发育成熟; 种皮具有较好的透性, 与休眠的关系不大; 种子不同部位均存在萌发抑制物, 胚乳中高含量的萌发抑制物是影响胚萌发的重要因素。内源激素ABA和IAA在巴东木莲种子休眠与萌发过程中起着重要作用, ABA是引起休眠的关键因素, IAA有助于种子的萌发, IAA/ABA相对含量的变化对种子的休眠和萌发产生重要影响。巴东木莲种子的休眠是由种子本身的形态和生理特点引起的综合休眠, 在4℃低温保湿条件下才能完成其形态和生理后熟过程, 而自然条件下, 巴东木莲种子成熟时正值秋季少雨, 很容易失水而不能完成其后熟过程而失去生活力, 这可能是导致该物种自然更新困难的重要原因。     

Regulation of dormancy in barley by blue light and after-ripening: effects on abscisic acid and gibberellin metabolism

White light strongly promotes dormancy in freshly harvested cereal grains, whereas dark and after-ripening have the opposite effect. We have analyzed the interaction of light and after-ripening on abscisic acid (ABA) and gibberellin (GA) metabolism genes and dormancy in barley (Hordeum vulgare 'Betzes'). Analysis of gene expression in imbibed barley grains shows that different ABA metabolism genes are targeted by white light and after-ripening. Of the genes examined, white light promotes the expression of an ABA biosynthetic gene, HvNCED1, in embryos. Consistent with this result, enzyme-linked immunosorbent assays show that dormant grains imbibed under white light have higher embryo ABA content than grains imbibed in the dark. After-ripening has no effect on expression of ABA biosynthesis genes, but promotes expression of an ABA catabolism gene (HvABA8'OH1), a GA biosynthetic gene (HvGA3ox2), and a GA catabolic gene (HvGA2ox3) following imbibition. Blue light mimics the effects of white light on germination, ABA levels, and expression of GA and ABA metabolism genes. Red and far-red light have no effect on germination, ABA levels, or HvNCED1. RNA interference experiments in transgenic barley plants support a role of HvABA8'OH1 in dormancy release. Reduced HvABA8'OH1 expression in transgenic HvABA8'OH1 RNAi grains results in higher levels of ABA and increased dormancy compared to nontransgenic grains.     

Sequential temperature control of multi-phasic dormancy release and germination of Paeonia corsica seeds

Aims The physiological responses during dormancy removal and multi-phasic germination were investigated in seeds of Paeonia corsica (Paeoniaceae).Methods Seeds of P. corsica were incubated in the light at a range of temperatures (10–25 and 25/10°C), without any pre-treatment, after W (3 months at 25°C), C (3 months at 5°C) and W + C (3 months at 25°C followed by 3 months at 5°C) stratification, and a GA 3 treatment (250 mg·l^-1 in the germination substrate). Embryo growth, time from testa to endosperm rupture and radicle emergence were assessed as separate phases. Epicotyl–plumule emergence was evaluated incubating the germinated seeds at 15°C for 2 weeks, at 5 and 25°C for 2 months on agar water before transplanting to the soil substrate at 10, 15 and 20°C and at 15°C for 2 months on the surface agar water with GA 3 .Important findings Embryo growth, testa rupture, endosperm rupture (radicle emergence) and growth of the epicotyl were identified as four sequential steps in seeds of P. corsica. Gibberellic acid alone and warm stratification followed by 15°C promoted embryo growth and subsequent seed germination. Cold stratification induced secondary dormancy, even when applied after warm stratification. After radicle emergence, epicotyl–plumule emergence was delayed for ca. 3 months. Mean time of epicotyl–plumule emergence was positively affected by cold stratification (2 months at 5°C) and GA 3. P. corsica seeds exhibited differential temperature sensitivity for the four sequential steps in the removal of dormancy and germination processes that resulted in the precise and optimal timing of seedling emergence.     

Weather,season, and daily stroke admissions in Hong Kong

Previous studies examining daily temperature and stroke incidence have given conflicting results. We undertook this retrospective study of all stroke admissions in those aged 35 years old and above to Hong Kong public hospitals from 1999 through 2006 in order to better understand the effects of meteorological conditions on stroke risk in a subtropical setting. We used Poisson Generalized Additive Models with daily hemorrhagic (HS) and ischemic stroke (IS) counts separately as outcomes, and daily mean temperature, humidity, solar radiation, rainfall, air pressure, pollutants, flu consultation rates, day of week, holidays, time trend and seasonality as predictors. Lagged effects of temperature, humidity and pollutants were also considered. A total of 23,457 HS and 107,505 IS admissions were analyzed. Mean daily temperature had a strong, consistent, negative linear association with HS admissions over the range (8.2-31.8°C) observed. A 1°C lower average temperature over the same day and previous 4 days (lags 0-4) being associated with a 2.7% (95% CI: 2.0-3.4%, P < .0.0001) higher admission rate after controlling for other variables. This association was stronger among older subjects and females. Higher lag 0-4 average change in air pressure from previous day was modestly associated with higher HS risk. The association between IS and temperature was weaker and apparent only below 22°C, with a 1°C lower average temperature (lags 0-13) below this threshold being associated with a 1.6% (95% CI:1.0-2.2%, P < 0.0001) higher IS admission rate. Pollutant levels were not associated with HS or IS. Future studies should examine HS and IS risk separately.     

Wheat seed proteins regulated by imbibition independent of dormancy status

Seed dormancy is an important trait in wheat (Trticum aestivum L.) and it can be released by germination-stimulating treatments such as after-ripening. Previously, we identified proteins specifically associated with after-ripening mediated developmental switches of wheat seeds from the state of dormancy to germination. Here, we report seed proteins that exhibited imbibition induced co-regulation in both dormant and after-ripened seeds of wheat, suggesting that the expression of these specific proteins/protein isoforms is not associated with the maintenance or release of seed dormancy in wheat.     

After-Ripening in Festuca idahoensis Seeds: Adaptive Dormancy and Implications for Restoration

Festuca idahoensis (Idaho fescue) was a common native perennial bunchgrass in the sagebrush steppe of the western United States until the introductions of domestic livestock and alien plants. Restoration of Idaho fescue to degraded sites will likely involve reseeding, and one of the factors affecting reseeding success is germinability of the seeds employed. We investigated effects of after-ripening and storage temperature on germinability of Idaho fescue seeds collected from a central Oregon site. Six months of after-ripening were required before maximum germination was obtained. Storage of dry seeds at either room temperature (20°C) or at cooler, alternating temperatures (5/15°C) did not alter the rate at which dormancy was lost. Storage at the warmer temperature promoted rapid germination in seeds that had broken dormancy. Seed longevity varied greatly from year to year. Seeds produced in a very dry year had poorer germination and shorter longevity than seeds produced during a year with near normal precipitation. Because seed dispersal occurs in late July and early August for Idaho fescue in central Oregon, a six-month after-ripening requirement ensures that the greatest potential germination coincides with the spring period most likely to provide sufficient moisture for seedling establishment.