A potential role for endogenous microflora in dormancy release,cytokinin metabolism and the response to fluridone in Lolium rigidum seeds

Background and Aims Dormancy in Lolium rigidum (annual ryegrass) seeds can be alleviated by warm stratification in the dark or by application of fluridone, an inhibitor of plant abscisic acid (ABA) biosynthesis via phytoene desaturase. However, germination and absolute ABA concentration are not particularly strongly correlated. The aim of this study was to determine if cytokinins of both plant and bacterial origin are involved in mediating dormancy status and in the response to fluridone.Methods Seeds with normal or greatly decreased (by dry heat pre-treatment) bacterial populations were stratified in the light or dark and in the presence or absence of fluridone in order to modify their dormancy status. Germination was assessed and seed cytokinin concentration and composition were measured in embryo-containing or embryo-free seed portions.Key Results Seeds lacking bacteria were no longer able to lose dormancy in the dark unless supplied with exogenous gibberellin or fluridone. Although these seeds showed a dramatic switch from active cytokinin free bases to O-glucosylated storage forms, the concentrations of individual cytokinin species were only weakly correlated to dormancy status. However, cytokinins of apparently bacterial origin were affected by fluridone and light treatment of the seeds.Conclusions It is probable that resident microflora contribute to dormancy status in L. rigidum seeds via a complex interaction between hormones of both plant and bacterial origin. This interaction needs to be taken into account in studies on endogenous seed hormones or the response of seeds to plant growth regulators.     

Green and blue light photoreceptors are involved in maintenance of dormancy in imbibed annual ryegrass (Lolium rigidum) seeds

Light plays an important role in two separate processes within the seeds of Lolium rigidum (annual ryegrass). Dormant seeds of L. rigidum remain dormant when imbibed in the light, but once seeds have lost dormancy through dark-stratification, light stimulates their germination. This study characterizes the light qualities and quantities which are effective in maintenance of dormancy. Dormant seeds were stratified under narrow- and broad-waveband light to identify the potential photoreceptors involved in dormancy maintenance, and to determine whether dark-induced dormancy loss is reversible by light. Blue and green light both mediated dormancy maintenance in a far-red-independent manner. Red light resulted in dormancy maintenance only when far-red wavelengths were excluded, suggesting a redundant function of phytochrome. At low fluence rates, white light was more effective than monochromatic light, suggesting the action of multiple photoreceptors in dormancy maintenance. By contrast, nondormant seeds did not germinate unless provided with red light. These results indicate that seed dormancy maintenance is potentially mediated through the actions of blue and green light photoreceptors. Seed dormancy could thus be added to the growing list of plant responses that may be mediated by green light in a cryptochrome-independent manner.     

Dormancy and longevity of annual ryegrass (Lolium rigidum) as affected by soil type, depth, rainfall, and duration of burial

Effects of soil type, time, depth of seed burial and rainfall pattern were investigated on the longevity of glyphosate resistant annual ryegrass (Lolium rigidum Gaudin) in the northern summer rainfall dominant grain region of Australia in a 16 month experiment conducted under polyhouse conditions. Lolium rigidum seeds placed in nylon bags were buried in pots at 5 and 10 cm depth in either Laureldale (clay) or Kirby (sandy loam) soil receiving simulated rainfall representing a Tamworth (summer) and Hamilton (winter) rainfall environment of Australia. The bags were exhumed every four months and tested for percent viable, germinable, dormant and dead seeds. In the short term (several months), factors such as soil type, rainfall and depth of burial affected the fate and condition of L. rigidum seeds and emergence pattern of seedlings. However, irrespective of treatment, all seeds lost their viability after 16 months of burial. Hence, in the longer term, L. rigidum behaviour in this summer dominant rainfall environment with different soils is likely to be similar to that where the weed occurs more commonly in the southern Mediterranean regions of Australia. Maximum emergence in the polyhouse occurred during mid autumn similar to that in the field. The results from this experiment will allow for the development of management strategies which may enhance the depletion of the soil seedbank of viable L. rigidum seeds in the shortest possible time.     

山楂种子体眠和萌发与营养物质植物生长物质关系的研究

对低温层积不同时期的山楂种仁中的氨基酸、植物生长物质等几项生理生化指标进行了定量分析的结果表明层积１５至１８个月时因缺水出现二次休眠,在种子内部发生一系列生理变化,从中得知脱落酸,吲哚乙酸为生长抑制剂,玉米素核苷,赤霉素为促进剂。脯氨酸对干旱比较明感,随着层积时间的延长营养物质趋于升高,酶的活性提高,为种子发芽提供了能量。     

Selection for low or high primary dormancy in Lolium rigidum Gaud seeds results in constitutive differences in stress protein expression and peroxidase activity

Seed dormancy in wild Lolium rigidum Gaud (annual ryegrass) populations is highly variable and not well characterized at the biochemical level. To identify some of the determinants of dormancy level in these seeds, the proteomes of subpopulations selected for low and high levels of primary dormancy were compared by two-dimensional polyacrylamide gel electrophoresis of extracts from mature, dry seeds. High-dormancy seeds showed higher expression of small heat shock proteins, enolase, and glyoxalase I than the low-dormancy seeds. The functional relevance of these differences in protein expression was confirmed by the fact that high-dormancy seeds were more tolerant to high temperatures imposed at imbibition and had consistently higher glyoxalase I activity over 0-42 d dark stratification. Higher expression of a putative glutathione peroxidase in low-dormancy seeds was not accompanied by higher activity, but these seeds had a slightly more oxidized glutathione pool and higher total peroxidase activity. Overall, these biochemical and physiological differences suggest that L. rigidum seeds selected for low dormancy are more prepared for rapid germination via peroxidase-mediated cell wall weakening, whilst seeds selected for high dormancy are constitutively prepared to survive environmental stresses, even in the absence of stress during seed development.     

Sucrose and light effects on in vitro cultures of potato,chokecherry and saskatoon berry during low temperature storage

Cultures of potato (Solanum tuberosum) cv. Atlantic, chokecherry (Prunus virginiana L.) cv. Garrington and saskatoon berry (Amelancher alnifolia Nutt.) cv. Northline grown in vitro for 3 weeks at 24/22 °C, 16-h photoperiod, 150 μmol m⁻² s⁻¹ photosynthetic photon flux density (PPFD) mixed fluorescent/incandescent light were stored for 6, 9 and 12 weeks at 4 °C under 0 (darkness) and 3 μmol m⁻² s⁻¹ PPFD (690 nm red light continuous illumination). Growth regulators free MSMO medium either with or without 30 g l⁻¹ sucrose was used to store the cultures. All cultures retained capacity to re-grow after storage. Tested factors, sucrose, light and the length of the storage period had an impact on shoot quality and re-growth capacity of the cultures. For either light treatment sucrose was essential for the low temperature maintenance of vigorous stock plants of potato, if stored for over 6 weeks. Chokecherry and saskatoon cultures stored well without sucrose; although chokecherry benefited from sucrose in the storage medium when the stock cultures were kept at the low temperature for 12 weeks. Low light significantly improved quality of the stored potato cultures, but had very little effect on both chokecherry and saskatoon berry cultures. The woody plant cultures grew during storage, and the longer the stock plants were stored, the more vigorous cultures they generated. The results indicate that growers can successfully use their existing facilities, small refrigerators and coolers with low light intensity, set at 4 °C, for short term storage of potato, chokecherry and saskatoon berry cultures. The potato cultures, which are known to be sensitive to prolonged low temperature storage, should be frequently monitored and subcultured as required. On the other hand, the woody plant stock cultures do not require any special attention when kept at 4 °C and re-grow the most vigorous shoots if stored for at least 12 weeks. This revised version was published online in August 2006 with corrections to the Cover Date.     

Role of relative humidity, temperature, and water status in dormancy alleviation of sunflower seeds during dry after-ripening

The effect of various combinations of temperature and relative humidity on dormancy alleviation of sunflower seeds during dry after-ripening was investigated. The rate of dormancy alleviation depended on both temperature and embryo moisture content (MC). Below an embryo MC of 0.1 g H(2)O g(-1) dw, dormancy release was faster at 15 °C than at higher temperatures. This suggests that dormancy release at low MC was associated with negative activation energy, supported by Arrhenius plots, and low Q(10) values. At higher MC, the rate of dormancy alleviation increased with temperature, correlating well with the temperature dependence of biochemical processes. These findings suggests the involvement of two distinct cellular mechanisms in dormancy release; non-enzymatic below 0.1 g H(2)O g(-1) dw and associated with active metabolism above this value. The effects of temperature on seed dormancy release above the threshold MC were analysed using a population-based thermal time approach and a model predicting the rate of dormancy alleviation is provided. Sunflower embryo dormancy release was effective at temperatures above 8 °C (the base temperature for after-ripening, Tb(AR), was 8.17 °C), and the higher the after-ripening temperature above this threshold value, the higher was the rate of dormancy loss. Thermodynamic analyses of water sorption isotherms revealed that dormancy release was associated with less bound water and increased molecular mobility within the embryonic axes but not the cotyledons. It is proposed that the changes in water binding properties result from oxidative processes and can, in turn, allow metabolic activities.     

Dormancy inDioscorea: Different temperature adaptation of seeds,bulbils and subterranean organs in relation to north-south distribution

Temperature dependencies of sprouting and germination were compared for subterranean perennating organs and seeds of ten closely related species of the genusDioscorea (Dioscoreaceae), a group of monocotyledonous summer perennials which are distributed from the tropics to the subarctic. The species used wereD. nipponica Makino,D. tokoro Makino,D. japonica Thunb.,D. tenuipes Franch. et Savat.,D. septemloba Thunb.,D. quinqueloba Thunb.,D. izuensis Akahori,D. bulbifera L. f.spontanea (Makino) Makino et Nemoto,D. pentaphylla. L. andD. alata L.; they are distributed from cold northern areas to warmer southern areas approximately in this order in and around Japan. Bulbil sprouting was also studied in those forming bulbils. Subterranean organs of the tropical species sprouted faster without any prior temperature treatment, whereas those of species from the more northern areas sprouted after prechilling. Northern species required longer, periods of prechilling for sprouting. On the other hand, with seeds or bulbils, the southern species required longer periods of prior temperature treatment for dormancy breaking. This difference in the length of dormant periods between seeds or bulbils and subterranean organs among the ten species may be related to their size and position of shedding; seeds or bulbils are small and are shed on the ground surface, whereas subterranean organs are large and are located below the surface. It is important to determine in other perennials whether the above relation between dormant features of seeds or bulbils and subterranean organs are common properties or not.     

Effect of light and temperature on the germination of lettuce seeds

A short period (15–30 min) at 30° C promotes germination of seeds of Lactuca sativa L. cv. Repolhuda in darkness. Far-red light reverses this stimulation, and the escape curves for phytochrome and high-temperature action are quite similar, indicating that the two factors act at a common point in the chain of events leading to germination. It is suggested that high temperature acts by decreasing the threshold of the active, far-red absorbing, form of phytochrome (Pfr) needed to promote germination.Abbreviations FR far-red light - Pfr far-red-absorbing form of phytochrome - R red light     

The influence of metabolic inhibitors and temperature on mutagenic activity during the storage of ethyl methanesulphonate-treated barley seeds

Anaerobic conditions, the inhibitor of aerobic respiration-sodium azide and the inhibitor of proteosynthesis-chloramphenicol did not influence the increase of toxic and mutagenic effects during the storage of ethyl methanesulphonate-treated barley seeds at 15 per cent water content. The storage of ethyl methanesulphonate-treated seeds at ? 20 °C or at 0 °C avoided all after-effects. With increasing storage temperature (5 °C, 15 °C and 25 °C) the increase of injury due to storage was more pronounced.     

Production and scavenging of reactive oxygen species in Fagus sylvatica seeds during storage at varied temperature and humidity

The accumulation of reactive oxygen species (ROS) in seed tissues plays an important role in the loss of seed viability during storage. In the present study, we examined whether the loss of germination capacity and viability of beech (Fagus sylvatica L.) seeds during storage under different temperatures (4, 20 and 30 degrees C) and relative humidity levels (45% and 75% RH) is associated with: (1) an increase in the level of ROS, such as superoxide radical (O2*-), oxygen peroxide (H2O2); and, (2) changes in low molecular antioxidants (ascorbate and glutathione) and enzymatic scavengers such as ascorbate peroxidase dehydroascorbate reductase, glutathione reductase, catalase, superoxide dismutase and guaiacol peroxidase. Beech seeds progressively lost their ability to germinate during 9 weeks of storage under the above conditions. The deleterious effects of temperature treatments increased with growing seed moisture content at higher humidity. The loss of seed viability was correlated with the generation of ROS during storage, which was more intensive at higher temperatures and humidity levels. The ascorbate content significantly increased in seeds stored in all temperature and humidity variants, when the seeds lost the ability to germinate to a large degree. At the same time, glutathione content dramatically decreased, but it was possible to observe a defensive reaction in seeds stored at 20 degrees C. Activities of all scavenging enzymes, measured after slow imbibition of seeds, significantly increased in comparison to the non-treated control (8-9% MC, -10 degrees C). This increase was higher in embryo axes than in cotyledons. Our results suggest that the loss of viability of beech seeds during storage at different temperatures, above zero, and at different humidity levels is closely related to ROS production, and that the antioxidative system is not sufficient to protect them.     

Dormancy in bulbils of several herbaceous plants: Effects of photoperiod,light, temperature,oxygen and gibberellic acid

Effects of some environmental conditions (photoperiod, white and colored lights, temperature, partial oxygen pressure) and growth regulators (gibberellic acid, 2-chloroethyltrimethylammonium chloride) on induction and release of dormancy of the bulbils ofDioscorea batalas, Laportea bulbifera, Elatostema involucratum andSedum bulbiferum were investigated. Bulbils were formed under short-day conditions inLaportea andElatostema, under long-day conditions inSedum, and irrespective of photoperiods inDioscorea. In all species exceptSedum, immature bulbils required light, particularly blue or far red, for sprouting (photo-sprouting stage), and mature bulbils required a cold treatment (thermo-sprouting stage). The duration of photo-sprouting and thermo-sprouting stages and the degree of dependency on light or low temperature of sprouting differed from species to species. Sprouting of chilled mature bulbils of these species was promoted by light, especially by red or green light. Both immature and mature bulbils ofSedum sprouted under short-day conditions. Continuous irradiation with blue, far-red and green light markedly inhibited their sprouting. Oxygen at high concentration inhibited the sprouting of immature bulbils inDioscorea; in the other species it promoted sprouting regardless of the maturation of the bulbils. Applications of gibberellic acid caused the sprouting of bulbils the absence of light, chilling or photoperiodic treatment in all species exceptDioscorea, in which gibberellic acid inhibited sprouting. Polyphenol oxidase activity was very high in the homogenates ofDioscorea bulbils, and increased further when the bulbils had been treated with gibberellic acid. In the other species, little or no such activity was observed.     

Externalization of membrane-bound activities during sheep reticulocyte maturation is temperature and ATP dependent

During the maturation of sheep reticulocytes in vitro, there is release of material that can be pelleted from the cell-free incubation medium by centrifugation at 100,000 X g. This pellet contains activities that are derived from both the plasma membrane and lysosomes. No evidence was obtained for the presence of mitochondrial activities or cytosolic enzyme activities. The release of these activities is ATP and temperature dependent, since reduction of either results in a greater retention of the activities by the cells and a lesser amount in the 100,000 X g pellet. The pelleted material is vesicular in nature, and the production and (or) release of the material are reduced upon ATP depletion or lowering of the temperature. It is concluded that the externalization of specific membrane components is a normal metabolic process that occurs during reticulocyte maturation and represents a means by which reticulocytes shed specific types of membrane-associated functions that are known to decrease during reticulocytes maturation.     

Oxylipin profile and antioxidant status of potato tubers during extended storage at room temperature

Potato tubers (cv. Bintje) (Solanum tuberosum L.) were stored under extreme conditions at 20 °C for 350 days without sprout inhibitors in order to assess whether aging- and/or senescence-related processes occurred. Under these extreme storage conditions, multiple sprouting followed by the formation of daughter tubers occurs. At the same time, an increase in respiration intensity, as evidenced by cytochrome c oxidase activity (E.C. 1.9.3.1), is observed, leading to a potential increase in reactive oxygen species (ROS) production. As polyunsaturated fatty acids are priority targets of oxidative attacks, the damage to lipids was assessed by oxylipin profiling in both free and esterified forms. Oxylipin profiling showed a predominance of linoleic acid-derived oxylipins and of 9-hydroxy and 9-hydroperoxy fatty acids in both free and esterified forms. No significant accumulation of individual oxylipin was observed 350 days after harvest. To further understand the absence of lipid breakdown products accumulation, the main enzymatic and non-enzymatic antioxidants were assessed. Antioxidant enzyme activities [superoxide dismutase (E.C. 1.15.1.1), catalase (E.C. 1.11.1.6.), ascorbate peroxidase (E.C. 1.11.1.11)] were enhanced during the advanced phase of aging. The main non-enzymatic antioxidant compound, ascorbate, decreased markedly in the early stages of storage, followed by a slower decline. Total radical scavenging activity was also maintained at the end of the storage period. Our results indicate that the enhanced aging process occurring during storage at room temperature does not seem to be associated with the changes classically encountered during leaf senescence or seed aging and that the observed degenerative processes do not surpass the protective potential of the tubers.     

Effects of seed maturation time and dry storage on light and temperature requirements during germination in invasive Prosopis juliflora

The effects of time of seed maturation and dry seed storage and of light and temperature requirements during seed incubation on final germination percentage and germination rate were assessed for the invasive shrub Prosopis juliflora (Sw.) D.C., grown under desert environmental conditions of the United Arab Emirates (UAE). Seeds were collected from Fujira on the northern coast of the UAE at different times during the growing seasons (autumn, winter and spring) and were germinated immediately and after 8 months of dry storage under room temperature (20±3 °C). Seeds were germinated at three temperatures (15, 25 and 40 °C) in both continuous light and darkness. The results showed significant effects for time of seed collection, seed storage, light and temperature of seed incubation and many of their interactions on both germination percentage and rate. Fresh seeds matured during autumn and winter germinated significantly greater at 40 °C and in light than at lower temperatures and in dark. Storage significantly increased germination percentage and rate; the increase was greater for seeds matured during winter than for seeds matured during spring. This indicates that dormancy breakage was greater in seeds of winter than seeds of spring. The need for high temperature to achieve greater germination was significantly reduced after seed storage, especially for seeds matured in autumn and winter.     

Effects of storage temperature on viability, germination and antioxidant metabolism in Ginkgo biloba L. seeds.

The behaviour of the Ginkgo biloba L. seeds was studied during storage at 4 and 25 degrees C. When stored at 25 degrees C, all the seeds died in 6 months. Cold temperatures preserved seed tissue viability for 1 year but did not preserve their capability to germinate, since such capability decreased after 6 months. A significant increase in lipid peroxidation occurred in the seed both in the embryo and in the endosperm. During storage a progressive deterioration of the endosperm tissues was evident. The two major water soluble antioxidants, ascorbate (ASC) and glutathione (GSH), showed different behaviour in the two conditions of storage and in the two main structures of the seed, the embryo and the endosperm. The ASC content of embryos and endosperms remained quite unchanged in the first 9 months at 4 degrees C, then increased. At 25 degrees C a significant decrease in the ASC content in the embryos was evident, whereas it remained more stable in the endosperm. The GSH pool decreased at both storage temperatures in the embryos. As far as the ASC-GSH redox enzymes are concerned, their activities decreased with storage, but changes appeared to be time-dependent more than temperature-dependent, with the exception of the endosperm ascorbate free radical (AFR) reductase (EC 1.6.5.4), the activity of which rapidly decreased at 25 degrees C. Therefore overall the antioxidant enzymes were scarcely regulated and unable to counteract oxidative stress occurring during the long-term storage.     

Endoreduplication in cucumber (Cucumis sativus) seeds during development, after processing and storage, and during germination

Flow cytometry was used to study endoreduplication in developing, stored and germinating seeds of cucumber ( Cucumis sativus ). Fruits growing in a commercial seed production field were collected every 7 days, starting 14 days after pollination (DAP) up to 63 DAP (commercial harvest time). Seeds were isolated and the proportion of nuclei with different DNA contents in the whole seeds and in the embryos was analysed. Germination capacity of fresh and dried seeds at 25°C was established. In addition, the same analyses were performed on the seeds after processing (fermentation, drying and cleaning), following 1 and 2 years of storage, and after imbibition for 3, 6 and 12 h. In the young developing seeds, endoreduplication up to 128C occurred but this decreased to 8C by maturity. The proportion of endosperm nuclei was the highest at 21 DAP (30%) and then decreased to below 14% at harvest and 8% after processing. In the mature processed seeds, the majority of embryo nuclei (about 80%) contained 2C DNA; however, about 2% of endoreduplicated (8C) nuclei were still present. Seeds did not show any germination capacity up to 21 DAP; then it gradually increased to reach 100% as early as 49 DAP, 2 weeks before commercial harvest time. The relationship between seed maturity, germination and cell cycle status is discussed. The mean C-value of the seed cells as well as the (4C + 8C + 16C)/2C ratio are recommended as markers of cucumber seed maturity and the advancement of germination/priming (the stage of germination sensu stricto ).