The Effects of Priming and Accelerated Ageing upon the Nucleic Acid Content of Leek Seeds and their Embryos

Using seed priming and accelerated ageing techniques, a singlelot of leek (Allium porrum) seeds was manipulated to producefour lots of seeds with different germination performance. Changesin content of the major nucleic acid species in whole seedsand embryos of two of these lots (primed and unprimed), weredetermined over the early stages of germination. The major effectof priming was an increased level of RNA species in the seedsand embryos, and this difference was maintained during germination.Comparison of nucleic acid levels in the dry seeds of thesetwo lots and two others (aged and aged then primed) indicatedthat there was no correlation with germination performance.Similar comparisons of the nucleic acid levels in the embryosof seeds imbibed for 1 d showed only a limited correlation betweenrRNA levels and germination performance. Analysis of these datasuggests that accelerated ageing has an adverse effect uponendosperm cells, which results in the degradation of their nucleicacids during priming. Furthermore, the viability of these agedseeds also falls during priming. The data also indicate thatratios of rRNA to DNA correlate with germination performanceof the four lots of seeds studied. It is proposed that sucha relationship is indicative of the efficiency of a primingtreatment, and may be useful in comparisons of naturally varyingseed lots. Key words: Leek, seed, germination, priming, nucleic acids     

Inter-nucleosomal DNA fragmentation and loss of RNA integrity during seed ageing

The germination of viable seeds is the basis for new plant growth and development. Seeds lose viability during storage, but the biochemical mechanisms of seed death are not fully understood. This study aimed to investigate degradation patterns of nucleic acids during seed ageing and subsequent water uptake. Seeds of Pisum sativum L. were artificially aged at 50°C and 12% seed water content (WC). Nucleic acids degradation was studied during ageing and during imbibition of four seed lots with differential viability from highly viable to dead. As seeds lost viability during ageing, DNA was gradually degraded into internucleosomal fragments, resulting in ‘DNA laddering’, in conjunction with disintegration of 18S and 28S rRNA bands. During imbibition, non-aged controls had high levels of DNA and RNA integrity through to radicle protrusion. In an aged seed lot with 85% total germination (TG) DNA fragmentation decreased upon imbibition probably due to nucleosome degradation, while rRNA integrity did not improve. In an aged seed lot with 44% TG, neither DNA nor rRNA integrity improved upon imbibition. Dead seeds showed DNA degradation as laddering throughout imbibition along with extensive degradation of rRNA. We present a model in which interlinked programmed and non-programmed events contribute to seed ageing, and suggest that protection of nucleic acids during ageing is key to seed longevity.     

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.     

萝卜种子萌发的逆温耐性诱导研究

以萝卜种子为试材,研究不同温度预处理对萌发期萝卜种子逆温耐性的影响。结果表明:萝卜种子在低于15℃和高于35℃下萌发,种子活力显著下降,而通过适度的温度胁迫处理,可提高萝卜种子在高温（35℃）下的萌发活力。15℃和25℃分别是预处理的适宜低温和高温,6 h是低温（15℃）预处理的适宜时间。萝卜种子经高温（30℃）预处理12 h后,在随后的低温（10℃）下萌发,也提高其种子活力,说明萝卜种子对高温和低温胁迫具有交叉适应性。     

Invigoration of Cotton Seedlings by Treatment of Seeds for Pregermination Activities

Incubation of cotton seeds (Gossypium hirsutum, cv. Acala S.J.2) at 20 °C and 95% r.h. for up to 12 d results in an increasein seed water content and in a higher rate of seedling development.When seed vigour is measured by tolerance to high temperaturestorage (aging conditions) or tolerance to low temperature andsalinity during germination, seeds incubated for up to 4 d aremore vigorous than the control seeds. Hence, such a presowingtreatment of seed could be used for seed invigoration. As theincubation proceeds, although a higher growth rate of seedlingsis obtained at optimal conditions, the tolerance to stress conditionsdeclines proportional to the duration of incubation. Results of chemical and biochemical examinations of the incubatedseeds are compared with those of seeds during normal imbibitionfor germination. The early stage of germination (up to 4 h)is accompanied by a decrease in reducing sugars and amino acids.At the later stage, an increase of these components and a decreasein the lipid fraction occur. During the first 4 h, an increaseand decrease of phosphoenolpyruvate carboxylase and of malicdehydrogenase occur. According to the chemical and biochemicalmarkers, it is evident that seed treatment at 20 °C and95% r.h. results in pregermination biochemical activities. Thefirst 4 d seem to be equivalent to the first 3 h of germinationunder normal conditons. At this period of time, degradationprocesses are taking place. If incubation is continued the resynthesisstage appears to be retarded by the low moisture available andtherefore degradation processes continue, resulting in lossof seed vigour. The resynthesis ability may therefore be a prerequisitefor the following steps in seed germination.     

Thermoperiodism Mechanism in the Germination of Cocklebur Seeds

In thermoperiodic germination of secondarily dormant cocklebur(Xanthium pennsylvanicum Wallr.) seeds, the extent of electronflow through the CN-sensitive, cytochrome path and the CN-resistant,alternative path differred in the cool and warm phases. Thecool phase triggered active engagement of the alternative pathduring the subsequent warm phase, and this led to an increasein the ratio of the alternative path flux to the cytochromepath flux. The cool and warm phases had different functionsin adenylate metabolism. The former acted mainly in the productionand accumulation of ATP, whereas the latter supplied ADP and,especially, AMP. Thus an increasing number of sequential thermoperiodiccycles augmented the size of the adenylate pool and the energycharge, both of which may be necessary for germination to takeplace. (Received September 16, 1981; Accepted November 5, 1982)     

An evaluation of the potential of low temperature pre-sowing treatments of tomato seeds as a means of improving germination performance

Low temperature pre-sowing treatment of tomato seeds substantially enhanced germination rates and was much more effective at improving uniformity of germination than equivalent osmotic treatments. Although radicle emergence was inhibited by the low temperature, imbibition studies showed that seeds were maintained at lag-phase moisture contents during treatment and some aspects of germinative metabolism began. No evidence was found for improved seedling growth rates per se as a result of pretreatment: in fact, initial axis growth may be temporarily reduced, probably as a consequence of depletion of reserves during the treatment period.     

Advantageous and Detrimental Effects of Osmotic Pre-Sowing Treatment on the Germination Performance of Agrostemma githago Seeds

Dormant and after-ripened seeds of Agrostemma githago (corn-cockle)were pretreated in polyethylene glycol 400 (PEG) solutions attemperatures which would have allowed germination if the seedshad been imbibed in water, viz. 4?C or 20?C for after-ripenedseeds, and 4?C for dormant seeds. Pretreated seeds germinatedfaster than untreated seeds. The maximum decrease of the T₅₀(time to 50% germination) was 66%. Furthermore, pretreated seedswere capable of germination at supra-optimal temperatures whichotherwise had inhibited germination completely (20?C for dormantseeds and 30?C for after-ripened seeds). The percentage germinationat a supra-optimal temperature was considerably higher whenthe seeds had been primed at a temperature at which they developedmore extension power. The advantageous effects of the osmotic pretreatment were lessthan might be expected when the osmoticum had inhibited onlycell elongation. This was largely, if not fully, due to a generaldetrimental effect of osmotic stress and not to a selectiveinhibition of the processes which occur during the pregerminativephase in preparation for growth. Thus, during priming seedscomplete all or almost all processes which occur in water-imbibedseeds prior to radicle emergence. Key words: Agroatemma githago, dormancy, germination, germination performance, osmotic stress, priming     

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.     

Changes in Nucleic Acid Levels Associated with Improved Germination Performance of Tomato Seeds After Low Temperature Presowing Treatment

Low temperature presowing treatment (LTPST) of tomato seeds,var. Moneymaker, increases their rate of germination. Duringthis treatment there is a large increase in nucleic acid content,especially rRNA, within the seeds. Denaturing gel electrophoresisindicates that the quality of this RNA improves during LTPST.Although replacement of fragmented rRNA may be an importantprerequisite for successful germination, the data show thatthis is unlikely to be the immediate cause of more rapid seedgermination. When compared with untreated controls during subsequentgermination, treated seeds show reduced rates of nucleic acidaccumulation and reduced RNA polymerase activity per unit DNA,implying that rRNA synthesis within these seeds is under somemeasure of stringent control. The association between nucleicacid metabolism and germination is discussed. tomato, Lycopersicon esculentum Mill., seed germination, presowing treatment, RNA, RNA polymerase     

Light and temperature action in germination of seeds of the empress tree (Paulownia tomentosa)

Seeds of the empress tree ( Paulownia tomentosa Steud.) were imbibed for two weeks in darkness at constant temperatures (18, 23 or 28°C), and then irradiated with red light for 5 min. Germination was poor if it took place at the same temperature as imbibition, but a high percentage was achieved if the seeds were exposed to higher or lower temperatures before they were irradiated. Maximum germination was obtained when the difference between pretreatment and imbibition was about 10°C. The effect increased with the duration of the pretreatment and was optimal at 24 h. The effect decreased as the time lapse between temperature pretreatment and red light irradiation increased, and it was lost after two days. If pretreatment was shorter than 24 h (12 h). a high percent of germination was obtained by alternating pretreatment and imbibition temperatures. The germination of seeds imbibed in 40% heavy water was also stimulated by temperature pretreatments. Light and temperature also exhibited an interactive effect in the germination of seeds that were imbibed in darkness for only 3 days. For each of the germination phases there was a temperature at which the time needed for 50% germination was the shortest, namely 35°C during imbibition, 37.5°C in the period of P_fr activity. and 32.5°C during radicle protrusion. The data obtained are shortly discussed in relation to the domestication of empress tree in Southern Europe.     

The Characteristics of Low Temperature Tolerance during Seed Germination of the Ephemeral Plant Lepidium apetalum (Cruciferae)

Lepidium apetalum was used to explore the characteristics and mechanismes of low temperature tolerance during seed germination. The results showed that pre treatment the seeds at 4℃ would improve the seeds viability after stressed by low temperature at -5℃ and -10℃ and the capability of seeds tolerance to cold temperature became weaker along with germination development. Although the growth of radicle was inhibited significantly at 4℃ during germination, the seeds could still survive and grow. With appropriate time of stratified at 4℃, germination pace would be improved and germination rate would not change, while both of which decreased if stratification lasted too long. Moreover, Lapetalum seeds were very sensible to the temperature during germination. It showed no germination at 4℃, while if seeds were stratificated for 48 hours at 4℃ and subsequently treated with 25℃ for more than 6 hours, seeds would germinate at 4℃ and the germination rate increased with the time of 25℃ treatment prolonged. If treatment at 25℃ was less than 5 houes, it showed no germinating. In addition, seeds with stratification for 10 days following 1 h treatment at 25℃ could also germinated at 4℃. The present study indicated that some pivotal genes that related with a key physiological stage before the onset of radicle protrusion might inhibite at 4℃, which resulted in Lapetalum seed could not germinate at 4℃. Except that physiological stage, the Lapetalum seed had a strong tolerance to low temperature at other germination stages. This experiment provided a new way to explore the germination mechanism of ephemerals plants seeds at low temperature.     

短命植物独行菜种子萌发过程对低温的耐受特性

以独行菜（Lepidium apetalum）为材料,研究了其种子在萌发过程中耐受低温的特性,并对耐受低温的机制作了初步的探讨。结果表明：1.萌发至I、II、III期的独行菜种子经过冷诱导处理后,对零下低温-5℃、-10℃胁迫具有较强的耐受性,这种耐受性随萌发发展相对有所降低。2.独行菜种子萌发过程中胚根生长速度明显受到低温抑制,但也仍然能够生长。3.适当时间的低温层积能使独行菜萌发势显著提高,对终萌发率影响不大;过长时间的低温层积会使独行菜种子萌发势和最终萌发率降低。4.独行菜种子在4℃条件下不能萌发,但4℃层积2d的种子经25℃处理6h以上后,可耐受4℃低温而萌发,且萌发率随25℃处理时间增加而升高。如果25℃处理时间少于5h则不能在4℃低温下萌发。经4℃层积10d、再经25℃萌发处理1h的独行菜种子就能在4℃低温下萌发。综合分析认为独行菜种子不能耐受4℃低温萌发,原因可能是在露白前存在一个关键的生理阶段,在4℃胁迫逆境中不能越过这个阶段,该阶段之前与之后的萌发过程都能耐受4℃低温,因此对低温胁迫有良好的耐受性。这为探索早春短命植物耐受低温萌发的机制提供了新的思路。     

Changes during light and dark osmotic treatment independently modulating germination and ribonucleic acid synthesis in Chenopodium bonus-henricus seeds

The content and synthesis of RNA in Chenopodium bonus-henricus L. seeds in different physiological states were evaluated. A moist-chilling treatment at 4°C removed the primary dormancy of seeds. A pretreatment of chilled seed in low osmotic potential (–8.6 bars) polyethylene glycol-6000 solution at 15°C in light led to an advancement in subsequent germination time in water while a treatment in darkness induced a secondary dormancy. The synthesis of total RNA and poly A (+) RNA was correlated with the capacity of seeds to germinate. Cordycepin or α-amanitin failed to inhibit the germination of unchilled seeds, chilled seeds or chilled seeds given a prior light osmotic treatment. Also, germination of chilled seeds was not affected by cordycepin applied during light osmotic treatment. However, cordycepin effectively depressed the synthesis of both total RNA and poly A (+) RNA in chilled seeds with or without a prior light osmotic treatment. These data suggest that germination per se may depend upon the activation of pre-existing mRNA, which might be functionally different from the newly synthesized mRNA including poly A (+) RNA.     

Biochemical changes in Pinus pinea seeds during storing

The changes in the germination-rate, the contents in germination-inhibitors and the biochemical differences in soluble proteins and nucleic acids in freshly harvested Pinus pinea seeds stored for various periods of time, up to 24 months, and at two different temperatures (room temperature and 4 degrees C), have been investigated. The present results show that the maturation or after-ripening process of this type of seeds might be induced during the first 6 and 12 months of storage. However, seeds stored for longer periods of time might also be thought to enter into the primary phases of the ageing process where early alterations occur, including the loss of germination-rate and germination-inhibitor contents in the seed coat, together with an incapacity for the seeds to increase their protein and nucleic acid levels during the germination process.     

Appearance of Nitrate in Soybean Seedlings and Chlorella Caused by Nitrogen Starvation

Disinfected seeds of soybean and actively growing cultures ofChlorella vulgaris were grown on nitrogen-free media. The nitratecontent of both the soybean seedlings and algal cultures substantiallyincreased during nitrogen starvation. The nitrate level in soybeanseedlings was at least eight times greater than seeds aftertwo to three weeks. Nitrogen starvation also caused an increasein the nitrate content of the algal cultures. Nitrate reductaseactivity also increased, and its appearance was sensitive tocycloheximide. Tungstate, added during starvation, inhibitedthe induction of nitrate reductase with a concomitant increasein the level of nitrate. These data suggest that oxidation ofreduced nitrogen compounds can occur in higher plants and algae. ¹ A contribution of the Texas Agricultural Experiment Station (Received June 6, 1981; Accepted August 25, 1981)     

盐生植物角果碱蓬种子二型性对环境的适应策略

角果碱蓬(Suaeda corniculata)是藜科一年生盐生植物, 在我国分布于北方盐碱滩涂和盐碱荒漠地区。角果碱蓬具有棕色和黑色两种异型体种子(简称棕色和黑色种子)。对采自内蒙古鄂托克前旗盐渍化生境的角果碱蓬二型种子的形态、休眠和萌发特性开展对比研究, 测定了二型种子休眠和萌发行为对温度、光照和盐分(NaCl)的响应, 以揭示盐生植物异型种子对温带盐漠生境的适应对策。结果表明: (1)二型性种子在大小、种皮特性和结实比例方面有显著差异。与黑色种子相比, 棕色种子个体较大, 种皮透水性强。黑色种子与棕色种子的结实比例约为5.6 : 1。(2)新成熟的棕色种子的萌发对各温度梯度和光照条件不敏感, 萌发率较高(84%-100%); 而新成熟的黑色种子萌发率较低(8%-78%), 萌发对光照敏感。(3)黑色种子具有浅度生理休眠, 种皮划破、赤霉素处理和低温层积均可有效地提高种子的萌发率。(4)二型种子萌发对土壤盐分的胁迫具有不同的响应。与黑色种子相比, 棕色种子对盐分胁迫不敏感, 在较高的盐分浓度下仍有较高的萌发率, 低温层积处理能够降低黑色种子对盐胁迫的敏感性, 有效地提高种子的初始萌发率、萌发恢复率和最终萌发率。角果碱蓬二型种子不同的形态、休眠和萌发特性, 提高了该物种在高度异质性生境中的适合度, 对种群成功地适应温带盐漠环境具有重要的意义。     

Significance of inhibition of seed germination under the leaf canopy in ash coppice

Abstract. The effect on seed germination of the red/far-red ratio of light (ζ) as modified by a leaf canopy was studied. The artificial light sources used and daylight filtered through leaves appeared to have identical effects in tests with lettuce seed.
The sensitivity of the seeds to ζ varied with the environmental conditions during pretreatment and during the actual germination test. Freshly harvested and dry-stored seeds of Cirsium palustre were strongly inhibited at low ζ; after stratification, the inhibition was much less. The fact that C. palustre emerges mainly after the coppice is felled could not be explained from these results combined with field data on ζ and the response of germination to temperature.
The ecological significance of inhibition at low ζ is to be found directly after dispersal. Seed germination will then be delayed until the following spring or later.
Geum urbanum showed primary dormancy, which was released by a stratification treatment. No inhibition of germination at low ζ was found in the spring and seeds germinated in light and darkness at low temperatures. These results explain why the species emerges early in spring regardless of the presence of the ash canopy.     

Decrease in sunflower (Helianthus annuus) seed viability caused by high temperature as related to energy metabolism, membrane damage and lipid composition

The aim of the present work was to investigate whether loss of germination ability and viability of sunflower (Helianthus annuus L.) seeds during incubation at a high temperature (45°C) was related to changes in energy metabolism, loss of membrane integrity, and/or changes in lipid composition. Pre‐treatment of seeds at 45°C progressively reduced subsequent germination at the optimal temperature (25°C). Seeds did not germinate at 45°C and almost all of them were dead after 72 h of soaking at this high temperature. This loss of seed viability was associated with a large increase in leakage of K⁺ and total electrolytes into the incubation medium, and with production of malondialdehyde in the embryonic axis and cotyledons, suggesting a loss of membrane integrity probably due to lipid peroxidation. ATP and ADP levels increased sharply during the first hours of imbibition at 45°C, remained high for about 24 h and then decreased. As a consequence, the energy charge followed a similar pattern. If the treatment at 45°C did not exceed 48 h, seeds recovered an apparently normal energy metabolism after transfer to 25°C, even though they lost their ability to germinate at this temperature. Therefore, energy metabolism at the whole embryo level cannot be considered as an indicator of germination ability. Incubation of seeds at 45°C resulted in an increase in triacylglycerols and diacylglycerols without a significant change in their fatty acid composition. It also induced a slight increase in phospholipid content with an increase in C_16:0, C_18:0 and C_18:1, but with no change in C_18:2. In phospholipids, the C_18:2/C_18:1 and (C_18:1 + C_18:2)/ (C_16:0 + C_18:0) ratios thus declined during treatment at 45°C. The results obtained suggest that deterioration of sunflower seeds during incubation at a high temperature is mainly related to membrane damage and alteration of energy metabolism, and that accumulation of malondialdehyde, which is an index of lipid peroxidation, does not correspond to a decrease in total lipids and phospholipids nor to a significant change in fatty acid composition, except in PL in which the C_18:2/C_18:1 and (C_18:1 + C_18:2)/ (C_16:0 + C_18:0) ratios slightly declined.