Response of “storage” effects in barley seeds to the dose of ethyl methanesulphonate and X-ray treatment

1. Ethyl methanesulphonate (BMS)-treated barley seeds stored at 30 % water content for 1 – 4 weeks are capable of recovering from the mutagen induced injury. The level and speed of the recovery depends on the EMS dose and storage period. Seeds treated with extremely severe EMS doses reducing the% of germination to 5 – 10 % cannot recover from the induced injury. 2. Due to a 1 to 6 weeks storage of EMS-treated seeds with 20 % water content the amount of induced injury increases in dependence on the applied EMS dose and storage period. Doses of EMS, reducing the seedling height in the M₁ generation to 90 – 95 % of the control reveal no storage after-effects. 3. Storage of X-ray exposed seeds at 20 % or 30 % water content did not result in any significant changes in the yield of induced genetic damage.     

Activity of three oxidases and total dehydrogenases during the recovery from methyl methanesulphonate-induced mutagenic damage in barley

Methyl methanesulphonate (MMS), applied on barley seeds, inhibits the seed respiration and thein vitro activity of cytochrome-oxidase and catalase, whereas doubles the activity of total dehydrogenases and has no influence on the activity of peroxidase. Storage of MMS-treated seeds for 10 days at 30% seed water content, thus under conditions favouring the recovery from the MMS-induced toxic and genetic effects, resulted in an enhancement of the cytochrome oxidase and catalase activity, which reached or exceeded the activity of the control. The activity of peroxidase, although increasing during storage, did not attain the value of the control and the activity of total dehydrogenases remained two times higher as compared to the control.     

Pre-replication recovery from methyl methanesulphonate induced chromosomal damage in Vicia faba seeds

Vicia faba seeds were treated with methyl methanesulphonate (MMS) and stored at 50 % water content for 0, 14 and 28 d. This water content prolongs the period between the mutagenic treatment and the onset of DNA synthesis. Storage of seeds after mutagen treatment at the selected water content led to a significant decrease in DNA damage, manifested as a reduction in the frequency of chromosomal aberrations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Storage of ethyl methanesulphonate-treated barley seeds with low moisture contents

Barley seeds were treated with ethyl methanesulphonate (EMS) for 3 h at 25° C, washed with tap water for 24 h at 25° C, redried at 40° C to different moisture contents below 15% and stored at 25° C in desiccators or in sealed plastic bags. The criteria used for expressing the effect of storage were the M₁ seedling height and the frequency of chromosomal aberrations. With 14·9% seed moisture a strong increase of biological injury occurred in the course of a 2-week storage, while storage of seeds having an initial moisture content of 11·7% led to a significant increase of injury only after 6 weeks. Superdry EMS-treated seeds with 5% or less moisture can be stored at 25° C without any changes in the biological effects. A method is recommended to avoid the EMS-storage effects.     

Water and Seed Survival

Between about –350 and –14 MPa the rate of lossof viability in orthodox seeds is a positive function of waterpotential. The relative effect of water potential has been analysedin an oily seed (lettuce) and a non-oily seed (barley) and foundto be more or less identical. The lower limit for the relationin various species coincides with a seed moisture content (wetbasis) between about 2 and 6%. Below this level there is littleor no improvement in longevity with reduction in moisture content.The upper limit coincides with moisture contents of between15 and 28%, depending on whether the seeds are oily or non-oily.A water potential of about –14 MPa is the threshold forrespiration which increases more-or-less linearly with increasein water potential above this level. Above this threshold, andproviding oxygen is available to sustain respiration, seed longevityincreases with increase in water potential except that, unlessthe seeds are dormant, germination may be initiated at a waterpotential of about –1·5 to –0·5 MPa.In the absence of oxygen there may be a slight further declinein longevity with increase in water potential above –14MPa before longevity reaches a minimum value Since they cannot be dried very much without immediate lossof viability, recalcitrant seeds survive longest in the presenceof oxygen at maximum water potential commensurate with preventinggermination. The threshold water potential for immediate lossof viability has not been determined for most species but itis probable that it is close to the water potential typicalof the permanent wilting point in these plants, say –2MPa Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

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.     

Recovery from ethyl methanesulphonate-induced genetical damage in barley. Independence of sodium azide treatment

Barley seeds were treated for 3 h at 25°C with 240 mM ethyl methanesulphonate (EMS), washed for 18 h, treated with various concentrations of unbuffered sodium azide (pH 6.7–7.3) for 3 h at 25°C, re-dried to 30% water content and either sown immediately or stored at 25°C for 12 days and then sown. The synergistic action of sodium azide post-treatment has been demonstrated only for the EMS-induced M₁ germination reduction, while the EMS-induced M₁ sterility and the yield of M₂ chlorophyll mutants were unaffected. The ?storage” recovery from EMS-induced mutagenic effects was insensitive to sodium azide post-treatment. The 12 day-seed storage at 25°C brought about an improvement of M₁ germination, M₁ survival, M₁ fertility and a decrease in the amount of M₂ mutants, regardless of whether sodium azide post-treatment was applied or not.     

Control of Seed Respiration and Growth in Vicia faba by Oxygen and Temperature: No Evidence for an Oxygen Diffusion Barrier

The rate of dry matter accumulation by seeds of Vicia faba L. cv. Minica increases with temperature in the range of 16 to 26°C. The duration of dry matter accumulation decreases with temperature, resulting in a decrease of final seed dry weight. In this study we test the hypothesis that a diffusion barrier for O₂, located in the seed coat, inhibits seed respiration and growth. The rate of O₂ uptake of intact seeds and of excised embryos and seed coats (separated seeds) was measured in air and buffer at 16, 20, and/or 26°C at various O₂ concentrations and developmental stages. Oxygen uptake rates of intact seeds in buffer were only 9 to 15% of those in air. In buffer, the respiration rate of intact seeds decreased at a pO₂ below air saturation (21 kilopascals), whereas separated seeds showed a decline of O₂ uptake only below 80% of air saturation. In air, embryo excision had no effect on the sensitivity of seed respiration to pO₂, at both 20 and 26°C. In air at 20°C, separated and intact seeds showed similar rates of O₂ uptake. Oxygen uptake by intact seeds, both halfway and beyond the linear growth phase, showed a temperature coefficient Q₁₀ of 2.3 and was insensitive to pO₂ in the range of 80 to 100% of ambient. These results indicate that V. faba seed respiration in air is not limited by the diffusion of O₂ into the seed.     

A comparison of mutation induction by 14 MeV fast neutrons and 137 Cs -rays in meiotic spermatocytes in the silkworm

Barley seeds were treated with methyl methanesulphonate (MMS) and ethyl methanesulphonate (EMS), stored at 15% water content and washed for 16–24 h. These treatments resulted in an increase of toxic and genetic effects. In teh DNA of embryos of such stored MMS- and EMS-treated seeds, a strong enhancement of the amount of single-strand breaks and/or alkali-labile sites took place. In contrast, the amount of alkylated sites, particularly of 7-methylguanine, was somewhat lower. It can be that the depurination and/or backbone breakage, which proceeds during the storage period, is responsible for the enhancement of toxic and genetic effects, whereas the influence of the alkylation of DNA during the storage period by the unreacted residual mutagen is negligible.     

Glassy State and Seed Storage Stability: The WLF Kinetics of Seed Viability Loss at T>Tgand the Plasticization Effect of Water on Storage Stability

The relationship between the glassy state in seeds and storage stability was examined, using the glass transition curve and a seed viability database from previous experiments. Storage data for seeds at various water contents were studied by Williams–Landel–Ferry (WLF) kinetics, whereas the glass transition curves of seeds with different storage stability were analysed by the Gordon–Taylor equation in terms of the plasticization effect of water on seed storage stability. It was found that the critical temperatures (T_c) for long-term storage of three orthodox seeds were near or below their glass transition temperatures (T_g), indicating the requirement for the presence of the glassy state for long-term seed storage. The rate of seed viability loss was a function of T-T_gat T>T_g, which fitted the WLF equation well, suggesting that storage stability was associated with the glass transition, and that the effect of water content on seed storage was correlated with the plasticization effect of water on intracellular glasses. A preliminary examination suggested a possible link between the glass transition curve and seed storage stability. According to the determined WLF constants, intracellular glasses in seeds fell into the second class of amorphous systems as defined by Slade and Levine (Critical Reviews in Food Science and Nutrition30: 115–360, 1991). These results support the interpretation that the glassy state plays an important role in storage stability and should be a major consideration in optimizing storage conditions.     

Theoretical basis of protocols for seed storage

The protocols presently established for optimum seed storage do not account for the chemical composition of different seed species, the physiological status of the seed, and the physical status of water within the seed. The physiological status of seeds from five species with varying chemical compositions was determined by measurements of rates of oxygen uptake and seed deterioration. The physical status of water was determined by water sorption characteristics. For each species studied, there was a specific moisture content for the onset of respiration, chemical reactions, and accelerated aging rates. The moisture contents at which these physiological levels were observed varied among the species and correlated with the lipid content of the seed. However, the changes in physiological activities and the physical status of water occurred at specific relative humidities: 91% for the onset of respiration, 27% for the increased rates of thermal-chemical reactions, and 19% for optimum longevity. Based on these observations, we propose that equilibrating seeds between 19 and 27% relative humidity provides the optimum moisture level for maintaining seed longevity during longterm storage.     

The Survival of Germinating Orthodox Seeds after Desiccation and Hermetic Storage

Seeds of barley (Hordeum vulgare L.) and mung bean (Vigna radiata(L.) Wilczek), with orthodox seed storage behaviour, were imbibedfor between 8 h and 96 h at 15 °C and 25 °C, respectively,while barley seeds were also maintained in moist aerated storageat 15 °C for 14 d. These seeds and seedlings, together withcontrols, were then dried to various moisture contents between3% and 16% (wet basis) and hermetically stored for six monthsat —20°C, 0°C or 15°C. In both species, neitherdesiccation nor subsequent hermetic storage of the control lotsresulted in loss in viability. The results for barley seedsimbibed for 24 h were similar to the control, but desiccationsensitivity increased progressively with duration of imbibitionbeyond 24 h in barley or 8 h in mung bean; these treatmentsalso reduced the longevity of the surviving seeds in air-drystorage. Loss in viability in barley imbibed for 48 h was mostrapid at the two extreme seed storage moisture contents of 3·6%and 14·3%, and in both these cases was more rapid at15 °C than at cooler temperatures. Similarly, for mung beanimbibed for 8 h, loss in viability was most rapid at the lowest(4·3%) moisture content, but in this case it was morerapid at –20 °C than at warmer temperatures. Thus,these results for the storage of previously imbibed orthodoxseeds conform with the main features of intermediate seed storagebehaviour Key words: Barley, Hordeum vulgare L., mung bean, Vigna radiata (L.) Wilczek, desiccation sensitivity, seed longevity, seed storage behaviour     

Subject index

Barley seeds were treated with ethyl methanesulphonate (EMS) for 5 h, washed for 18 h, exposed for 3 h to caffeine, re-dried to 30% water content and either immediately sown or stored for 10 days at 25°. The caffeine post-treatment considerably potentiated the EMS-induced M₁ germination and the M₁ seedling height reduction, but had no influence on the EMS-induced M₁ sterility or percentage of M₂ chlorophyll mutants. The caffeine post-treatment also had no significant influence on the “storage” recovery from the EMS-induced genetic effects.     

The Effect of Low Temperature during Imbibition on Germination Characteristics of Tetracentron sinense (Tetracentraceae) Seeds

In this paper, the effects of low temperature, soil moisture and freezing time on the germination of Tetracentron sinense Oliv. seeds were studied by simulating wild environment conditions, and the factors influencing the natural regeneration of Tsinense population were discussed. The results are as follows: (1) The imbibition process of Tsinense seed can be divided into four stages, the water absorbing capacity of Tsinense seed after imbibition would increase with the increase of soil moisture, and the seeds would be fully imbibed under the condition of 30% soil moisture; (2) The germination rate, germination force and water content of Tsinense seed increased with the increase of storage time, and the vigor index gradually decreased after increasing before; (3) At different low temperature, the germination rate of Tsinense seed reached the highest when the soil moisture is 20% or 30%; (4) When treated at 20% soil moisture for 2-4 days or 21-28 days, 0℃ is best for the germination rate of Tsinense seed; when treated at 30% soil moisture for 14-28 days, -7℃ is best fort he germination rate of Tsinense seed; (5) When treated at the same condition of freezing time, the germination rate of seeds in fallen leaves had no significant difference with the decrease of low temperature; under the same condition of low temperature (0℃ low temperature except), the germination rate increased after frozen 28 days. The results showed that there is no significant influence of low temperature during imbibition on germination characteristics of Tetracentron sinense seeds and indicating that low temperature during imbibition is not the main factor influencing the natural regeneration of Tsinense population.     

Studies on the Peanut Seed Vigor Stored in Different Gases

Peanut seeds (water content 6.21% ) were stored at 38–40 ℃ for artificial ageing storage with different gases. N2 or CO2 delayed peanut seed ageing in comparison with air. The rate of seed germination was not affected by storing for 26 weeks under N2 or CO2, but the vitality of seed lost when stored under air. Determination of peanut seed vigor (root length+ axial length)×percentage of germination may precisely show the change in seed quality during ageing. The results indicate the positive correlation between the respiration and subsequent growth in peanut seeds, and the significantly negative correlation between vigor and electrical conductivity or sugar content in the seed extract.     

PEG预处理对青稞种子萌发和幼苗生理特性的影响

选择国内外28份青稞品种材料幼苗第一片展开叶,分别测定其相对含水量和失水率,并选择其中对水分胁迫敏感与不敏感的材料各1份,研究不同浓度(5%～30%)聚乙二醇(PEG)预处理对青稞种子萌发、幼苗生长和生理特性的影响,探讨短期水分胁迫对青稞生长发育的调节作用。结果显示:(1)28个青稞材料中‘旱地紫青稞’幼苗叶片的相对含水量最高(60.16%)、‘大麻青稞’最低(38.98%),而离体失水率‘旱地紫青稞’最低(8.80%)、‘大麻青稞’最高(20.20%)。说明‘大麻青稞’对水分胁迫最敏感,而‘旱地紫青稞’最不敏感。(2)随着胁迫程度的增加,青稞种子的萌发率和生根率,幼苗的根长、苗高和鲜重均呈先增加后降低的趋势,‘旱地紫青稞’和‘大麻青稞’种子分别在15%和10%PEG处理下发芽和生根最佳且均与对照差异显著(P<0.05),两品种的幼苗根长、苗高和鲜重均在10%PEG处理下表现最佳,但‘大麻青稞’与对照差异不显著。(3)‘旱地紫青稞’幼苗叶片可溶性蛋白和叶绿素含量随PEG处理浓度增加而逐渐增加,而丙二醛含量和相对电导率则逐渐降低,并在30%PEG处理下效果差异极显著(P<0.01);‘大麻青稞’叶片可溶性蛋白和叶绿素含量随PEG处理浓度增加呈先增加后降低的趋势,丙二醛含量和相对电导率呈先降低后增加的趋势,并在20%PEG处理下最佳。研究表明,短时间低浓度PEG处理对青稞种子萌发、幼苗生长及可溶性蛋白、叶绿素、丙二醛含量和相对电导率等生理指标的改善均有一定的促进作用;高浓度PEG处理却具有抑制作用,且高浓度PEG胁迫条件下,耐旱性强比耐旱性弱品种的自我调控能力更强。     

矮沙冬青种子特性和萌发影响因素的研究

 对矮沙冬青(Ammopiptanthus nanus)种子的特性和萌发影响因素进行了初步研究,结果表明：种子不易传播;虫蛀率高,室温贮藏60 d的种子虫害率为38%;易形成硬实,含水量为7.68%的种子在30 ℃温水中浸泡90 h,只有33.33%能吸水膨胀。种子萌发时不需光,在15～30 ℃和室温（18～32 ℃）条件下,经9 d的萌发,发芽率均可达80%以上,30℃时萌发最快;在1～2 cm深的沙壤中,种子出苗率可达75%以上,超过3 cm显著降低,超过6 cm则低于20%;种子在沙壤中萌发时,沙壤的适宜湿度为19.35%～28.75%,高于32.43%或低于3.85%,很少有种子萌发;含水量分别为19.36%、10.64% 和7.68%的种子发芽率无显著差异,在-10 ℃和5 ℃下贮藏7个月,发芽率也无显著降低,但在室温和35 ℃下贮藏7个月则显著下降,发芽率下降的速度与种子本身的含水量和贮藏温度正相关;在湿度分别为7.41%、13.79%和28.57%的沙壤中播种育苗,幼苗死亡率高达77.49%、81.25%和89.49%,即使用三唑酮拌种,死亡率亦高达50.27%、69.53%和76.03%,幼苗死亡率与沙壤湿度正相关。     

Influence of post-flowering temperature on seed development, and subsequent performance of crisp lettuce

Crisp lettuce plants cv. Saladin were grown from the time they started flowering, at 20/10°C (16 h day, 8 h night), 25/15°C and 30/20°C in glasshouses on two occasions in 1985. Yields of seed increased from, on average, 15 g to 27 g and then fell to 20 g per plant with progressive increases in temperature. The number of mature florets per plant increased with temperature but the number of seeds per mature floret was lower at 20/10°C and 30/20°C than at 25/15°C. An increase in temperature reduced mean seed weight by up to 45%, seed volume by 15%, cell numerical volume density (N_v) by 27% and the number of cells per seed by 39%. Percentage seed germination reached a maximum early in seed development at the stage when the pappus appeared through the involucral bracts. Differences in percentage germination and vigour of seeds (slope test) from different temperatures were accounted for largely by the effects on mean seed weight. However, when germinated at 30°C seeds produced at 30/20°C germinated more readily than those produced at 25/15°C or 20/10°C. Seed vigour gradually increased with an increase in the length of storage after harvest, reaching a maximum after 260 days. In general, seeds produced at 25/15°C exhibited a greater variation in numbers of seeds per floret, N_v, seed weight, times of seedling emergence, seedling and mature head weight than seeds produced at lower or higher temperatures.     

Involvement of ABA in induction of secondary dormancy in barley (Hordeum vulgare L.) seeds

At harvest, barley seeds are dormant because their germination is difficult above 20 degrees C. Incubation of primary dormant seeds at 30 degrees C, a temperature at which they do not germinate, results in a loss of their ability to germinate at 20 degrees C. This phenomenon which corresponds to an induction of a secondary dormancy is already observed after a pre-treatment at 30 degrees C as short as 4-6 h, and is optimal after 24-48 h. It is associated with maintenance of a high level of embryo ABA content during seed incubation at 30 degrees C, and after seed transfer at 20 degrees C, while ABA content decreases rapidly in embryos of primary dormant seeds placed directly at 20 degrees C. Induction of secondary dormancy also results in an increase in embryo responsiveness to ABA at 20 degrees C. Application of ABA during seed treatment at 30 degrees C has no significant additive effect on the further germination at 20 degrees C. In contrast, incubation of primary dormant seeds at 20 degrees C for 48 and 72 h in the presence of ABA inhibits further germination on water similarly to 24-48 h incubation at 30 degrees C. However fluridone, an inhibitor of ABA synthesis, applied during incubation of the grains at 30 degrees C has only a slight effect on ABA content and secondary dormancy. Expression of genes involved in ABA metabolism (HvABA8'OH-1, HvNCED1 and HvNCED2) was studied in relation to the expression of primary and secondary dormancies. The results presented suggest a specific role for HvNCED1 and HvNCED2 in regulation of ABA synthesis in secondary seed dormancy.     

Effects of Moisture Content on Germination of Seeds of Hancornia speciosa Gom. (Apocynaceae)

Seeds of Hancornia speciosa germinated best at a temperatureof 20–30 °C. The viability of the seeds during storagewas short and the best storage conditions for viability entailedkeeping the seeds in polyethylene bags. Seed viability was maintainedonly when the seeds were stored at a moisture content above30%; storage conditions which allowed dehydration resulted ina rapid loss of viability (the seeds showed recalcitrant behaviour). Low temperature during storage did not improve longevity. Arelationship between germination and moisture content was established,but when the moisture content fell below 25% there was a drasticreduction of germination. After 9 weeks of storage, even athigh moisture content, seeds lost viability. Loss of seed viability during seed dehydration was associatedwith increased leakage of electrolytes and organic solutes,and reduced tetrazolium staining during subsequent imbibition. Hancornia speciosa, germination, recalcitrant seeds, storage, moisture