Lipid peroxidation and peroxide-scavenging enzymes associated with accelerated aging and hydration of watermelon seeds differing in ploidy

Hydration offers an effective means for raising seed performance in many crop species. The objective of this study was to evaluate the effect of vermiculite hydration on germinability and several physiological activities related to vigor in artificially aged watermelon seeds differing in ploidy. Aging was achieved by incubating the seeds at 45°C and 79% relative humidity for 6 days, then the seeds were air-dried to their original moisture level (4.7%). Hydration was achieved by mixing the untreated and aged seeds with moist vermiculite No. 3 at 25°C for 24 h. The partially hydrated seeds were air-dried at 25°C for 36 h to 4.7% moisture level. Significant differences existed between unaged and aged seeds, with lower germination percentage and slower germination speed in the latter. Aging also increased lipid peroxidation and reduced the activity of peroxide-scavenging enzymes. The germinability of aged watermelon seed was restored partially by vermiculite hydration. The activities of protein synthesis and peroxide-scavenging enzymes in axis and cotyledon portions of the seeds were also increased by hydration treatment. The changes in germination and related physiological responses in relation to aging and hydration are similar in seeds differing in ploidy, despite differences in their germination performance, seed leakage, extent of lipid peroxidation and activities of peroxide-scavenging enzymes.     

Can antioxidant’s reactive oxygen species (ROS) scavenging capacity contribute to aged seed recovery? Contrasting effect of melatonin,ascorbate and glutathione on germination ability of aged maize seeds

It is well known that antioxidants such as AA (reduced ascorbate), glutathione (GSH) (reduced glutathione) and melatonin can delay seed ageing. Can they recover aged seed? Artificial aged maize seeds were obtained and their reduced germination rate (GR) and high lipid peroxidation were recorded. Exogenous melatonin was applied on these aged seeds and enhanced GR was observed. However, treatment with other antioxidants such as AA, GSH or DMTU (dimethyl thiourea) did not significantly improve or even reduce the GR of aged seeds. In addition, melatonin improved germination ability of theses aged seeds can be significantly impaired by DDC (diethyldithiocarbamic acid, a specific inhibitor of superoxide dismutase or superoxide dismutase (SOD)) and ATZ (aminotriazol, a specific inhibitor of catalase or CAT). In a further study, we found that melatonin but not other antioxidants (AA, GSH and DMTU) significantly induced CAT and SOD activities of aged seeds after imbibition. Accordingly, melatonin significantly reduced lipid peroxidation in aged seeds than that of other antioxidants. Taken together, these data suggest that melatonin induced antioxidant enzyme but not its direct reactive oxygen species (ROS) scavenging capacity contributing to recovery of aged maize seeds.     

西瓜种子内源β-半乳甘露聚糖酶活性与分布以及与萌发速度的关系

利用单粒种子凝胶扩散法研究了β-半乳甘露聚糖酶在西瓜种子萌发过程中的分布以及与西瓜种子萌发速率的关系。结果发现,在胚根尖突破种皮前吸胀的西瓜种子中,内源β-半乳甘露聚糖酶主要分布于种子的胚膜套中,并起到减弱外种皮和胚膜套细胞壁对胚根伸出的机械阻力的作用。对具有不同萌发速率的品种以及引发处理和未处理的西瓜种子中酶活性的检测证明,β-半乳甘露聚糖酶活性与西瓜种子萌发速度相关。固体基质引发三倍体西瓜种子过程中β-半乳甘露聚糖酶的活化和种皮阻力的减弱,是引发种子提高了萌发速度和萌发能力的原因之一。     

Antioxidant role of glutathione associated with accelerated aging and hydration of triploid watermelon seeds

Hydration offers an effective means for raising seed performance in many crop species. The objective of the present study was to evaluate the effect of hydration in vermiculite on germinability and antioxidant capacity of glutathione in artificially aged triploid seeds of two watermelon cultivars. Aging was achieved by incubating the seeds at 45°C and 79% relative humidity for 6 days and then air-drying them to their original moisture level (4%). Hydration was achieved by mixing untreated and aged seeds with moist vermiculite No. 3 at 25°C for 24 h. The partially hydrated seeds were air-dried at 25°C for 36 h to 4.7% moisture level. Aged seeds germinated more slowly and poorly than unaged seeds. Aging also decreased the level of glutathione and reduced the activity of several enzymes involved in glutathione antiperoxidation. The germinability of aged triploid seeds was restored partially by hydration in vermiculite. The glutathione content and the activity of antiperoxidative enzymes using glutathione as substrate were also increased by hydration. The changes in germination and glutathione level in relation to aging and hydration were similar for the two cultivars, despite differences in their germination performance and related physiological traits.     

Ce^3+和La^3+对人工老化沙葱种子活力及生理特性的影响

以未老化和人工老化后的沙葱(Allium mongolicum Regel.)种子为材料,采用氯化铈(Ce3+)和氯化镧(La3+)浸种,测定种子萌发和生理指标,探讨Ce3+和La3+浸种对种子萌发、老化种子活力和生理特性的影响。结果显示:(1)在老化0~5 h时,Ce3+和La3+处理可显著促进沙葱种子萌发,提高种子活力;在老化5 h后,Ce3+和La3+处理对种子萌发无明显促进作用。(2)在老化0~15 h时,Ce3+和La3+处理的沙葱种子中抗氧化酶活性和抗坏血酸(AsA)含量提高,其超氧阴离子自由基(O2-·)产生速率、过氧化氢(H2O2)含量和丙二醛(MDA)含量显著降低;在老化15 h后,Ce3+和La3+处理的种子抗氧化酶活性提高、AsA含量降低,O2-·产生速率和MDA含量提高。(3)在老化5 h时,沙葱种子呼吸速率发生跃变达到最大,Ce3+和La3+处理显著降低了种子呼吸速率。(4)Ce3+和La3+处理在老化0~5 h时提高了沙葱种子超弱发光(UWL)强度,但在老化5 h后沙葱种子的UWL强度降低。研究认为,在沙葱种子人工老化初期,Ce3+和La3+浸种处理可以诱导增强种子抗氧化酶活性和提高AsA含量,有效清除因老化产生积累的过量活性氧(ROS),减轻过氧化伤害,提高种子活力;种子老化中后期,其内部ROS产生与清除系统发生紊乱,加剧了ROS对种子结构的损伤,Ce3+和La3+浸种处理的缓解效应丧失。     

Age-induced changes in cellular membranes of imbibed soybean seed axes

The physical and chemical properties of microsomal membranes and cellular antioxidant systems were investigated in imbibed soybean ( Glycine max L. Merr. cv. Maple Arrow) seeds following aging for 5 years at room temperature. The loss of germination capacity in aged seeds was associated with increased solute leakage during imbibition and with a loss of membrane phospholipid. Higher levels of free fatty acids were observed in the microsomal membranes from aged seeds. However, there was no change in fatty acid saturation. Wide angle X-ray diffraction studies indicated the presence of gel phase in addition to liquid-crystalline phase lipid domains in the membranes of aged seeds. Those from fresh seeds were exclusively liquid-crystalline. Fluorescence depolarization, using diphenylhexatriene, suggested that the microviscosity of the membrane bilayer was increased by aging. Aged seeds had a lower antioxidant potential in the lipid fraction, lower tocopherol content, and reduced ascorbate:dehydroxyascorbate ratio indicating that the aging process was associated with exposure to an oxidative stress.     

Free radical scavenging as affected by accelerated ageing and subsequent priming in sunflower seeds

Lipid peroxidation resulting from loss of free radical scavenging is thought to be involved in deterioration of sunflower (Helianthus annuus L.) seeds during accelerated ageing. In other respects, presoaking of seeds in a solution of low water potential (osmopriming) has been demonstrated to reinvigorate aged seeds. The aim of the present work was to study the effect of osmopriming on the germination of aged sunflower seeds and to investigate whether this effect was associated with the restoration of antioxidant defence systems. Seeds were aged for 5 days at 45°C and 100% relative humidity and then primed for various durations up to 7 days at 15°C in a solution of polyethylene glycol 6000 at ?2 MPa. Lipid peroxidation was estimated by measuring malondialdehyde (MDA) and conjugated diene contents, and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) were measured throughout the treatments. Accelerated ageing resulted in a marked decrease in the germination rate, and was associated with an increase in the levels of MDA and conjugated dienes, thus indicating lipid peroxidation. Ageing was also characterized by a decrease in the activities of CAT and GR. The activities of SOD and DHAR were much less altered. No APX activity was detected whatever the seed treatment. Priming of aged seeds progressively restored the initial germinative ability and resulted in a marked decrease in the levels of MDA and conjugated dienes, indicating a fall in lipid peroxidation processes. These effects of priming were also well correlated to the recovery of SOD, CAT and GR activities. Priming treatment for 7 days led to full restoration of the cell detoxifying mechanisms which were strongly altered during ageing. Glutathione content showed the same changes as GR activity. There existed a clear-cut relationship between seed germinative energy, expressed as the germination rate, and the efficiency of free radical scavenging systems, in particular CAT and GR activities and glutathione content. The results suggest that the antioxidant defence systems might play a key role in seed vigour.     

Lipid peroxidation and peroxide-scavenging in soybean seeds during aging

The possible role of lipid peroxidation in seed deterioration was investigated during natural aging and accelerated aging of seeds of edible soybean ( Glycine max [L], Merr. cv. Kaohsiung Selection No. 1). Natural aging was achieved by sealing the seeds in aluminum foil bags coated with polyethylene and storing the seeds at room temperature for 3 to 12 months. Accelerated aging was obtained by incubating the seeds at 45°C and close to 100% relative humidity for 3 to 12 days, after which the seeds were air dried to their original moisture level (8%). The results indicate that both natural and accelerated aging enhanced lipid peroxidation, as germination was depressed. Aging also inhibited the activity of superoxide dismutase, catalase, ascorbate peroxidase and peroxidase. The changes in germination and physiological activities, expressed as a function of aging duration, were somewhat similar in the two aging treatments.     

人工老化处理对结球甘蓝种子生理生化特性的影响

以结球甘蓝品种‘冬升’种子为材料,研究高温(40℃)高湿(相对湿度100%)人工老化处理过程中种子的萌发特性、种子浸出液相对电导率和丙二醛、可溶性蛋白、可溶性糖含量以及抗氧化酶活性的变化,以揭示种子劣变的机理。结果表明:(1)人工老化处理甘蓝种子的含水量和不正常苗率均随着老化时间的延长逐渐增加,而种子发芽势、发芽率、发芽指数和活力指数的增加均逐渐降低。(2)随着处理天数的延长,老化处理甘蓝种子的浸出液电导率显著增大,浸出液可溶性糖含量逐渐升高,可溶性蛋白含量表现出显著下降趋势,而种子MDA含量呈先升高后逐渐下降的趋势。(3)在结球甘蓝种子老化进程中,其种子中SOD、POD、CAT活性变化的趋势相似,均随老化程度的加深而逐渐降低;而APX活性在老化处理的最初2d显著增加,第6天显著降低。研究发现,在结球甘蓝种子老化进程中,种子活力和萌发率显著降低,其种子浸出液电导率、可溶性蛋白含量、可溶性糖含量、保护酶活性变化与种子老化及劣变程度密切相关,膜脂过氧化作用可能是引起或加剧种子老化劣变的重要原因之一。     

GA3浸种对大白杜鹃种子萌发的影响

用不同浓度的GA3浸种大白杜鹃种子并测定其萌发结果表明,不同浓度GA3处理均能增强大白杜鹃种子活力,极显著提高种子发芽率和发芽势,其中500mg·L～GA3浸种处理的效果最明显。GA3处理使种子的可溶性蛋白质含量降低,MDA含量下降,而SOD、POD和CAT活性增强。这表明GA3浸种后能增强抗氧化酶的活性,降低膜脂过氧化程度,从而促进种子提前萌发。     

Variation in temperature and light but not salinity invokes antioxidant enzyme activities in germinating seeds of Salsola drummondii

Seeds with efficient antioxidant defence system show higher germination under stress conditions; however, such information is limited for the halophyte seeds. We therefore studied lipid peroxidation and antioxidant responses of a leaf-succulent halophyte Salsola drummondii during seed germination under different salinity levels (0, 200 and 800 mM NaCl), temperature (10/20, 20/30 and 25/35°C) and light regimes. Seeds absorbed water and germinated in less than 1 h in non-saline control while increases in salinity decreased the rate of water uptake as well as seed germination. Non-optimal temperatures (10/20 and 25/35°C) and complete dark condition reduced seed germination in comparison to those seeds germinated under optimal temperature (20/30°C) and 12-h photoperiod, respectively. Generally, higher lipid peroxidation and antioxidant enzyme activities were observed in seeds at non-optimal temperature and in those seeds germinated in dark. Decrease in reduced ascorbic acid content was found in highest salinity and temperature treatments, while reduced glutathione content did not change significantly with changes in salinity, temperature and light regimes. These results indicate variation in temperature and light but not salinity enhances antioxidant enzyme activities in germinating seeds of Salsola drummondii.     

Changes in germinability,lipid peroxidation,and antioxidant enzyme activities in pear stock (<Emphasis Type="Italic">Pyrus betulaefolia</Emphasis> Bge.) seeds during room- and low-temperature storage

The aim of this study was to determine if loss of germinability in Pyrus betulaefolia seeds stored at 4°C and at room temperature is associated with a loss of membrane lipid peroxidation or changes in antioxidant enzyme activities. The results indicated that germination percentage clearly decreased when seeds were stored at room temperature rather than at 4°C from 6 to 12 months. Room-temperature storage of the pear stock seed for 12 months decreased germination to 15.52%, but germination percentage was not changed when seed was stored at 4°C for 12 months. MDA, a marker for membrane lipid peroxidation, increased significantly under room-temperature storage conditions. Antioxidant enzyme (SOD, POD, and CAT) activities were a good indicator of germination percentage in pear stock seeds. Antioxidant enzyme activities of pear stock seeds at 4°C were higher than antioxidant enzyme activities in seeds stored at room temperature from 6 to 12 months. Antioxidant enzyme activities of the pear stock seed decreased markedly under conditions of room-temperature storage from 6 to 12 months. The results of this study showed that long-term room-temperature storage was detrimental for maintaining the vigor of P. betulaefolia seeds. The mechanisms responsible for this outcome are a higher level of membrane lipid peroxidation and a lower level of activity of antioxidant enzymes.     

Changes in malondialdehyde content and in superoxide dismutase, catalase and glutathione reductase activities in sunflower seeds as related to deterioration during accelerated aging

Sunflower ( Helianthus annuus L.) seeds progressively lost their ability to germinate at 25°C, the optimal temperature for germination, after accelerated aging was carried out at 45°C (a temperature too high to permit germination) in water or at 76 or 100% relative humidity (RH). The deleterious effects of the high-temperature treatment increased with increasing seed moisture content. Incubation of seeds at 45°C in water resulted in electrolyte leakage, which indicated a loss of membrane integrity. A relationship between leakage and loss of seed viability could not be assumed, since no increase in electrolyte efflux occurred after aging al 100% RH. Accelerated aging induced accumulation of malondialdehyde, suggesting that seed deterioration was associated with lipid peroxidation. However, there was no direct relationship between lipid peroxidation and deterioration in membrane integrity. Loss of seed viability was also associated with a decrease in superoxide dismutase, catalase and glutathione reductase activities. Finally, the results obtained suggest that sunflower seed deterioration during accelerated aging is closely related to a decrease in the activities of detoxifying enzymes and to lipid peroxidation.     

Accelerated ageing and subsequent imbibition affect seed viability and the efficiency of antioxidant system in macaw palm seeds

Accelerated ageing is an accurate test indicator of seed vigor and storability that helps to understand the mechanisms of cellular and biochemical deterioration that occur during seed ageing. This study was carried out to elucidate the mechanisms of ageing in macaw palm embryos. Seeds were artificially aged during 4, 8 and 12 days at 45 °C and 100% relative humidity. After ageing, seeds were tested for viability (tetrazolium), electrical conductivity, lipid peroxidation (MDA) and hydrogen peroxide (H₂O₂) content. Part of the aged seeds was imbibed for 8 days and then determined the hydrogen peroxide content and the activity of antioxidant system enzymes (superoxide dismutase, catalase and glutathione reductase). Ageing reduced the embryo viability from 8 days of treatment and increased malondialdehyde content (MDA) and solute leakage. Hence, membrane permeability correlated with both loss of viability and lipid peroxidation. Imbibition after ageing significantly increased H₂O₂ content along with superoxide dismutase activity. Catalase activity was significantly higher than control in embryos aged from 8 days and imbibed, and glutathione reductase activity did not change. Our results suggest that macaw palm seed deterioration during accelerated ageing is closely related to lipid peroxidation, and that enzymatic antioxidant system is not completely efficient in reducing reactive oxygen species after imbibition, a critical phase to germination. Moreover, accelerated ageing test can be used as a reliable model to understand the mechanisms involved in palm seeds deterioration.     

Lipid peroxidation and peroxide-scavenging enzymes associated with accelerated aging of peanut seed

Accelerated aging is known to reduce seed viability and vigor in many crop species. The phenomenon is due in part to aging-induced lipid peroxidation, which has the potential to damage membranes of the seed tissues. This study was undertaken to evaluate the effect of accelerated aging on germinability and several physiological characteristics related to peroxidation in the seed of two peanut cultivars. Accelerated aging was achieved by incubating seed at 45°C and 79% relative humidity in a closed chamber for 3, 6, or 9 days. The results indicate that accelerated aging inhibited seed germination and seedling growth. Enhanced lipid peroxidation and increased peroxide accumulation were observed in the axis and cotyledons of aged seed. Accelerated aging also inhibited the activity of superoxide dismutase, peroxidase, ascorbate peroxidase, and lipoxygenase. Seed axes appeared to be more susceptible to aging than cotyledons. The changes in germination and physiological activities, expressed as a function of aging duration, were similar in the two cultivars, despite differences in their seed weight.     

Drought tolerance of Periploca sepium during seed germination: antioxidant defense and compatible solutes accumulation

Periploca sepium Bunge is a native and widespread shrub on the Loess Plateau, an arid and semi-arid region in China. To understand the adaptability of its seed germination to dry environments, we investigated the germination rate, water relations, lipid peroxidation, antioxidant capacity and accumulation of major organic solutes during seed germination under water deficit conditions. Results showed that seeds pre-treated by hydration–dehydration or ?0.9 MPa PEG germinated faster than control seeds, indicating strong resistance of P. sepium to drought condition. The re-dried seeds showed higher proline, total free amino acids (TFAA) and soluble proteins (SP) contents than control dry seeds, indicating the maintenance of physiological advancement when dehydrated. Osmotic stress made seed germination stay on the plateau phase (phase II). However, germinating seeds moved into phase III immediately once transferred into distilled water. Large increases in SP and soluble sugars (SS) of both re-dried and osmotic stressed seeds help themselves to resist drought stress. The re-hydrated seeds showed significantly higher levels of proline, TFAA, SP and SS than control seeds. The largely accumulated SS during osmotic stress declined sharply when transferred into distilled water. Our data demonstrate that P. sepium’s tolerance to drought stresses during germination is associated with enhanced activity of antioxidant enzymes and accumulation of some compatible solutes. Seed physiological advancement progressed slowly under low water conditions and it was maintained when seeds were air dried. This strategy ensures high and more rapid seed germination of P. sepium under drying and wetting conditions in drought-prone regions.     

Change in the reaction of the antioxidant system of wheat sprouts after UV-irradiation of seeds

The response of the antioxidant system of sprouts of wheat Triticum aestivum L. to preliminary irradiation of seeds with UV light was studied. The dependence of lipid peroxidation and the extent of antioxidant activity on the duration of irradiation was studied. It was shown that low doses of UV radiation (5-15 min) stimulate the antioxidant protection of green wheat sprouts grown for eight days. Increasing the irradiation time to 30-60 min leads to the inhibition of lipid peroxidation by the antioxidant system. A more prolonged irradiation of seeds with UV light (for 1-6 h) led to an increase in the level of lipid peroxidation in sprouts. However, 1-2-day-old sprouts from seeds irradiated for 5-6 h, adapted themselves to the influence due to the compensatory mechanisms. By the 8th day of germination of preliminarily irradiated seeds, the content of antioxidants and malone dialdehyde returned to the norm. The dynamics of activity of peroxidase in seeds irradiated with low doses of UV light for 30 min was studied. It was found that on the third day of seed germination, a decrease in peroxidase activity followed by its slight increase occurred. The maximum activity of the enzyme in the endosperm was observed on day 5-6, and in roots and green sprouts, on day 3-5 of germination. It was concluded that antioxidants and peroxidase are involved in the compensatory mechanisms of inhibition of free radicals formed upon UV irradiation of seeds.     

Germination and ROS detoxification in bell pepper (Capsicum annuum L.) under NaCl stress and treatment with microalgae extracts

We evaluated the salt tolerance of hybrids of pepper (Capsicum annuum L.) during germination. Treatments were applied at 0, 25, and 50 mM NaCl with preparations of supplemental extracts of the microalgae Dunaliella salina and Phaeodactylum tricornutum to determine the percentage germination rate as well as measured indicators of oxidative stress caused by the salt treatments during seed germination. We found that root growth was favorably influenced by the microalgae leading to increased germination rate. Tissues were analyzed in terms of superoxide radical production, lipid peroxidation, and activity of antioxidant enzymes viz. superoxide dismutase, catalase, and glutathione peroxidase. Our results suggest that application of microalgae extracts significantly reduced (p?<?0.05) superoxide radical production, as well as lower lipid peroxidation in comparison to plants without extracts of microalgae. The antioxidant enzymes increased in the presence of microalgae showing a significant difference (p?<?0.05). The results suggest differences in oxidative metabolism in response to the magnitude of salt stress and concentrations of microalgae help mitigate salt stress in plants during the germination process.     

链带藻源生物刺激剂提高小麦对盐胁迫的生理适应

盐胁迫是影响小麦萌发、生长和生产的最重要环境因素。探究链带藻（Desmodesmus Sp.）生物刺激剂对盐胁迫条件下小麦种子和早期幼苗抗盐、生长和生理的缓解效应以及最佳施用浓度,可为其应用于缓解小麦盐胁迫影响提供理论依据。【方法】通过室内培养皿培养法,将小麦种子置于100 mmol/L NaCl胁迫下,外源添加25,50,100,200 mg/L的链带藻提取物（DAE）,处理7 d后测量各项萌发和生长参数。【结果】外源添加DAE处理缓解了盐胁迫对小麦种子萌发和早期幼苗生长的抑制作用,提高了盐胁迫下小麦种子的萌发率和叶片含水量,促进了生物量的积累;提高了幼苗叶片超氧化物歧化酶、过氧化物酶、过氧化氢酶、抗坏血酸过氧化物酶活性以及脯氨酸、可溶性总糖、可溶性蛋白质和叶绿素的含量;降低了脂质过氧化作用,减少了丙二醛含量和膜透性。在100 mmol/L NaCl胁迫条件下,25 mg/L DAE对盐胁迫下小麦种子萌发及早期幼苗生长抑制作用的缓解效果最佳。【结论】链带藻细胞提取物通过促进小麦种子早期萌发的启动,提高小麦幼苗叶绿素含量、抗氧化酶活性和渗透调节能力,增强小麦种子及早期幼苗对盐胁迫的适应性,提升了小麦的耐盐能力。