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.     

Priming bitter gourd seeds with selenium solution enhances germinability and antioxidative responses under sub-optimal temperature

Priming offers an effective means for counteracting sub-optimal temperature induced oxidative injury and raising seed performance in many crop species. In Taiwan, bitter gourd is frequently seeded when ambient temperatures are below the optimum. The objectives of the present study were to evaluate the effect of priming on germinability and antioxidative capacity in two bitter gourd cultivars germinated under 20 and 25°C conditions. Sodium selenite was also added to the priming solution. Priming was achieved by moistening the seeds with 1, 2, 5, or 10 mg l−1 sodium selenite solutions on germination paper at 25°C for 48 h and then air-drying them to their original moisture levels. A sub-optimal temperature of 20°C decreased seed performance by enhancing peroxidative injury. The sub-optimal temperature germinability was partially restored by priming the seeds with selenium solutions up to the 2 mg l−1 level. The improved seed performances were linked to the priming-enhanced free radical and peroxide-scavenging activities linked to the ascorbate-glutathione cycle. However, glutathione peroxidase was the only enzyme showing positive response with increasing selenium level up to 10 mg l−1, suggesting that this enzyme is selenium-inducible. The changes in germination and antioxidative activities in relation to selenium priming and sub-optimal temperature were similar for the two cultivars, despite differences in their germination performance and related antioxidative traits.     

Changes in oxidative stress enzymes during artificial ageing in cotton (Gossypium hirsutum L.) seeds

The present study was carried out to elucidate the mechanism of seed deterioration in two cotton (Gossypium hirsutum L.) cultivars (HS6 and H1098). The seeds were artificially aged at 40 +/- 1 degree C and 100% relative humidity for 4 days. In both cultivars, germinability decreased, whereas membrane deterioration, as assayed by electrical conductivity of the seed leachates, increased progressively with artificial ageing. The decrease in germinability was well correlated with increased accumulation of total peroxide and malondialdehyde content and decreased activities of antioxidant enzymes peroxidase, catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase. Hydropriming for 2 h and ascorbic acid priming for 12 h partially maintained germination and the activities of various antioxidant enzymes under artificial ageing and the accumulation of peroxide and MDA content was decreased. The results suggest that cotton seed deterioration during accelerated ageing is closely related to a decrease in activities of various peroxide scavenging enzymes and to lipid peroxidation.     

Prolonged aerated hydration for improvement of seed quality in Brassica oleracea L.

Seeds of cauliflower cv. Hipop and Brussels sprouts cv. Asmer Aries were aged at 20% moisture content for 24 h; all seeds retained a germination of over 70% after ageing although the mean germination time increased. Prolonged aerated hydration for up to 32 h at 20°C followed drying resulted in improved performance of both unaged and aged cauliflower seeds and aged Brussels sprouts. Thus, all seed showed reductions in the mean germination time to the extent that after 32 h hydration the aged cauliflower seeds performed as well as high quality unaged seed. The improvement of aged seeds was also revealed an increase in germination after the controlled deterioration test following up to 24 h (cauliflower) or 32 h (Brussels sprouts) aerated hydration. This increase was indicative of a decrease in the extent of deterioration present after aerated hydration. Deleterious effects of prolonged hydration were observed in Brussels sprouts after 32 h although these may be explained desiccation injury after treatment since radicle emergence had occurred during hydration. The improvements in seed performance may be explained the activation of metabolic repair occurring during the early part of the hydration period therereducing the extent of deterioration that has been sustained during ageing, with further improvements due to the advancement of the germination process.     

Increasing low-temperature resistance of soybean,Glycine max (L.) merr., by exposure of seeds to water saturated atmosphere

Hydration level of seeds determines vigour of soybean,Glycine max (L.) Merr. The seeds should contain at least 20% of moisture for a good performance at low temperatures. This hydration level can be reached by pre-sowing exposure of seeds in water saturated atmosphere (WSA). The exposure decreases both visible symptoms of chilling injury and leaching electrolytes during imbibition, and increases both germination and growth of seedlings at 10 to 12 °C. It is concluded that the cell membrane integrity is re-established in a course of the WSA exposure.     

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.     

Function of the ascorbate-glutathione cycle in aged sunflower seeds

The function of the ascorbate-glutathione (AsA/GSH) cycle was analyzed in seeds of sunflower ( Helianthus annuus L. cv. Peredovik) subjected to accelerated ageing at 43°C and 75% relative humidity for 1 to 11 days. The study was performed using dry seeds and seeds hydrated by imbibition in distilled water for 4 h at 25 °C. Lipid peroxidation was also determined by measuring the malondialdehyde (MDA) level. As the ageing period increased, a progressive loss of seed viability became increasingly evident. Even though high levels of MDA were delected, the MDA level did not change during accelerated ageing, suggesting that lipid peroxidation might occur to some extent. The study of the ascorbate/glutathione (AsA/GSH) cycle revealed that the GSH system is the major detoxifying mechanism in both dry and imbibed sunflower seeds. The GSH system is mainly located in the embryo, and its protective role is mediated by reactions that consume the GSH pool and, thereby, minimize the increase of the oxidized form (GSSG). Seed imbibition activates cellular metabolism and allows some antioxidant enzymes like glutathione reductase (EC 1,6,4,2) to act upon toxic agents. These reactions provide a reducing status, so that repair of damage becomes possible. However, prolonged ageing conditions (11 days) result in an irreversible damage, as evidenced by the appearance of dead seeds when the germination period ended. Multiple regression analysis revealed the effectiveness of the GSH system in aged seeds, especially upon imbibition and until the AsA/GSH cycle became completely functional.     

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.     

Ageing-induced changes in glutathione system of sunflower seeds

The glutathione system is thought to be involved in defence mechanisms present in plant tissues. The efficacy of this system was evaluated in large seeds of sunflower ( Helianthus annuus L. cv. Peredovik) in response to accelerated ageing (43°C/75% relative humidity from 1 to 11 days). Differences between the embryo axis and cotyledons in relation to the glutathione system were also investigated. Additionally, lipid peroxidation was determined by measuring the malondialdehyde (MDA) content. All assays were performed using dry seeds and seeds subsequently hydrated by imbibition in distilled water for 12 h at 25°C. Accelerated ageing caused a marked decrease in seed viability, accompanied by an increase in mean germination time. There were no changes in total glutathione in dry seeds. However, the distribution in its reduced (GSH) and oxidized (GSSG) forms revealed that ageing produced a slow conversion from GSH to GSSG. As the ageing period increased, this effect was accompanied by a decrease in glutathione reductase (GR, EC 1.6.4.2) activity. The results also indicated that the GSH system exerts a different response in the embryo axis as compared with the cotyledon: (1) the GSH levels decreased less in the cotyledons than in axes of aged seeds, and (2) the GSSG level in cotyledons was independent of ageing, while its amount increased in aged embryo axes. These different responses, in conjunction with the lower MDA levels in large as compared with small seeds, indicate a possible protective role of the reserve lipids. The efficacy of the GSH system in aged seeds was associated with seed viability, as revealed by multiple regression analysis. Upon imbibition, aged seeds were able to restore their GSH levels, reaching values approximating those of unaged seeds.     

Reversal of the effects of aging in soybean seeds

Accelerated aging predisposed seeds to imbibition injury. Slowing the rate of hydration prevented the loss of germinability due to imbibition injury. Germinability of accelerated aged seeds (50 hours) was increased from 10 to 90% by controlling the rate of imbibition. Slow hydration also prevented seed electrolyte leakage. This may indicate that cell membrane permeability or rupture was a major factor contributing to the loss of germinability after aging.

Reversal of the effects of aging (repair) was accomplished by slowly inbibing and then redrying seeds (priming). This treatment lowered steep water conductivity by a factor of 2 to 5. Priming also increased the per cent germination of low vigor seeds. The mechanism of this reversal was probably metabolic because it depended on temperature, seed moisture, and treatment duration.

Priming doubled the survival of seeds in the accelerated aging vigor test. The `rejuvenation' was accepted as evidence for metabolic repair. Since the `vigor' of seeds was increased by priming, metabolic repair probably included other subcellular components as well as the plasma membrane.

     

Germination Responses of Taraxacum platycarpum Seeds to Temperature

Abstract Germination responses of Taraxacum platycarpum seeds to temperature were examined under laboratory conditions to investigate the emergence-season choice mechanism of the seeds. Almost all the newly collected seeds were non-dormant. Under constant temperature conditions, maximum percentage germination (approximately 90%) was attained at temperatures 6–16°C, where simple linear relationships were observed between the temperature and the rates of germination, i.e. the reciprocals of the time taken to germinate by seed subpopulations with 10–80% germination. Thermal time required for germination of the subpopulations ranged from 600 Kh (degree Kelvin × hours) to 1500 Kh with a relatively constant base temperature of about 2.5°C. Lower limit temperature for germination was slightly below 6°C. Higher limit temperature for germination has the normal distribution with the mean ±SD of 19±2.5°C. Pre-exposure of imbibed seeds to temperatures higher than the higher limit temperature for germination, 25 and 30°C, had no effect on the germinability and the rate of germination at a circa-optimum temperature. Moist chilling treatment at 4°C caused an increase in the variation of germination rate within the seed population, but no evidence for dormancy-inducing or breaking effects was obtained.     

Osmoconditioning and Ageing of Pepper Seeds During Storage

The effects of osmoconditioning on the germination at 15 and25 °C of pepper (Capsicum annuum L.) seeds were studiedover a 3-year period with respect to temperature of storage.Untreated seeds stored at 5 °C showed high germinabilitythroughout the entire storage period, whereas untreated seedsstored at 25 °C showed a progressive decline in germinability,especially when assayed at 15 °C. Seeds that had been osmoconditionedprior to storage retained a high level of germinability irrespectiveof either storage or germination temperatures. When seeds thathad been stored at 25 °C were osmoconditioned after storage,there was a significantly higher germinability (assayed at 15 °C) in comparison with the corresponding untreated seeds.Seeds that were osmoconditioned twice (prior to and after storage)germinated in a similar way to those that had been osmoconditionedonce only Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

Correlation between seed longevity and RNA integrity in the embryos of rice seeds

The mature embryos of rice seeds contain translatable mRNAs required for the initial phase of germination. To clarify the relationship between seed longevity and RNA integrity in embryos, germinability and stability of embryonic RNAs were analyzed using the seeds of japonica rice cultivars subjected to controlled deterioration treatment (CDT) or long periods of storage. Degradation of RNA from embryos of a japonica rice cultivar “Nipponbare” was induced by CDT before the decline of the germination rate and we observed a positive relationship between seed germinability and integrity of embryonic RNAs. Moreover, this relationship was confirmed in the experiments using aged seeds from the “Nipponbare”, “Sasanishiki” and “Koshihikari” rice cultivars. In addition, the RNA integrity number (RIN) values, calculated using electrophoresis data and Agilent Bioanalyzer software, had a positive correlation with germinability (R²=0.75). Therefore, the stability of embryonic RNAs required for germination is involved in maintaining seed longevity over time and RIN values can serve as a quantitative indicator to evaluate germinability in rice.     

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.     

Ageing of tomato seeds involves glutathione oxidation

The effect of seed ageing on the oxidation of reduced glutathione (GSH) and the role of GSH oxidation in ageing-induced deterioration were studied in seeds of tomato ( Lycopersicon esculentum Mill. cv. Lerica, Moneymaker and Cromco). Both long-term storage at 15°C/30% relative humidity (RH) and artificial ageing at 20°C/75% RH, 30°C/45% RH and 60°C/45% RH resulted in a marked loss of GSH and a simultaneous, though not proportional, increase in its oxidized form GSSG. The glutathione thiol-disulfide status shifted towards a highly oxidized form, while the total glutathione pool decreased. The extent of GSH oxidation differed between ageing conditions and was not directly related to the extent of seed deterioration. Thiobarbituric acid-reactive substances did not increase in ageing tomato seeds, suggesting that lipid peroxidation did not take place. Hydration of seeds, either upon imbibition in water or by priming in an osmotic solution, resulted in a rapid decrease in GSSG, a shift of the glutathione redox couple to a mainly reduced status and an increase in the glutathione pool, in both control and aged seeds. The results indicate that, in tomato seeds, (1) seed ageing involves GSH oxidation into GSSG, which is indicative of oxidative stress, (2) ageing does not affect the GSSG reduction capacity upon subsequent imbibition, and (3) the lowered viability of aged seeds cannot directly be ascribed to the decreased GSH pool or To the highly oxidized glutathione redox status.     

Reduced mitochondrial and ascorbate–glutathione activity after artificial ageing in soybean seed

The effect of artificial ageing on the relationship between mitochondrial activities and the antioxidant system was studied in soybean seeds (Glycine max L. cv. Zhongdou No. 27). Ageing seeds for 18 d and 41 d at 40 °C reduced germination from 99% to 52% and 0%, respectively. In comparison to the control, malondialdehyde content and leachate conductivity in aged seeds increased and were associated with membrane damage. Transmission electron microscopy and Percoll density gradient centrifugation showed that aged seeds mainly contained poorly developed mitochondria in which respiration and marker enzymes activities were significantly reduced. Heavy mitochondria isolated from the interface of the 21% and 40% Percoll were analyzed. Mitochondrial antioxidant enzymes activities including superoxide dismutase, ascorbate peroxidase, glutathione reductase, monodehydroascorbate reductase, and dehydroascorbate reductase were significantly reduced in aged seeds. A decrease in total ascorbic acid (ASC) and glutathione (GSH) content as well as the reduced/oxidized ratio of ASC and GSH in mitochondria with prolonged ageing showed that artificial ageing reduced ASC–GSH cycle activity. These results suggested an elevated reactive oxygen species (ROS) level in the aged seeds, which was confirmed by measurements of superoxide radical and hydrogen peroxide levels. We conclude that mitochondrial dysfunction in artificially aged seeds is due to retarded mitochondrial and ASC-GSH cycle activity and elevated ROS accumulation.     

Seed Germination Traits of Loquat (<i>Eriobotrya japonica</i> Lindl.) as Affected by Various Pre-Sowing Treatments (Cutting of Cotyledons,Removal of Perisperm,Moist Chilling and/or Exogenous Application of Gibberellin)

The purpose of this study was to investigate the effects of various presowing treatments on the germinability (final germination percentage) and germination rate of loquat seeds in order to increase seedling production in nurseries (applied research) as well as provide answers for important physiological issues related to loquat seeds and their seed coat (basic research). Three experiments were carried out with various pre-sowing treatments. These treatments included full or partial removal of seed coat (perisperm), partial cutting of cotyledons as well as moist chilling at 5°C for 13 days and/or soaking the seeds in water or 250 ppm gibberellic acid (GA3) solution for 24 h. According to the results, cotyledons excision resulted in delayed germination, regardless of the presence or absence of the seed coat in comparison with the decoated seeds that demonstrated the highest germination rate amongst them. In addition, even the partial excision of seed coats affected positively both the germinability and the germination rate, compared to the control-intact seeds. Furthermore, control-intact seeds had a higher germination percentage when exposed to moist chilling independently of the application or not of gibberellin; while the combination of gibberellin application and moist chilling improved both the percentage and the rate of germination of decoated seeds. In conclusion, the role of perisperm (seed coat) in the germination procedure of loquat seeds seems to be important, indicating the existence of seed coat-imposed dormancy on loquat seeds. Finally, the existence of a mild endogenous embryo-dormancy on loquat is also discussed.     

The effects of phosphate supply on uptake of potassium ions, 2,4-D and atrazine by wheat and maize

The germination percentage of peach [ Prunus persica (L.) Batsch cv. Halford] seeds at 20°C was low (< 20%) after incubation at 5°C for as long as 35 days, but then increased considerably (> 40%) when the seeds were maintained at 5°C for longer than 42 days. Four zones of gibberellin-like activity were found in partially purified seed extracts. Gibberellin-like activity remained low in seeds incubated at 5°C for as long as 28 days, but increased significantly in three of these zones after 35 days, and in the fourth zone after 49 days. The increase in gibberellin-like activity was evident prior to the transfer of the seeds to 20°C. Moreover, seeds maintained at 5°C germinated at this temperature after 63 days. For seeds incubated and germinated at 20°C, both the germination percentage and the gibberellin-like activity remained low throughout the experimental period. Application of the growth retardant paclobutrazol to seeds after 28 days of a 49 day total incubation period at 5°C did not substantially reduce seed germination, although the increase in gibberellin-like activity was prevented. Seeds did, however, require a longer time to germinate after transfer to 20°C and were dwarfed in appearance. Application of GA₃ to seeds prior to stratification increased the percentage germination of seeds only when they had been incubated at 5°C for at least 35 days. The major changes in gibberellin-like activity are, therefore, associated not so much with the processes which allow germination to take place in peach, but more with those processes which allow normal growth and development of the seedling.     

The prevention of dehydration injury in celery seeds by polyethylene glycol, abscisic acid, dark and high temperature

Celery seeds ( Apium graveolens L.) given a germination induction period (3 days imbibition at 17°C in the light) could be prevented from germinating by up to 14 days subsequent exposure to high temperature (32°C), polyethylene glycol (PEG), abscisic acid (ABA) or dark (22°C). When the seeds were returned to 17°C in the light, germination occurred and, except for the high temperature treatment, was more rapid compared to seeds given a germination induction period only.
Celery seeds incubated for 3 days at 17°C in the light and then air-dried at 20°C germinated slowly when re-sown at 17°C in the light, and achieved only 19% germination after 21 days. Exposing the seeds to high temperature, PEG, ABA or dark for up to 14 days before drying maintained seed viability and subsequent germination was faster. The longer treatment periods gave increased benefit, and PEG was the most effective treatment. It is suggested that the effectiveness of the treatments in inducing dehydration tolerance relates to their ability to inhibit germination possibly via their prevention of cell expansion.     

The Maillard reaction and oxidative stress during aging of soybean seeds

The chemical reactions that may lead to the loss of seed viability were investigated both during the accelerated aging and natural aging of soybeans ( Glycine max Merrill cv. Chippewa 64). Under conditions of accelerated aging (36°C and 75% RH), fluorescence of soluble proteins accumulated, which was closely correlated with the loss of seed germinability and vigor. We were able to show this correlation by using partially purified proteins for the assay. Fluorescence also increased in seeds under good storage conditions (5°C for up to 21 years), although there was a less significant correlation between seed viability and the accumulation of fluorescent products during the time of natural aging. The rise in protein fluorescence is interpreted as an increase of Maillard products. The carbonyl content of soluble proteins (a measure of the oxidative damage) did not change significantly during either accelerated aging or natural aging: however the elimination of carbonyls during germination seemed to be hindered in seeds that had poor germination. The Maillard reaction may be a consequence of the formation of reducing sugars through a gradual hydrolysis of oligosaccharides during aging. Preliminary evidence from the natural aging study showed that, when seeds were in the glassy state, the sugar hydrolysis was inhibited. These results suggest that the Maillard reaction and oxidative reaction may play an important role in seed deterioration.