Effects of gibberellic Acid and gold light on germination, enzyme activities, and amino Acid pool size in a dwarf strain of watermelon

Gibberellic acid (GA₃) promotes and continuous gold light inhibits germination of seeds of a dwarf strain (WB-2) of watermelon [Citrullus lanatus (Thunb.) Matsu. and Nakai]. Osmotic inhibition of germination with mannitol in light-grown seeds of WB-2 was only slightly reversed by GA₃ at the concentrations used, whereas, GA₃ substantially relieved osmotic inhibition in dark-grown seeds.

The effects of GA₃ and gold light on development of catalase and invertase activities and on levels of free amino acids in germinating seeds of WB-2 were examined. Light depressed development of catalase and invertase activity. Levels of free amino acids increased more slowly in embryonic axes of light- than dark-incubated seeds, but in cotyledons higher levels of amino acids were maintained in light-grown seeds. GA₃ accelerated the development of catalase activity in whole embryos and invertase activity in embryonic axes, but did not significantly affect invertase activity in cotyledons during germination. GA₃ had little effect on amino acid pools in cotyledons and embryonic axes.

     

Enzyme Profiles in Seedling Development and the Effect of Itaconate, an Isocitrate Lyase-directed Reagent

Changes in levels of isocitrate lyase, malate synthase, and catalase have been investigated during germination of flax (Linum usitatissimum L.) in the presence and absence of itaconate. Germination was accompanied by a rapid increase in these enzymes during the first 3 days. The presence of 38 millimolar itaconate inhibited the incidence of seed germination and the growth of embryo axes as well as the appearance of isocitrate lyase but did not alter the levels of malate synthase, catalase, or NADP⁺-isocitrate dehydrogenase. The specific activity for the latter enzyme was constant throughout germination. Oxalate or succinate, each at 38 millimolar, had no effect upon germination of flax seeds. Itaconate did not inhibit the activities of malate synthase, catalase, or NADP⁺-isocitrate dehydrogenase in vitro but was a potent noncompetitive inhibitor of isocitrate lyase (K_i:17 micromolar at 30 C, pH 7.6). Itaconate (at 38 millimolar) did not alter the appearance of malate synthase but reduced the incidence of germination, onset of germination, and growth of the embryo axis as well as the specific activity of isocitrate lyase in seedlings of Zea mays, Vigna glabra, Glycine hispida, Vigna sinensis, Trigonella foenumgraecum, Lens culinaris, and Medicago sativa. The incidence and onset of germination of wheat seeds were unaltered by the same concentration of itaconate but seedlings did not contain isocitrate lyase or malate synthase. The data suggest that itaconate may be isocitrate lyase-directed in inhibiting the germination of fatty seeds.     

Exogenous Diethyl Aminoethyl Hexanoate,a Plant Growth Regulator,Highly Improved the Salinity Tolerance of Important Medicinal Plant <Emphasis Type="Italic">Cassia obtusifolia</Emphasis> L.

The purpose of the present study was to investigate the mechanism of DA-6 in alleviating the salinity inhibition of Cassia obtusifolia L. seeds and seedlings. NaCl (100 mM) was used to mimic salinity stress in a series of experiments. Varying combinations of DA-6 were added to seeds and seedlings under salinity stress. Seed germination indices and seedling parameters were investigated. Seed germination and seedling growth were significantly inhibited under salinity stress. NaCl-induced inhibitory effects on seed germination and seedling growth were ameliorated by DA-6 with different concentrations. Addition of DA-6 to seeds (50 µM) and seedlings (100 µM) significantly alleviated damage to the plant cells under salinity stress. DA-6 (regardless of the presence or absence of NaCl) enhanced chlorophyll concentration, total soluble sugars, free proline, and soluble protein, and improved photosystem II (PSII) photochemical efficiency levels (F _v/F _m), PSII actual photochemical efficiency (ΦPSII), and the photochemical quench coefficient. In contrast, the initial fluorescence (F _o) and the non-photochemical quenching coefficient decreased. Application of DA-6 also enhanced the activities of superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POD; EC 1.11.1.7), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), and glutathione reductase (GR; EC 1.6.4.2), thus alleviating oxidative damage, as indicated by decreases in thiobarbituric acid-reactive substances, hydrogen peroxide concentration (H₂O₂), relative conductivity, and lipoxygenase activity (LOX; EC 1.13.11.12). Based on the experimental results, we conclude that DA-6 induces advantageous effects on the attenuation of salt-stress inhibition of C. obtusifolia seeds and seedlings and alleviates oxidative damage by conferring beneficial cytoprotection and activating antioxidant enzymes. DA-6 can be used as an effective plant growth regulator to alleviate salinity stress.     

Hydrogen peroxide scavenging regulates germination ability during wheat (Triticum aestivum L.) seed maturation

Hydrogen peroxide (H₂O₂) promotes seed germination of cereal plants and ascorbic acid which acts as antioxidant suppresses the germination of wheat seeds, but the role of H₂O₂ scavenging on germination during seed maturation has not been demonstrated. We investigated relationship of germination, ascorbate, H₂O₂ scavenging enzymes and sensitivity to ascorbic acid (AsA) maturing seeds of two typical wheat (Triticum aestivum L.) cultivars, cvs. Shirogane-Komugi and Norin61. Shirogane-Komugi had marked high germination ability than Norin61 during seed maturation. Although the H₂O₂ content had no difference in the two culti-vars, sensitivity to AsA of Norin61 seeds was higher than that of Shirogane-Komugi seeds during seed maturation. The sensitivity to AsA closely correlated with germination characteristic in the two cultivars. Especially, at 28 days after pollination (DAP), sensitivity to AsA in Norin61 seeds was remarkably high. At that stage, no significant differences were observed in endogenous AsA level, ascorbate peroxidase (APX, EC 1.11.1.11) and dehydroascorbate reductase (DHAR, EC 1.8.5.1) activities in the two cultivars. However, catalase (CAT, EC 1.11.1.6) activity and CAT mRNA in Norin61 were remarkably higher than in Shirogane-Komugi. Sensitivity to AsA at 35 and 42 DAPs kept high levels in Norin61, and endogenous AsA and CAT activity in the seeds were significantly higher than in Shirogane-Komugi. These results revealed a direct correlation between germination and antioxidant sensitivity during the developmental stages of wheat seeds.Key words: ascorbic acid, germination, hydorogen peroxide, maturation, wheat seed     

Reduced pyridine nucleotide-dependent N-hydroxy amine oxidase and reductase activities of hepatic microsomes

The metabolism of a number of primary and secondary hydroxylamines by hepatic microsomes is described. A cyanide-insensitive, reduced pyridine nucleotide-dependent hydroxylamine reductase activity that is independent of oxygen concentration catalyzes the reduction of hydroxylamine and a number of its mono- and disubstituted derivatives to the parent amines. At the pH optimum of 6.3 for the reductase, NADH is the preferred cofactor. The enzyme does not catalyze the reduction of 4-hydroxyaminoquinoline-1-oxide (HAQO) or of 1- or 2-naphthylhydroxylamine, the only known carcinogenic hydroxylamines tested. A hydroxylamine oxidase activity that requires both oxygen and reduced pyridine nucleotide oxidizes only disubstituted hydroxylamines, and the apparent initial product is the corresponding nitrone. Most nitrones undergo immediate hydrolysis in aqueous solution. At the pH optimum of 7.6 for the oxidase, NADPH is the preferred cofactor. NADPH cannot be replaced by a hydrogen peroxide-generating system, and the reaction is not affected by the addition of large amounts of exogenous catalase. Of the various organs which were assayed, the liver contained the greatest amount of both the reductase and oxidase activities; and the major portion of both activities in liver homogenates was found in the microsomal fraction. The two activities respond differently to agents such as deoxycholate, n-octylamine, and sulfhydryl inhibitors, indicating that the reduction and oxidation of the hydroxylamines are catalyzed by different enzymes or enzyme systems. Both activities are insensitive to carbon monoxide and N,N′-diphenyl-p-phenylenediamine (DPPD), an inhibitor of lipid peroxidation.     

Activities of Hydrogen Peroxide-Scavenging Enzymes in Germinating Wheat Seeds

During imbibition and germination of wheat (Triticum aestivum)in the dark over 72 h, activities of the enzymes of the ascorbate(AsA)-dependent H₂O₂-scavenging pathway, AsA peroxidase, monodehydroascorbate(MDAsA) reductase, dehydroascorbate (DHAsA) reductase and glutathione(GSSG) reductase as well as superoxide dismutase (SOD), catalaseand guaiacol peroxidase were determined both in whole grainsand in isolated embryos and endosperm. With the exception of DHAsA reductase, activities of the otherenzymes assayed increased in germinating seeds, especially duringradicle emergence (between 24–48 h of imbibition). Theseincreases, particularly for AsA peroxidase, were much higherin the embryo than in the endosperm. Within 72 h of imbibition,activities per seed increased 116-fold for AsA peroxidase, 19-foldfor guaiacol peroxidase, 5-fold for catalase and only 1·4-foldfor SOD. In contrast to the decreases in DHAsA reductase, theother AsA recycling enzyme, MDAsA reductase, increased 5-foldwithin 72 h. The results indicate that, in wheat seeds, imbibition and germinationis associated with enhanced cellular capacity to detoxify H₂O₂.For this detoxification the operation of AsA peroxidase togetherwith the AsA-regenerating enzymes appears to be of particularimportance. Key words: Ascorbate peroxidase, germination, hydrogen peroxide detoxification, inhibition, wheat     

Interactions of light and a temperature shift on seed germination

Germination of Rumex obtusifolius L. seeds is potentiated to an observable degree in 2 minutes by a single shift in temperature from 20 to 35 C. Half-maximal potentiation requires less than 32 minutes at the higher temperature. Similar sensitivities to shifts in temperature were observed for seeds of Barbarea vulgaris, R.Br. B. verna (Mill.) Asch., and Lepidium virginicum L. A shift in temperature interacts strongly with change in form of phytochrome induced by light on germination of the four kinds of seeds. The potentiated effects for R. obtusifolius are only moderately affected by 40 μm cycloheximide. Both the temperature shift and light actions are apparently independent of processes of synthesis necessary for growth.     

Aspects of nitrogen metabolism in the rice seedling

The effects of nitrogen source NO₃⁻ or NH₄⁺ on nitrogen metabolism during the first 2 weeks of germination of the rice seedling (Oryza sativa L., var. IR22) grown in nutrient solution containing 40 μg/ml N were studied. Total, soluble protein, and free amino N levels were higher in the NH₄⁺-grown seedling, particularly during the 1st week of germination. Asparagine accounted for most of the difference in free amino acid level, in both the root and the shoot. Nitrate and nitrite reductase activities were present mainly in the shoot and were higher in the NO₃⁻-grown seedling, whereas the activity of glutamate dehydrogenase and glutamine synthetase in the root tended to be lower than that of the NH₄⁺-grown seedling during the 1st week of germination. Glycolate oxidase and catalase activities were present mainly in the shoot. Maximum activity of the above five enzymes occurred 7 to 10 days after germination. Differences in the zymograms of nitrate reductase, glutamate dehydrogenase, and catalase were mainly between shoot and root and not from N source. Nitrite reductase bands were observed only in plants grown in plants grown in NO₃⁻.     

Nitrate and nitrate reductase in Erythrina caffra seeds: Enhancement of induction by anoxia and possible role in germination

The nitrate level in seed embryonic axes of Erythrina caffra Thunb. which is capable of anaerobic germination, was about 2.5 times higher than in seed axes of Pisum sativum L. a species incapable of anaerobic germination. Nitrate levels in E. caffra seeds decreased during germination and this was not due to leaching. Both NADH- and NADPH-dependent nitrate reductase (NAR) activities increased during germination. The increase was prevented by cycloheximide. The activity of NADH-NAR (EC 1.6.6.1) was higher than that of NADPH-NAR (EC 1.6.6.3). Both NAR activities were higher in anoxia than in air during germination. The NAR activities in Pisum seeds were very much lower than in Erythrina seeds. Anoxia (N₂ or argon) enhanced the induction of NAR by KNO₃ in germinated E. caffra axes. The NADH- and NADPH-NAR activities were induced to equally high levels by KNO₃ under anoxia. The enhancement was depressed by cycloheximide. It is concluded that nitrate and NAR activity may play a role in the anaerobic germination of E. caffra seeds.Abbreviation NAR nitrate reductase Financial support was obtained from the University of the Orange Free State and the Foundation for Research Development     

Exogenous 5-Aminolevulenic Acid Promotes Seed Germination in Elymus nutans against Oxidative Damage Induced by Cold Stress

The protective effects of 5-aminolevulenic acid (ALA) on germination of Elymus nutans Griseb. seeds under cold stress were investigated. Seeds of E. nutans (Damxung, DX and Zhengdao, ZD) were pre-soaked with various concentrations (0, 0.1, 0.5, 1, 5, 10 and 25 mg l⁻¹) of ALA for 24 h before germination under cold stress (5°C). Seeds of ZD were more susceptible to cold stress than DX seeds. Both seeds treated with ALA at low concentrations (0.1–1 mg l⁻¹) had higher final germination percentage (FGP) and dry weight at 5°C than non-ALA-treated seeds, whereas exposure to higher ALA concentrations (5–25 mg l⁻¹) brought about a dose dependent decrease. The highest FGP and dry weight of germinating seeds were obtained from seeds pre-soaked with 1 mg l⁻¹ ALA. After 5 d of cold stress, pretreatment with ALA provided significant protection against cold stress in the germinating seeds, significantly enhancing seed respiration rate and ATP synthesis. ALA pre-treatment also increased reduced glutathione (GSH), ascorbic acid (AsA), total glutathione, and total ascorbate concentrations, and the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), whereas decreased the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), and superoxide radical (O₂ ^•−) release in both germinating seeds under cold stress. In addition, application of ALA increased H⁺-ATPase activity and endogenous ALA concentration compared with cold stress alone. Results indicate that ALA considered as an endogenous plant growth regulator could effectively protect E. nutans seeds from cold-induced oxidative damage during germination without any adverse effect.     

The system modulating ROS content in germinating seeds of two Brazilian savanna tree species exposed to As and Zn

The effects of increasing arsenic (0, 10, 50, 100 mg L^?1) and zinc (0, 50, 80, 120, 200 mg L^?1) doses on germination and oxidative stress markers (H₂O₂, MDA, SOD, CAT, APX, and GR) were examined in two Brazilian savanna tree species (Anadenanthera peregrina and Myracrodruon urundeuva) commonly used to remediate contaminated soils. The deleterious effects of As and Zn on seed germination were due to As- and Zn-induced H₂O₂ accumulation and inhibition of APX and GR activities, which lead to oxidative damage by lipid peroxidation. SOD and CAT did not show any As- and Zn-induced inhibition of their activities as was seen with APX and GR. We investigated the close relationships between seed germination success under As and Zn stress in terms of GR and, especially, APX activities. Increased germination of A. peregrina seeds exposed to 50 mg L^?1 of Zn was related to increased APX activity, and germination in the presence of As (10 mg L^?1) was observed only in M. urundeuva seeds that demonstrated increased APX activity. All the treatments for both species in which germination decreased or was inhibited showed decreases in APX activity. A. peregrina seeds showed higher Zn-tolerance than M. urundeuva, while the reverse was observed with arsenic up to exposures of 10 mg L^?1.     

Calcium-dependent signaling pathway in the heat-induced oxidative injury in <Emphasis Type="Italic">Amaranthus lividus</Emphasis>

Heat caused reduction in membrane protein thiol content, increased accumulation of thiobarbituric acid reactive substances and reduced germination rate and early growth in germinating Amaranthus lividus seeds. Imposition of heat stress during early germination also causes accumulation of reactive oxygen species like superoxide and hydrogen peroxide while activities of antioxidative enzymes catalase, ascorbate peroxidase, and glutathione reductase decreased. Calcium chelator (EGTA), calcium channel blocker (LaCl₃) and calmodulin inhibitor (trifluroperazine) aggravated these effects. Added calcium reversed the effect of heat, implying that protection against heat induced oxidative damage and improvement of germination requires calcium and calmodulin during the recovery phase of post-germination events in Amaranthus lividus.     

Heat and chilling induced disruption of redox homeostasis and its regulation by hydrogen peroxide in germinating rice seeds (Oryza sativa L., Cultivar Ratna)

Extremes of temperature (both heat and chilling) during early inbibitional phase of germination caused disruption of redox-homeostasis by increasing accumulation of reactive oxygen species (superoxide and hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of total thiol content and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) in germinating tissues of rice (Oryza sativa L., cultivar Ratna). Imbibitional heat and chilling stress also induced oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation and protein oxidation [measured in terms of thiobarbituric acid reactive substances (TBARS), free carbonyl content (C = O groups) and membrane protein thiol level (MPTL)]. Treatment with standardized low titer hydrogen peroxide during early imbibitional phase of germination caused significant reversal in oxidative damages to the newly assembled membrane system imposed by heat and chilling stress [evident from the data of TBARS, C = O, MPTL, ROS accumulation, membrane permeability status, membrane injury index and oxidative stress index] in seedlings of experimental rice cultivar. Imbibitional H₂O₂ pretreatment also caused up-regulation of antioxidative defense (activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and total thiol content) in the heat and chilling stress-raised rice seedlings. When the parameters of early growth performances were assessed (in terms of relative growth index, biomass accumulation, relative germination performance, mean daily germination, T₅₀ value), it clearly exhibited significant improvement of early growth performances of the experimental rice cultivar. The result proposes that an ‘inductive pulse’ of H₂O₂ is required to switch on some stress acclimatory metabolism through which plant restores redox homeostasis and prevents or repairs oxidative damages to newly assembled membrane system caused by unfavorable environmental cues during early germination to the rice cultivar Ratna. The importance of mitigating oxidative damages to membrane lipid and protein necessary for post-germinative growth under extremes of temperature is also suggested.     

Arginine catabolism in the cotyledons of developing and germinating pea seeds

Arginine is the predominant free amino acid in the cotyledons of developing seeds of Pisum sativum L. cv Marzia. Breakdown of arginine was measured by injecting l-[guanido-¹⁴C]arginine into detached cotyledons. Cotyledons of developing seeds showed a low rate of ¹⁴CO₂ evolution whereas a much higher rate of ¹⁴CO₂ evolution was measured from cotyledons of seeds 4 days after the onset of germination. The activities of the catabolic enzymes arginase, urease, and ornithine aminotransferase were measured throughout development and germination. Arginase and ornithine aminotransferase were present at an early stage of development. Urease activity appeared later as the seeds started to desiccate. During germination, all three enzymes were present. The different course of activity of these enzymes indicates that they are controlled separately.     

Effects of hematin and carbon monoxide on the salinity stress responses of Cassia obtusifolia L. seeds and seedlings

Aims

The purpose of the present study was to investigate the mechanism of carbon monoxide (CO) and hematin in alleviating the inhibition of Cassia obtusifolia seeds and seedlings. NaCl (100?mM) was used to mimic salinity stress in a series of experiments.

Methods

Varying combinations of CO in a saturated aqueous solution and hematin (1.0?μM) were added to seeds and seedlings under salinity stress. Seed germination indices and seedling parameters were investigated.

Results

Seed germination and seedling growth were significantly inhibited under salinity stress. NaCl-induced inhibitory effects on seed germination and seedling growth were ameliorated by hematin or the CO aqueous solution. Addition of 1.0?μM hematin or 5?% CO-saturated aqueous solution to seeds and seedlings significantly alleviated damage to the plant cells under salinity stress. Hematin and the CO aqueous solution enhanced chlorophyll concentration, total soluble sugars, free proline, and soluble protein, and improved photosystem II (PSII) photochemical efficiency levels, PSII actual photochemical efficiency, and the photochemical quench coefficient. In contrast, the non-photochemical quenching coefficient decreased. Hematin and the CO aqueous solution also enhanced the activities of superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, and glutathione reductase, thus alleviating oxidative damage, as indicated by decreases in hiobarbituric acid reactive substances, hydrogen peroxide concentration, relative conductivity, and lipoxygenase activity. Heme oxygenase (HO) activity was increased by hematin treatment. Hematin may contribute to endogenous HO-derived CO, since the addition of zinc protoporphyrin IX or hemoglobin reversed the protective effects conferred by hematin specified above.

Conclusions

Based on the experimental results, we conclude that hematin and CO induce advantageous effects on the attenuation of salt-stress inhibition of C. obtusifolia seeds and seedlings and alleviate oxidative damage by conferring beneficial cytoprotection and activating anti-oxidant enzymes.     

Catalase is a key enzyme in seed recovery from ageing during priming

Ageing induces seed deterioration expressed as the loss of seed vigour and/or viability. Priming treatment, which consists in soaking of seeds in a solution of low water potential, has been shown to reinvigorate aged seeds. We investigate the importance of catalase in oxidation protection during accelerated ageing and repair during subsequent priming treatment of sunflower (Helianthus annuus L.) seeds. Seeds equilibrated to 0.29 g H₂O g⁻¹ dry matter (DM) were aged at 35 °C for different durations and then primed by incubation for 7 days at 15 °C in a solution of polyethylene glycol 8000 at −2 MPa. Accelerated ageing affected seed germination and priming treatment reversed partially the ageing effect. The inhibition of catalase by the addition of aminotriazol during priming treatment reduced seed repair indicating that catalase plays a key role in protection and repair systems during ageing. Ageing was associated with H₂O₂ accumulation as showed by biochemical quantification and CeCl₃ staining. Catalase was reduced at the level of gene expression, protein content and affinity. Interestingly, priming induced catalase synthesis by activating expression and translation of the enzyme. Immunocytolocalization of catalase showed that the enzyme co-localized with H₂O₂ in the cytosol. These results clearly indicate that priming induce the synthesis of catalase which is involved in seed recovery during priming.     

Impact of dormancy regulating chemicals on salinity induced dormancy in Lasiurus scindicus and Panicum turgidum: two desert glycophytic grasses

The indigenous forage grasses Lasiurus scindicus and Panicum turgidum are candidate species for the restoration of degraded desert rangelands. The impact of five dormancy regulating chemicals on overcoming salinity-induced germination inhibition was assessed under the best germination conditions in the two species. Seeds were germinated in a series of NaCl concentrations: 0–200 mM NaCl for P. turgidum, and 0–300 mM NaCl for L. scindicus. Lasiurus scindicus seeds were more tolerant to salinity than those of P. turgidum. Twenty percent of P. turgidum seeds germinated in 100 mM NaCl and none in the higher levels, but 47.5% and 8.8% of L. scindicus seeds germinated in 100 and 200 mM NaCl, respectively. The five studied chemicals (fusicoccin, GA₃, kinetin, nitrate and thiourea) did not succeed in improving germination of non-saline treated seeds of the two species, compared to the control, except thiourea in P. turgidum. The salinity-induced germination inhibition in P. turgidum was completely alleviated by the application of gibberellic acid (GA₃), partially alleviated by the application of fusicoccin, kinetin and thiourea, but not affected by nitrate. In L. scindicus, the germination inhibition was completely alleviated by fusicoccin, GA₃, nitrate and thiourea, but partially alleviated by kinetin. For using the two grass species in restoration of degraded rangelands affected by higher salinity, the results suggest using fusicoccin, GA₃, nitrate and thiourea with L. scindicus and GA₃ with P. turgidum seeds as a preseeding treatment can overcome the problem of reduced germination.     

Alleviation of osmotic stress in Brassica napus,B. campestris,and B. juncea by ascorbic acid application

The roles of ascorbic acid (AsA, 1 mM) under an osmotic stress [induced by 15 % (m/v) polyethylene glycol, PEG-6000] were investigated by examining morphological and physiological attributes in Brassica species. The osmotic stress reduced the fresh and dry masses, leaf relative water content (RWC), and chlorophyll (Chl) content, whereas increased the proline (Pro), malondialdehyde (MDA), and H₂O₂ content, and lipoxygenase (LOX) activity. The ascorbate content in B. napus, B. campestris, and B. juncea decreased, increased, and remained unaltered, respectively. The dehydroascorbate (DHA) content increased only in B. napus. The AsA/DHA ratio was reduced by the osmotic stress in all the species except B. juncea. The osmotic stress increased the glutathione (GSH) content only in B. juncea, but increased the glutathione disulfide (GSSG) content and decreased the GSH/GSSG ratio in all the species. The osmotic stress increased the activities of ascorbate peroxidase (APX) (except in B. napus), glutathione reductase (GR) (except in B. napus), glutathione S-transferase (GST) (except in B. juncea), and glutathione peroxidase (GPX), and decreased the activities of catalase (CAT) and monodehydroascorbate reductase (MDHAR) (only in B. campestris). The osmotic stress decreased the glyoxalase I (Gly I) and increased glyoxalase II (Gly II) activities. The application of AsA in combination with PEG improved the fresh mass, RWC, and Chl content, whereas decreased the Pro, MDA, and H₂O₂ content in comparison with PEG alone. The AsA addition improved AsA-GSH cycle components and improved the activities of all antioxidant and glyoxalase enzymes in most of the cases. So, exogenous AsA improved physiological adaptation and alleviated oxidative damage under the osmotic stress by improving the antioxidant and glyoxalase systems. According to measured parameters, B. juncea can be recognized as more drought tolerant than B. napus and B. campestris.     

Changes in activities of antioxidant enzymes during <Emphasis Type="Italic">Chenopodium murale</Emphasis> seed germination

The activities and isoenzyme pattern of catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) have been studied during germination of Chenopodium murale seeds. CAT and SOD activities were similar in dry seeds and during first 2 d of imbibition. CAT activity increased during radicle protrusion and early seedling development. The maximum SOD activity was found at final stages of germination and early seedling development. POD activity was not detected until the 6^th day of germination, indicating POD involvement not until early seedling development. Gibberellic acid (GA₃, 160 μM) delayed and synchronized C. murale germination.