Antioxidant Enzyme Activities during in vitro Morphogenesis of Gladiolus and the Effect of Application of Antioxidants on Plant Regeneration

Activity of antioxidant enzymes was evaluated during somatic embryogenesis and shoot organogenesis from cultured leaf segments of Gladiolus hybridus Hort. The effect of exogenous antioxidants on somatic embryogenesis and shoot organogenesis has also been monitored. Activity of superoxide dismutase (SOD) gradually increased during somatic embryogenesis. while activities of catalase (CAT) and peroxidase (POX) decreased. In contrast, increase in CAT and POX activity and a concomitant decrease in SOD activity were noted during shoot organogenesis. Exogenous application of antioxidants such as glutathione (GSH), α-tocopherol and ascorbate (AA) inhibited somatic embryogenesis but stimulated shoot organogenesis. The frequency of somatic embryogenesis increased with the addition of H₂O₂. However, H₂O₂ inhibited shoot organogenesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Antioxidative responses to different altitudes in leaves of alpine plant Polygonum viviparum in summer

Mountain environmental stresses result in increased formation of hydrogen peroxide (H₂O₂) and accumulation of malondialdehyde (MDA) in leaves of Polygonum viviparum. The activities of several antioxidative system enzymes such as superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7), glutathione reductase (GR, EC 1.6.4.2), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and the contents of several non-enzymatic antioxidants such as reduced form of ascorbate (ASC), dehydroascorbate (DHA), reduced glutathione (GSH), and oxidized glutathione (GSSG) were investigated in leaves of P. viviparum, which were collected from three altitudes (2,200, 3,200, and 3,900 m) of Tianshan Mountain in China. The activities of these four antioxidative enzymes were accompanied by increases of H₂O₂ levels from 2,200 to 3,200 m. However, the activities of CAT and POD were decreased, whereas the activities of SOD and GR continually increased at 3,900 m. Analyses of isoforms of SOD, CAT, POD, and GR showed that the leaves of P. viviparum exposed different altitude conditions are capable of differentially altering the intensity. Additionally, two new isoforms of SOD were detected at 3900 m. A continual increase in the ASC, ASC to DHA ratio, GSH and GSH/[GSH + GSSG] ratio, and the activity of DHAR were observed in leaves of P. viviparum with the elevation of altitude. These results suggest that the higher contents of ASC, GSH as well as an increase in reduced redox state may be essential to antioxidation processes in the leaves of P. viviparum, whereas antioxidant enzymes system is a cofactor in the processes.     

Effects of NaCl and Mycorrhizal Fungi on Antioxidative Enzymes in Soybean

The effects of different concentrations of NaCl on the activities of antioxidative enzymes in the shoots and roots of soybean (Glycine max [L.] Merr cv. Pershing) inoculated or not with an arbuscular mycorrhizal fungus, Glomus etunicatum Becker & Gerdemann, were studied. Furthermore, the effect of salt acclimated mycorrhizal fungi on the antioxidative enzymes in soybean plants grown under salt stress (100 mM NaCl) was investigated. Activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were increased in the shoots of both mycorrhizal (M) and nonmycorrhizal (NM) plants grown under NaCl salinity. Salinity increased SOD activity in the roots of M and NM plants, but had no effect on CAT and polyphenol oxidase activities in the roots. M plants had greater SOD, POD and ascorbate peroxidase activity under salinity. Under salt stress, soybean plants inoculated with salt pre-treated mycorrhizal fungi showed increased SOD and POD activity in shoots, relative to those inoculated with the non pre-treated fungi.     

Enzymes regulating the accumulation of active oxygen species during the hypersensitive reaction of bean to Pseudomonas syringae pv. phaseolicola

Changes in the activities of superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POD; EC 1.11.1.7) and catalase (CAT; EC 1.11.1.6) which regulate the persistence of active oxygen species (AOS) were examined in leaves of bean (Phaseolus vulgaris L. cv. Tendergreen) undergoing compatible and incompatible interactions to race 6 and race 3 strains, respectively, of the halo-blight bacterium Pseudomonas syringae pv. phaseolicola. Resistance of cv. Tendergreen to race 3 is determined by the R3 gene and was expressed by a hypersensitive reaction (HR) which was associated with a rapid increase in lipid peroxidation between 8 and 12 h after inoculation. Five main isoforms of SOD were resolved by native polyacrylamidegel electrophoresis (PAGE). Major changes were found in the activities of the cytosolic Cu, Zn-SOD3 and Cu, ZnSOD5 isoforms, which increased by 6 h after inoculation with race 3, and the possibly peroxisomal MnSOD2 isoform, which decreased rapidly in tissue undergoing the HR. Three further minor isoforms of SOD showed a strong increase in activity during the HR. A low level of extracellular SOD activity was also resolved; two isoforms, one of which increased dramatically in activity during the HR, were detected within intercellular fluids recovered from inoculation sites. Fewer changes in SOD activities were found during the compatible interaction to race 6, and they did not occur until 16 h after inoculation. In tissue around infiltration sites, no decrease in the activity of Mn-SOD2 was observed but slight increases in some other isoforms were found. Four groups of POD isoforms were detected in both 3,3-diaminobenzidine/H₂O₂-and o-dianisidine/H₂O₂-stained PAGE gels. Significant changes in activity were again associated with development of the HR. In particular, by 2 h after inoculation, increases in POD3a, b and c isoforms were detected within total soluble extracts and also in POD3c within intercellular fluids (no other isoform was found in the apoplasm). By contrast, POD1 and POD2 activities generally declined following inoculation. The principal change in activity in tissues surrounding infiltration sites was an increase in POD3 isoforms following inoculation with race 3. Measurements of total activity showed a decrease in CAT activity as early as 2 h after inoculation, followed by a recovery after 8 h and a further decrease as infiltrated tissue collapsed during the HR. A more-gradual decline in CAT activity was observed at sites undergoing the compatible interaction and also in tissue surrounding inoculation sites. The spatial and temporal changes detected in activities of CAT and isoforms of SOD and POD clearly demonstrate the complexity and potential subtlety of control of the production and persistence of AOS in bean following microbial challenge. The generation of AOS through HR-specific, early increases in extra-cellular POD and SOD isoforms is discussed.This work was supported in part by the scientific Research Foundaation (OTKA F 5082), the foundation for Hungarian science, a british council scolership to A.L.A and the U.K. Agricultural and food Reaserch council.     

盐胁迫对大米草幼苗某些生理指标的影响

研究了大米草幼苗在不同培养盐度(0、20、30、50、100mmol/LNaCl)下,MDA、游离脯氨酸、可溶性糖、可溶性蛋白质含量以及保护酶(SOD、POD、CAT)活性等生理指标的变化情况。结果表明:保护酶(SOD、POD、CAT)活性在盐胁迫40d前逐渐上升且达显著差异。随着胁迫时间延长,MDA含量与CK相比逐渐降低。随着盐分胁迫浓度的增加及盐胁迫时间延长,大米草叶片中游离脯氨酸、可溶性糖、可溶性蛋白质含量呈上升趋势。在盐胁迫下,渗透调节物质的积累作用是大米草对盐胁迫的主要响应过程,其体内的抗氧化保护酶在此过程中也发挥了重要的作用。     

Investigations on the antioxidative defence responses to NaCl stress in a mangrove, Bruguiera parviflora: Differential regulations of isoforms of some antioxidative enzymes

Two-month-old healthy seedlings of a true mangrove, Bruguiera parviflora, raised from propagules in normal nursery conditions were subjected to varying concentrations of NaCl for 45 d under hydroponic culture conditions to investigate the defence potentials of antioxidative enzymes against NaCl stress imposed oxidative stress. Changes in the activities of the antioxidative enzymes catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POX), glutathione reductase (GR) and superoxide dismutase (SOD) were assayed in leaves to monitor the temporal regulation. Among the oxidative stress triggered chemicals, the level of H₂O₂ was significantly increased while total ascorbate and total glutathione content decreased. The ratio of reduced to oxidized glutathiones, however, increased due to decreased levels of oxidized glutathione in the leaf tissue. Among the five antioxidative enzymes monitored, the APX, POX, GR and SOD specific activities were significantly enhanced at high concentration (400 mM NaCl), while the catalase activities declined, suggesting both up and downregulations of antioxidative enzymes occurred due to NaCl imposed osmotic and ionic stress. Analysis of the stress induced alterations in the isoforms of CAT, APX, POX, GR and SOD revealed differential regulations of the isoforms of these enzymes. In B. parviflora one isoform of each of Mn-SOD and Cu/Zn-SOD while three isoforms of Fe-SOD were observed by activity staining gel. Of these, only Mn-SOD and Fe-SOD2 content was preferentially elevated by NaCl treatment, whereas isoforms of Cu/Zn-SOD, Fe-SOD1 and Fe-SOD3 remained unchanged. Similarly, out of the six isoforms of POX, the POX-1,-2,-3 and -6 were enhanced due to salt stress but the levels of POX-4 and -5 remained same as in control plants suggesting preferential upregulation of selective POX isoforms. Activity staining gel revealed only one prominent band of APX and this band increased with increased salt concentration. Similarly, two isoforms of GR (GR1 and GR2) were visualized on activity staining gel and both these isoforms increased upon salt stress. In this mangrove four CAT-isoforms were identified, among which the prominent CAT-2 isoform level was maximally reduced again suggesting differential downregulation of CAT isoforms by NaCl stress. The results presented in this communication are the first report on the resolutions of isoforms APX, POX and GR out of five antioxidative enzymes studied in the leaf tissue of a true mangrove. The differential changes in the levels of the isoforms due to NaCl stress may be useful as markers for recognizing salt tolerance in mangroves. Further, detailed analysis of the isoforms of these antioxidative enzymes is required for using the various isoforms as salt stress markers. Our results indicate that the overproduction of H₂O₂ by NaCl treatment functions as a signal of salt stress and causes upregulation of APX, POX, GR and deactivations of CAT in B. parviflora. The concentrations of malondialdehyde, a product of lipid peroxidation and lipoxygenase activity remained unchanged in leaves treated with different concentrations of NaCl, which again suggests that the elevated levels of the antioxidant enzymes protect the plants against the activated oxygen species thus avoiding lipid peroxidation during salt stress.     

结球白菜离体子叶不定芽再生过程中的组织学及生理变化

以日本引进品种爱知结球白菜(Brassica rapa ssp．pekinensis CV．AiehiHakusai,C1)为试材,对离体子叶不定芽再生过程中的组织学和生理变化进行了研究。结果表明,子叶在离体培养过程中,不定芽发生方式为器官直接发生。在不定芽形成前,可溶性蛋白质含量、POD和SOD的活性均呈上升趋势。随着细胞的脱分化,代谢活动逐渐旺盛,酶活性增强,可溶性蛋白质含量增加,表明不定芽形成过程中形态变化与生理变化紧密相联。培养基中添加AgNO3对酶活性有促进作用,并促进不定芽的分化。     

淹水胁迫对铺地黍几种保护酶活性的影响

目的:探讨淹水胁迫对铺地黍几种保护酶活性的影响.方法:通过模拟试验,研究不同淹水时间对铺地黍体内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)及多酚氧化酶(PPO)活性的影响.结果:在淹水胁迫前期,铺地黍体内SOD、CAT和PPO活性总体表现为升高的变化特征.之后,随着胁迫时间的延长,3种酶活性均逐渐降低,在淹水5w时,SOD、CAT和PPO活性分别为对照的38.7%、50.3%和178.2%.而在整个淹水期间POD活性则一直呈升高的趋势.结论:几种保护酶对淹水胁迫的响应能力有差异,其中SOD、CAT活性的变化可作为铺地黍对淹水反应的生理指标.     

Induction of the activities of antioxidative enzymes and the levels of malondialdehyde in cucumber seedlings as a consequence of Bemisia tabaci (Hemiptera: Aleyrodidae) infestation

The activities of antioxidative enzymes and the concentration of malondialdehyde were assayed in cucumber leaves (Cucumis sativus L.) 0, 6, 12, 24, 48, 72, and 96 h after the cucumber seedlings were infested by Bemisia tabaci (Gennadius). The results indicated that the activities of antioxidative enzymes were increased after herbivore infestation, including superoxide dismutase (SOD), and peroxidase (POD), and the levels of malondialdehyde (MDA), which is a product of membrane lipid peroxidation in the leaves. The enzymes and MDA showed peaks of different activity levels at 24 and 48 h after the infestation. SOD activity reached the highest peak, 10.4% higher than control, at 24 h, POD activity reached the highest peak, 213.2% higher than control, at 6 h, catalase (CAT) activity was not statistically significant compared with the control, and MDA content reached the highest peak, 59.9% higher than control, at 48 h. The results suggested that the enhanced activities of antioxidative enzymes and MDA content may contribute to bioprotection of cucumber plants against B. tabaci infestation. Handling editor: Henryk Czosnek     

Effects of metalaxyl enantiomers stress on root activity and leaf antioxidant enzyme activities in tobacco seedlings

The objective of this experiment was to study the effects of metalaxyl enantiomers on the activity of roots and antioxidative enzymes in tobacco seedlings. Water culture experiment was conducted to analyze the effects of different concentrations of metalaxyl enantiomers (30 and 10 mg L^?1) on root activity and leaf superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities and malondialdehyde (MDA) content of tobacco seedlings. The results showed that metalaxyl significantly inhibited root activity and significantly improved leaf SOD, POD, and CAT activities and MDA content. A better physiological response in tobacco seedlings was observed at 30 mg L^?1 than at 10 mg L^?1 metalaxyl. The stereoselectivity for different enantiomers had no obvious effect on root activity and the leaf POD activity, but it affected significantly the SOD and CAT activities and MDA content. The SOD activity was promoted more by R‐enantiomer than by S‐enantiomer at 30 mg L^?1 metalaxyl, and the same effect was observed on CAT activity from the beginning to the end of the stress period. The MDA content under the stress by R‐enantiomer was higher than that under the stress by S‐enantiomer at 10 mg L^?1 metalaxyl.     

Differential biochemical responses of wheat shoots and roots to nickel stress: antioxidative reactions and proline accumulation

Wheat (Triticum aestivum L. cv. ‘Zyta’) seedlings were treated with 10, 100 and 200 μM Ni. Tissue Ni accumulation, length, relative water content (RWC), proline and H₂O₂ concentrations as well as the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and glutathione S-transferase (GST) were studied in the shoots and roots after 6 days of Ni exposure. Treatment with Ni, except for its lowest concentration, resulted in a significant reduction in wheat growth. In comparison to the shoots, the roots showed greater inhibition of elongation, which corresponded with higher accumulation of Ni in these organs. Both shoots and roots responded to Ni application with a decrease in RWC and enhancement in proline concentration. Greater dehydration of the shoot tissue was accompanied by more intense accumulation of proline. Treatment of the wheat seedlings with the highest concentration of Ni led to about 60% increase in H₂O₂ concentration in both studied organs. Apart from CAT, constitutive activities of antioxidative enzymes were much higher in the roots than in the shoots. Exposure of the seedlings to Ni resulted in SOD activity decline, which was more marked in the roots. While the shoots showed a substantial decrease (up to 30%) in CAT activity, in the roots the activity of this enzyme remained unchanged. After Ni application APX, POD and GST activities increased several-fold in the shoots, whereas in the roots they were not significantly altered. The results suggest that differential antioxidative responses of the shoots and roots of wheat seedlings to Ni stress might be related to diverse constitutive levels of antioxidant enzyme activities in both organs.     

Response of antioxidant enzymes in rice (<Emphasis Type="Italic">Oryza sauva</Emphasis> L. cv. Dongjin) under mercury stress

We studied the effects of different concentrations of mercury (0.0 to 100 μM) on growth and photosynthetic efficiency in rice plants treated for 21 d. In addition, we investigated how this metal affected the malondialdehyde (MDA) content as well as the activity of five antioxidant enzymes — superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol peroxidase (POD), and catalase (CAT). Photosynthetic efficiency (Fμ/F_m) and seedling growth decreased as the concentration of Hg was increased in the growth media. Plants also responded to Hg-induced oxidative stress by changing the levels of their antioxidative enzymes. Enhanced lipid peroxidation was observed in both leaves and roots that had been exposed to oxidative stress, with leaves showing higher enzymatic activity. Both SOD and APX activities increased in treatments with up to 50 μM Hg, then decreased at higher concentrations. In the leaves, both CAT and POD activities increased gradually, with CAT levels decreasing at higher concentrations. In the roots, however, CAT activity remained unchanged while that of POD increased a bit more than did the control for concentrations of up to 10 μM Hg. At higher Hg levels, both CAT and POD activities decreased. GR activity increased in leaves exposed to no more than 0.25 μM Hg, then decreased gradually. In contrast, its activity was greatly inhibited in the roots. Based on these results, we suggest that when rice plants are exposed to different concentrations of mercury, their antioxidative enzymes become involved in defense mechanisms against the free radicals that are induced by this stress.     

小叶锦鸡儿抗沙埋生长与抗氧化酶及同工酶变化的关系

小叶锦鸡儿(Caraganas tenophylla L.)是广泛应用于流动沙丘治理的优良固沙植物。然而关于其抗沙埋生理机理目前尚不清楚。选择生长在科尔沁沙地的小叶锦儿为试验材料,依据株高对其进行不同程度沙埋(轻度、中度、重度沙埋), 并通过测定沙埋过程中植株高度、不同部位叶片丙二醛(MDA)含量、抗氧化酶活力、抗氧化酶同工酶谱变化, 以揭示其抗沙埋生理适应机理和基因调控机理。结果表明:沙埋6d,植株各部位生长加快,尤其是顶部和基部生长更快。叶片MDA含量降低、整株植物叶片平均过氧化氢酶(CAT)、过氧化物酶(POD)、过氧化物歧化酶(SOD)活力增加,但重度沙埋使抗氧化酶活力下降。沙埋12d,植株各部位生长继续加大, 沙下叶片凋落。与对照相比,沙上叶片MDA含量成倍增加,并与叶片POD、SOD和CAT活力的大幅度提高呈正相关,并与对照差异显著(P < 0.01)。同时,不同厚度沙埋6d,叶片CAT同工酶出现两新带CAT III和CATII;POD同工酶谱带(6条酶带)随沙埋厚度增加,叶片PODII区带加宽、色加深,POD I 和POD III酶带消失。但是,不同厚度沙埋下,沙上和沙下叶片CAT、SOD和POD酶谱带数和活力均相同。这表明在沙埋应激适应反应期(6d),叶片抗氧化酶活力的增强与抗氧化酶基因表达增强和基因启动有关。受到沙埋重力胁迫的成熟叶可能将胁迫信号传递给沙上没有沙埋的叶子及生长点,导致整株叶片产生整体适应性反应,激活抗氧化酶系统,以致加速生长。因此,小叶锦鸡儿萌蘖生物学特性和抗氧化酶对沙埋胁迫快速响应在维护氧自由基代谢平衡和植株快速恢复生长中起重要保护作用。     

Study of Antioxidant Enzymes Activity during Organogenesis and In Vitro Propagation of Asiatic Hybrid Lily

A protocol for indirect differentiation of shoots / roots from leaf callus of Asiatic hybrid lily was developed through in vitro methods. The involvement of antioxidant enzymes, like, SOD, POX and CAT, and their isoenzymes during organogenesis in the morphogenetic callus was stud ied.The activity of these enzymes was increased during early development and differentiation of callus. SOD activity increased significantly as compared to POX and CAT during root formation, while it decreased in shoot formation and the decrease was significant in POX and CAT enzymes. The results indicate that the organogenesis is a very complicated biological process involving up and down regulation of a number of antioxidant enzymes, which seem to play an important role during organogenesis of Lilium callus.     

Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhiza uralensis Fisch)

The effects of salinity and drought on the antioxidative system (SOD, POD, CAT) were studied in liquorice seedlings (Glycyrrhiza uralensis Fisch). The results showed that both salt and drought stresses could induce oxidative stress, as indicated by the increase level of lipid peroxidation. The activities of SOD and POD were up-regulated by salt and drought stress, while CAT activity decreased. An additional MnSOD isoenzyme was detected in liquorice subjected to 2%NaCl stress. The data also showed that although the activity of SOD was differentially influenced by drought and salinity, the changes of antioxidant enzyme activities subjected to drought stress follow a pattern similar to that subjected to salt stress, indicating that similar defensive systems might be involved in the oxidative stress injury in liquorice.     

Activity of antioxidant enzyme during <Emphasis Type="Italic">in vitro</Emphasis> organogenesis in <Emphasis Type="Italic">Crocus sativus</Emphasis>

The effect of various hormonal combinations on regeneration of shoots and roots from meristem-derived callus of Crocus sativus L. and activities of antioxidant enzymes have been studied. The most efficient regeneration occurred with 1.0 mg dm⁻³ 1-naphthaleneacetic acid (NAA) + 1.0 mg dm⁻³ thidiazuron and 1.0 mg dm⁻³ NAA + 2.0 mg dm⁻³ kinetin. For sprouting, regenerated shoot were subcultured on Murashige and Skoog medium containing 1.0 mg dm⁻³ NAA + 1.0 mg dm⁻³ benzylaminopurine (BAP). Protein content and superoxide dismutase activity decreased in regenerated shoots and roots and increased in sprouting shoots, while catalase (CAT), peroxidase (POX) and polyphenol oxidase (PPO) activities increased during organogenesis and decreased in sprouting shoots. High CAT and PPO activities were detected in regenerated roots, whereas high POX activity was observed in regenerated shoot.     

艳婀珍蝶取食对薇甘菊叶片生理指标的影响

艳婀珍蝶取食后,对薇甘菊叶片的超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)的活性,总酚含量,有机自由基(DPPH·)清除能力进行分析。结果表明,取食后3h内所测各量即发生变化,但与对照差异不显著。取食4d中,叶片的SOD、POD活性总体上分别高于对照,CAT活性总体上低于对照,说明在艳婀珍蝶胁迫过程中,SOD和POD所起的作用比CAT大。取食叶SOD、POD活性均在48h时达到最大值,之后下降;CAT活性在24h时达到最大值,之后也迅速下降。取食叶PPO活性动态变化程度较大,表现出3个峰值,分别为对照的1·83,1·92倍和2·17倍;总酚含量表现为先上升后下降的趋势;对DPPH·的清除力一直显著性甚至极显著性低于对照。取食叶的SOD与CAT和POD均呈正相关性,且相关性大于对照;PPO与总酚含量在取食叶与对照中也均呈弱正相关性。实验结果表明,薇甘菊的保护酶对艳婀珍蝶胁迫的应激效应是短暂而有限的,艳婀珍蝶的取食破坏了薇甘菊叶片功能,较大程度的干扰了薇甘菊保护酶系统的防御代谢,薇甘菊的总抗氧化能力降低。薇甘菊也不能通过改变酚类物质含量来抵御艳婀珍蝶的取食,艳婀珍蝶取食对薇甘菊有较明显的控制作用。     

Effects of the infestation by Actinote thalia pyrrha (Fabricius) on the physiological indexes of Mikania micrantha leaves

Mikania micrantha is a dangerous exotic weed now found throughout Guangdong Province, China. Introduced by the Indonesian Oil Palm Institute, lepidopteran defoliator Actinote thalia pyrrha (Fabricius) is a potential biological control factor for M. micrantha. Changes in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), polyphenoldoxidase (PPO), the total phenolics content, and the capacity of scavenging 1,1-diphenyl-2-picrylhydazyl (DPPH) free radicals in M. micrantha leaves were analyzed after infestation by A. thalia pyrrha larvae. The results showed that a slight change of all the indexes in the damaged leaves appeared within 3 h. After four days of infestation by A. thalia pyrrha , SOD and POD activities in the damaged leaves were higher than those of the control. But CAT activity was lower than that of the control, indicating that the SOD and POD functions were impeded more than those of CAT. The SOD and POD activities reached their highest values after 48 h of infestation by A. thalia pyrrha and then decreased, whereas the highest activity of CAT occurred after 24 h and then decreased. PPO activity fluctuated greatly with its three peak values being 1.83, 1.92, and 2.17 times that of the control. The total phenolics content increased initially and then decreased after 72 h of infestation by A. thalia pyrrha . The capacity of scavenging DPPH was significantly lower than that of the control (p<0.05). SOD activity positively correlated with CAT and POD activities for both damaged and normal leaves. However, the value of the correlation coefficient in the damaged leaves was higher than that in the normal leaves. The total phenolics content also showed a weak positive correlation with PPO activity. It is suggested that the protective response of M. micrantha to infestation by A. thalia pyrrha was short and limited, but resulted in a reduced function of the leaves, disturbed the metabolism in the protective enzyme system, and decreased the antioxidative capacity. M. micrantha was unable to resist the feeding stress by altering the content of the total phenolics.     

Differential response of antioxidative enzymes in embryonic axes and cotyledons of germinating lupine seeds

Germination of lupine (Lupinus luteus L.) seeds was accompanied by an increase in concentration of free radicals with g ₁ and g ₂ values of 2.0056 ± 0.0003 and 2.0033 ± 0.0005, respectively. The highest intensity of free radical signal was observed in embryo axes immediately after radicle protruded through the seed coat. Hydrogen peroxide accumulated in embryonic axes and cotyledons during imbibition before the onset of germination in the seed population. The activities of superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) rose progressively in embryo axes. In cotyledons SOD activity did not change significantly, while that of CAT increased during germination. The enhancement of Cu, Zn-SODs and Mn-SOD isoforms in embryonic axes was observed. A new isoform of catalase was synthesized, suggesting that it plays a relevant role during germination. SOD and CAT activities were detected in dry seeds. Free radical generation and response of antioxidative enzymes differed between embryo axes and cotyledons during the germination timecourse.