离子注入对萝卜过氧化物酶、淀粉酶和蛋白酶同工酶的影响

研究了氮离子和氩离子注入萝卜种子对萝卜幼苗蛋白含量、过氧化物酶活性及过氧化物酶、淀粉酶和蛋白酶同工酶的影响。结果表明 :离子注入后 ,减低萝卜过氧化物酶活性和蛋白含量。萝卜不同生长时期同工酶变化不一样。在子叶时期 ,过氧化物酶同工酶谱带无明显变化 ,淀粉酶同工酶有酶带的消失 ;而在真叶时期 ,过氧化物酶在负极区减少一条酶带 ,Rf为 0 .2 2 ,正极区增加一条酶带 ,Rf为 0 .6 ,且随剂量增加 ,酶带着色增强 ;淀粉酶同工酶在注入剂量为 5× 10 5N+ / cm2 )时 ,有同工酶带增加 ,Rf为 0 .6 1。低剂量时蛋白酶活性增强 ,谱带增多 ,大剂量则减弱。因此 ,N+和 Ar+注入后 ,可影响萝卜过氧化物酶和淀粉酶的表达及蛋白质的合成或降解。     

Peroxidase Activity and Isoperoxidase Pattern in Tobacco Leaves Infected with Tobacco Necrosis Virus and Other Viruses Inducing Necrotic and Non-Necrotic Alterations

The level of peroxidase activity was greatly enhanced in tobacco leaves infected by tobacco necrosis virus (TNV) and other viruses which induce necrotic symptoms (TMV, ToMV and PVY^N). The intensity was related to the age of the leaves infected: absent or neglible in mature leaves and very pronounced in young growing infected leaves. On the contrary, changes in peroxidase activity were negligible when the infection was provoked by viruses which do not produce necrotic reactions (TMV and PVY^O). Analysis of the peroxidase isoenzymes, pattern in tobacco leaves infected by TNV and other necrosis-inducing viruses revealed in all cases, a slight increase in anionic (pl 3.5–3.7) and a considerable increase in moderately anionic isoenzymes particularly the pl 4.6 isoenzyme which in TNV and PVY^N-infected leaves reached levels up to 21 and 72 times the healthy control values. A considerable increase in the cationic (pl9.3–8.8) isoenzymes and the appearance of one moderately cationic isoenzyme (pl 8.2) was also detected. In leaf extracts from-virus-infected tobacco leaves with nonnecrotic response, no, or negligible alterations on the isoenzyme pattern were detected. However, infection by a fungal parasite (Erisyphe cichoracearum), which established a fully compatible, non-necrotic, interaction with tobacco leaves, like the necrosis-inducing viruses, changed the isoperoxidase pattern. The data suggest the necrotic alterations and associated changes in the peroxidase activity and isoperoxidase pattern in virus-infected leaves are not clearly related.     

Overexpression of the endogenous peroxidase-like gene spi 2 in transgenic Norway spruce plants results in increased total peroxidase activity and reduced growth

Peroxidases constitute a large family of proteins found in all higher plants. Owing to the complexity of the peroxidase isoenzyme family it has been difficult to assess the precise function of individual peroxidase enzymes. In this work we have studied the effects of an endogenous peroxidase-like gene from Norway spruce [Picea abies (L.) Karst], spi 2, on the development and growth of Norway spruce somatic embryo plants. Embryogenic cells of Norway spruce transformed with spi 2 under control of the maize ubi-1 promoter showed up to 40 times higher total peroxidase activity than the control cells; regenerated plants overexpressing spi 2 showed an increased total peroxidase activity. Based on these results and the overall sequence similarity with cationic peroxidases we conclude that spi 2 encodes a peroxidase. Overexpression of spi 2 resulted in increased sensitivity to stress, leading to a reduction in epicotyl formation and in height growth compared with control plants. The plants overexpressing spi 2 also showed a deeper phloroglucinol staining but similar levels of Klason lignin.     

Variability of antioxidant enzyme activity and isoenzyme profile in needles of Serbian spruce (Picea omorika (Panč.) Purkinye)

Variations were studied of the activity and isoenzyme patterns of soluble peroxidase, catalase, catechol oxidase and superoxide dismutase, in needles of the Balkan endemic conifer Serbian spruce, Picea omorika (Pan?.) Purkinye. The samples were collected from the natural habitat of the species, Mt. Tara. Seasonal changes were found to affect enzymatic activities and isoenzyme profiles. Total protein content was significantly lower in the summer than in other seasons. Several isoforms of peroxidase, catechol oxidase and superoxide dismutase (SOD), as well as two catalase isoenzymes were detected. The number of peroxidase isoenzymes was greatest during the vegetative season. Catalase and catechol oxidase peaked in summer and spring, respectively. Total SOD and Mn-SOD activities were significantly higher in the winter samples than the summer ones.     

Stress-induced changes in glutamate dehydrogenase activity imply its role in adaptation to C and N metabolism in lupine embryos

The modifying effect of sucrose on glutamate dehydrogenase (GDH) activity and isoenzyme pattern was investigated in isolated embryos of lupine ( Lupinus luteus L.), cultured in vitro in a medium with sucrose (+S) or without sucrose (−S) and exposed to cadmium (Cd) and lead (Pb) stress. Sucrose starvation of lupine embryos led to a rapid increase in the specific activity of GDH, immunoreactive β-polypeptide and it was accompanied by appearance of new cathodal isoforms of enzyme. This suggests that isoenzymes induced in lupine embryos by sucrose starvation combine into GDH hexamers with the predominance of β-GDH subunits synthetized under GDH1 gene control. The addition of sucrose to the medium caused an opposite effect. Along with upregulation of catabolic activity of GDH by sucrose starvation, activity of proteolytic enzymes was also induced. These data can point to regulatory mechanism implying a sucrose dependent repression of the GDH1 gene according to the mechanism of catabolic repression. Treatment of embryos with Cd ²⁺ or Pb²⁺ resulted in ammonium accumulation in the tissues, accompanied by an increase in anabolic activity of GDH and activity of anodal isoenzymes, in both (+S) and (−S) embryos without new de novo synthesis of α subunit proteins. Thus, GDH isoenzyme profiles may reflect the physiological function of GDH, which appears to be an important link of metabolic adaptation in cells, aimed at using carbon sources other than sugar during carbohydrate starvation (catabolic activity of GDH) and protecting plant tissues against ammonium accumulated because of heavy metal stress (anabolic activity of GDH).     

Peroxidase Isoenzymes in Strawberry Roots Infected with Binucleate Rhizoctonia spp. and their Implication in Disease Resistance

Infection of strawberry plants with binucleate Rhizoctonia spp. results in an increase in peroxidase activity and the appearance of new isoforms of the enzyme. In healthy and diseased roots of two different strawberry genotypes seven peroxidase isoenzymes were found. In healthy strawberry cv. Senga Sengana, which was moderately resistant to infection, four isoenzymes (1, 2, 5, and 6) were found. Moreover the activity of these isoenzymes was increased and three new isoenzymes (3, 4, and 7) were found in infected roots. In the strawberry hybrid 3/2/86/88/R, which is very susceptible to infection, only isoenzyme 2 was present in the roots of healthy plants. Following infection, the activity of isoenzyme 2 was increased and five new isoenzymes (1, 4, 5, 6, and 7) were detectable. The results obtained indicate that strawberry resistance to binucleate Rhizoctonia may be correlated with peroxidase isoenzyme profile with particular reference to isoform 3, which is only present in infected roots of the moderately resistant cv., Senga Sengana.     

Effect of drought on proteins and isoenzymes in rice during germination

Two drought tolerant varieties TKM-1 and TKM-2 and two drought susceptible varieties Jaya and Improved Sabarmati of rice were studied for soluble protein pattern and isoenzymes of malate dehydrogenase, glutamate dehydrogenase, esterase and peroxidase during germination at different water stress. MDH, GDH and esterase patterns were not affected, but the soluble proteins were changed. Peroxidase isoenzyme pattern from drought tolerant and susceptible varieties showed characteristic differences. The intensity of bands with higher electrophoretic mobility decreased in Jaya and Improved Sabarmati while in TKM-1 and TKM-2 the intensity of these bands did not change much after 72 hr water stress. In shoots of Jaya and Improved Sabarmati, the activity of the peroxidase isoenzymes decreased more than in TKM-1 and TKM-2 shoots with increase in water stress.     

Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of <Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) czern

Metabolic adaptations to heavy metal toxicity in plants are thought to be related with developmental growth stage and the type of metal by which plant is affected. In the present study, changes in ionically bound CWP, soluble peroxidase activity, H(2)O(2) level and Malonaldehyde content in roots of cadmium and copper stressed seedlings and cadmium stressed 3-4 leaf stage plants of Brassica juncea were investigated. Cadmium inhibits root growth and reduces fresh biomass. The reduction in root growth and fresh biomass is correlated with increased lipid peroxidation and reduced tolerance. Treatment with cadmium resulted in an increase in ionically bound CWP activity in roots of seedlings but no significant change in its activity was found in roots of 3-4 leaf stage plants. Increased level of H(2)O(2) in roots of cadmium and copper treated seedlings, show a direct correlation with increased activity of ionically bound CWP. H(2)O(2) level in 3-4 leaf stage plant roots was found to be very low. Soluble peroxidase activity decreased in cadmium (50 and 100 mu-icroM) treated seedlings but it was ineffective to cause any change in its activity in 3-4 leaf stage plants. Copper treated seedlings showed an increase in ionically bound CWP activity, H(2)O(2) level and MDA content. Ascorbic acid (50 mM) pretreated seedlings shows significant decrease in ionically bound CWP activity when exposed to 50 muM cadmium. Hence, it is concluded that inhibition of root growth in Brassica juncea seedlings by cadmium, is associated with CWP catalyzed H(2)O(2) dependent reactions which are involved in metabolic adaptations to heavy-metal stress.     

Cadmium and copper induction of oxidative stress and antioxidative response in tomato (<Emphasis Type="Italic">Solanum lycopersicon</Emphasis>) leaves

We compare cadmium and copper induced oxidative stress in tomato leaves and the antioxidative enzyme response during a time course of 96 h. Plants were subjected to 25 μM of CdCl₂ or CuSO₄ and malondialdehyde (MDA) level and activity of guaiacol peroxidase, superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were determined. The results showed that there was an early increase in the MDA level and in the guaiacol peroxidase activity more pronounced with copper exposure during almost all the time course of the experiment. The activity of superoxide dismutase and catalase was induced very early after cadmium and copper treatment, reached a maximal value after 12 h and then declined but it remained always slightly higher than the control at the end of the experiment. Ascorbate peroxidase activity pathway was similar to superoxide dismutase or catalase with a maximal activity after 48 h of heavy metal exposure. Induction of glutathione reductase activity observed only under copper exposure is maintained during almost all the experimental time. The antioxidative activity developed by tomato leaves is more induced by copper treatment. This can be related to the ability of this metal to induce more than cadmium an accumulation of reactive oxygen species (ROS) at the cellular level. Decline in the antioxidative enzymes activity at the end of the experiment can be a consequence of cadmium- and copper-inducing a further ROS formation that might affect enzymes activity.     

Peroxidase Ontogeny in a Dwarf Pea Stem as Affected by Gibberellin and Decapitation

The ontogeny of peroxidase activity and isoenzyme pattern wasinvestigated in the stem of dwarf pea plants. Peroxidase activityper unit soluble protein was a given internode is highest inthe youngest growth stage, drops during elongation, remainsconstant upon cessation of growth, and increase at senescence.The lower the internode on the stem the higher is its peroxidaseactivity. These developmental differences are already apparentat the youngest growth stage of the internodes adn increaseduring elongation. Several anodic and five cathodic isoperoxidasesare apparent after starch gel electrophoresis. This patternis constant for all internodes at all growth stages, but therelative importance of particular isoenzymes changes with time. Gibberellic acid (GA₃) treatment causes greatly elongated internodes,decreased soluble protein, and inhibition of the rise in peroxidaseactivity within 4–8 h. Application of GA₃ to young internodesleads to a persistent depression in peroxidase activity, whiletreated older internodes suffer only a temporary depression.GA₃ causes no qualitative changes in the isoenzyme pattern butproduces some quantitative alterations in internodes in whichits influence on peroxidase activity is persistent. Decapitation of untreated and GA₃-treated dwarfs has littleinfluence on internode elongation, causes an increase in peroxidaseactivity, especially in the upper internodes, and alters therelative activity of particular isoenzymes. By contrast, decapitationinhibits elongation of young internodes in genetically tallpea plants.     

Antioxidative responses of wheat treated with realistic concentration of cadmium

Two cvs. of wheat differently sensitive to many stress factors (cv. Ofanto less sensitive than cv. Adamello) were grown in a controlled environment with cadmium near threshold concentrations supplying the metal at equal-effect concentrations. Cd excess determined in both cvs. a reduction in water and turgor potential but a maintenance of relative water content. Cv Ofanto showed a higher capacity of Cd exclusion from roots but a higher translocation to shoots in comparison with cv. Adamello. Notwithstanding the higher metal concentration in leaves of cv. Ofanto, K⁺ leakage was more pronounced in Adamello suggesting that mechanisms of Cd detoxification and tolerance such as vacuolar compartmentalisation were activated in the first one. In Adamello plants, ethylene rose at the lowest metal concentration and the activation in roots of the antioxidative enzymes catalase, ascorbate peroxidase and guaiacol peroxidase came into play whereas in Ofanto ethylene and catalase did not change. Following cadmium treatment, superoxide dismutase activity was reduced or remained at the control value in roots and in leaves. For both cultivars ascorbate peroxidase, syringaldazine peroxidase and guaiacol peroxidase activities were always higher in roots than in leaves. These activities were induced by Cd in Ofanto leaves, whereas in Adamello leaves they remained at control levels or increased somewhat at the highest metal concentration. Cadmium changed the peroxidase isozyme pattern in both cultivars. Cv. Ofanto showed, as for other stress such as drought, salinity, nickel and copper, a co-tolerance towards Cd. Analogies in the response to other metals such as copper could be found in activation of catalase at the lower metal concentration in cv. Adamello and in the induction of ascorbate peroxidase in leaves of cv. Ofanto.     

Phenols and phenol oxidases are involved in cadmium accumulation in the water plants Nymphoides peltata (Menyanthaceae) and Nymphaeae (Nymphaeaceae)

This comparative study investigates the mechanism of cadmium accumulation in the semiaquatic plant Nymphoides peltata (Menyanthaceae) and the aquatic plant Nymphaea (Nymphaeaceae). It was conducted as part of an ongoing study of the use of water plants for phytoremediation. Epidermal structures, known as hydropotes, are located on the abaxial epidermis of the leaf laminae of Nymphoides peltata and are shown to contain phenols, peroxidase and polyphenol oxidase activities. When plants are subjected to 50 mg/l of cadmium in the growth medium, these hydropotes accumulate cadmium. Cadmium-induced increases in phenols, peroxidase and polyphenol oxidase activities were determined in plant extracts. Cadmium binding by polymerized phenols was demonstrated in vivo. In comparison with Nymphaeae epidermal glands, N. peltata hydropotes are larger, open, and create bigger crystal, the latter principally composed of calcium and, proportionally, less cadmium. Although both plants showed similar levels of cadmium accumulation, N. peltata was sensitive while Nymphaeae was resistant to this cadmium level. It is suggested that in these water plants the main mechanism for cadmium accumulation is based on the trapping of cadmium crystals by polymerized phenols in specialized epidermal structures and this is due to peroxidase and polyphenol oxidase activities. Nymphaeae, with greater peroxidase activity and more polyphenols, is more resistant to this heavy metal than N. peltata.     

模拟微重力条件下几种植物的过氧化物酶同工酶谱分析

对模拟微重力条件下的人参果、马铃著和草莓处理后进行了过氧化物酮同工酶谱分析实验。结果表明,在模拟微重力条件下过氧化物同工酶的活性明显增强。     

Effect of cadmium on growth and the activity of H2O2 scavenging enzymes in Colocassia esculentum

Cadmium is a non-essential heavy metal that can be harmful even at low concentrations to plants. Colocassia esculentum (Araceae) plant was studied to know its tolerance capability to cadmium. Colocassia esculentum plants grown in pots containing different concentrations of cadmium (Cd) were analyzed for dry matter, fresh weight and total metal content. Cadmium depressed dry matter production of the plant up to 33%. Plant accumulated larger portion of the heavy metal in the roots followed by stem and leaf. Chlorophyll content of the plant declined on treatment with heavy metal while the activity of antioxidative enzymes catalase and peroxidase increased. Colocassia esculentum also showed an increase in total protein along with greater A₂₅₀/A₂₈₀ value suggesting an increase in metal protein complexes. Cadmium elicited anatomical changes in the root of C. esculentum. These changes under heavy metal stress indicate the adaptive properties of this plant species.     

光质对石刁柏愈伤组织培养中生长和过氧化物酶的影响

报道了在石刁柏愈伤组织培养中,采用５种不同光质处理,对愈伤组织生长及其过氧化物酶（ＰＯＤ）活性及同工酶的影响,实验表明,不同光质处理对生长有不同的效应,蓝光、红光下的培养物生长较快。不同光质对培养物质的ＰＯＤ活性亦有不同效应,ＰＯＤ同工酶谱亦有不同,蓝光、红光下的培养物,ＰＯＤ活性较高。ＰＯＤ活性变化与愈伤组织生长呈正相关。     

Optimalization of the peroxidase production by tissue cultures of horseradish <Emphasis Type="Italic">in vitro</Emphasis>

Tissue cultures of Armoracia rusticana L., both transformed with Agrobacterium rhizogenes and nontransformed, were screened for peroxidase activity. Most of the derived and tested strains exhibited 20 times higher activity [from 99 to 723 U g⁻¹(d.m.)] than the root of the intact plant [(30 U g⁻¹ (d.m.)]. The highest peroxidase activity was found in tumour culture growing on the medium without growth regulators. The influence of the addition of sugars and heavy metal ions in the medium on peroxidase production was tested. Increase in peroxidase activity was observed after cultivation of horseradish culture with cadmium, cobalt, nickel or lead ions.This work is supported by Grant Agency of Czech Republic Project No. 526/04/0135.     

Chemical composition and biochemical changes in needles of Scots pine (Pinus sylvestris L.) stands at different stages of decline in Bulgaria

Water content was decreased in the needles of Scots pines (Pinus sylvestris L.) with moderate and slight levels of stress-induced decline. In pines with moderate decline a misbalance of foliar nutritional elements was observed. In 1-year-old needles of afflicted pines, the content of potassium and calcium was decreased, but the content of phosphorus, magnesium, iron, manganese and of the heavy metals cadmium and lead was increased. In 2-year-old needles of damaged pines, the content of calcium, iron and of the heavy metal cadmium was decreased, but the content of phosphorus, magnesium, manganese and zinc was increased. Potassium deficiency and zinc levels above toxicity tolerance characterized the nutrient status of declined Scots pines. In the same pines chlorophyll a/b ratio was diminished in 2-year-old needles and the protein content was increased in 1-year-old needles but was decreased in 2-year-old needles.In the second phase of the study 25- and 40-year-old pines with slight decline were examined. An increase in the protein content of 2-year-old needles of 25-year-old declined pines and of both 1- and 2-year-old needles of older 40-year-old pines was observed. A 280% increase of peroxidase activity in 1-year-old and a 178% increase in 2-year-old needles of 25-year-old damaged pines were shown. In older 40-year-old damaged pines a slight increase only in 1-year-old needles was seen. The observed age-related decrease in peroxidase activity in older 40-year-old symptomatic pines might be responsible for the lower resistance to decline in older pines. Our results show that clear but complex chemical and biochemical changes are observed in the needles of Scots pines in Bulgaria and that these changes hold promise for use as indicators of stress.     

The vacuolar localization of a basic peroxidase isoenzyme responsible for the synthesis of α-31,41-anhydrovinblastine in Catharanthus roseus (L.) G. Don Leaves

Partially purified protein extracts of Catharanthus roseus leaves were able to couple catharanthine and vindoline to produce α-3′,4′-anhydrovinblastine (AVLB) in a reaction strictly dependent on H₂O₂. This result, and the co-purification of peroxidase with AVLB synthetase activity, strongly suggest a peroxidase-like nature for the coupling enzyme. Only one peroxidase isoenzyme was detected in C. roseus leaves, and it was shown that this isoenzyme consists of a molecularly-heterogeneous basic peroxidase (EC 1-11-1-7) mainly located in the vacuole. These results suggest that a basic peroxidase located in the vacuole may be the main enzyme responsible for AVLB synthesis in C. roseus leaves. This isoenzyme was also found in cell walls where a peroxidase inhibitor was detected.     

Peroxidase isoenzymes in Linum

Summary Isoenzymes of peroxidase were separated on acrylamide gels in 2 genotypes of Linum usitatissimum L. and their F ₁, F ₂ and first backcross progeny. Active extracts were obtained from homogenates of main stem tissue; activity was measured both before and after electrophoretic separation. The relationship of isoenzyme activity to gross (prior to electrophoretic separation) activity was investigated, as well as the relative behaviour of isoenzyme activity in the various genotypes and generations. Gross activity was correlated with isoenzyme activity; there was also evidence of maternal as well as genetic effects on isoenzyme activity.     

大别山区六种黄精属植物的五种同工酶分析

采用聚丙烯酰胺凝胶电泳法对皖西大别山区六种黄精属植物的抗坏血酸氧化酶(AOD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)、酯酶(EST)和过氧化物酶(POD)五种同工酶进行了比较分析。结果表明:(1)五种同工酶共显示出66条酶带,其中有3条是黄精属的特征酶带;并且来源于不同种的同一种酶的谱带数、相对迁移率、酶活性均不相同,呈现多样性;(2)五种同工酶谱的模糊聚类分析结果与形态学分类的结果一致,利用酶谱差异可以将六种黄精初步区分;(3)在六种黄精属植物中,湖北黄精、轮叶黄精、多花黄精和金寨黄精是较进化类型,玉竹和长梗黄精是较原始类型。