Effects of short-term ozone fumigation on tobacco plants: response of the scavenging system and expression of the glutathione reductase

The role that the constituents of the ascorbate–glutathione cycle play in the mechanism of contrasting ozone sensitivities was examined in mature and old tobacco leaves after acute ozone-fumigation (150 p.p.b., 5 h). Levels of the enzyme activities associated with the detoxifying system were lower in ozone-sensitive Bel W3 control plants than in unfumigated ozone-tolerant Bel B plants. In particular, the endogenous activities of ascorbate peroxidase (APX) and glutathione reductase (GR), and the metabolites ascorbic acid (AA) and reduced glutathione (GSH) were more abundant in Bel B than Bel W3 control plants. These results suggest that the higher tolerance of Bel B to O₃ is associated with a greater initial content of the antioxidant enzymes or metabolites. Only in the mature leaves of the ozone-tolerant Bel B cv. did fumigation trigger activation of APX and, weakly, of dehydroascorbate reductase (DHAR). The activity of these enzymes was significantly lower after ozone treatment in both mature and old leaves of Bel W3 than in control plants. Fumigation had little effect on the ascorbate content. Its main effects on the glutathione pool were that it boosted the oxidized form and lowered the reduced form, particularly in mature Bel W3 leaves. Extractable GR activity remained unchanged in both Bel B and Bel W3 immediately after fumigation, but increased slightly 24 h later, particularly in mature leaves of Bel W3. Exposure to O₃ caused a sharp decline in chloroplastic GR mRNA levels in both cultivars. However, as Western blot analysis failed to detect any major changes in GR protein content at this time, the protein must be highly stable. There is therefore a good correlation between tolerance to O₃ and high endogenous levels of antioxidant metabolites such as AA and GSH in tobacco. In addition, the degree of inducibility of the system discriminates the two cultivars investigated.     

CO2 assimilation, xanthophyll cycle pigments and PSII efficiency in pumpkin plants as affected by ozone fumigation

CO₂ assimilation, xanthophyll cycle pigments and PSII efficiency were analyzed in two different ages of pumpkin leaves (Cucurbita pepo L. cv. Ambassador) exposed to 150 nmol mol^-1 of ozone (5 days, 5 h day^-1). Gas-exchange measurements revealed a reduction in CO₂ assimilation and stomatal conductance, accompanied by an increase in the intercellular CO₂ concentration both in young and in mature leaves as compared to their respective controls. In both leaves, F₀ remained unchanged, while F_m and the F_v/F_m ratio decreased after O₃ fumigation, indicating that ozone may induce an alteration in the capability of photosystem II (PSII) to reduce the primary acceptor Q_A. In the mature leaves the photochemical quenching (q_p) was significantly lowered by the pollutant, but this was not the case in the young leaves where q_p did not change. In both mature and young ozonated pumpkin leaves, the development of non-photochemical quenching caused a decrease in the PSII photochemical rate, as shown by the correlation between F_v/F_m and the de-epoxidation state of dark-adapted leaves. Decreases in the F_v/F_m ratio are generally attributed to damage to the PSII reaction centre, apart from the down-regulation of the capacity of PSII electron transport. While in young ozonated leaves the decrease in the F_v/F_m ratio was not associated with damage to the D1 protein, in mature ozonated pumpkin leaves, the decrease in the F_v/F_m was accompanied by a significant decline in the D1 content. In conclusion, ozone exposure induces alterations in the light reactions of photosynthesis in both young and mature leaves. However, in young leaves the engagement of the xanthophyll cycle appears to counteract ozone effects against the photosynthetic apparatus as demonstrated by the absence of damage to the D1 protein. On the other hand, the loss of D1 protein in mature fumigated leaves suggests that the activation of the xanthophyll cycle is not sufficient to prevent photoinhibition, probably because a physiological state of senescence adds to the oxidative stress.     

Peroxidase Release Induced by Ozone in Sedum album Leaves: Involvement of Ca

The effect of ozone was studied on the peroxidase activity from various compartments of Sedum album leaves (epidermis, intercellular fluid, residual cell material, and total cell material). The greatest increase following a 2-hour ozone exposure (0.4 microliters O(3) per liter) was observed in extracellular peroxidases. Most of the main bands of peroxidase activity separated by isoelectric focusing exhibited an increase upon exposure to ozone. Incubation experiments with isolated peeled or unpeeled leaves showed that leaves from ozone-treated plants release much more peroxidases in the medium than untreated leaves. The withdrawal of Ca(2+) ions reduced the level of extracellular peroxidase activity either in whole plants or in incubation experiments. This reduction and the activation obtained after addition of Ca(2+) resulted from a direct requirement of Ca(2+) by the enzyme and from an effect of Ca(2+) on peroxidase secretion. The ionophore A23187 promoted an increase of extracellular peroxidase activity only in untreated plants. The release of peroxidases by untreated and ozone-treated leaves is considerably lowered by metabolic inhibitors (3-(3,4-dichlorophenyl)-1,1-dimethylurea and sodium azide) and by puromycin.     

Involvement of the thylakoidal NADH-plastoquinone-oxidoreductase complex in the early responses to ozone exposure of barley (Hordeum vulgare L.) seedlings

A possible implication of the plastid NADH-plastoquinone-oxidoreductase (Ndh) complex in the response against ozone-mediated oxidative stress in barley (Hordeum vulgare L.) leaves was investigated. After a 4 h treatment, exposure of barley seedlings to moderate ozone concentrations produced leaf-age-dependent increases in lipid peroxidation, peroxidase, and Ndh complex activities in the thylakoid membranes. A significant amount and activity of the Ndh complex were detected in mature barley leaves, but not in young barley leaves. In fact, young barley leaves behaved like ndh-deficient leaves in most of the aspects studied. When plants were exposed to photo-oxidative light after ozone fumigation, the recovery of Fv/Fm was lower in young leaves than in mature leaves. Ozone treatment significantly decreased non-photochemical quenching (qN) in young leaves, but not in mature leaves. Mature leaves showed higher levels of the energy (DeltamuH+) dependent (qE) component of qN. Treatment with antimycin A, an inhibitor of cyclic electron flow, increased the decay of qN produced by ozone in young leaves, but not in mature ones. The reduction state of plastoquinone increased after ozone treatment in mature dark-adapted leaves and was strongly quenched by far red light. It is proposed that the function of the Ndh complex helps the maintenance of qN, probably through the poising of the redox steady-state level of the intersystem carriers and then by optimizing the rate of cyclic electron flow. This should constitute an age-dependent early response in barley leaves, by contributing to minimize photoinhibition in the presence of ozone and high light.     

Expression of arabidopsis cytosolic ascorbate peroxidase gene in response to ozone or sulfur dioxide

The effects of ozone or sulfur dioxide on antioxidant enzymes were investigated in Arabidopsis thaliana. Plants were fumigated with 0.1–0.15 ppm ozone or sulfur dioxide up to about 1 week in an environment-controlled chamber. Both pollutants increased the activities of ascorbate peroxidase and guaiacol per-oxidase in leaves, but had little effect on the activities of superoxide dismutase, catalase, monodehydroascorbate reductase, dehydroascorbate reductase or glutathione reductase. Ozone was more effective than sulfur dioxide in increasing the activities of the peroxidases. Ascorbate peroxidase activity increased 1.8-fold without a lag period during fumigation with 0.1 ppm ozone, while guaiacol peroxidase activity increased 4.4-fold with a 1-day lag. Expression of the APX1 gene encoding cytosolic ascorbate peroxidase was further investigated. Its protein levels in leaves exposed to 0.1 ppm ozone for 4 or 8 days were 1.5-fold higher than in controls. Both ozone and sulfur dioxide elevated APX1 mRNA levels in leaves at 4 and 7 days, whereas at 1 day only ozone was effective. The induction of APX1 mRNA levels by ozone (3.4- to 4.1-fold) was more prominent than that by sulfur dioxide (1.6-to 2.6-fold). The APX1 mRNA level increased by day and decreased by night. Exposure of plants to 0.1 ppm ozone enhanced the APX1 mRNA level within 3 h, which showed a diurnal rhythm similar to that of the control. These results demonstrate that near-ambient concentrations of ozone as well as similar concentrations of sulfur dioxide can induce APX1 gene expression in A. thaliana.Environmental Biology Division     

Role of Ascorbate in Detoxifying Ozone in the Apoplast of Spinach (Spinacia oleracea L.) Leaves

Both reduced and oxidized ascorbate (AA and DHA) are present in the aqueous phase of the extracellular space, the apoplast, of spinach (Spinacia oleracea L.) leaves. Fumigation with 0.3 [mu]L L-1 of ozone resulted in ozone uptake by the leaves close to 0.9 pmol cm-2 of leaf surface area s-1. Apoplastic AA was slowly oxidized by ozone. The initial decrease of apoplastic AA was <0.1 pmol cm-2 s-1. The apoplastic ratio of AA to (AA + DHA) decreased within 6 h of fumigation from 0.9 to 0.1. Initially, the concentration of (AA + DHA) did not change in the apoplast, but when fumigation was continued, DHA increased and AA remained at a very low constant level. After fumigation was discontinued, DHA decreased very slowly in the apoplast, reaching control level after 70 h. The data show that insufficient AA reached the apoplast from the cytosol to detoxify ozone in the apoplast when the ozone flux into the leaves was 0.9 pmol cm-2 s-1. The transport of DHA back into the cytosol was slower than AA transport into the apoplast. No dehydroascorbate reductase activity could be detected in the apoplast of spinach leaves. In contrast to its extracellular redox state, the intracellular redox state of AA did not change appreciably during a 24-h fumigation period. However, intracellular glutathi-one became slowly oxidized. At the beginning of fumigation, 90% of the total glutathione was reduced. Only 10% was reduced after 24-h exposure of the leaves to 0.3 [mu]L L-1 of ozone. Necrotic leaf damage started to become visible when fumigation was extended beyond a 24-h period. A close correlation between the extent of damage, on the one hand, and the AA content and the ascorbate redox state of whole leaves, on the other, was observed after 48 h of fumigation. Only the youngest leaves that contained high ascorbate concentrations did not exhibit necrotic leaf damage after 48 h.     

Balance between anionic and cationic extracellular peroxidase activities in Sedum album leaves after ozone exposure. Analysis by high-performance liquid chromatography

Peroxidase activity was assayed with different electron donors (guaiacol, ascorbate, syringaldazine) in the intercellular fluid of Sedum album L. leaves after ozone exposure. Anionic and cationic peroxidases were separated and purified by high performance ion-exchange and gel permeation chromatography. Both isoperoxidases were tested as regards their molecular weight and apparent kinetic constants with different substrates. Ascorbate peroxidase activity was rapidly stimulated after ozone exposure, whereas syringaldazine peroxidase activity reached its maximum 24 h later. Increases in ascorbate and syringaldazine peroxidase activities occurred simultaneously with increases in cationic and anionic peroxidase activities, respectively. Apparent K_m values indicate a high affinity of cationic peroxidases for ascorbate and of anionic peroxidases for syringaldazine. The metabolic role of this balance between cationic and anionic peroxidases after ozone exposure is discussed.     

Iron deficiency differently affects peroxidase isoforms in sunflower

The response of both specific (ascorbate peroxidase, APX) and unspecific (POD) peroxidases and H(2)O(2) content of sunflower plants (Helianthus annuus L. cv. Hor) grown hydroponically with (C) or without (-Fe) iron in the nutrient solution were analysed to verify whether iron deficiency led to cell oxidative status. In -Fe leaves a significant increase of H(2)O(2) content was detected, a result confirmed by electron microscopy analysis. As regards extracellular peroxidases, while APX activity significantly decreased, no change was observed in either soluble guaiacol or syringaldazine-dependent POD activity following iron starvation. Moreover, guaiacol-dependent POD activity was found to decrease in both ionically and covalently-cell-wall bound fractions, while syringaldazine-POD activity decreased only in the covalently-bound fraction. At the intracellular level both guaiacol-POD and APX activities underwent a significant decrease. The overall reduction of peroxidase activity was confirmed by the electrophoretic separation of POD isoforms and, at the extracellular level, by cytochemical localization of peroxidases by diaminobenzidine staining. The electrophoretic separation, besides quantitative differences, also revealed quantitative changes, particularly evident for ionically and covalently-bound fractions. Therefore, in sunflower plants, iron deficiency seems to affect the different peroxidase isoenzymes to different extents and to induce a secondary oxidative stress, as indicated by the increased levels of H(2)O(2). However, owing to the almost completely lack of catalytic iron capable of triggering the Fenton reaction, iron-deficient sunflower plants are probably still sufficiently protected against oxidative stress.     

Thiol-dependent degradation of protoporphyrin IX by plant peroxidases

Protoporphyrin IX (PP) is the last porphyrin intermediate in common between heme and chlorophyll biosynthesis. This pigment normally does not accumulate in plants because its highly photodynamic nature makes it toxic. While the steps leading to heme and chlorophylls are well characterized, relatively little is known of the metabolic fate of excess PP in plants. We have discovered that plant peroxidases can rapidly degrade this pigment in the presence of thiol-containing substrates such as glutathione and cysteine. This thiol-dependent degradation of PP by horseradish peroxidase consumes oxygen and is inhibited by ascorbic acid.     

Species- and age-dependent sensitivity to ozone in young plants of pea,wheat and spinach: Effects on acyl lipid and pigment content and metabolism

Acyl lipids and pigments were analyzed in young plants of garden pea, spring wheat and spinach exposed to < 5 or 65 nl l^?1 ozone 12 h per day for 6 days. In one set of experiments, the plants were exposed to ¹⁴CO₂ for 2 h 3 days prior to ozone exposure. The plants responded differently to the moderately enhanced level of ozone used Spinach was not at all sensitive while in both pea and wheat, leaves of different ages differed in ozone sensitivity. In pea, ozone sensitivity increased with leaf age. In the second and third oldest leaves, the amounts of galactolipids per leaf area and the proportions of 18:3 of the total lipid extract and of phosphatidylglycerol decreased. In the second oldest leaf, ozone also caused a decreased proportion of 18:3 of monogalactosyldiacylglycerol. In the fourth oldest leaf, lipid composition and galactolipid unsaturation was unaffected, but ozone caused decreased leaf expansion resulting in increased acyl lipid content per leaf area. In both the first and second leaves of wheat, ozone fumigation caused a marked decrease in the content of monogalactosyldiacylglycerol and in the first leaf, the contents of phosphatidylcholine and phosphatidylethanolamine increased. The proportion of 18:3 in phosphatidylcholine was larger in ozone-fumigated than in control plants, while the reverse applied for phosphatidylglycerol. In the oldest sampled leaves of pea and wheat, ozone caused an increase in the radioactivity associated with β-carotene, indicating increased turnover. Thus, while spinach was unaffected, in both pea and wheat ozone caused a decrease in the proportion of chloroplast membrane lipids to non-chloroplast membrane lipids in older leaves while younger leaves were less sensitive.     

Antioxidant metabolite levels in ozone-sensitive and tolerant genotypes of snap bean

Ozone-sensitive and tolerant genotypes of snap bean ( Phaseolus vulgaris L.) were compared for differences in leaf ascorbic acid (vitamin C), glutathione and α -tocopherol (vitamin E) content to determine whether antioxidant levels were related to ozone tolerance. Seven genotypes were grown in pots under field conditions during the months of June and July. Open top chambers were used to establish either a charcoal filtered (CF) air control (36 nmol mol⁻¹ ozone) or a treatment where CF air was supplemented with ozone from 8:00 to 20:00 h with a daily 12 h mean of 77 nmol mol⁻¹. Fully expanded leaves were analyzed for ascorbic acid, chlorophyll, glutathione, guaiacol peroxidase (EC 1.11.1.7) and α -tocopherol. Leaf ascorbic acid was the only variable identified as a potential factor in ozone tolerance. Tolerant genotypes contained more ascorbic acid than sensitive lines, but the differences were not always statistically significant. Genetic differences in glutathione and α -tocopherol were also observed, but no relationship with ozone tolerance was found. Guaiacol peroxidase activity and leaf α -tocopherol content increased in all genotypes following a one week ozone exposure, indicative of a general ozone stress response. Ozone had little effect on the other variables tested. Overall, ozone sensitive and tolerant plants were not clearly distinguished by differences in leaf antioxidant content. The evidence suggests that screening for ozone tolerance based on antioxidant content is not a reliable approach.     

二氧化硫污染对植物体酶活性的影响

通过用６种浓度的ＳＯ２气体对６种植物进行熏气处理,然后测定植物叶片内的过氧化物酶,抗坏血酸氧化酶及多酚氧化酶的活性,为环境监测提供理论依据。通过试验,发现ＳＯ２气体对植物叶内不同酶的活性的影响有别。６种植物受不同浓度的ＳＯ２污染,叶内的过氧化物酶活性随ＳＯ２浓度增加而增加,促进叶片的衰老;而对其他的２种酶,则６种植物表现不同     

小麦与条锈菌互作中过氧化物酶的细胞化学研究

采用细胞化学方法对小麦与条锈菌互作过程中过氧化物酶的分布及其活性大小进行了研究,结果表明：过氧化物酶主要分布于细胞壁和细胞间隙中;在未行接种的小麦叶片中,抗病品种和感病品种的过氧化物酶活性均比较低;条锈菌侵染后,诱导抗、感病品种叶片中的过氧化物酶活性升高,且抗病品种升高的幅度明显大于感病品种;感病品种中过氧化物酶活性在侵染位点附近细胞壁上表现升高,而抗病品种中该酶的活性在侵染点细胞以及远离侵染点的叶肉细胞的细胞壁和细胞间隙中均显著升高。高活性的过氧化物酶是小麦抗条锈性的生化标记和重要机制之一。     

Ozone detoxification in the apoplasm and symplasm of spinach,broad bean and beech leaves at ambient and elevated concentrations of ozone in air

Spinach (Spinacia oleracea L.), broad bean (Vicia faba L.) and beech (Fagus sylvatica L.) plants were exposed to ozone at concentrations often measured in air during the summer months (120–300 g·m^–3) and antioxidants were determined in the leaf tissue and in the aqueous phase of the cell wall, the apoplasm. Concentrations of both reduced ascorbate (AA) and its oxidized form, dehydroascorbate (DHA), showed the tendency to increase transiently in the apoplasm of spinach leaves 6–24 h after starting fumigation with ozone. In beech leaves, apoplasmic AA and DHA increased 3–7 d after beginning of treatment. At the very high concentration of 1600 g O₃·m^–3, an increase of apoplasmic AA was already measured after 1 d in beech leaves. Apparently, spinach and beech leaves respond to oxidative stress by increasing AA transport into the apoplasm and by accelerating DHA export. In contrast to these observations, DHA accumulated during 3 d of fumigation with only 120 g O₃·m^–3 in the apoplasm of broad bean leaves, while AA contents did not increase. After termination of fumigation, the extracellular redox state of ascorbate normalized within 1 d. Glutathione could not be detected in the apoplasm of any of the three leaf species. Intracellular AA changed its redox state in response to exposure to elevated concentrations of ozone. After 4–6 weeks of fumigation with 200–300 g O₃·m^–3 an increase of intracellular DHA was measured in beech leaves. At the same time, chlorophyll contents decreased and characteristic symptoms of ozone damage could be observed. However, no significant change in the redox state of apoplasmic ascorbate could be detected in beech leaves. Evidently, detoxification of ozone by apoplasmic AA was insufficient to protect the leaf tissue. Fumigation with a high ozone concentration (1600 g·m^–3) caused an appreciable increase in the cellular contents of the oxidized forms of ascorbate and glutathione in beech leaves. Whereas in spinach leaves intracellular antioxidant contents and redox states were not altered during fumigation with 120–240 g O₃·m^–3, in broad bean leaves the intracellular DHA concentration increased and intracellular ascorbate became more oxidized after fumigation of the plants with 120 g O₃·m^–3. Apparently, broad bean leaves are more sensitive to ozone than beech and spinach leaves.Abbreviations AA ascorbate, reduced form - DHA ascorbate, oxidized form (dehydroascorbate) - FW fresh weight - GSH glutathione, reduced form - GSSG glutathione, oxidized form - IWF intercellular washing fluid - V_air intercellular air space volume of leaves - V_apo apoplasmic water volume of leaves This work was supported within the Sonderforschungsbereich 251 of the University of Würzburg.     

The fungitoxicities of cuticular and cellular components of citrus lime leaves

Cuticular and cellular components of leaves of Citrus aurantifolia Swingle were tested for toxicity to Gloeosporium limetticola Clausen, responsible for the withertip disease. Cuticular waxes isolated from mature, resistant leaves showed little activity. A dimethoxy furocoumarin, isopimpinellin, isolated from the cellular components, was highly toxic, and ester and acidic fractions obtained from both young susceptible and mature resistant leaves showed some activity. The contents in the leaves and the toxicities in vitro of the active constituents suggest that they may contribute, to an important degree, to the defence of the leaf against G. limetticola.     

Changes in phenol and peroxidase in the leaves ofJava citronella infected withCurvularia andropogonis

The contet of phenols, o—dihydroxyphenols and peroxidase activity in healthy andCurvularia andropogonis (Zimm.) Boedijn infected leaves ofJava citronella were determined. As a result of infection, the content of phenols and peroxidase increased two- and four-fold, respectively in necrotic lesions compared to healthy leaves. In surrounding tissue of lesions, their increase was one-and half fold only. The peroxidase activity decreased with the maturity of the necrotic lesions. Necrotic lesions produced in response to infection appear to be the consequence of higher accumulation of phenols and their oxidation by peroxidase.     

Effects of 2-month ozone exposure in spinach leaves on photosynthesis,antioxidant systems and lipid peroxidation

The photosynthesis response, antioxidant systems and lipid peroxidation were studied in leaves from spinach plants (Spinacia oleracea L.) in response to ozone fumigation, ambient air and charcoal filtered air treatments. The photosynthetic activity was tested through gas exchange and chlorophyll a fluorescence measurements. Ambient air and ozone fumigation caused a decrease in the photosynthetic rate (25% and 63%, respectively) mainly due to a reduced mesophyll activity, as evidenced by the increased intercellular CO₂ concentration. These data agree with a large reduction in the non-cyclic electron flow (7% and 16%), a lower capacity to reduce the quinone pool and a higher development of non-photochemical quenching upon high O₃ concentration. The results suggest that the oxidative stress produced, together with the stimulation of superoxide dismutase (SOD, EC 1.15.1.1) and ascorbate peroxidase (APX, EC 1.11.1.11) activities and the increase in lipid peroxidation (20% and 36%, respectively), generated an alteration of the membrane properties.     

Differential distribution of leaf chemistry in eucalypt seedlings due to variation in whole-plant nutrient availability

We investigated the effects of whole-plant nutrient-availability on the degree of distribution of some plant primary and secondary (nitrogen, fibre, flavonols, gallotannins and cineole) chemicals across young, mature and old leaves of seedlings of Eucalyptus nitens. Four treatments that ranged from low to high nutrient-application rates resulted in mean whole-plant foliar concentrations of 0.63%, 0.85%, 1.11% and 1.82% nitrogen dry matter (N%DM) for treatments A, B, C and D, respectively. Within-plant distribution (across the leaf age profile of young, mature and old leaves within a eucalypt seedling) of N%DM ranged from zero in treatment A to a wide range of distribution in treatment D (low N%DM concentrations in old leaves to high N%DM concentrations in young leaves). Similarly, the distribution of fibre ranged from zero in treatment A to a wide range of distribution in treatment D, but with high concentrations in old leaves and low concentrations in young leaves. In contrast, flavonols (weakly) and gallotannins had a wide range of distribution in treatment A (low concentrations in old leaves to high concentrations in young leaves) but were little or not distributed in the other treatments. Again in contrast, cineole was strongly distributed between old and young leaves (low concentrations in old leaves to high concentrations in young leaves) across all treatments while concentrations in mature leaves reflected one or other leaf age depending on treatment. Protein precipitable phenols in treatments A, B and C were high in young, and low in old leaves; whereas in treatment D they were low and similar between leaves of different ages.     

夏季南亚热带森林演替中后期优势种幼叶花色素苷的光保护作用

为探讨夏季南亚热带森林演替过程中优势树种幼叶的光保护机制,以演替中期优势树种木荷(Schima superba)、黧蒴(Castanopsis fissa)、锥栗(C.chinensis)和演替后期优势种华润楠(Machilus chinensis)、厚壳桂(Cryptocarya chinensis)、黄果厚壳桂(C.concinna)为材料,分析了2种生长光强(全光照和30%全光照)下6种优势种幼叶和成熟叶的叶片表型、光合色素含量、花色素苷含量、抗氧化能力、类黄酮含量、总酚含量和最大量子产量(Fv/Fm)恢复效率间的差异。结果表明,两个演替阶段幼叶的叶绿素含量(Chl a+b)、Chl a/b比成熟叶低,但光保护物质比成熟叶多;演替中期幼叶的花色素苷含量和总抗氧化能力比演替后期的高,而类黄酮和总酚含量比演替后期的低;全光照下幼叶的总酚、类黄酮、总抗氧化能力及Fv/Fm恢复效率都要比30%全光照的高,并且含有花色素苷的幼叶恢复得更快。因此,植物的光合能力与自身的光保护潜力成反比关系,演替中期优势种幼叶的光保护在很大程度上是因为花色素苷的积累而演替后期优势种是因为自身抗氧化物质(类黄酮、总酚)的共同作用。     

Apoplastic peroxidases and ascorbate are involved in manganese toxicity and tolerance of Vigna unguiculata

Excessive manganese (Mn) supply induced the formation of brown spots on leaves as typical Mn toxicity symptoms in cowpea ( Vigna unguiculata L. Walp.) grown in hydroponics. Differences in Mn resistance between cv. TVu 91 (Mn-sensitive) and cv. TVu 1987 (Mn-tolerant) expressed in the density of brown spots in older leaves were due to higher Mn tissue tolerance. Apoplastic water-soluble peroxidase (POD) in the apoplastic washing fluid (AWF) was enhanced by increasing Mn leaf content and generally significantly higher in leaves of cv. TVu 91 than in cv. TVu 1987. Electrophoresis of AWF revealed the presence of several water-soluble POD isoenzymes. At toxic Mn supply, the activities of these and additional POD isoenzymes increased more in the Mn-sensitive cultivar. Levels of ascorbic acid in the apoplast and cytoplasm of the Mn-sensitive cv. TVu 91 decreased with increasing leaf Mn contents, whereas Mn-tolerant cv. TVu 1987 was not affected. Mn treatment lead to a stimulation of the enzymes of the ascorbic acid regeneration system (monodehydroascorbic acid reductase and glutathione reductase) in both cultivars, but the activation of glutathione reductase was clearly more enhanced in the Mn-tolerant cultivar TVu 1987. The results provide circumstantial evidence that apoplastic ascorbate and peroxidases are involved in the expression of Mn toxicity and genotypic Mn tolerance.