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1.
Adverse conditions often induce an increase in active oxygen species (AOS) such as hydrogen peroxide (H2O2). H2O2 is converted to water, and thus becomes detoxified by enzymes such as ascorbate peroxidase (APX; EC 1.11.1.11). APX activity is estimated by the disappearance rate of ascorbic acid, which becomes oxidized. However, ascorbate is also a substrate of guaiacol peroxidase (POX; EC 1.11.1.7). POX oxidizes phenols (including flavonoids), whereby ascorbate accepts an electron from phenoxyl or flavonoid radicals. Ascorbate becomes thereby converted to the monodehydroascorbate radical, which subsequently can become converted to dehydroascorbate. POX isozymes therefore convert hydrogen peroxide to water and oxidize ascorbate, just as APX does. POX activity is usually estimated by monitoring the formation of tetraguaiacol from guaiacol. This reaction is not specific, as some APX isozymes show rather high activity when using guaiacol or similar phenols as a substrate. It is concluded that APX activity can easily be confounded with POX activity, and vice versa. Proper methods should be used to separate the two enzyme activities.  相似文献   

2.
Pheophytinase (PPH) activity and gene expression of chlorophyll (Chl)-degrading enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica Group) florets were determined. PPH is involved in the dephytylation of Mg-free Chl a, pheophytin (Phy) a. However, in vitro chlorophyllase (Chlase, EC.3.1.1.14) also uses Phy a as a substrate to produce pheophorbide (Pheide) a by dephytylation. For an accurate determination of PPH activity, the PPH protein fraction was separated from Chlase protein by ammonium sulfate precipitation. The protein precipitated by 45-60% saturated ammonium sulfate included a little bit of Chlase activity and was suitable for PPH determination. PPH activity in broccoli florets treated with a UV-B dose of 19 kJ m−2 was repressed for the first 2 d of storage at 15 °C, whereas it increased gradually with senescence of control broccoli florets. The expression level of BoCLH1 was reduced in broccoli florets on day 4 of storage, while BoCLH2 and BoCLH3 were up-regulated with UV-B treatment. A high BoPAO expression level was found in senescent broccoli florets, and the up-regulation of this gene was delayed by UV-B treatment. The highest expression level of BoPPH was found in the control, and its expression was clearly repressed by UV-B treatment on day 2 of storage. We suggest that the up-regulation of Chl-degrading enzyme genes could be delayed by UV-B treatment, resulting in the suppression of floret yellowing in stored broccoli.  相似文献   

3.
4.
The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g?1 DW at 10 μm Cu and a value of 4415 μg g?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H2O2, O2˙? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.  相似文献   

5.
Postharvest yellowing of broccoli (Brassica oleracea L. Italica Group) is an important indicator of quality deterioration and occurs with chlorophyll (Chl) degradation. Postharvest ethanol vapor treatment could delay the yellowing of broccoli florets, through the suppression of Chl degradation. As the first step in identifying the mechanism of this delay, the effects of postharvest ethanol vapor treatment on activities and gene expression of the Chl catabolic enzymes in broccoli were determined. Broccoli branchlets were placed in a perforated polyethylene bag with or without (control) an ethanol pad and stored at 20 °C in darkness. The Chl contents of the control broccoli florets decreased remarkably after three days in storage, whereas the contents of the ethanol-treated broccoli showed no significant changes except at day one. Changes in chlorophyllase activity in the ethanol-treated broccoli were similar to those of the control until three days in storage, but then the activity tended to decease. Mg-dechelatase and Chl-degrading peroxidase activities, which increased greatly with senescence in the controls, remained unchanged in the ethanol-treated broccoli. BoCLH1 expression showed changes almost similar to those of chlorophyllase activity. BoPAO expression in the control broccoli increased greatly at day one in storage and was maintained at a high level until three days. In contrast, this expression in ethanol-treated broccoli was suppressed until two days. BoRCCR expression in the control broccoli increased until two days in storage, while the expression in the ethanol-treated broccoli showed no change during storage. These results show that postharvest ethanol vapor treatment suppressed the activities and gene expression of Chl catabolic enzymes, resulting in delayed yellowing of broccoli florets.  相似文献   

6.
Leaf senescence is an oxidative process, and most of the catabolic processes involved in senescence are propagated irreversibly once initiated. An experiment was conducted to test the hypothesis that nitrophenolates (Atonik, a plant growth regulator) spray can delay the leaf senescence through reduced oxidative damage. Atonik 3.75 g a.i. ha?1 was sprayed during boll filling stage on cotton, and the senescence process was evaluated by quantifying total chlorophyll contents, photosynthetic rate, Fv/Fm ratio, various reactive oxygen species (ROS) content, antioxidant content and antioxidant enzyme activity from 90 days after sowing (DAS) to 130 DAS. The result indicated that nitrophenolate spray reduced the hydrogen peroxide (H2O2), superoxide anion (O2?) accumulation, lipid peroxidation (malondialdehyde), lipoxygenase activity and membrane permeability over unsprayed control. The antioxidant enzyme activity (superoxide dismutase, SOD; ascorbate peroxidase, APX; peroxidase, POX; glutathione peroxidase, GSH‐Px) were significantly increased by the nitrophenolate spray. POX (118.1 %) and GSH‐Px (143.3 %) activities were enhanced to a higher level compared to APX (8.5 %) activity at 130 DAS. Enhanced accumulation of ascorbate (144.9 %), phenol (154.7 %) and proline (50 %) was seen in nitrophenolate‐sprayed plants compared with unsprayed control plants at 130 DAS. Ascorbate content is increased by higher dehydroascorbate reductase enzyme activity. Ascorbate was thus able to replenish reducing equivalents to phenoxyl radicals resulting in an increase in phenolic compounds. The increased phenolic acid content may be involved in scavenging the ROS produced during senescence process. The higher level of reduced ascorbate and low level of endogenous H2O2 in the leaves may be the prerequisite for delayed leaf senescence in the nitrophenolate‐sprayed plants. Based on the present work, it can be concluded that nitrophenolate‐sprayed plants can postpone the leaf senescence by peroxide/phenolic/ascorbate system which is involved in scavenging the ROS produced during leaf senescence.  相似文献   

7.
8.
Degreening caused by chlorophyll degradation is the most important feature that determines postharvest loss of quality in broccoli. Chlorophyll molecules are assembled to several thylakoid proteins, from which chlorophylls must be released in order to be catabolized. Stay-Green (SGR), a chloroplast-located protein, specifically interacts with light harvesting complex subunits helping toward their destabilization and to the release of chlorophylls. In this work, a fragment of a gene encoding a SGR from broccoli (BoSGR) was cloned. The expression of BoSGR was analyzed and detected an important increment during postharvest senescence, simultaneously with chlorophyll degradation. In order to analyze the effect of different growth regulators, different groups of broccoli heads were treated with cytokinins, ethylene and 1-MCP. Cytokinins and 1-MCP delayed the increment of BoSGR expression whereas ethylene accelerated the process. In addition, several postharvest treatments that delay degreening in broccoli florets were applied to evaluate their effects on BoSGR expression. Samples treated with modified atmosphere, hot air, UV-C or white lights showed a delay in chlorophyll degradation and degreening. In most cases, the treatments also delayed the increment of BoSGR expression during senescence, reaffirming the relationship between the expression of this gene and chlorophyll degradation.  相似文献   

9.
The effects of edible coatings and mild heat shocks on quality aspects of refrigerated broccoli were studied. Minimally processed broccoli was coated with either chitosan or carboxymethyl-cellulose with or without a previous application of a mild heat shock of 1.5 min at 50 °C. Product was packaged in multilayered polyolefin bags and stored at 5 °C for 18 d. Quality parameters such as weight loss, texture, colour, ascorbic acid content, total chlorophyll content, oxygen concentration inside the bags, browning potential, mesophilic aerobic counts, and sensory quality, were evaluated during storage. Edible coatings exhibited a beneficial impact on broccoli quality. The weight loss in uncoated broccoli was found to be between 2 and 5 times higher compared to coated samples. During storage, coated florets from both thermally and non-thermally treated samples, presented higher retention of the (?a*/b*) ratio indicating better green colour retention and a reduced rate of floret yellowing. Chitosan coating always presented the lower ascorbic acid degradation rates (twofold lower compared with control samples). Broccoli texture for uncoated samples increased significantly during storage. However, for carboxymethyl-cellulose coated broccoli a slight increase in texture was observed while for chitosan coated broccoli no significant changes in texture were observed throughout the storage period. After the edible coating application the microbial broccoli load dropped by around 1.5 and 0.9 logarithmic units in chitosan and carboxymethyl-cellulose films, respectively. During storage, the application of chitosan coating significantly reduced total microbial counts in the thermally and non-thermally treated uncoated samples. Among the assayed edible coatings, chitosan effectively maintained quality attributes and extended shelf life of minimally processed broccoli. The single application of a mild heat shock had a measurable influence in reducing weight loss, enzymatic browning in broccoli stems, and in delaying yellowing of broccoli florets. Moreover, chitosan coating combined with a mild heat shock showed the best performance for long-term refrigerated storage of minimally processed broccoli.  相似文献   

10.
Maize (Zea mays L. cv. 777) plants grown in hydroponic culture were treated with 50 μm CdSO4. Growth and metabolic parameters indicative of oxidative stress and antioxidant responses were studied in leaves of plants treated with Cd. Apart from increasing lipid peroxidation and H2O2 accumulation, supply of Cd suppressed growth, fresh and dry mass of plants and decreased the concentrations of chloroplastic pigments. The activities of catalase (CAT; EC 1.11.1.6), peroxidase (POD; EC 1.11.1.7), ascorbate peroxidase (APX; EC 1.11.1.11) and superoxide dismutase (SOD; EC 1.15.1.1) were increased in plants supplied 50 μm Cd. Localization of activities of isoforms of these enzymes (POD, APX and SOD) on native gels also revealed increase in the intensities of pre‐existing bands. Stimulated activities of CAT, POD, APX and SOD in maize plants supplied excess Cd do not appear to have relieved plants from excessive generation of reactive oxygen species (ROS). It is, therefore, concluded that supply of 50 μm Cd induces oxidative stress by increasing production of ROS despite increased antioxidant protection in maize plants.  相似文献   

11.
Lipoxygenases (LOX) belong to a large family of plant enzymes that catalyze the hydroperoxidation of polyunsaturated fatty acids. Most of them are expressed during senescence and contribute to membrane deterioration and biosynthesis of jasmonic acid, a known senescence enhancer. In this work, we cloned a fragment of a gene encoding a LOX from broccoli (BoLOX1). The analysis of the sequence revealed that BoLOX1 is closely related to other LOX from higher plants. Furthermore, we analyzed the expression of BoLOX1 and detected a larger increase during postharvest senescence. A slight increase of total lipoxygenase activity was also found during senescence. In other sets of experiments, broccoli heads were treated with plant hormones, such as cytokinin and ethylene, as a way to assess the effect of such compounds on the expression of BoLOX1. Cytokinin treatment delayed the increase of BoLOX1 expression and lipoxygenase activity whereas ethylene accelerated both processes. Also, several postharvest treatments were applied in order to delay senescence in broccoli florets and to evaluate their effects on BoLOX1 expression. Samples treated with modified atmosphere, hot air, UV-C or white light showed a delay in chlorophyll degradation and degreening. In most cases, the treatments also delayed the increase of BoLOX1 expression, reaffirming the relationship between the expression of this gene and senescence. However, treatments like modified atmospheres and visible light markedly increased lipoxygenase activity, which suggests a lack of correlation between BoLOX1 expression and lipoxygenase activity.  相似文献   

12.
Watercore usually spreads in mature ‘Fuji’ apples and may induce flesh browning disorders through unknown mechanisms. We analyzed H2O2 and ascorbic acid (AA) contents and the activities of four enzymes involved in the AA–glutathione (AA–GSH) cycle in watercore and watercore-free tissues in ‘Fuji’ apple fruit during storage. H2O2 levels always were higher in watercore than in watercore-free tissues. AA levels were lower in watercore tissues at harvest and decreased in both tissue types during the storage period. However, AA was completely absent from watercore tissue but not watercore-free tissue three months after harvest. Ascorbate peroxidase (APX) activities were always higher in watercore than in watercore-free tissues, and the activities of dehydroascorbate reductase (DHAR) decreased continuously after harvest in both tissue types. These results suggested that higher production of H2O2 caused by anaerobic conditions in watercore activated APX which acted as a redox signal; the concomitant net consumption of AA was not balanced by the decline of DHAR activity, leading to decreased antioxidant levels. On the other hand, the gradual increases in monodehydroascorbate reductase (MDAR) and glutathione reductase (GR) activity observed during storage accompanied by low AA levels and DHAR activity might indicate a declining efficiency of the AA–GSH cycle.  相似文献   

13.
Effects of hydrogen peroxide (H2O2), as exogenous reactive oxygen, on browning and active oxygen metabolism in pericarp of harvested ‘Fuyan’ longan fruit were investigated. The results showed that as compared with the control fruit, there was a higher browning index in pericarp of H2O2-treated fruit. The fruit treated with H2O2 resulted in increased rate of superoxide anion (O2) production, reduced activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), decreased amounts of ascorbic acid (AsA), glutathione (GSH) and carotenoid, and increased malondialdehyde (MDA) content. These results indicated that H2O2-induced browning in pericarp of harvested longan fruit might be due to a reducing capacity of active oxygen scavenging and an increase of accumulation of O2, which might stimulate membrane lipid peroxidation, disrupt cellular membrane structure, and cause the loss of cellular compartmentalization, in turn, resulting in the contact of polyphenol oxidase (PPO) and peroxidase (POD) with phenolic substrates and subsequently oxidation phenolics to form brown polymers.  相似文献   

14.
Two chilling‐tolerant genotypes, that is, weedy rice WR03‐45 and cultivated rice Lijiangxintuanheigu and two chilling‐sensitive genotypes, that is, weedy rice WR03‐26 and cultivated rice Xiuzinuo were used in this study to investigate the effects of exogenous abscisic acid (ABA) on protection against chilling damage as well as on changes in physiological features. The results showed that under chilling stress the increased levels of superoxide radical (), hydrogen peroxide (H2O2) and malondialdehyde (MDA) in WR03‐45 and Lijiangxintuanheigu were lower than those in WR03‐26 and Xiuzinuo. Activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR)) and non‐enzymatic antioxidants (ascorbate acid (AsA) and reduced glutathione (GSH)) were enhanced in WR03‐45 and Lijiangxintuanheigu, whereas they were decreased significantly in WR03‐26 and Xiuzinuo. Application of exogenous ABA reduced the chilling damage in the four genotypes. The pre‐treatment with ABA decreased the levels of , H2O2 and MDA caused by chilling stress in the four genotypes through increasing the activities of SOD, CAT, APX, GR and the contents of AsA and GSH in the four genotypes under chilling stress. Moreover, pre‐treatment with Fluridone, the ABA biosynthesis inhibitor, prohibited the effects of ABA through enhancing the oxidative damages and suppressing the antioxidant defence systems under chilling stress. The results indicate the mechanism for rice with chilling tolerance is to enhance the capacity of antioxidant defence systems under chilling stress. Furthermore, ABA plays important roles in the tolerance of rice against chilling stress for it could induce the capacity of whole antioxidant defence systems including enzymatic and non‐enzymatic constitutions under chilling stress.  相似文献   

15.
Central broccoli heads (cv. de Cicco) were harvested and treated with UV-C light (4, 7, 10, or 14 kJ m−2). All treatments delayed yellowing and chlorophyll degradation at 20 °C but the irradiation dose of 10 kJ m−2 allowed retaining the highest chlorophyll content yet had lower amounts of pheophytins than every treatment other than 7 kJ m−2. This dose was selected to analyze the effect of UV-C on postharvest broccoli senescence at 20 °C. The UV-C treatment delayed yellowing, chlorophyll a and b degradation, and also the increase in pheophytins during storage. The activity of chlorophyll peroxidase and chlorophyllase was lower in UV-C treated broccoli. Instead, Mg-dechelatase activity increased immediately after the treatment, but after 4 and 6 d this activity was lower in UV-C treated florets than in controls. Treated broccoli also displayed lower respiration rate, total phenols and flavonoids, along with higher antioxidant capacity. The results suggest that UV-C treatments could be a useful non-chemical method to delay chlorophyll degradation, reduce tissue damage and disruption, and maintain antioxidant capacity in broccoli.  相似文献   

16.
The effects of neutral electrolysed water (NEW), ultraviolet light C (UV-C) and superatmospheric O2 packaging (HO), single or combined, on the quality of fresh-cut kailan-hybrid broccoli for 19 days at 5 °C were studied. As controls, washing with water and sanitation with NaClO were both used. Electrolyte leakage, sensory, microbial and nutritional quality changes throughout shelf-life were studied. At day 15, the combined treatments achieved lower mesophilic and psychrophilic growth compared to the single ones. Single treatments produced higher ascorbate peroxidase (APX) reductions just after its application, while superoxide dismutase (SOD) showed the opposite behaviour. After 5 days at 5 °C, a great increase of APX and guaiacol peroxidase (GPX) activity was observed, NEW + UV-C + HO and HO-including treatments achieving the highest and the lowest APX increases, respectively. UV-C-including treatments produced the highest α-linolenic acid (ALA) decreases ranging 35–38% over control contents on the processing day. NEW-including treatments greatly reduced, throughout shelf-life, ALA and stearic acid (SA) content by 27–44% and 31–61%, respectively. Total phenolic content and antioxidant capacity (1415 mg ChAE kg−1 fw and 287 mg AAE kg−1 fw, respectively) remained quite constant during shelf-life. In general, the treatments and their possible combinations seem to be promising techniques to preserve, or even enhance, the quality of fresh-cut kailan-hybrid broccoli and, probably, other vegetables.  相似文献   

17.
Shiitake (Lentinus edodes) mushrooms were exposed to UV-C light (4 kJ/m2) and stored in modified atmosphere packaging (MAP) prior to 15 days at 1 ± 1 °C and 95% relative humidity plus 3 days at 20 °C. Mushroom firmness, total phenolics, total flavonoids, ascorbic acid and H2O2 contents, superoxide anion (O2) production rate and activities of catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) were measured. UV-C treatment resulted in maintenance of a high level of firmness during 15 days at low temperature and reduced the decrease in firmness during shelf-life storage. Furthermore, treated samples showed higher total flavonoids, ascorbic acid, and delayed the increases in both O2 production rate and H2O2 contents. However, no clear treatment effects were seen in total phenolics contents. The treatment also increased the antioxidant enzyme activities of CAT, SOD, APX and GR throughout the storage period. These results indicate that postharvest application of UV-C radiation can delay softening and enhance antioxidant capacity in shiitake mushrooms.  相似文献   

18.
江力  李俊 《农产品加工.学刊》2006,(10):108-110,114
以山药为材料,研究了鲜切加工加速组织衰老与活性氧代谢的关系。结果表明,鲜切加工提高了山药抗氧化酶:超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和抗坏血酸-过氧化物酶(APX)的活性,促进了HO和丙二醛22(MDA)的积累,加速了抗坏血酸(ASA)的损失,加强了膜脂过氧化作用。统计分析表明,HO含量,MDA含量22和ASA含量三者之间存在显著相关性。  相似文献   

19.
转codA基因提高番茄植株的耐热性   总被引:3,自引:0,他引:3  
以野生型番茄(cv. Moneymaker)和转codA番茄为材料,用不同温度(25、30、35、40、45和50℃)分别处理2 h,测定叶片净光合速率(Pn)、PSII最大光化学效率(Fv/Fm)、过氧化氢(H2O2)含量、丙二醛(MDA)含量、相对电导率(REC)和抗氧化酶活性等生理指标;42℃高温处理0、3和6 h后,检测热响应基因的表达量以及D1蛋白的含量,研究高温胁迫对上述参数的影响,探讨转codA基因提高番茄叶片耐热性的机制。。结果表明,高温胁迫下,转codA基因番茄叶片Pn和Fv/Fm的抑制程度明显低于野生型,H2O2、MDA的积累量以及REC均低于野生型,而且明显增强了过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)的活性。此外,转codA基因番茄叶片中抗氧化酶基因和热胁迫基因的表达水平均高于野生型,而D1蛋白的降解水平低于野生型。转codA基因番茄体内合成的甜菜碱提高了转基因番茄的耐热性,这与提高和维持较高的抗氧化酶活性、促进热激响应基因的表达及减缓D1蛋白的降解等有关。  相似文献   

20.
小麦近等基因系与白粉病菌互作的生理指标研究   总被引:3,自引:0,他引:3  
小麦品种百农3217感白粉病,它的一个近等基因系(Xbd/百农3217(BC7F6))含抗病基因xbd,具有抗白粉病功能。以这两个品系为试验材料,接种白粉菌,分析了0~6 d两个品系叶片中过氧化氢(H2O2)的含量和7种代谢酶的活性,这些代谢酶包括超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)、谷胱甘肽过氧化物酶(GPX)、脱氢抗坏血酸还原酶(DHAR)、抗坏血酸过氧化物酶(APX)和谷胱甘肽转硫酶(GST)等。目的是探讨在白粉菌和小麦植物的互作过程中,感病、抗病植物生理指标之间的差异。试验结果表明:接种白粉病菌后,H2O2含量在感病品系中显著下降,在抗病品系中显著上升;在白粉菌侵染进程中,CAT酶活性,感病与抗病品系无明显差异;感病品系叶片内SOD、GR、GPX、APX和GST活性在接种后期显著上升,高于抗病品系;DHAR活性变化则无明显规律。在白粉病侵染小麦植株过程中,H2O2具有重要作用,多种代谢机制共同参与,各种酶的总体表现影响了H2O2含量的变化。  相似文献   

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