Ascorbate levels and the activity of key enzymes in ascorbate biosynthesis and recycling in the leaves of 22 Chinese persimmon cultivars

The objective of this study was to determine diversities of ascorbic acid (AsA) and activities of enzymes involved in AsA metabolism, and to determine their relationships to AsA content in the leaves of Chinese persimmon (Diospyros kaki) cultivars. Results showed that there were huge differences not only in the contents of AsA and glutathione (GSH) but also in the enzyme activities of l-galactono-1,4-lactone dehydrogenase (GLDH, EC 1.3.2.3), dehydroascorbate reductase (DHAR, EC 1.8.5.1), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) in the leaves of 22 Chinese persimmon cultivars. Control of AsA levels mainly depended on the capacity for biosynthesis and GLDH activity played a main role in determining AsA levels in the leaves of persimmon. Both MDHAR and DHAR, MDHAR in particular, also played an important role in maintaining AsA redox state by reducing oxidized AsA back to AsA. But contents of GSH and hydrogen peroide (H₂O₂) and activity of APX were not significantly correlated with AsA content in the leaves of persimmon.     

低温胁迫对草莓叶片SOD和AsA-GSH循环酶系统的影响

以草莓试管苗为试材,研究低温胁迫对草莓离体叶片抗坏血酸、脱氢抗坏血酸含量和SOD以及抗坏血酸 -谷胱甘肽（AsA-GSH）循环中4种酶活性的影响。结果表明,低温胁迫导致超氧化物歧化酶、抗坏血酸过氧化物酶和单脱氢抗坏血酸还原酶活性上升,脱氢抗坏血酸还原酶和谷胱甘肽还原酶活性下降;同时使还原态抗坏血酸含量下降而脱氢抗坏血酸含量上升。随着低温处理时间延长,过多活性氧不能被防御体系有效清除,植物受到伤害。试验结果还表明,抗坏血酸过氧化物酶具有较强的氧化还原抗坏血酸的能力,单脱氢抗坏血酸还原酶是AsA-GSH循环再生抗坏血酸的主要酶。     

Melatonin alleviates cold-induced oxidative damage by regulation of ascorbate–glutathione and proline metabolism in melon seedlings (Cucumis melo L.)

Cold stress is one of the most detrimental environmental factors affecting plant growth and development. Melatonin (MEL), a natural indoleamine compound, responds to various environmental cues. To explore the role of MEL in the response of melon (Cucumis melo L.) seedlings to cold stress, the effects of exogenous MEL on the ascorbate–glutathione (AsA–GSH) cycle and proline metabolism were investigated. Melon seedlings were sprayed with various concentrations of MEL (0, 50, 100, 200, or 400 μM), then exposed to cold stress, 12/6°C (day/night) for 7 d, followed by recovery at 28/18°C for another 7 d. The results showed that MEL, especially the 200 μM treatment, dramatically alleviated growth inhibition caused by cold stress, manifested by increased plant growth and decreased O₂^?¯ production rate and malondialdehyde content. Importantly, exogenous application of MEL enhanced the ratios of reduced and oxidized forms of AsA (AsA/DHA) and GSH (GSH/GSSG), and the activities of ascorbate peroxidase (APX), glutathione reductase (GR), and dehydroascorbate reductase (DHAR) involved in the ascorbate–glutathione (AsA–GSH) cycle in melon seedings grown under cold stress. Besides, MEL pretreatment further increased the contents of proline and soluble protein under cold stress. The results reveal that protective effects of MEL against cold stress in melon seedlings are most likely associated with the regulation of the AsA–GSH cycle and proline metabolism as an effective antioxidant system.     

铅胁迫对姜叶片活性氧代谢的影响

以‘莱芜大姜’为试材,采用污灌方式研究不同水平铅（Pb）胁迫（土壤Pb 含量分别为0、150、450 和1 350 mg · kg-1）对姜叶片活性氧代谢的影响。结果表明,随Pb 胁迫水平的升高及胁迫时间的延长,姜叶片中生成速率和H2O2 含量显著升高,但过度Pb 胁迫则使其降低,而丙二醛（MDA）含量和电解质渗漏率则持续增加。Pb 胁迫40 d 时,超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）、谷胱甘肽还原酶（GR）、抗坏血酸过氧化物酶（APX）、单脱氢抗坏血酸还原酶（MDHAR）、脱氢抗坏血酸还原酶（DHAR）等抗氧化酶活性随Pb 胁迫水平的升高显著增强,还原型抗坏血酸（AsA）、总抗坏血酸（AsA + DHA）及还原型谷胱甘肽（GSH）、总谷胱甘肽（GSH + GSSG）等抗氧化物质含量显著升高,AsA/DHA、GSH/GSSG 比值显著增加;随胁迫时间的增加,抗氧化酶活性及抗氧化物质含量则多以150 mg · kg-1 的轻度Pb 胁迫的较高,过度的Pb 胁迫则使其显著降低,表明AsA-GSH 循环遭受严重破坏。     

CaCl_2对NaCl胁迫下酸枣苗AsA-GSH循环的影响

以酸枣(Ziziphus acidojujuba C.Y.Cheng et M.J.Liu)水培实生幼苗为试材,研究了CaCl_2对NaCl胁迫下其根、茎、叶中抗坏血酸—谷胱甘肽循环(AsA-GSH)的影响。结果表明:与对照(日本园试配方营养液)相比,NaCl处理下根和叶中AsA含量升高,MDHAR活性升高,APX活性下降;根、茎、叶中GSH含量升高。NaCl+CaCl_2处理与NaC处理相比,根和茎中AsA含量进一步升高,叶中AsA含量下降,叶中APX下降,根中MDHAR活性进一步升高,茎和叶中DHAR活性升高;根和叶中GSH含量下降。外源CaCl_2可能通过提高酸枣实生苗根、茎和叶中AsA再生关键酶MDHAR和DHAR活性,促进根和茎中AsA的再生与生物合成,从而提高根、茎清除过量H_2O_2的能力,缓解NaCl胁迫对酸枣幼苗根和茎部的伤害。     

Exogenous paraquat changes antioxidant enzyme activities and lipid peroxidation in drought-stressed cucumber leaves

In order to examine whether paraquat modifies the functioning of antioxidants and oxidative stress levels in drought-stressed plants, a cucumber cultivar (Cucumis sativus cv. Yuexiu no. 3) was grown hydroponically for 2 days. Drought stress, which was induced by polyethylene glycol (PEG), increased the contents of malonaldehyde (MDA), superoxide radical (O₂⁻) and hydrogen peroxide (H₂O₂) in cucumber leaves, while pretreatment of paraquat decreased them. Under drought stress induced by PEG, we observed the decreased contents of MDA, H₂O₂ and O₂⁻ in paraquat-pretreated plants in comparison to unpretreated stressed plants. Drought stress and paraquat both increased the activities of antioxidants such as superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), guaiacol peroxidase (GPX, EC 1.11.1.7), ascorbate peroxidase (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1), monodehydroascorbate reducatase (MDHAR, EC 1.6.5.4), glutathione reductase (GR, EC 1.6.4.2), reduced glutathione (GSH) and reduced ascorbate (AsA). But the combined effect of paraquat application and drought stress resulted in the highest activities of antioxidants. So paraquat is able to moderate the activities of scavenging system enzymes and to influence oxidative stress intensity under drought stress induced by PEG.     

外源抗坏血酸对离体苹果叶片衰老的影响

 以苹果(Malus domestica Borkh. ) 品种‘嘎拉’秋梢成熟叶片为试材, 在5 mmol·L ^{- 1}抗坏血酸(AsA) 培养条件下, 研究了外源AsA 对离体苹果叶片衰老过程( 0 ～72 h) 中抗坏血酸—谷胱甘肽(AsA - GSH) 循环各组分和AsA合成关键酶L - 半乳糖酸- 1, 4 - 内酯脱氢酶(GalLDH) 的影响。结果表明, 外源AsA部分抑制了离体苹果叶片衰老过程中膜脂过氧化, 降低了H2O2含量, 但对表示膜损伤的相对膜透性影响不大。对AsA - GSH循环各组分来说, 外源AsA维持了叶片衰老过程中抗坏血酸过氧化物酶(APX) 和脱氢抗坏血酸还原酶(DAHR) 的高活性, 部分抑制了单脱氢抗坏血酸还原酶(MDHAR) 活性的下降, 同时降低了GSH的氧化程度, 从而提高了AsA含量和再生能力。外源AsA 在衰老前期抑制了GalLDH活性, 降低了AsA的合成能力, 而后期维持了GalLDH的高活性。     

低温对仁用杏雌蕊抗坏血酸-谷胱甘肽循环的影响低温对仁用杏雌蕊抗坏血酸-谷胱甘肽循环的影响

 利用人工智能模拟霜箱,设8 个温度（18、–1、–2、–3、–4、–5、–6、–7 ℃）,对 抗寒性不同的两个仁用杏（Prunus armeniaca Linn.）品种雌蕊的过氧化氢（H₂O₂）含量和抗坏血酸-谷胱 甘肽（AsA-GSH）循环进行了研究。结果表明,低温胁迫增加了仁用杏雌蕊H₂O₂ 含量,–7 ℃时抗晚霜 品种‘围选1 号’和晚霜敏感品种‘龙王帽’分别为对照（18 ℃）的 5.49 倍和7.22 倍。低温期间,‘围 选1 号’雌蕊AsA 等酶含量及APX 等酶活性均高于‘龙王帽’,–7 ℃时,‘围选1 号’雌蕊AsA、MDHA、 DHA、GSH 和GSSG 含量比对照分别下降了66%、69%、48%、52%、53%,‘龙王帽’下降了78%、79%、 59%、66%、65%,–7 ℃时,‘围选1 号’雌蕊APX、MDHAR、DHAR、GPX、GR 活性、AsA/DHA、 GSH/GSSG 分别是对照的2.87 倍、1.07 倍、1.42 倍、1.60 倍、1.20 倍、65%和1.03 倍,‘龙王帽’是对照 2.41 倍、96%、1.13 倍、1.08 倍、98%、54%、97%。结果表明‘围选1 号’雌蕊在低温期间有较高的AsA-GSH 循环效率,可有效抑制H₂O₂ 的积累,这可能是其抗晚霜能力较强的重要原因之一。     

红肉脐橙发育过程中抗氧化系统与活性氧的变化

 以红肉脐橙果肉为材料, 比较了不同时期谷胱甘肽(GSH) 、抗坏血酸(AsA) 含量及相关酶活性和活性氧(AOS) 产生速率的变化。结果表明: 果肉在整个发育过程中, 脱氢抗坏血酸还原酶(DHAR) 活性在很大程度上可以促进AsA积累, 两者呈显著的正相关( r = 0.870, P <0.05) , 并且与抗坏血酸过氧化物酶(AsA-POD) 活性在动态上彼此消长; 谷胱甘肽还原酶(GR) 的变化趋势与GSH相同,呈单峰型的变化, 两者的相关系数为r = 0.978 ( P < 0.01) ; AsA /DHAsA和GSH /GSSG比率在整个发育过程中呈下降的趋势, 而活性氧(H₂O₂ , O₂^·) 水平一直呈上升变化。     

油菜素内酯对Ca(NO_3)_2胁迫下黄瓜叶绿体AsA-GSH循环及叶黄素稳态的影响

研究了外源油菜素内酯(24–表油菜素内酯,EBL)对Ca(NO_3)_2胁迫下黄瓜光合器官的调控作用。在Ca(NO_3)_2胁迫条件下,外源EBL显著提高了叶绿体中还原型抗坏血酸和谷胱甘肽的含量,分别是对照的2.44倍和1.40倍。AsA-GSH循环中的抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性比胁迫处理增加了36.27%和105.83%。外源EBL的施用显著增加了紫黄质(V)、玉米黄质(Z)和总叶黄素含量,减少了花药黄质(A)的积累,叶黄素脱环氧化态比胁迫处理增加了9.60%。胁迫条件下,EBL显著提高了植株叶片的非光化学猝灭(NPQ),缓解了PSⅡ实际光化学效率(Φ_(PSⅡ))的下降。EBL处理后,叶绿体膜多种不饱和脂肪酸的水平比对照明显增加。试验结果表明,外源EBL能够通过调控AsA-GSH循环及热耗散,增强叶绿体抗氧化能力,从而抵抗Ca(NO_3)_2胁迫。     

外源褪黑素对硝酸盐胁迫条件下黄瓜幼苗抗氧化系统的影响

以‘津春4号’黄瓜为试验材料,采用灌根的方法研究了外源褪黑素（MT）对硝酸盐胁迫下幼苗抗氧化系统的影响,为MT在设施蔬菜生产中的应用提供参考。结果表明：硝酸盐胁迫处理后,黄瓜幼苗叶片中超氧阴离子（）产生速率、过氧化氢（H₂O₂）和丙二醛（MDA）含量显著增加;抗坏血酸（AsA）和谷胱甘肽（GSH）含量及活性均下降;超氧化物歧化酶（SOD）、过氧化物酶（POD）、抗坏血酸过氧化物酶（APX）、谷胱甘肽还原酶（GR）活性在硝酸盐胁迫3 d时,过氧化氢酶（CAT）活性在胁迫6 d时,脱氢抗坏血酸还原酶（DHAR）活性在胁迫9 d时均达最高,之后又逐渐下降;单脱氢抗坏血酸还原酶（MDHAR）活性则持续降低。MT处理可有效抑制硝酸盐胁迫条件下黄瓜幼苗中  的产生速率及H₂O₂和MDA的积累,提高AsA和GSH含量及活性,增强SOD、POD、CAT、APX、MDHAR、DHAR、GR活性,提高叶片的光合速率及干物质积累。这些结果说明,MT可减缓硝酸盐胁迫的危害,提高黄瓜幼苗对硝酸盐的耐性,且100 μmol · L^-1 MT处理效果最佳。     

猕猴桃果实发育过程中AsA代谢产物积累及相关酶活性的变化

 以美味猕猴桃品种‘秦美’果实为材料, 研究了其生长发育过程中与AsA代谢循环系统相关的物质抗坏血酸(AsA) 、谷胱甘肽(GSH) 、草酸(OA) 、酒石酸( TA) 和过氧化氢(H₂O₂ ) 的含量及相关酶活性的变化及其相互关系。结果表明: 在果实生长发育过程中, 花后AsA含量明显增加, 花后30 d达到最高后开始下降, 花后75 d后基本保持不变。就整个果实中总的AsA积累量而言, 花后开始显著增加, 到45 d达到最大值后至成熟基本保持不变。这表明猕猴桃果实的AsA积累主要发生在幼果期。GSH随着果实发育在花后120 d前其含量及积累量均有增加, 但积累也主要发生在幼果期。OA含量的变化与H₂O₂含量和抗坏血酸过氧化物酶(APX) 活性相似, 均在花后开始显著下降, 到花后30 d后变化不大; 而TA含量的变化趋势与AsA一致。抗坏血酸氧化酶(AO) 、单脱氢抗坏血酸还原酶(MDHAR) 和脱氢抗坏血酸还原酶(DHAR) 的活性变化基本一致, 均在花后开始显著升高, 60 d达到最大后迅速下降, 在90 d后至成熟基本保持不变。     

镉对姜叶片活性氧代谢影响的时间和剂量效应

为探讨姜对镉（Cd）胁迫的生理响应特性,通过外施CdCl2 ? 2.5H2O溶液,研究不同Cd水平对姜叶片活性氧代谢的影响。结果表明,Cd胁迫下,姜叶片中  生成速率和H2O2含量显著升高,且呈剂量和时间效应,同时丙二醛（MDA）含量和电导率持续增加。姜幼苗期,超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）、谷胱甘肽还原酶（GR）、抗坏血酸过氧化物酶（APX）、单脱氢抗坏血酸还原酶（MDHAR）、脱氢抗坏血酸还原酶（DHAR）、抗坏血酸氧化酶（AAO）等抗氧化酶活性均被Cd胁迫所诱导增强,还原型抗坏血酸（AsA）、总抗坏血酸（AsA + DHA）、酸溶性SH、还原型谷胱甘肽（GSH）、植物络合素等抗氧化物质含量显著升高,AsA/DHA、GSH/GSSG比值显著增加;随Cd胁迫浓度和时间的增加,相关保护酶活性及抗氧化物质含量均显著降低,表明姜对Cd较短期（40 d）胁迫或较低浓度Cd胁迫具有适应性,但胁迫120 d或过高浓度的Cd胁迫严重损伤了姜叶片抗氧化系统,使活性氧代谢失调,叶片膜系统遭到严重伤害。     

Antioxidant activity and oxygen-scavenging system in orange pulp during fruit ripening and maturation

The antioxidant activity (ferric reducing/antioxidant power, FRAP) and oxygen-scavenging system of pulps during the fruit ripening and maturation were investigated in three cultivars of sweet orange (Citrus sinensis (L.) Osbeck). The highest FRAP values and activities of antioxidant enzymes and contents of nonenzymes were detected in ‘Red Flesh’ navel orange. The activities of oxygen-scavenging enzymes, superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (G-POD), decreased with ripening and maturation of fruit. Ascorbate peroxidase (AsA-POD) and dehydroascorbate reductase (DHAR) activities were also stably declined, whereas glutathione reductase (GR) activity and nonenzymes in the ascorbate–glutathione cycle displayed a single-peak pattern which paralleled the significant changes in FRAP values. In addition, the ratios of ascorbate/dehydroascorbate (AsA/DHAsA), reduced glutathione/oxidized glutathione (GSH/GSSG) were also decreased. Changes in the activities of antioxidant enzymes and the contents of nonenzymes during ripening indicated that the antioxidant system plays a fundamental role in the ripening of orange fruits.     

低温胁迫对嫁接黄瓜叶片抗坏血酸-谷胱甘肽循环的影响

 为了探讨嫁接增强黄瓜抗冷性的分子机制，本试验以黑籽南瓜为砧木，‘津绿3号’黄瓜为接穗，对低温胁迫下嫁接和自嫁黄瓜幼苗叶片抗坏血酸-谷胱甘肽循环中抗氧化酶基因的表达和相关酶活性及AsA、GSH和 H₂0₂含量变化进行了研究。结果表明: 低温胁迫下, 嫁接黄瓜叶片DHAR mRNA和GR mRNA相对表达量均大于自嫁黄瓜，GalLDH mRNA和APX mRNA相对表达量均与自嫁黄瓜差异无显著差异； DHAR、GR、GalLDH和APX活性均高于自嫁黄瓜；AsA和GSH含量与AsA/DHA和GSH/GSSG比值均高于自嫁黄瓜， DHA、GSSG和H₂O₂含量均低于自嫁黄瓜。嫁接黄瓜叶片DHAR和GR较高的转录水平维持的较高DHAR和GR活性是AsA和GSH含量明显高于自嫁黄瓜的重要原因， 而嫁接黄瓜叶片较高的GalLDH和APX活性与GalLDH mRNA和APX mRNA相对表达量无关。     

Hydrogen peroxide pretreatment alters the activity of antioxidant enzymes and protects chloroplast ultrastructure in heat-stressed cucumber leaves

To elucidate the physiological mechanism of heat stress mitigated by exogenous hydrogen peroxide (H₂O₂) further, seedlings of Cucumis sativus cv Lvfeng no. 6 were subjected to two temperatures (42/38 and 25/18 °C) after pretreatment with 1.5 mM H₂O₂. We investigated whether exogenous H₂O₂ could protect chloroplast ultrastructure under heat stress (42/38 °C) and whether the protective effect was associated with the regulation of antioxidant enzymes. The heat condition disintegrated the membranes of 71.4% chloroplasts in the leaf cells and resulted in the elevated levels of malondialdehyde (MDA) and endogenous H₂O₂. When H₂O₂ pretreatment was combined with the heat stress, the abnormal chloroplasts occurred at a rate of 33.3%, and the contents of MDA and endogenous H₂O₂ were decreased. Heat stress and exogenous H₂O₂ both increased the activities of antioxidant enzymes such as glutathione peroxidase (GSH-Px, EC 1.11.1.9), dehydroascorbate reductase (DHAR, EC 1.8.5.1), monodehydroascorbate reducatase (MDHAR, EC 1.6.5.4), and glutathione reductase (GR, EC 1.6.4.2). The combined effect of heat stress and H₂O₂ pretreatment led to higher activity of antioxidant enzymes including superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), GSH-Px, DHAR, MDHAR and GR in comparison to the heat treatment alone. We propose that exogenous H₂O₂ increases antioxidant enzyme activities in cucumber leaves, decreases lipid peroxidation, and thus protects the ultrastructure of chloroplasts under heat stress.     

硒对菠菜抗氧化系统及过氧化氢含量的影响

 研究了不同浓度硒对菠菜的生长、超氧化物歧化酶(SOD) 、抗坏血酸过氧化物酶(APX) 、过氧化氢酶(CAT) 、脱氢抗坏血酸还原酶(DHAR) 、谷胱甘肽过氧化物酶(GSH-PX) 等酶活性以及抗坏血酸(AsA) 、过氧化氢(H₂O₂) 含量的影响。结果表明: 低浓度硒( ≤0. 1 mg·L^-1) 促进了菠菜的生长, 而高浓度硒(1. 0 mg·L^-1) 抑制生长; 低浓度硒提高了SOD、CAT及GSH2PX活性, 而APX和DHAR 活性则随硒浓度的升高而降低; AsA 含量随硒浓度的增加而减少, H2O2 含量先随硒浓度的增加而下降, 但在高硒浓度时又上升。总之, 适量的硒处理能增强菠菜的抗氧化能力, 增加产量。