植物抗坏血酸过氧化物酶研究进展(综述)

本文从酶的作用机制、酶学特征、分子生物学等方面综述植物抗坏血酸过氧化物酶（APX）的研究进展。     

植物抗坏血酸过氧化物酶的作用机制、酶学及分子特性

介绍叶绿体中H2O2的产生和清除,抗坏血酸过氧化物酶(APX)的酶学和分子特性,APX同工酶在植物体内的分布和功能及其相互之间的区别,APX与细胞色素C过氧化酶(CPX)和谷胱苷肽过氧化物酶(GPX)等一些在不同生物中的H2O2清除酶的异同之处,以及有关APX基因工程的研究进展.     

植物抗坏血酸过氧化物酶

植物抗坏血酸过氧化物酶沈文飚黄丽琴徐朗莱（南京农业大学理学院应用化学系,南京２１００９５）关键词抗坏血酸过氧化物酶植物抗坏血酸过氧化物酶（ＡＰＸ,ＥＣ１．１１．１．１１）的发现至今已有２０多年了。Ｆｏｙｅｒ和Ｈａｌｉｗｅｌ［１］首先于１９７６年发现以...     

逆境胁迫下植物体内活性氧代谢及调控机理研究进展

活性氧(reactive oxygen species, ROS)在植物生长发育中扮演着十分重要的角色。适当浓度的ROS是植物所必需的,而在逆境胁迫下ROS会大量积累,从而抑制植物的生长发育甚至杀死植物。为了维持体内ROS的动态平衡,植物进化出了一系列的ROS产生及清除机制。本文对近年来植物在逆境下的ROS产生、清除及其调节机制的研究进展予以综述,重点介绍转录及翻译后水平的ROS清除及其调节机制,并对植物ROS代谢及调控机理的研究提出了进一步展望。     

龙眼体胚发生过程中抗坏血酸过氧化物酶活性的变化

在龙眼体胚发生的阶段性同步化材料中,均检测到与体胚发生可能有密切关系的抗坏血酸过氧化物酶(APX)活性;龙眼体胚中至少存在有胞浆型APX和叶绿体型APX;APX的转录与其酶活性变化并不一致。     

低温胁迫对黄瓜子叶抗坏血酸过氧化物酶活性和谷胱甘肽含量的影响

低温胁迫下黄瓜幼苗子叶抗坏血酸过氧化物酶活性和GSH含量显著下降,下降幅度随低温胁迫程度增加而递增。不同低温胁迫下酶活性和GSH含量变化与子叶电解质泄漏和MDA的增加呈负相关。幼苗用MV和MDA预处理可加剧由低温引起的抗坏血酸过氧化物酶活性和GSH含量降低,而用苯甲酸钠和α-生育酚预处理则可减轻这种降低,显示出过量的活性氧及其引发的膜脂过氧化产物MDA对抗坏血酸过氧化物酶和GSH均有伤害影响。     

朝鲜碱茅抗坏血酸过氧化物酶的基因克隆和表达

构建到酵母表达载体pYES2-PutAPx,并导入酵母IVSC1菌株后在半乳糖的诱导下分析具有抗氧化性.本研究成功克隆了抗坏血酸过氧化物酶基因(PutA Px)编码区,并做了初步分析,为进一步研究逆境诱导的氧化胁迫的作用机理研究奠定了基础.     

植物过氧化物酶研究进展

过氧化物酶 [peroxidase,POD,EC1 .1 1 .1 .7(X) ]是广泛存在于各种动物、植物和微生物体内的一类氧化酶。催化由过氧化氢参与的各种还原剂的氧化反应 :RH2 H2 O2 →2 H2 O R。植物过氧化物酶的研究可追溯到 1 80 9年用愈创树脂为底物进行的颜色反应。但直到一个世纪之后才开展此酶的分离和命名。已知的催化反应底物超过 2 0 0种 ,以及多种过氧化物和辅助因子。迄今被研究最深入的应首推辣根过氧化物酶 (horseradish pero-xidase,HRP)。早在 1 94 0年 ,Thorell即用电泳方法从部分纯化的辣根组织中区分出 2种不同的 HRP,之后此酶…     

植物抗坏血酸的合成和代谢以及相关酶基因的调控

本文对植物抗坏血酸的生物合成与代谢途径以及相关酶基因调控的研究进展作介绍。     

箭舌豌豆根系抗坏血酸及相关酶对镉胁迫的响应

以箭舌豌豆(Vicia sativa L.)品种L3(耐镉)和ZM(镉敏感)为材料,研究了不同程度镉胁迫下箭舌豌豆幼苗根系抗坏血酸(AsA)含量、脱氢抗坏血酸还原酶(DHAR)同工酶活性、抗坏血酸过氧化物酶(APX)同工酶活性以及APX基因表达的变化。结果显示:(1)2个箭舌豌豆品种根系AsA和脱氢抗坏血酸(DHA)含量在镉胁迫下显著升高;AsA/DHA比值在镉耐性品种L3中显著升高,在敏感品种ZM中显著下降;相同镉处理浓度下,L3根系AsA含量和AsA/DHA比值显著大于ZM。(2)2个品种根系DHAR的活性电泳共显示4条同工酶条带,它们的活性均随镉处理浓度的升高而升高;其中DHAR1只在L3显示,DHAR4只在ZM显示;相同镉处理浓度下,品种L3的DHAR的总活性大于品种ZM。(3)2个品种根系APX的活性电泳共显示11条同工酶条带,其中的APX1、2、4仅在敏感品种ZM中受镉胁迫诱导,APX 8在耐性品种L3中受到比敏感品种ZM更显著的诱导;克隆得到1个箭舌豌豆APX基因,荧光定量RT-PCR结果显示该基因的转录在L3和ZM根系均受镉处理诱导。研究表明,镉胁迫下2个箭舌豌豆品种根系AsA含量,AsA代谢相关酶DHAR和APX的活性以及APX的转录水平均显著升高;镉耐性品种L3较敏感品种ZM能更有效地促进AsA循环,维持更高的AsA水平,从而更有效地缓解镉胁迫诱导产生的氧化胁迫,这可能是L3较ZM具有更高镉耐性的重要机制之一。     

Cytosolic ascorbate peroxidase 1 protects organelles against oxidative stress by wounding- and jasmonate-induced H2O2 in Arabidopsis plants

Background

Reactive oxygen species (ROS) are not only cytotoxic compounds leading to oxidative damage, but also signaling molecules for regulating plant responses to stress and hormones. Arabidopsis cytosolic ascorbate peroxidase 1 (APX1) is thought to be a central regulator for cellular ROS levels. However, it remains unclear whether APX1 is involved in plant tolerance to wounding and methyl jasmonate (MeJA) treatment, which are known to enhance ROS production.

Methods

We studied the effect of wounding and MeJA treatment on the levels of H₂O₂ and oxidative damage in the Arabidopsis wild-type plants and knockout mutants lacking APX1 (KO-APX1).

Results

The KO-APX1 plants showed high sensitivity to wounding and MeJA treatment. In the leaves of wild-type plants, H₂O₂ accumulated only in the vicinity of the wound, while in the leaves of the KO-APX1 plants it accumulated extensively from damaged to undamaged regions. During MeJA treatment, the levels of H₂O₂ were much higher in the leaves of KO-APX1 plants. Oxidative damage in the chloroplasts and nucleus was also enhanced in the leaves of KO-APX1 plants. These findings suggest that APX1 protects organelles against oxidative stress by wounding and MeJA treatment.

General significance

This is the first report demonstrating that H₂O₂-scavenging in the cytosol is essential for plant tolerance to wounding and MeJA treatment.     

Silicon-mediated alleviation of Mn toxicity in Cucumis sativus in relation to activities of superoxide dismutase and ascorbate peroxidase

The effects of exogenous silicon (Si) on plant growth, activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and catalase, and concentrations of ascorbate and glutathione were investigated in cucumber (Cucumis sativus L.) plants treated with excess manganese (Mn) (600 microM). Compared with the treatment of normal Mn (10 microM), excess Mn significantly increased H2O2 concentration and lipid peroxidation indicated by accumulation of thiobarbituric acid reactive substances. The leaves showed apparent symptoms of Mn toxicity and the plant growth was significantly inhibited by excess Mn. The addition of Si significantly decreased lipid peroxidation caused by excess Mn, inhibited the appearance of Mn toxicity symptoms, and improved plant growth. This alleviation of Mn toxicity by Si was related to a significant increase in the activities of SOD, APX, DHAR and GR and the concentrations of ascorbate and glutathione.     

Reactive oxygen intermediates and glutathione regulate the expression of cytosolic ascorbate peroxidase during iron-mediated oxidative stress in bean

Excess of free iron is thought to harm plant cells by enhancing the intracellular production of reactive oxygen intermediates (ROI). Cytosolic ascorbate peroxidase (cAPX) is an iron-containing, ROI-detoxifying enzyme induced in response to iron overload or oxidative stress. We studied the expression of cAPX in leaves of de-rooted bean plants in response to iron overload. cAPX expression, i.e., mRNA and protein, was rapidly induced in response to iron overload. This induction correlated with the increase in iron content in leaves and occurred in the light as well as in the dark. Reduced glutathione (GSH), which plays an important role in activating the ROI signal transduction pathway as well as in ROI detoxification, was found to enhance the induction of APX mRNA by iron. To determine whether cAPX induction during iron overload was due to an increase in the amount of free iron, which serves as a co-factor for cAPX synthesis, or due to iron-mediated increase in ROI production, we tested the expression of APX in leaves under low oxygen pressure. This treatment, which suppresses the formation of ROI, completely abolished the induction of cAPX mRNA during iron overload, without affecting the rate of iron uptake by plants. Taken together, our results suggest that high intracellular levels of free iron in plants lead to the enhanced production of ROI, which in turn induces the expression of cAPX, possibly using GSH as an intermediate signal. We further show, using cAPX-antisense transgenic plants, that cAPX expression is essential to prevent iron-mediated tissue damage in tobacco.     

外源SOD和APX基因在转基因烟草中的表达与遗传

分析转超氧化物歧化酶基因（SOD）或抗坏血酸过氧化物酶基因（APX）烟草及其自交和杂交后代的叶片中超氧化物歧化酶（SOD）和过氧化物酶（POD）活性的结果表明：转基因烟草的SOD和POD活性在终花期最强,不同叶位叶中SOD活性差异不明显,POD活性以下部叶为最高;转基因烟草的SOD或POD活性显著高于近等基因的非转基因品系。杂交后代（F1、F2）的SOD活性能保持稳定,略高于亲本;自交后代（S1～S3）与自交亲本的SOD和POD活性相当。     

Thylakoid membrane-bound ascorbate peroxidase is a limiting factor of antioxidative systems under photo-oxidative stress

To evaluate the physiological importance of thylakoid membrane-bound ascorbate peroxidase (tAPX) in the active oxygen species-scavenging system of chloroplasts, the level of tAPX in tobacco plants was altered by expression of the tAPX cDNA in both sense and antisense orientation. The tobacco plants transformed with constructs of antisense tAPXs from spinach and tobacco could not be obtained, suggesting that the suppression of tAPX in higher plants had a severe effect on the growth even under normal conditions. In contrast, the transgenic tobacco plants (TpTAP-12) overexpressing tAPX, which had approximately 37-fold higher activity than that of the wild-type plants, were generated. The TpTAP-12 plants showed increased tolerance to oxidative stress caused by application of methylviologen (MV, 50 microm) under light intensity (300 and 1600 microE m(-2) sec(-1)) and by chilling stress with high light intensity (4 degrees C, 1000 microE m(-2) sec(-1)). At 24 h after the MV treatment under illumination at 300 microE m-2 sec-1, destruction of chlorophyll was observed in the wild-type plants, but not in the TpTAP-12 plants. The activities of thiol-modulated enzymes in the Calvin cycle, the level and redox status of ascorbate (AsA), and the activity of tAPX in the wild-type plants significantly decreased, while those in the TpTAP-12 plants were hardly changed. These observations suggest that tAPX is a limiting factor of antioxidative systems under photo-oxidative stress in chloroplasts, and that the enhanced activity of tAPX functions to maintain the AsA content and the redox status of AsA under stress conditions.