铅胁迫下三叶鬼针草内源一氧化氮的生成及其对氧化损伤的缓解效应

对不同浓度铅(Pb)胁迫下三叶鬼针草(Bidens pilosa L.)叶、茎和根中内源一氧化氮(NO)和活性氧(ROS)的生成机制及根系活力的变化,内源NO对Pb胁迫下三叶鬼针草幼苗氧化损伤的缓解效应进行了研究。结果显示,在0~1000 mg/L范围内,随着Pb浓度的增加,叶片中NO含量呈升高趋势,根中NO含量呈先升高后降低的趋势,但仍高于对照,Pb浓度在0~400 mg/L范围内,茎中NO含量与对照持平,Pb浓度大于600 mg/L时,茎中NO含量低于对照;600 mg/L Pb处理能显著增强叶、茎和根中一氧化氮合成酶(NOS)和硝酸还原酶(NR)活性,显著增加叶和茎中亚硝酸根离子(NO_2~-)和类胡萝卜素(Car)含量,NOS、NR、NO_2~-和Car均能促进叶片中内源NO的生成,NOS是根中内源NO生成的主要途径。Pb胁迫使超氧阴离子(O_2~(·-))产生速率、过氧化氢(H_2O_2)含量、丙二醛(MDA)含量和相对电导率(REC)显著升高,从而造成幼苗严重的膜脂过氧化损伤,而胁迫诱发产生的NO能降低根中ROS的产生,促进幼苗根系活力,进而缓解胁迫造成的膜脂过氧化损伤。     

NO对盐胁迫下苜蓿根系生长抑制及氧化损伤的缓解效应

以"甘农4号"苜蓿品种为材料,采用水培法,用NO供体硝普钠(SNP)、硝普钠类似物亚铁氰化钠(不产生NO)、NO特异清除剂c-PTIO、一氧化氮合酶(NOS)活性抑制剂N-硝基-L-精氨酸甲脂(L-NAME)、硝酸还原酶(NR)活性抑制剂钨酸盐处理苜蓿植株,研究NO对盐胁迫下苜蓿幼苗根系生长、根系活力、根系中渗透调节物质、膜脂过氧化、活性氧含量及抗氧化酶活性等的影响,探讨NO调控苜蓿幼苗根系耐盐性的生理机制。结果表明:盐胁迫下SNP处理提高了根系活力,促进了苜蓿幼苗根系生长,降低游离脯氨酸含量,促进可溶性蛋白含量增加;增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、愈创木酚过氧化物酶(GPX)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性,提高还原型抗坏血酸(As A)和还原型谷胱甘肽(GSH)含量,降低过氧化氢(H2O2)、羟自由基(OH·)含量、超氧阴离子(O·-2)产生速率和膜脂过氧化产物丙二醛(MDA)含量;同时,SNP处理显著促进了苜蓿幼苗根系内源NO的积累。NO供体SNP的类似物亚铁氰化钠对盐胁迫下苜蓿根系各项生理生化指标无明显影响;盐胁迫下添加c-PTIO、L-NAME和钨酸盐进一步降低了苜蓿幼苗根系活力和根系生长,抑制了根系抗氧化系统活性,加剧了根系膜脂过氧化作用,降低了内源NO积累,添加SNP则能缓解该抑制效应;表明外源SNP处理能明显缓解盐胁迫对苜蓿幼苗根系生长的抑制和氧化损伤,且通过NOS和NR途径产生的内源NO也可能在苜蓿根系适应盐胁迫的调节中起关键作用;该研究结果为苜蓿耐盐机制及NO在苜蓿耐盐育种、化学调控和盐碱地栽培利用等提供了理论依据。     

水杨酸对锌胁迫下小麦幼苗生长抑制的缓解效应

以小麦品种‘新麦18’为材料,采用室内水培实验研究了不同浓度水杨酸(SA)处理对300 mg.L-1锌胁迫下小麦种子萌发和幼苗生长的影响。结果表明:在Zn2+胁迫下,小麦种子的发芽势和发芽率、幼苗根长、芽长以及幼苗叶片的可溶性蛋白含量、根系活力显著降低,而脯氨酸和丙二醛(MDA)含量显著增加(P<0.05);外施SA显著提高了Zn2+胁迫下小麦种子的发芽势和发芽率,同时也使Zn2+胁迫7 d后的小麦幼苗的根长、芽长,幼苗叶片的脯氨酸和可溶性蛋白含量以及根系活力显著升高,膜脂过氧化产物MDA含量却显著降低(P<0.05)。由此可见,外施SA可通过提高小麦幼苗根长和芽长来增加幼苗根系活力,通过提高小麦幼苗可溶性蛋白含量、脯氨酸含量来维持细胞膜的稳定性,降低膜脂过氧化伤害程度,从而缓解了Zn2+胁迫对幼苗生长的抑制,并以14 mg.L-1外源水杨酸缓解效果最好。     

γ-氨基丁酸浸种对番茄种子及幼苗耐盐性调节的生理机制

以番茄‘金棚一号’为材料,研究了外源γ-氨基丁酸(GABA)浸种处理对NaCl胁迫下种子萌发及幼苗生长和生理代谢的影响。结果显示:(1)NaCl胁迫显著抑制了番茄种子的萌发和胚根生长,同时导致番茄幼苗体内活性氧(O2.-、H2O2)大量积累,膜脂过氧化程度加重,幼苗叶片光合系统Ⅱ活性显著降低,幼苗的生长受到严重抑制。(2)外源GABA浸种能够显著提高盐胁迫下番茄种子的萌发和胚根的生长,并以10.00mmol.L-1 GABA浸种处理效果最好。(3)外源GABA浸种处理显著提高了NaCl胁迫下番茄幼苗根系和叶片抗氧化酶(SOD、POD和CAT)活性,降低了活性氧(O2.-、H2O2)的产生和膜脂过氧化程度,通过维持较高的光合系统Ⅱ活性,促进了幼苗的生长及生物量积累,但GABA的缓解效应存在较大的浓度差异,其中以10.00mmol.L-1 GABA处理效果较好。研究表明,10.00mmol.L-1 GABA浸种处理能够通过促进番茄种子萌发和幼苗生长来缓解盐胁迫的伤害。     

钙对高温胁迫下烟草幼苗抗氧化代谢的影响

用10mmol/LcaCl2溶液处理烟草幼苗,能减缓高温胁迫所引发的一些胁变反应,如叶绿素破坏、积累膜脂质过氧化产物MDA,增加细胞膜相对透性,降低内源抗氧化剂（AsA,GSH）和脯氨酸含量,在高温胁迫下,SOD和CAT等抗氧化酶活性呈现先升后降的变化趋势。恢复生长以后,钙处理和未处理的烟草幼苗其内源抗氧化剂含量和抗氧化酶活性均有不同程度的回升。由此认为,高温胁迫后烟草幼苗体内抗氧化酶活性受抑,活性氧积累以及由此引发的膜脂过氧化作用是烟草幼苗高温伤害的原因之一。     

钙对烟草叶片热激忍耐和活性氧代谢的影响

热胁迫导致烟草叶片细胞膜系统显著受损,表现为SOD活性降低和MDA含量明显升高,叶片叶绿素含量下降,活性氧增加。10mmol/LcaCl2溶液处理烟草幼苗后,能有效降低热胁迫下叶片细胞膜透性,维持较高的SOD和CAT等抗氧化酶活性,减缓O2^-形成和膜脂过氧化反应。研究结果表明,CaCl2处理提高了烟草叶片膜稳定性和膜保护酶活性,有利于保护细胞膜结构,降低高温对烟草幼苗的伤害。钙离子螯合剂EGTA能在一定程度上降低烟草叶片的抗热性。     

镉胁迫对芥蓝根系质膜过氧化及ATPase活性的影响

水培条件下,以“香港白花”芥蓝品种为供试材料,研究4种不同浓度镉（0、1.0、2.0、4.0、8.0 mg/L Cd）处理对芥蓝幼苗根系质膜过氧化及ATPase活性的影响。结果表明,与对照相比,随着Cd处理浓度的增加,芥蓝根系活力呈现降低的变化趋势,而根系超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性以及丙二醛（MDA）、H2O2含量和O2-产生速率表现出升高的趋势,表明芥蓝受到活性氧物质的胁迫。1.0、2.0 mg/L Cd浓度处理下的H2O2含量与对照差异不显著,而O2-产生速率则在1.0 mg/L浓度处理下与对照差异不显著。随着Cd处理浓度的增加,芥蓝根系质膜H -ATPase和Ca2 -ATPase活性呈现出先增后减的变化趋势。1.0 mg/LCd浓度处理时,H -ATPase和Ca2 -ATPase活性与对照差异不显著（P > 0.05）,而在2.0、4.0和8.0 mg/LCd处理时,两种ATPase活性显著降低（P < 0.05）,并且与膜脂过氧化水平呈极显著的负相关（R2 > 0.969）。因此,低浓度Cd处理对芥蓝根系质膜两种ATPase活性影响较小,较高浓度Cd处理使芥蓝根系活力和质膜ATPase的损伤加重。     

钙对烟草叶片热激忍耐和活性氧代谢的影响

热胁迫导致烟草叶片细胞膜系统显著受损,表现为SOD活性降低和MDA含量明显升高,叶片叶绿素含量下降,活性氧增加。10 mmol/L CaCl2溶液处理烟草幼苗后,能有效降低热胁迫下叶片细胞膜透性,维持较高的SOD和CAT等抗氧化酶活性,减缓 O-·2 形成和膜脂过氧化反应。研究结果表明,CaCl2处理提高了烟草叶片膜稳定性和膜保护酶活性,有利于保护细胞膜结构,降低高温对烟草幼苗的伤害。钙离子螯合剂EGTA能在一定程度上降低烟草叶片的抗热性。     

牛磺酸对小麦幼苗生长的生理效应

采用牛磺酸溶液培育小麦幼苗,测定10、100、500、1 000、5 000 mg/L的牛磺酸对小麦幼苗的光合作用PS Ⅱ光化学效率、细胞膜相对透性和膜脂过氧化以及生长的影响.结果表明,与对照组相比,适宜浓度的牛磺酸处理可促进小麦幼苗的生长,使其根长、株高、单株幼苗的干重和鲜重增加,并在一定程度上提高叶片的光化学效率,降低细胞膜相对透性和膜脂过氧化产物的含量;最适处理浓度约为500 mg/L.说明牛磺酸对小麦幼苗细胞膜有一定的保护作用.     

镉胁迫对平邑甜茶脂肪酸构成及脂质过氧化的影响

以平邑甜茶幼苗为试材,研究了镉胁迫下幼苗叶片和根系膜脂肪酸构成、活性氧、脂氧合酶和丙二醛含量的变化.结果表明：氯化镉处理后7～12 h,脂肪酸种类及其相对含量变化最为明显.处理后7 h,叶片和根系脂肪酸不饱和水平升至最高,含量分别达8282%和7243%;叶片可检测到的脂肪酸在处理后12 h由11种增至14种,根系则在处理17 h后由4种增至6种.O₂^.-产生速率在处理3 h、H₂O₂含量在处理7 h时升至最高,丙二醛含量和脂氧合酶活性则随着处理时间的延长逐渐增加.镉胁迫通过诱导活性氧和脂氧合酶来改变平邑甜茶脂肪酸构成,并引起脂质过氧化;镉处理12 h前,脂质过氧化是活性氧和脂氧合酶的共同结果;但处理12 h后,脂质过氧化加剧主要在于脂氧合酶活性的持续增加.     

Arsenic-induced oxidative stress in the common bean legume,Phaseolus vulgaris L. seedlings and its amelioration by exogenous nitric oxide

The adverse effects of arsenic (As) toxicity on seedling growth, root and shoot anatomy, chlorophyll and carotenoid contents, root oxidizability (RO), antioxidant enzyme activities, H₂O₂ content, lipid peroxidation and electrolyte leakage (EL%) in common bean (Phaseolus vulgaris L.) were investigated. The role of exogenous nitric oxide (NO) in amelioration of As-induced inhibitory effect was also evaluated using sodium nitroprusside (100 μM SNP) as NO donor and 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (200 μM PTIO) as NO scavenger in different combinations with 50 μM As. As-induced growth inhibition was associated with marked anomalies in anatomical features, reduction in pigment composition, increased RO and severe perturbations in antioxidant enzyme activities. While activity of superoxide dismutase and catalase increased, levels of ascorbate peroxidase, dehydroascorbate reductase and glutathione reductase decreased significantly and guaiacol peroxidase remained normal. The over-accumulation of H₂O₂ content along with high level of lipid peroxidation and electrolyte leakage indicates As-induced oxidative damage in P. vulgaris seedlings with more pronounced effect on the roots than the shoots. Exogenous addition of NO significantly reversed the As-induced oxidative stress, maintaining H₂O₂ in a certain level through balanced alterations of antioxidant enzyme activities. The role of NO in the process of amelioration has ultimately been manifested by significant reduction of membrane damage and improvement of growth performance in plants grown on As + SNP media. Onset of oxidative stress was more severe after addition of PTIO, which confirms the protective role of NO against As-induced oxidative damage in P. vulgaris seedlings.     

WGA reduces the level of oxidative stress in wheat seedlings under salinity

Protective effect of exogenous wheat germ agglutinin (WGA) on wheat seedling (Triticum aestivum L.) during salinity stress was studied. In particular, we examined the state of pro- and antioxidant systems as well as the level of peroxide oxidation of lipids and electrolyte leakage under control conditions and when stressed with NaCl. Generation of superoxide anions and activity of both superoxide dismutase (SOD) and peroxidase increased during saline stress. Accumulation of O₂ ^·− resulted in peroxide oxidation of lipids and electrolyte leakage in response to stress. The injurious effect of salinity on root growth of seedlings was manifested by a decreased mitotic index (MI) in apical root meristem. This study show that WGA pretreatment decreased salt-induced superoxide anion generation, SOD and peroxidase activities, levels of lipid peroxidation and electrolytes leakage as well as correlating with a reduction in the inhibition of root apical meristem mitotic activity in salt-treated plants. This suggests that exogenous WGA reduced the detrimental effects of salinity-induced oxidative stress in wheat seedlings. Thus WGA effects on a balance of reactive oxygen species (ROS) and activities of antioxidant enzymes may provide an important contribution to a range of the defense reactions induced by this lectin in wheat plants.     

Antioxidant response of wheat roots to drought acclimation

Wheat (Triticum aestivum L.) seedlings of a drought-resistant cv. C306 were subjected to severe water deficit directly or through stress cycles of increasing intensity with intermittent recovery periods. The antioxidant defense in terms of redox metabolites and enzymes in root cells and mitochondria was examined in relation to membrane damage. Acclimated seedlings exhibited higher relative water content and were able to limit the accumulation of H₂O₂ and membrane damage during subsequent severe water stress conditions. This was due to systematic up-regulation of superoxide dismutase, ascorbate peroxidase (APX), catalase, peroxidases, and ascorbate–glutathione cycle components at both the whole cell level as well as in mitochondria. In contrast, direct exposure of severe water stress to non-acclimated seedlings caused greater water loss, excessive accumulation of H₂O₂ followed by elevated lipid peroxidation due to the poor antioxidant enzyme response particularly of APX, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and ascorbate–glutathione redox balance. Mitochondrial antioxidant defense was found to be better than the cellular defense in non-acclimated roots. Termination of stress followed by rewatering leads to a rapid enhancement in all the antioxidant defense components in non-acclimated roots, which suggested that the excess levels of H₂O₂ during severe water stress conditions might have inhibited or down-regulated the antioxidant enzymes. Hence, drought acclimation conferred enhanced tolerance toward oxidative stress in the root tissue of wheat seedlings due to both reactive oxygen species restriction and well-coordinated induction of antioxidant defense.     

24-Epibrassinolide alleviated zinc-induced oxidative stress in radish (Raphanus sativus L.) seedlings by enhancing antioxidative system

This article encompasses the results on the effects of 24-epibrassinolide (EBR) on the changes in reactive oxygen species (ROS) and activities of antioxidative enzymes in radish (Raphanus sativus L.) seedlings subjected to zinc (Zn) stress. Zn toxicity resulted in significant enhancement in the level of membrane lipid peroxidation, protein oxidation, contents of hydrogen peroxide (H₂O₂) and hydroxyl radical (^·OH), the production rate of superoxide radicals (O ₂ ^·? ) and the activities of lipoxygenase and NADPH oxidase in radish seedlings indicating the induction of oxidative stress. However, Zn-mediated enhancement in indices of oxidative stress was considerably decreased by EBR treatment. EBR application enhanced the activities of catalase, superoxide dismutase, guaiacol peroxidase, glutathione peroxidase, and peroxidase in radish seedlings under Zn stress. EBR treatment reduced the activity of ascorbic acid oxidase in Zn stressed seedlings. Further, EBR application also enhanced the free proline and phenol levels under Zn stress. From the results obtained in this study, it can be inferred that EBR application alleviated oxidative damage caused by over production of ROS through the up regulation of antioxidative capacity in Zn stressed radish seedlings.     

Antioxidant responses of citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), exposed to thermal stress

Relatively low or high temperatures are responsible for a variety of physiological stress responses in insects and mites. Induced thermal stress was recently associated with increased reactive oxygen species (ROS) generation, which caused oxidative damage. In this study, we examined the time-related effect of the relatively low (0, 5, 10, and 15 °C) or high (32, 35, 38, and 41 °C) temperatures on the activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidases (POX), and glutathione-S-transferase (GST), and the total antioxidant capacity (TEAC) of the citrus red mite, Panonychus citri (McGregor). The malondialdehyde (MDA) concentration, as a marker of lipid peroxidation in organisms, was also measured in the citrus red mite under thermal stress conditions. Results showed that SOD and GST activities were significantly increased and play an important role in the process of antioxidant response to thermal stress. Lipid peroxidation levels increased significantly (P < 0.001) and changed in a time-dependent manner. CAT and POX activity, as well as TEAC, did not vary significantly and play a minor role to remove the ROS generation. These results suggest that thermal stress leads to oxidative stress and antioxidant enzymes play an important role in reducing oxidative damage in the citrus red mite.     

Zinc induces distinct changes in the metabolism of reactive oxygen and nitrogen species (ROS and RNS) in the roots of two Brassica species with different sensitivity to zinc stress

Background and Aims Zinc (Zn) is an essential micronutrient naturally present in soils, but anthropogenic activities can lead to accumulation in the environment and resulting damage to plants. Heavy metals such as Zn can induce oxidative stress and the generation of reactive oxygen and nitrogen species (ROS and RNS), which can reduce growth and yield in crop plants. This study assesses the interplay of these two families of molecules in order to evaluate the responses in roots of two Brassica species under high concentrations of Zn.Methods Nine-day-old hydroponically grown Brassica juncea (Indian mustard) and B. napus (oilseed rape) seedlings were treated with ZnSO₄ (0, 50, 150 and 300 µm) for 7 d. Stress intensity was assessed through analyses of cell wall damage and cell viability. Biochemical and cellular techniques were used to measure key components of the metabolism of ROS and RNS including lipid peroxidation, enzymatic antioxidants, protein nitration and content of superoxide radical (O2·−), nitric oxide (NO) and peroxynitrite (ONOO⁻).Key Results Analysis of morphological root damage and alterations of microelement homeostasis indicate that B. juncea is more tolerant to Zn stress than B. napus. ROS and RNS parameters suggest that the oxidative components are predominant compared with the nitrosative components in the root system of both species.Conclusions The results indicate a clear relationship between ROS and RNS metabolism as a mechanism of response against stress caused by an excess of Zn. The oxidative stress components seem to be more dominant than the elements of the nitrosative stress in the root system of these two Brassica species.     

南方根结线虫对不同砧木嫁接番茄苗活性氧清除系统的影响

采用盆栽人工接种方法,对番茄嫁接苗进行了抗性评价,研究了番茄嫁接苗叶片中抗氧化酶活性和活性氧代谢的动态变化。结果表明,接种南方根结线虫(J2)后,砧木嫁接苗表现为高抗,自根嫁接苗为高感。通过嫁接换根,与自根嫁接苗相比,砧木嫁接苗明显提高了接穗叶片的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性,降低了超氧阴离子(O.2-)产生速率以及过氧化氢(H2O2)和丙二醛(MDA)含量。表明番茄植株体内的活性氧水平和抗氧化酶活性的高低与其抗根结线虫的能力密切相关,较低的活性氧水平和较高的抗氧化酶活性有利于减轻对膜系统的伤害,提高番茄植株的抗根结线虫能力。     

The golden root, Rhodiola rosea, prolongs lifespan but decreases oxidative stress resistance in yeast Saccharomyces cerevisiae

The effect of aqueous extract from R. rosea root on lifespan and the activity of antioxidant enzymes in budding yeast Saccharomyces cerevisiae have been studied. The supplementation of the growth medium with R. rosea extract decreased survival of exponentially growing S. cerevisiae cells under H₂O₂-induced oxidative stress, but increased viability and reproduction success of yeast cells in stationary phase. The extract did not significantly affect catalase activity and decreased SOD activity in chronologically aged yeast population. These results suggest that R. rosea acts as a stressor for S. cerevisiae cells, what sensitizes yeast cells to oxidative stress at exponential phase, but induces adaptation in stationary phase cells demonstrating the positive effect on yeast survival without activation of major antioxidant enzymes.     

Cytoprotective propensity of Bacopa monniera against hydrogen peroxide induced oxidative damage in neuronal and lung epithelial cells

Hydrogen peroxide (H₂O₂), a major reactive oxygen species (ROS) produced during oxidative stress, is toxic to the cells. Hence, H₂O₂ has been extensively used to study the effects of antioxidant and cytoprotective role of phytochemicals. In the present investigation H₂O₂ was used to induce oxidative stress via ROS production within PC12 and L132 cells. Cytoprotective propensity of Bacopa monniera extract (BME) was confirmed by cell viability assays, ROS estimation, lipid peroxidation, mitochondria membrane potential assay, comet assay followed by gene expression studies of antioxidant enzymes in PC12 and L132 cells treated with H₂O₂ for 24 h with or without BME pre-treatment. Our results elucidate that BME possesses radical scavenging activity by scavenging 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), superoxide radical, and nitric oxide radicals. The IC₅₀ value of BME against these radicals was found to be 226.19, 15.17, 30.07, and 34.55 µg/ml, respectively). The IC₅₀ of BME against ROS, lipid peroxidation and protein carbonylation was found to be 1296.53, 753.22, and 589.04 µg/ml in brain and 1137.08, 1079.65, and 11101.25 µg/ml in lung tissues, respectively. Further cytoprotective potency of the BME ameliorated the mitochondrial and plasma membrane damage induced by H₂O₂ as evidenced by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase leakage assays in both PC12 and L132 cells. H₂O₂ induced cellular, nuclear and mitochondrial membrane damage was restored by BME pre-treatment. H₂O₂ induced depleted antioxidant status was also replenished by BME pre-treatment. This was confirmed by spectrophotometric analysis, semi-quantitative RT-PCR and western blot studies. These results justify the traditional usage of BME based on its promising antioxidant and cytoprotective property.