首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到16条相似文献,搜索用时 140 毫秒
1.
以白菜型油菜‘陇油6号’和‘天油2号’为试验材料,经MAPK抑制剂U0126、H2O2清除剂DMTU、NADPH氧化酶抑制剂DPI和IMD预处理后再分别进行盐胁迫、PEG-6000模拟干旱胁迫,研究其对两种油菜幼苗活性氧、抗氧化酶活性和RbohCRbohF基因表达的影响.结果表明: 盐胁迫和PEG-6000模拟干旱胁迫下,两种白菜型油菜中H2O2积累量上升,O2积累量下降,抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT、抗坏血酸过氧化物酶APX和谷胱甘肽还原酶GR)活性和RbohCRbohF基因表达均升高.与单独胁迫处理相比,两种油菜O2积累、抗氧化酶活性和RbohCRbohF基因的表达量均明显降低,经DMTU、DPI和IMD预处理后再分别进行盐和干旱胁迫,H2O2积累量下降,但U0126预处理后再进行胁迫处理,H2O2积累量上升.说明NADPH氧化酶、MAP激酶级联途径、H2O2参与了盐、干旱胁迫下活性氧产生、抗氧化酶活性变化和RbohCRbohF基因表达的调控.  相似文献   

2.
利用cDNA末端快速分离(RACE)技术从陇油6号油菜中克隆得到一个新的谷胱甘肽还原酶基因GR2,全长2073 bp,开放阅读框1692 bp,编码563个氨基酸,预测蛋白质分子量为60.7 kDa,等电点7.9.实时荧光定量PCR分析表明:GR2基因在油菜根、茎、叶中均有表达,其中在叶中表达量最高.GR1和GR2基因的转录以及谷胱甘肽还原酶(GR)活性受到低温、高温、干旱、高盐胁迫的诱导,表明油菜谷胱甘肽还原酶在抵御低温、高温、干旱、高盐胁迫过程中发挥重要作用.脱落酸(ABA)预处理后再进行上述胁迫处理,与单独上述胁迫相比,GR1和GR2基因的转录以及GR活性水平明显上升,表明ABA可以诱导GR1和GR2基因表达和GR酶活性.MAPKK抑制剂U0126预处理后再进行上述胁迫处理,与单独上述胁迫相比,GR1和GR2基因的转录以及GR活性水平明显下降,表明U0126对GR1、GR2基因表达以及GR酶活性有抑制作用.  相似文献   

3.
利用RACE技术从‘陇油6号’油菜中克隆得到1个新的CBF(C-repeat binding factor)基因(GenBank登录号为KP974691),全长1 000bp,其中包括5′-UTR 114bp,3′-UTR 241bp,开放阅读框645bp,编码215个氨基酸,推测的蛋白质分子量23.8kD,理论等电点为4.86。在氨基酸序列水平上,该基因与多种植物具有较高的一致性,其中与萝卜、紫茎泽兰、卷果涩荠和拟南芥的一致性分别为93.5%、83.7%、81.0%、79.0%。实时荧光定量PCR分析表明,CBF基因在油菜的茎、叶和下胚轴中均有表达,没有组织特异性;该基因表达受低温、ABA、H2O2诱导,MAPKK抑制剂U0126预处理12h再经低温、ABA、H2O2诱导,与单独处理结果相比,CBF基因表达明显降低,表明该基因在油菜适应低温、ABA、H2O2胁迫过程中发挥作用。  相似文献   

4.
报道了干旱胁迫下外源24-表油菜素内酯(EBR)对辣椒幼苗叶片H2O2和MDA含量,抗氧化酶活性,以及耐旱相关基因表达的影响。结果表明,0.1 μmol·L-1 EBR处理诱导了辣椒幼苗叶片H2O2含量的增加,并提高了SOD、APX、CAT、DHAR、MDAR和GR活性;干旱胁迫下,EBR处理显著诱导了辣椒叶片抗氧化酶活性的增加,并抑制了H2O2和MDA含量的上升;EBR处理也促进了cAPX和MDAR等抗氧化酶基因的表达,以及WRKY3、WRKY6和MYB等转录因子的表达。由此认为,适宜浓度的外源EBR可能是通过信号分子H2O2调控辣椒叶片中WRKY和MYB等转录因子的表达以调控相关耐旱基因表达,增强细胞的抗氧化酶活性,减轻干旱造成的膜质过氧化伤害,从而增强了辣椒幼苗的耐旱性。  相似文献   

5.
以白菜型油菜‘陇油6号’和‘天油2号’为试验材料,经MAPK抑制剂U0126、H_2O_2清除剂DMTU、NADPH氧化酶抑制剂DPI和IMD预处理后再分别进行盐胁迫、PEG-6000模拟干旱胁迫,研究其对两种油菜幼苗活性氧、抗氧化酶活性和RbohC、RbohF基因表达的影响.结果表明:盐胁迫和PEG-6000模拟干旱胁迫下,两种白菜型油菜中H_2O_2积累量上升,O_2积累量下降,抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT、抗坏血酸过氧化物酶APX和谷胱甘肽还原酶GR)活性和RbohC、RbohF基因表达均升高.与单独胁迫处理相比,两种油菜O_2积累、抗氧化酶活性和RbohC、RbohF基因的表达量均明显降低,经DMTU、DPI和IMD预处理后再分别进行盐和干旱胁迫,H_2O_2积累量下降,但U0126预处理后再进行胁迫处理,H_2O_2积累量上升.说明NADPH氧化酶、MAP激酶级联途径、H_2O_2参与了盐、干旱胁迫下活性氧产生、抗氧化酶活性变化和RbohC、RbohF基因表达的调控.  相似文献   

6.
以“陇油7号”油菜为实验材料,研究了外源ATP对油菜幼苗耐寒性的影响。结果表明:与单独低温胁迫相比,外源ATP预处理再进行低温胁迫后,油菜幼苗MDA含量、O2-.含量均显著降低,而叶绿素含量、抗氧化酶活性(SOD、POD、CAT、APX)和RBOHD、RBOHF、CPK4、CPK5基因表达均增加;与外源ATP+低温相比,EGTA+外源ATP+低温处理下,MDA含量显著增加,总叶绿素含量、T-AOC酶活性、RBOHD、RBOHF基因表达均显著下降,DMTU+外源ATP+低温处理下,MDA含量显著增加,总叶绿素含量、Ca2+-ATPase酶活性、CPK4、CPK5基因表达均下降,表明外源ATP通过Ca2+和H2O2依赖性机制影响油菜幼苗的耐寒性。  相似文献   

7.
以烟草悬浮细胞BY-2(Nicotiana tabacum L.cv.Bright Yellow-2)为材料,探讨了在铜离子胁迫下植物细胞死亡发生过程中胞外H2O2及NADPH氧化酶所扮演的角色。实验结果表明,随着外源CuCl2浓度的上升(从0~700 μmol·L-1),细胞死亡水平不断上升,且胞外H2O2的水平也不断增加。在300 μmol·L-1的CuCl2诱导细胞死亡的过程中,加入H2O2清除剂N-N-二甲基硫脲(DMTU)降低了胞外CuCl2胁迫下H2O2含量增加的同时也降低了细胞死亡水平的上升,这一观察表明了铜离子胁迫所导致的细胞死亡的发生和胞外H2O2的增加有关。进一步的研究表明,300 μmol·L-1 CuCl2的胁迫导致了NADPH氧化酶活性的显著性上升,而加入NADPH氧化酶的抑制剂(二亚苯基碘,DPI,)则降低了CuCl2胁迫所导致的细胞死亡和胞外H2O2含量的上升。上述结果表明,胞外H2O2和NADPH氧化酶参与了CuCl2对植物细胞死亡的诱导作用。  相似文献   

8.
过氧化氢和氯化钙对香蕉幼苗抗寒性的影响   总被引:15,自引:0,他引:15  
用H2O2和CaCl2单独或混合使用的方法喷洒香蕉幼苗,并置于低温培养箱中进行冷胁迫处理,发现它们可提高香蕉幼苗冷胁迫期间叶片POD活性,降低细胞质泄漏,增加可溶性糖含量及减缓叶绿素降解,从而减轻冷伤害程度。H2O2和CaCl2混合处理的效果优于单独处理,二者有协同效应。  相似文献   

9.
苯丙氨酸解氨酶在诱导黄瓜幼苗抗寒性中的作用   总被引:2,自引:2,他引:0  
为了探讨苯丙氨酸解氨酶(PAL)在诱导黄瓜幼苗抗寒性中的作用,采用喷施特异抑制剂(AOPP)的方法控制PAL活性,测定幼苗抗寒性的变化.结果表明: 低温可以诱导黄瓜幼苗叶片中PAL的基因表达和活性升高;喷施AOPP显著抑制了叶片中PAL活性,减少了酚类和类黄酮物质的积累.低温对黄瓜幼苗造成显著伤害,AOPP预处理加剧了低温对幼苗的损伤,幼苗抗寒性降低.与对照相比,幼苗叶片中相对电解质渗漏率和丙二醛(MDA)含量显著升高,PSII最大光化学效率(Fv/Fm)降低,光化学猝灭参数Y(NO)升高,胁迫相关基因(PR1-1a、COR47、P5CS、HSP70)的诱导表达受到抑制.低温导致黄瓜幼苗叶片中H2O2积累,还原型抗坏血酸(AsA)含量降低,脱氢抗坏血酸(DHA)含量升高,AsA∶DHA减小;喷施AOPP的幼苗中抗氧化酶(过氧化氢酶CAT、抗坏血酸过氧化物酶APX)活性显著低于对照,H2O2过量积累,AsA∶DHA更低.施用H2O2清除剂可以有效缓解喷施AOPP引起的低温损伤加剧,而施用CAT抑制剂的幼苗对低温胁迫更敏感.表明低温诱导了PAL活性升高,促进了苯丙烷类次生代谢产物的合成,提高了胞内抗氧化酶活性,可有效清除活性氧分子,维持AsA氧化还原状态,缓解低温引起的光损伤和氧化损伤.  相似文献   

10.
H2O2介导的H2S产生参与干旱诱导的拟南芥气孔关闭   总被引:1,自引:0,他引:1  
以野生型拟南芥(Arabidopsis thaliana)及其突变体(atrbohD、atrbohF、atrbohD/F、atl-cdes、atd-cdes)和过表达株系(OEL-CDes、OED-CDes)为材料, 利用药理学实验, 结合分光光度法和激光共聚焦显微技术, 探讨硫化氢(hydrogen sulfide, H2S)在干旱诱导的拟南芥气孔关闭中的作用及其与过氧化氢(hydrogen peroxide, H2O2)的关系。结果表明, H2S清除剂次牛磺酸(hypotaurine, HT)及合成抑制剂氨氧基乙酸(aminooxy acetic acid, AOA)、羟胺(hydroxylamine, NH2OH)和丙酮酸钾(potasium pyruvate, C3H3KO3)+氨水(ammonia, NH3)均可不同程度抑制干旱诱导的气孔关闭; 干旱对OEL-CDes和OED-CDes植株气孔关闭的诱导作用明显, 而atl-cdes和atd-cdes叶片气孔对干旱胁迫反应的敏感性下降; 干旱胁迫能明显增加拟南芥保卫细胞中H2O2水平及叶片中H2S含量, 提高D-/L-半胱氨酸脱巯基酶活性及基因表达量, 而对突变体atrbohD、atrbohF和atrbohD/F没有显著影响。清除H2O2可减弱干旱胁迫对H2S含量和D-/L-半胱氨酸脱巯基酶活性的诱导效应。研究 结果表明H2S位于H2O2下游参与干旱诱导拟南芥气孔关闭的信号转导过程。  相似文献   

11.
Hydrogen peroxide, produced by inflammatory and vascular cells, induces oxidative stress that may contribute to endothelial dysfunction. In smooth muscle cells, H2O2 induces production of O2 by activating NADPH oxidase. However, the mechanisms whereby H2O2 induces oxidative stress in endothelial cells are poorly understood. We examined the effects of H2O2 on O2 levels on porcine aortic endothelial cells (PAEC). Treatment with 60 μmol/L H2O2 markedly increased intracellular O2 levels (determined by conversion of dihydroethidium to hydroxyethidium) and produced cytotoxicity (determined by propidium iodide staining) in PAEC. Overexpression of human manganese superoxide dismutase in PAEC reduced O2 levels and attenuated cytotoxicity resulting from treatment with H2O2. L-NAME, an inhibitor of nitric oxide synthase (NOS), and apocynin, an inhibitor of NADPH oxidase, reduced O2 levels in PAEC treated with H2O2, suggesting that both NOS and NADPH oxidase contribute to H2O2-induced O2 in PAEC. Inhibition of NADPH oxidase using apocynin and NOS rescue with L-sepiapterin together reduced O2 levels in PAEC treated with H2O2 to control levels. This suggests interaction-distinct NOS and NADPH oxidase pathways to superoxide. We conclude that H2O2 produces oxidative stress in endothelial cells by increasing intracellular O2 levels through NOS and NADPH oxidase. These findings suggest a complex interaction between H2O2 and oxidant-generating enzymes that may contribute to endothelial dysfunction.  相似文献   

12.
Hydrogen peroxide (H2O2) is known to both induce and inhibit apoptosis, however the mechanisms are unclear. We found that H2O2 inhibited the activity of recombinant caspase-3 and caspase-8, half-inhibition occurring at about 17 μM H2O2. This inhibition was both prevented and reversed by dithiothreitol while glutathione had little protective effect. 100–200 μM H2O2 added to macrophages after induction of caspase activation by nitric oxide or serum withdrawal substantially inhibited caspase activity. Activation of H2O2-producing NADPH oxidase in macrophages also caused catalase-sensitive inactivation of cellular caspases. The data suggest that the activity of caspases in cells can be directly but reversibly inhibited by H2O2.  相似文献   

13.
水杨酸(salicylic acid,SA)处理可诱导丹参悬浮培养细胞内H2O2产生及其培养基碱化。利用NADPH氧化酶抑制剂咪唑(imidazole,IMD)、H2O2淬灭剂二甲基硫脲(dimethylthiourea,DMTU)、质膜H+-ATPase抑制剂钒酸钠(Na3VO4)及激活剂壳梭孢菌素(fusicoccin,FC)处理丹参悬浮培养细胞,探讨SA诱导的H2O2迸发与培养基碱化之间的关系。结果表明,H2O2可促发培养基碱化,IMD和DMTU抑制SA诱发的培养基碱化,说明H2O2参与SA诱发的培养基碱化过程;SA抑制质膜H+-ATPase活性,Na3VO4引发培养基碱化并使H2O2迸发时间提前,FC处理逆转了SA诱导的培养基碱化及H2O2迸发,说明质膜H+-ATPase调控培养基pH值变化,培养基碱化促进了H2O2产生。因此,丹参悬浮培养细胞内H2O2水平与其培养基碱化程度之间相互关联、共同作用,协同响应SA的诱导。  相似文献   

14.
Excessive generation of reactive oxygen species (ROS) in the central nervous system (CNS) is a leading cause of neuronal injury. Despite yet unknown mechanisms, oxidant compounds such as H2O2 have been shown to stimulate the release of arachidonic acid (AA) in a number of cell systems. In this study, H2O2 and menadione, a compound known to release H2O2 intracellularly, were used to examine the phospholipases A2 (PLA2) responsible for AA release from primary murine astrocytes. Both H2O2 and menadione dose-dependently stimulated AA release, and the release mediated by H2O2 was completely inhibited by catalase. H2O2 also stimulated phosphorylation of extracellular signal-regulated kinases (ERK1/2) and cytosolic phospholipase A2 (cPLA2). However, complete inhibition of cPLA2 phosphorylation by U0126, an inhibitor for mitogen-activated protein kinase kinase (MEK) and GF109203x, a nonselective PKC inhibitor preferring the conventional and novel isoforms, only reduced H2O2-stimulated AA release by 50%. MAFP, a selective, active, site-directed, irreversible inhibitor of both cPLA2 and the Ca2+-independent iPLA2, nearly completely inhibited H2O2-mediated AA release; but, HELSS, a potent irreversible inhibitor of iPLA2, only inhibited H2O2-mediated AA release by 40%. Along with the observation that H2O2-mediated AA release was only partially inhibited upon chelating intracellular Ca2+ by BAPTA, these results indicate the involvement of both cPLA2 and iPLA2 in H2O2-mediated AA release in murine astrocytes.  相似文献   

15.
为了解活性氧(reactive oxygen species,ROS)在香菇菌丝后熟转色形成中的作用及其自噬细胞学特征,以香菇工厂化菌株KS11为研究材料,分析其在菌丝后熟转色过程中4个时间点(30、45、60、75 d)的活性氧含量(ROS)、丙二醛(MDA)含量、NADPH氧化酶浓度、抗氧化酶活性以及外源活性氧和DPI对其影响的表型试验,利用透射电镜观察该过程菌丝细胞自噬特征变化,并运用实时荧光定量PCR对自噬基因Atg8的表达水平进行比较分析。结果表明:(1) H2O2作为主要的活性氧因子在菌丝后熟转色形成中呈现显著动态变化,后熟转色过程中不断升高,并在转色中第60天呈高峰值。(2) NADPH氧化酶浓度与H2O2含量变化呈紧密正相关。(3)外源施加一定浓度H2O2显著促进香菇菌丝后熟转色,且DPI作为NADPH氧化酶抑制剂显著抑制了香菇菌丝后熟转色的发生。(4)香菇菌丝后熟转色过程中,细胞自噬特征逐渐增强,并在转色中后期最显著。上述结果表明以H  相似文献   

16.
The role of H2O2 in abscisic acid (ABA)-induced rice leaf senescence is investigated. ABA treatment resulted in H2O2 production in rice leaves, which preceded the occurrence of leaf senescence. Dimethylthiourea, a chemical trap for H2O2, was observed to be effective in inhibiting ABA-induced senescence, ABA-increased malondialdehyde (MDA) content, ABA-increased antioxidative enzyme activities (superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase), and ABA-decreased antioxidant contents (ascorbic acid and reduced glutathione) in rice leaves. Diphenyleneiodonium chloride (DPI) and imidazole (IMD), inhibitors of NADPH oxidase, and KCN and NaN3, inhibitors of peroxidase, prevented ABA-induced H2O2 production, suggesting NADPH oxidase and peroxidase are H2O2-generating enzymes in ABA-treated rice leaves. DPI, IMD, KCN, and NaN3 also inhibited ABA-promoted senescence, ABA-increased MDA contents, ABA-increased antioxidative enzyme activities, and ABA-decreased antioxidants in rice leaves. These results suggest that H2O2 is involved in ABA-induced senescence of rice leaves.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号