Relationship between chloroplastic H2O2 and the salicylic acid response

Reactive oxygen species (ROS) act as signaling molecules for regulating plant responses to abiotic and biotic stress and there exist source- and kind-specific pathways for ROS signaling. Recently, we created a novel system for producing H₂O₂ in Arabidopsis chloroplasts by chemical-dependent thylakoid membrane-bound ascorbate peroxidase (tAPX) silencing using an estrogen-inducible RNAi method. Microarray analysis revealed that the expression of a large set of genes was altered in response to tAPX silencing, some of which are known to be involved in pathogen response/resistance. Furthermore, we found that tAPX silencing enhances the levels of salicylic acid (SA) and the response to SA, a central regulator for biotic stress response. In this addendum, we describe the relationship between chloroplastic H₂O₂ and SA in stress response, and discuss the function of the kind- and source-specific ROS signaling in SA-mediated stress response.     

Controlled levels of salicylic acid are required for optimal photosynthesis and redox homeostasis

Sudden exposure of plants to high light (HL) leads to metabolic and physiological disruption of the photosynthetic cells. Changes in ROS content, adjustment of photosynthetic processes and the antioxidant pools and, ultimately, gene induction are essential components for a successful acclimation to the new light conditions. The influence of salicylic acid (SA) on plant growth, short-term acclimation to HL, and on the redox homeostasis of Arabidopsis thaliana leaves was assessed here. The dwarf phenotype displayed by mutants with high SA content (cpr1-1, cpr5-1, cpr6-1, and dnd1-1) was less pronounced when these plants were grown in HL, suggesting that the inhibitory effect of SA on growth was partly overcome at higher light intensities. Moreover, higher SA content affected energy conversion processes in low light, but did not impair short-term acclimation to HL. On the other hand, mutants with low foliar SA content (NahG and sid2-2) were impaired in acclimation to transient exposure to HL and thus predisposed to oxidative stress. Low and high SA levels were strictly correlated to a lower and higher foliar H(2)O(2) content, respectively. Furthermore high SA was also associated with higher GSH contents, suggesting a tight correlation between SA, H(2)O(2) and GSH contents in plants. These observations implied an essential role of SA in the acclimation processes and in regulating the redox homeostasis of the cell. Implications for the role of SA in pathogen defence signalling are also discussed.     

Thylakoid-bound ascorbate peroxidase mutant exhibits impaired electron transport and photosynthetic activity

In chloroplasts, stromal and thylakoid-bound ascorbate peroxidases (tAPX) play a major role in the removal of H(2)O(2) produced during photosynthesis. Here, we report that hexaploid wheat (Triticum aestivum) expresses three homeologous tAPX genes (TaAPX-6A, TaAPX-6B, and TaAPX-6D) mapping on group-6 chromosomes. The tAPX activity of a mutant line lacking TaAPX-6B was 40% lower than that of the wild type. When grown at high-light intensity photosystem II electron transfer, photosynthetic activity and biomass accumulation were significantly reduced in this mutant, suggesting that tAPX activity is essential for photosynthesis. Despite the reduced tAPX activity, mutant plants did not exhibit oxidative damage probably due to the reduced photochemical activity. This might be the result of a compensating mechanism to prevent oxidative damage having as a consequence a decrease in growth of the tAPX mutant plants.     

胶原蛋白肽经由Nrf2信号通路对H_2O_2诱导的肝细胞氧化损伤的保护作用(英文)

目的:氧化应激在肝脏疾病中扮演着重要的角色。胶原蛋白肽是天然的抗氧化剂,其在动物实验中已经被证实有抑制氧化应激的作用。最新研究证实胶原蛋白肽将有可能被应用在肝脏疾病的预防中,但是很少有研究报道其分子作用机制。因此本研究在胶原蛋白肽是对H2O2诱导的正常人的肝细胞系HL7702氧化损伤有保护作用的基础上,并探索其分子作用机制。方法:实验设空白对照组,H2O2模型组,胶原蛋白肽低、中、高剂量组(10,100,200μg/ml)。胶原蛋白肽各组加入相应浓度的药物预处理12 h后,与模型组一起加入300μM H2O2的H2O2共同培养12 h,空白对照组正常培养。细胞毒性是由CCK8和乳酸脱氢酶(LDH)的释放检测。抗氧化试剂盒检测细胞内活性氧的水平,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量的变化。Western blot检测细胞内Nrf2蛋白的表达水平。结果:胶原蛋白肽对H2O2诱导的正常人的肝细胞系HL7702氧化损伤有保护作用。胶原蛋白肽能够及时清除细胞内的活性氧,增加Nrf2的蛋白表达水平,提高超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性,减轻脂质过氧化反应,从而保护正常人的肝细胞系HL7702。结论:总之,胶原蛋白肽通过增加Nrf2的蛋白表达水平,提高抗氧化活性,对H2O2诱导损伤的肝细胞发挥保护作用。本研究为胶原蛋白肽的分子作用机制提供了新的证据,将有助于预防氧化应激所致的肝损伤。     

植物胁迫信号转导过程中活性氧和促细胞分裂原活化蛋白激酶的调节作用

以H2O2为代表的活性氧(reactive oxygen species,ROS)和以促细胞分裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)为代表的蛋白激酶广泛存在于植物细胞并参与各种生理反应过程.生物胁迫条件下,一些MAP激酶特异性地调节氧化猝发(oxidative burst,OXB)和过敏反应(hypersensitive response,HR),水杨酸(salicylic acid,SA)诱导的MAP激酶(SA-induced protein kinase,SIPK)和ROS共同参与系统获得性抗性(systemic acquired resistance,SAR)的建立;SIPK、P38 MAPK等分别与H2O2共同调节臭氧、受伤和渗透胁迫等多种非生物胁迫生理反应.ROS和MAP激酶共同调节植物胁迫信号转导,但其机制尚需进一步的研究.     

Nrf2/HO-1信号通路在人诱导性多能干细胞氧化应激中的作用

氧化应激是诱导性多能干细胞(induced pluripotent stem cell, iPSC)在培养和应用中遇到的一个关键问题,探讨其作用机制具有重要的理论和实践意义。目前有关iPSC氧化应激的研究相对较少,Nrf2/HO-1信号通路在其中的作用尚不明了。因此,本研究以不同浓度的H2O2(100、200、300、400 μmol/L)处理人iPSC(hiPSC),分别在4 h和24 h于倒置显微镜下观察hiPSC及其饲养层细胞SNL氧化损伤的程度,通过碱性磷酸酶（alkaline phosphatase, AP）试剂盒和超氧化物阴离子荧光探针,分别检测hiPSC多能性和细胞活性氧(reactive oxygen species, ROS)水平,并通过qRT-PCR检测H2O2处理4 h后早期应激状态下Nrf2和HO 1 mRNA的表达水平,免疫细胞化学和Western印迹检测p-Nrf2和HO-1蛋白质的表达量。结果表明：hiPSC和SNL细胞的ROS水平呈H2O2剂量依赖性升高。除了100 μmol/L H2O2组hiPSC的细胞形态和多能性保持较好外,其余浓度H2O2均导致hiPSC出现不同程度损伤和死亡。但与SNL细胞相比,hiPSC中ROS水平相对较低,细胞状态也相对较好。SNL细胞中Nrf2和HO-1-mRNA表达的变化幅度与H2O2浓度呈线性相关,而hiPSC中Nrf2和HO-1表达的变化幅度与H2O2浓度之间并未呈现线性相关,其中Nrf2在100 μmol/L H2O2组表达量最高,而HO-1在200 μmol/L H2O2组表达量最高,意味着hiPSC氧化应激调控机制的复杂性。综上结果表明,hiPSC具有较好的抗氧化能力,其相关机制与Nrf2/HO-1信号通路有关,同时也可能涉及到其它相关通路的交互作用。     

SB202190 调节蚕豆保卫细胞中SA 诱导H2O2 产生

运用激光共聚焦扫描技术, 在p38 MAP激酶专一抑制剂SB202190处理下, 探索植物促分裂原活化蛋白激酶(mitogenactivated protein kinase, MAP激酶)介导蚕豆(Vicia faba)保卫细胞中H2O2为代表的活性氧(reactive oxygen species, ROS)信号机制, 发现: p38 MAP激酶专一抑制剂SB202190处理没有导致蚕豆保卫细胞中H2O2和Ca2+探针荧光强度增强, 与水杨酸 (salicylic acid, SA) 或脱落酸 (abscisic acid, ABA) 迅速加强2种探针荧光强度形成鲜明对比; 而该抑制剂分别与SA和ABA共同处理, 前者H2O2探针荧光强度没有增加, 而后者荧光强度仍然能够增加; 而进一步使用Ca2+螯合剂BAPTA和SB202190 ＋SA共同处理, H2O2探针荧光强度没有增加。这些结果初步表明: 无论胞质Ca2+浓度高低, SB202190调节蚕豆保卫细胞中SA诱导H2O2产生, 但是不调节植物逆境信使分子ABA 此类的反应。因此推测, 植物细胞中可能有类似动物和酵母细胞中的p38MAP激酶类, 并可能专一调节植物保卫细胞中H2O2信号通路。据我们所知, 这是首次报道SB202190和SA共同调节植物保卫细胞中ROS信号过程。     

过量表达番茄类囊体膜抗坏血酸过氧化物酶基因（StAPX）提高了烟草种苗的抗氧化能力

为了探讨叶绿体类囊体膜抗坏血酸过氧化物酶（tAPX）与其抗氧化性的关系,从番茄叶片中分离了叶绿体类囊体膜抗坏血酸过氧化物酶基因（StA跚并转入到烟草中。以野生型（WT）、转正义StAPX烟草株系T3-3和T3-6为试材,测定了外源过氧化氢诱导的氧化胁迫条件下APX酶活性、过氧化氢酶（CAT）活性、过氧化氢（H2O2）含量、叶绿素荧光参数及叶绿素含量等。Northern杂交显示StAPX因的表达受外源H2O2氧化胁迫的诱导。氧化胁迫下转基因烟草的APX酶活性和清除H2O2的能力都显著高于野生型,并且转基因烟草比野生型具有更高的PSII最大光化学效率及叶绿素含量。结果表明,．刚尸舶勺过量表达有助于提高外源H2O2诱导的转基因烟草的抗氧化能力。     

Ozone-induced ethylene production is dependent on salicylic acid,and both salicylic acid and ethylene act in concert to regulate ozone-induced cell death

Ethylene is known to influence plant defense responses including cell death in response to both biotic and abiotic stress factors. However, whether ethylene acts alone or in conjunction with other signaling pathways is not clearly understood. Ethylene overproducer mutants, eto1 and eto3, produced high levels of ethylene and developed necrotic lesions in response to an acute O3 exposure that does not induce lesions in O3-tolerant wild-type Col-0 plants. Treatment of plants with ethylene inhibitors completely blocked O3-induced ethylene production and partially attenuated O3-induced cell death. Analyses of the responses of molecular markers of specific signaling pathways indicated a relationship between salicylic acid (SA)- and ethylene-signaling pathways and O3 sensitivity. Both eto1 and eto3 plants constitutively accumulated threefold higher levels of total SA and exhibited a rapid increase in free SA and ethylene levels prior to lesion formation in response to O3 exposure. SA pre-treatments increased O3 sensitivity of Col-0, suggesting that constitutive high SA levels prime leaf tissue to exhibit increased magnitude of O3-induced cell death. NahG and npr1 plants compromised in SA signaling failed to produce ethylene in response to O3 and other stress factors suggesting that SA is required for stress-induced ethylene production. Furthermore, NahG expression in the dominant eto3 mutant attenuated ethylene-dependent PR4 expression and rescued the O3-induced HR (hypersensitive response) cell death phenotype exhibited by eto3 plants. Our results suggest that both SA and ethylene act in concert to influence cell death in O3-sensitive genotypes, and that O3-induced ethylene production is dependent on SA.     

SB202190调节蚕豆保卫细胞中SA诱导H2O2产生

运用激光共聚焦扫描技术,在p38MAP激酶专一抑制剂SB202190处理下,探索植物促分裂原活化蛋白激酶（mitogen-activated protein kinase,MAP激酶）介导蚕豆（Vicia faba）保卫细胞中H2O2为代表的活性氧（reactive oxygen species,ROS）信号机制,发现：p38MAP激酶专一抑制剂SB202190处理没有导致蚕豆保卫细胞中H2O2和Ca^2＋探针荧光强度增强,与水杨酸（salicylic acid,SA）或脱落酸（abscisic acid,ABA）迅速加强2种探针荧光强度形成鲜明对比;而该抑制剂分别与SA和ABA共同处理,前者H2O2探针荧光强度没有增加,而后者荧光强度仍然能够增加;而进一步使用Ca^2＋螯合剂BAPTA和SB202190＋SA共同处理,H2O2探针荧光强度没有增加。这些结果初步表明：无论胞质Ca^2＋浓度高低,SB202190调节蚕豆保卫细胞中SA诱导H2O2产生,但是不调节植物逆境信使分子ABA此类的反应。因此推测,植物细胞中可能有类似动物和酵母细胞中的p38MAP激酶类,并可能专一调节植物保卫细胞中H2O2信号通路。据我们所知,这是首次报道SB202190和SA共同调节植物保卫细胞中ROS信号过程。