活性氧与植物防卫反应

目的：总结分析活性氧在植物防卫反应中的作用。方法：根据近10年内公开发表的20篇中英文文献对活性氧的种类、特性，以及在植物抗病过程中所起的作用进行较全面的总结。结果：介绍活性氧的种类、性质，着重讨论了活性氧的产生和消除机理；并对当前该领域今后的研究方向进行了展望。结论：活性氧在植物防卫反应过程中起双重作用，即高浓度时对植物起伤害作用，低浓度时起信号传递作用诱导植物抗性基因和防卫基因的表达。     

S-nitrosylation of NADPH oxidase regulates cell death in plant immunity

Changes in redox status are a conspicuous feature of immune responses in a variety of eukaryotes, but the associated signalling mechanisms are not well understood. In plants, attempted microbial infection triggers the rapid synthesis of nitric oxide and a parallel accumulation of reactive oxygen intermediates, the latter generated by NADPH oxidases related to those responsible for the pathogen-activated respiratory burst in phagocytes. Both nitric oxide and reactive oxygen intermediates have been implicated in controlling the hypersensitive response, a programmed execution of plant cells at sites of attempted infection. However, the molecular mechanisms that underpin their function and coordinate their synthesis are unknown. Here we show genetic evidence that increases in cysteine thiols modified using nitric oxide, termed S-nitrosothiols, facilitate the hypersensitive response in the absence of the cell death agonist salicylic acid and the synthesis of reactive oxygen intermediates. Surprisingly, when concentrations of S-nitrosothiols were high, nitric oxide function also governed a negative feedback loop limiting the hypersensitive response, mediated by S-nitrosylation of the NADPH oxidase, AtRBOHD, at Cys 890, abolishing its ability to synthesize reactive oxygen intermediates. Accordingly, mutation of Cys 890 compromised S-nitrosothiol-mediated control of AtRBOHD activity, perturbing the magnitude of cell death development. This cysteine is evolutionarily conserved and specifically S-nitrosylated in both human and fly NADPH oxidase, suggesting that this mechanism may govern immune responses in both plants and animals.     

盐胁迫对两种忍冬属植物活性氧平衡的影响

以红白忍冬和郁香忍冬盆栽苗为试验材料,研究不同浓度的NaCl胁迫对两者生理指标的影响,并比较它们的耐盐性差异。结果表明,在盐胁迫条件下,两种植物叶片的电导率、MDA含量均不断上升。胁迫期间,红白忍冬的H2O2含量相对于对照没有明显变化,O.2-产生速率在处理前期明显上升,上升出现的时间迟于郁香忍冬;而郁香忍冬的H2O2含量一直维持较高水平,O.2-产生速率在处理前期也有所增加。红白忍冬的SOD、APX和CAT活性总体表现出先上升后下降的趋势,而POD活性则一直上升。郁香忍冬在胁迫期间,其SOD活性有小幅增加;APX和CAT的活性均呈现先上升后下降的趋势;POD活性前期没有明显增加,在处理终期时也仅在100mmol/L胁迫下才显著增加。表明两种植物在遭受盐胁迫时,均能启动抗氧化酶系统来清除活性氧,降低氧化胁迫的伤害。同时,相对于郁香忍冬,红白忍冬通过快速上调抗氧化酶活性,降低盐胁迫对膜脂过氧化作用,保持膜的完整性,从而更好地抵御氧化损害更耐受盐胁迫。     

活性氧与植物抗病性的关系

病原菌侵入植物后可诱导活性氧的出发，并且在非寄主互作中比寄 互作中活性氧的积累更明显。我们在禾谷类作物与白粉菌互作的研究中，也得到了同样的结论。活性氧迸发已被认为是寄主防卫反应之一，在植物的抗病性中具有很重要的作用。     

Reactive oxygen species have a causal role in multiple forms of insulin resistance

Insulin resistance is a cardinal feature of type 2 diabetes and is characteristic of a wide range of other clinical and experimental settings. Little is known about why insulin resistance occurs in so many contexts. Do the various insults that trigger insulin resistance act through a common mechanism? Or, as has been suggested, do they use distinct cellular pathways? Here we report a genomic analysis of two cellular models of insulin resistance, one induced by treatment with the cytokine tumour-necrosis factor-alpha and the other with the glucocorticoid dexamethasone. Gene expression analysis suggests that reactive oxygen species (ROS) levels are increased in both models, and we confirmed this through measures of cellular redox state. ROS have previously been proposed to be involved in insulin resistance, although evidence for a causal role has been scant. We tested this hypothesis in cell culture using six treatments designed to alter ROS levels, including two small molecules and four transgenes; all ameliorated insulin resistance to varying degrees. One of these treatments was tested in obese, insulin-resistant mice and was shown to improve insulin sensitivity and glucose homeostasis. Together, our findings suggest that increased ROS levels are an important trigger for insulin resistance in numerous settings.     

Wangzaozin A调节NADPH氧化酶源性活性氧诱导HL-60细胞分化

利用台盼蓝排染法、吉姆萨染色及流式细胞术对对映-贝壳杉烷二萜wangzaozin A影响人早幼粒白血病细胞HL-60生长、细胞形态、NBT还原能力、细胞吞噬及细胞表面抗原CD11b表达进行了检测.结果显示,0.2~0.8μmol/L wangzaozin A抑制HL-60细胞生长,0.6和0.8μmol/L处理组细胞显示了明显的G1期周期阻滞.随着wangzaozin A浓度升高及处理时间延长,细胞核质比减小,肾形核、杆状核和分叶核细胞增多及胞质中颗粒状物质增加;同时,细胞NBT还原能力、吞噬能力及细胞表面抗原CD11b表达显著增强,表明wangzaozin A可诱导HL-60细胞向成熟粒细胞分化.进一步利用荧光探针DCF检测显示0.6和0.8μmol/L wangzaozin A处理细胞12h后细胞内ROS显著升高;抗氧化剂NAC及NADPH氧化酶抑制剂APO显著抑制wangzaozin A诱导HL-60细胞上调CD11b表达,表明wangzaozin A可通过上调NADPH氧化酶源性的活性氧浓度诱导HL-60细胞分化.     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide （H2O2） induced by abscisic acid （ABA） were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H2O2, which was mainly derived from super-oxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane （PM） NADPH oxidase activity, which coin- cided on time and magnitude with the elevation in ABA-induced accumulation of H2O2. Moreover, these enhanced effects were pro- nouncedly inhibited by two NADPH oxidase inhibitors, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of H2O2 in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H2O2 in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in sus-pension culture cells of tobacco, and that NADPH oxidase and H2O2 appear to be important components in ABA signal transduction pathway in plants.     

木霉菌产生的葡聚糖酶和植物病害防治

木霉菌能够产生不同类型的葡聚糖酶，包括α-1，3-、β-1，3-、β-1，4-和β-1，6-葡聚糖酶，酶的产生受葡萄糖抑制，而为葡萄糖聚合体如真菌细胞壁所诱导。已经克隆了几个萄聚糖酶的编码基因。该酶是木霉菌产生的重要抑菌蛋白，能够与几丁质酶或抗生素协同抗菌，用于改良植物病害生物防治菌和植物。     

Loosening of plant cell walls by expansins

Plant cell walls are the starting materials for many commercial products, from lumber, paper and textiles to thickeners, films and explosives. The cell wall is secreted by each cell in the plant body, forming a thin fibreglass-like network with remarkable strength and flexibility. During growth, plant cells secrete a protein called expansin, which unlocks the network of wall polysaccharides, permitting turgor-driven cell enlargement. Germinating grass pollen also secretes an unusual expansin that loosens maternal cell walls to aid penetration of the stigma by the pollen tube. Expansin's action has puzzling implications for plant cell-wall structure. The recent explosion of gene sequences and expression data has given new hints of additional biological functions for expansins.     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide (H₂O₂) induced by abscisic acid (ABA) were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H₂O₂, which was mainly derived from superoxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane (PM) NADPH oxidase activity, which coincided on time and magnitude with the elevation in ABA-induced accumulation of H₂O₂. Moreover, these enhanced effects were pronouncedly inhibited by two NADPH oxidase inhibitor, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of ₂O₂ in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H₂O₂ in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in suspension culture cells of tobacco, and that NADPH oxidase and H₂O₂ appear to be important components in ABA signal transduction pathway in plants.     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide (H₂O₂) induced by abscisic acid (ABA) were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H₂O₂, which was mainly derived from superoxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane (PM) NADPH oxidase activity, which coincided on time and magnitude with the elevation in ABA-induced accumulation of H₂O₂. Moreover, these enhanced effects were pronouncedly inhibited by two NADPH oxidase inhibitor, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of ₂O₂ in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H₂O₂ in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in suspension culture cells of tobacco, and that NADPH oxidase and H₂O₂ appear to be important components in ABA signal transduction pathway in plants.     

盐和过氧化氢胁迫下交替氧化酶调节根生长和细胞死亡

用不同浓度的NaCl或外源过氧化氢(H_2O_2)溶液处理野生型(WT)拟南芥和交替氧化酶(alternative oxidase,AOX)基因反义抑制突变体(AS-12)拟南芥,发现随NaCl和过氧化氢浓度的增加,两种拟南芥幼根生长速率降低,且出现了明显的细胞死亡。在50和100 mM NaCl或在50 mM H_2O_2处理下,与WT植株相比,AS-12植株幼根的生长速率更低,且组织的细胞死亡程度也更加明显。150 mM NaCl或100及200 mM H_2O_2处理导致AS-12与WT幼根的生长停止和严重的细胞死亡,但两种植株幼根在生长速率和细胞死亡程度上无明显差别。以上实验结果说明交替氧化酶在一定水平的盐胁迫和氧化压力下能够影响幼根的生长和细胞死亡的发生。