The effect of reactive oxygen species on ethylene production induced by osmotic stress in etiolated mungbean seedling

After 10 h osmotic stress in 25% polyethylene glycol (PEG6000) solution (–1.8 MPa) at 25 °C in darkness, the etiolated mungbean seedlings were transferred to pure water for recovery. The ethylene release rate and the level of reactive oxygen species (ROS), including superoxide radical (O₂^–) and hydrogen peroxide (H₂O₂), were investigated during the recovery process. The results showed that ethylene production rate and amount of ROS increased dramatically after osmotic stress, and a close correlation was observed between ethylene release rate and concentrations of ROS. Inhibitors of ethylene biosynthesis, aminoethoxyvinylglycine (AVG) or aminooxyacetic acid (AOA), could reduce the ethylene release rate, but had no significant influence to the content of O₂^– and H₂O₂. As well as, silver thiosulfate (STS), an inhibitor of ethylene action, exhibited no obvious effect to the concentration of ROS, showing stress-inducible ethylene was not the cause for the increase of stress-inducible ROS. On the other hand, exogenous generator of superoxide radical (methylviologen, MV, or sodium dithionite, Na₂S₂O₄) could enhance the ethylene production evidently, which could be inhibited by exogenous scavenger of superoxide radical (superoxide dismutase, SOD, or 1, 4-diazabicyclo (2,2,2) octane, DABCO). However, either exogenous H₂O₂ or catalase (CAT) had no significant influence on ethylene production. The results suggested that it was superoxide radical but not H₂O₂which was involved directly in osmotic stress-inducible ethylene biosynthesis. The dual-role of superoxide radical on stress ethylene biosynthesis was also discussed.     

根系渗透胁迫时杨树光合作用光抑制与活性氧的关系

为更多地了解自然条件下活体叶片的光抑制,研究了渗透胁迫时杨树无性系幼苗叶片的光抑制与活性氧代谢的关系．结果表明,随胁迫时间的延长和胁迫强度的增大,杨树叶片O2^-生成加快,H2O2和丙二醛(MDA)含量增多,超氧物歧化酶(SOD)活性升高,过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性降低,活性氧代谢失衡,光合作用的光抑制加剧．用二乙基二硫代氨基甲酸铜盐抑制SOD活性,或用甲基紫精加速O2^-的生成,亦可使杨树叶片发生光抑制．渗透胁迫时杨树无性系幼苗清除H2O2能力降低,限制了叶片通过Mehler反应耗散过剩光能,防御光破坏作用的发挥;光抑制的发生与活性氧的积累有关．     

Evidence for the involvement of nitric oxide and reactive oxygen species in osmotic stress tolerance of wheat seedlings: Inverse correlation between leaf abscisic acid accumulation and leaf water loss

Nitric oxide (NO) and reactive oxygen species (ROS) play important roles in both abscisic acid (ABA) signaling and stress-induced ABA accumulation. However, little is known about their physiological roles in the whole plant. In this study, the effects of NO and ROS on leaf water control and the roles of ABA were determined using wheat (Triticum aestivum L.) seedlings. As compared with the control, osmotic stress reduced leaf water loss (LWL) while it increased leaf ABA content. The effects of osmotic stress on LWL and ABA contents were partially reversed by NO scavengers or NO synthase (NOS) inhibitors. Furthermore, sodium nitroprusside (SNP) at concentrations between 0.01 and 10 mM all reduced LWL efficiently and induced ABA accumulation in a dose-dependent manner. When ABA synthesis was inhibited by fluridone or actidione, the effects of SNP on LWL were partially reversed. These results suggest that NO is involved in leaf water maintenance of wheat seedlings under osmotic stress, and one of the possible mechanisms is by stimulating ABA synthesis. The ROS scavengers used in our experiments had no effects on either LWL or ABA accumulation induced by osmotic stress. However, all ROS induced LWL reduction and ABA accumulation significantly. Hydrogen peroxide had the same effects as SNP on LWL and induced ABA accumulation in a dose-dependent manner but had a maximal effect at 1 mM. Fluridone reversed the effects of H₂O₂ on both LWL reduction and ABA accumulation, while actidione had no effect. These results suggest that ROS are also involved in leaf water maintenance of wheat seedlings by stimulating ABA biosynthesis, but with a different mechanism to that of NO. The ABA-independent mechanism in NO/ROS regulation of leaf water balance is discussed, in relation to our results.     

6-苄（基）腺嘌呤和抗坏血酸对渗透胁迫时杨树光合作用光抑制的影响

研究了6-BA和A5A对渗透胁迫时杨树幼苗叶片光合作用光抑制和活性氧代谢的影响．结果表明,渗透胁迫时杨树叶片净光合速率(Pn)和表观量子效率(AQY)降低,光合作用光抑制加剧,超氧化物歧化酶(SOD)活性升高,抗坏血酸过氧化物酶(APX)活性降低,O2产生加快,H2O2和膜脂过氧化产物丙二醛(MDA)含量升高．6-BA和A5A预处理使胁迫时叶片SOD和APx活性升高。O2生成减少。H2O22和MDA含量降低,同时缓解了光合作用的光抑制．相关分析表明,杨树叶片活性氧水平和MDA含量与Pn和AQY呈负相关．胁迫时杨树叶片活性氧的积累与光合作用光抑制有一定关系,6-BA和A5A对光抑制的缓解作用与其对活性氧清除系统的促进作用有关。     

Post-harvest alterations in polyamines and ethylene in two diverse rose species

Changes in the concentrations of endogenous polyamines and ethylene were determined in two diverse species of rose viz. Rosa damascena and Rosa bourboniana during post-harvest periods. At full bloom, the concentrations of free putrescine was significantly higher than rest of the polyamines, i.e. spermine and spermidine in both the species. The concentrations of all the polyamines decreased during subsequent periods upto 48 h after full bloom. Similar situation was also observed in conjugated fraction but in bound fraction, during full bloom, the concentration of spermine was higher than rest of the polyamines. In both the species, ethylene showed higher levels during full bloom with maximum in R. damascena, which increased, during post-harvest periods. The possible significance of polyamines, ethylene and their interactions is discussed during post-harvest periods in flowers.     

CO2加富对盐胁迫下黄瓜幼苗叶片光合特性及活性氧代谢的影响

以‘津优35号’黄瓜(Cucumis sativus L.)水培苗为试材,采用裂区设计,主区因素为CO_2浓度处理,设大气CO_2浓度(Symbol{B@400μmol/mol)和CO_2加富[(800±40)μmol/mol]2个CO_2浓度水平,裂区因素为盐胁迫处理,用NaCl模拟盐胁迫,设对照(0 mmol/L NaCl)、盐胁迫(80 mmol/L NaCl)2个盐分水平,研究了CO_2加富对盐胁迫下黄瓜幼苗生长、光合特性及活性氧代谢的影响。结果表明:盐胁迫显著抑制黄瓜幼苗的生长,并降低了叶绿素含量、ETR、Φ_(PSII)、核酮糖-1,5-二磷酸羧化酶(RuBPCase)活性及净光合速率;盐胁迫增加了丙二醛及活性氧的累积,与此同时也提高了脯氨酸含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,但降低了过氧化物酶(POD)活性。CO_2加富显著提高了盐胁迫下黄瓜幼苗的株高、茎粗、叶面积及地上部鲜重,降低了叶绿素a、叶绿素b、类胡萝卜素及叶绿素(a+b)含量,但显著提高了净光合速率和RuBPCase活性,同时降低了气孔导度及蒸腾速率,并且使其具有较高的表观电子传递速率及PSII实际光化学效率;CO_2加富显著提高了盐胁迫下黄瓜幼苗叶片脯氨酸含量及SOD、POD、CAT活性,丙二醛、过氧化氢含量和超氧阴离子产生速率显著降低。综上所述,CO_2加富可通过提高幼苗叶片净光合速率、脯氨酸含量及抗氧化酶活性,降低蒸腾速率、减少丙二醛含量及活性氧的积累,从而缓解盐胁迫对黄瓜植株造成的伤害。     

Overexpression of phytoene synthase gene from Salicornia europaea alters response to reactive oxygen species under salt stress in transgenic Arabidopsis

A phytoene synthase gene SePSY was isolated from euhalophyte Salicornia europaea L. The 1655 bp full-length SePSY has an open reading frame of 1257 bp and encodes a 419-amino acid protein. The overexpression of SePSY enhanced the growth of transgenic Arabidopsis. When the plants were exposed to 100 mM NaCl, the photosynthesis rate and photosystem II activity (Fv/Fm) increased from 92% to 132% and from 9.3% to 16.6% in the transgenic lines than in the wild-type, respectively. The transgenics displayed higher activities of SOD and POD and lower contents of H(2)O(2) and MDA than the WT. In conclusion, the transgenic lines showed higher tolerance to salt stress than WT plants by increased photosynthesis efficiency and antioxidative capacity. This is the first report about improving the salt tolerance by genetic manipulation of carotenoid biosynthesis.     

Accumulation of reactive oxygen species in arbuscular mycorrhizal roots

We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H₂O₂ accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl₃, intracellular accumulation of H₂O₂ was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H₂O₂ was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.     

Biochar coupling with phosphorus fertilization modifies antioxidant activity,osmolyte accumulation and reactive oxygen species synthesis in the leaves and xylem sap of rice cultivars under high-temperature stress

Increasing temperature poses a serious threat to rice productivity. This study investigated the impact of various biochar treatments and phosphorous (P) fertilization on osmolyte accumulation, ROS development, and antioxidant activity in two rice cultivars (IR-64 and Huanghuazhan) under high-temperature stress. All plants of both cultivars were grown in a controlled environment under ambient temperatures (AT), high day temperatures (HDT) or high night temperatures (HNT). The different fertilization treatments were biochar alone, P alone and biochar + P with control. In the leaves and xylem sap of both rice cultivars, particularly in the susceptible cv. IR-64, high-temperature stress increased the production of MDA and H₂O₂. HDT and HNT decreased total soluble sugars, protein, and proline levels in both rice cultivars. HNT was observed as more harmful compared to HDT during most of the studied characteristics. The response of antioxidant enzyme activities, viz, SOD, POD, CAT, APX, ASC, GSH, GR, and GSSC activities, to the temperature treatments varied between the two cultivars. Antioxidant activities decreased in the leaves and xylem sap of IR-64 but increased in those of Huanghuazhan upon exposure to high-temperature stress. Huanghuazhan exhibited better heat tolerance compared to IR-64, which was linked to its increased antioxidant enzyme activation and metabolite synthesis. As compared to the control, all soil fertilization treatments considerably reduced the adverse impacts of high temperature on the rice cultivars. The combination of biochar and P resulted in better performance compared to the other treatments in terms of all studied attributes.     

Ethylene-induced overproduction of reactive oxygen species is responsible for the development of watersoaking in immature cucumber fruit

Watersoaking is an ethylene-induced disorder observed in some members of the Cucurbitaceae including cucumber (Cucumis sativus L.), watermelon (Citrullus lanatus Thunb. Matsum and Nakai), and tropical pumpkin (Cucurbita moschata Duch.). Previous studies have found that immature beit-alpha cucumber (cv. Manar) exhibit watersoaking after 6 d of continuous exposure to 10 μL L⁻¹ ethylene in air (21 kPa O₂). The present study was designed to investigate the early dynamics of ethylene responses in immature cucumber fruit in order to provide insight into the watersoaking triggering mechanism. Changes in respiration, epidermal color, firmness, reactive oxygen species (ROS) production and electrolyte leakage were evaluated as a function of time under different ethylene concentrations and exposure duration. Ethylene concentrations exceeding 10 μL L⁻¹ did not accelerate changes in any of the evaluated responses. The first detectable change was a significant rise in respiration on day 2, followed by a significant rise in ROS on day 4, and significant degreening, mesocap softening, and increased electrolyte leakage on day 6; the latter responses coincident with incipient watersoaking. Varying the duration of exposure to ethylene indicated that the critical exposure time is between 2 and 4 d. Notably, all deleterious responses to ethylene were suppressed under a hypoxic atmosphere. A model is proposed in which ethylene induces a sharp increase in respiration with a concomitant sharp rise in ROS, which the immature fruit is incapable of quenching. The resulting production of excess ROS leads to discoloration and membrane deterioration, leading to the release of cytoplasmic content, rapid softening, and the visual symptom of watersoaking.     

The complex interplay of iron metabolism,reactive oxygen species,and reactive nitrogen species: Insights into the potential of various iron therapies to induce oxidative and nitrosative stress

Production of minute concentrations of superoxide (O₂⁻) and nitrogen monoxide (nitric oxide, NO) plays important roles in several aspects of cellular signaling and metabolic regulation. However, in an inflammatory environment, the concentrations of these radicals can drastically increase and the antioxidant defenses may become overwhelmed. Thus, biological damage may occur owing to redox imbalance—a condition called oxidative and/or nitrosative stress. A complex interplay exists between iron metabolism, O₂⁻, hydrogen peroxide (H₂O₂), and NO. Iron is involved in both the formation and the scavenging of these species. Iron deficiency (anemia) (ID(A)) is associated with oxidative stress, but its role in the induction of nitrosative stress is largely unclear. Moreover, oral as well as intravenous (iv) iron preparations used for the treatment of ID(A) may also induce oxidative and/or nitrosative stress. Oral administration of ferrous salts may lead to high transferrin saturation levels and, thus, formation of non-transferrin-bound iron, a potentially toxic form of iron with a propensity to induce oxidative stress. One of the factors that determine the likelihood of oxidative and nitrosative stress induced upon administration of an iv iron complex is the amount of labile (or weakly-bound) iron present in the complex. Stable dextran-based iron complexes used for iv therapy, although they contain only negligible amounts of labile iron, can induce oxidative and/or nitrosative stress through so far unknown mechanisms. In this review, after summarizing the main features of iron metabolism and its complex interplay with O₂⁻, H₂O₂, NO, and other more reactive compounds derived from these species, the potential of various iron therapies to induce oxidative and nitrosative stress is discussed and possible underlying mechanisms are proposed. Understanding the mechanisms, by which various iron formulations may induce oxidative and nitrosative stress, will help us develop better tolerated and more efficient therapies for various dysfunctions of iron metabolism.     

Mitochondria in homeostasis of reactive oxygen species in cell, tissues, and organism

The recent knowledge on mitochondria as the substantial source of reactive oxygen species, namely superoxide and hydrogen peroxide efflux from mitochondria, is reviewed, as well as nitric oxide and subsequent peroxynitrite generation in mitochondria and their effects. The reactive oxygen species formation in extramitochondrial locations, in peroxisomes, by cytochrome P450, and NADPH oxidase reaction, is also briefly discussed. Conditions are pointed out under which mitochondria represent the major ROS source for the cell: higher percentage of non-phosphorylating and coupled mitochondria, in vivo oxygen levels leading to increased intensity of the reverse electron transport in the respiratory chain, and nitric oxide effects on the redox state of cytochromes. We formulate hypotheses on the crucial role of ROS generated in mitochondria for the whole cell and organism, in concert with extramitochondrial ROS and antioxidant defense. We hypothesize that a sudden decline of mitochondrial ROS production converts cells or their microenvironment into a “ROS sink” represented by the instantly released excessive capacity of ROS-detoxification mechanisms. A partial but immediate decline of mitochondrial ROS production may be triggered by activation of mitochondrial uncoupling, specifically by activation of recruited or constitutively present uncoupling proteins such as UCP2, which may counterbalance the mild oxidative stress.     

干旱胁迫下多裂骆驼蓬(Peganum multisectum Bobr)叶片乙烯释放和多胺含量变化与活性氧积累的关系

通过盆栽土培试验研究了干旱胁迫下多裂骆驼蓬(Peganum multisectum Bobr)叶片乙烯释放、多胺含量、活性氧水平及细胞膜透性的动态变化.结果表明,随着土壤干旱强度的增加和胁迫时间的延长,多裂骆驼蓬叶片内丙二醛(MDA)含量和细胞膜透性呈递增变化趋势;轻度胁迫下,乙烯产生速率、多胺(Put、Spm、Spd)、活性氧(O2-·、H2O2)和MDA含量及多胺氧化酶(PAO)活性随胁迫时间延长无明显变化;中度和重度胁迫初期(胁迫后0～30d)腐胺(Put)、精胺(Spm)和亚精胺(Spd)含量快速提高,乙烯生成速率下降,而O2-·、H2O2、MDA含量和PAO活性相对平稳;胁迫后期(胁迫后45～75d)乙烯生成速率增高,Put、Spm和Spd含量下降, O2-·、H2O2和MDA含量大幅度增加,膜透性增大,且乙烯释放与O2-·、H2O2含量呈显著正相关.表明中度和重度胁迫下多裂骆驼蓬叶片存在乙烯和多胺合成的相互制约,乙烯释放增加诱导活性氧积累导致膜透性急剧增大.     

Longevity of animals under reactive oxygen species stress and disease susceptibility due to global warming

The world is projected to experience an approximate doubling of atmospheric CO_2 concentration in the next decades. Rise in atmospheric CO_2 level as one of the most important reasons is expected to contribute to raise the mean global temperature 1.4 ℃-5.8 ℃ by that time. A survey from 128 countries speculates that global warming is primarily due to increase in atmospheric CO_2 level that is produced mainly by anthropogenic activities. Exposure of animals to high environmental temperatures is mostly accompanied by unwanted acceleration of certain biochemical pathways in their cells. One of such examples is augmentation in generation of reactive oxygen species(ROS) and subsequent increase in oxidation of lipids, proteins and nucleic acids by ROS. Increase in oxidation of biomolecules leads to a state called as oxidative stress(OS). Finally, the increase in OS condition induces abnormality in physiology of animals under elevated temperature. Exposure of animals to rise in habitat temperature is found to boost the metabolism of animals and a very strong and positive correlation exists between metabolism and levels of ROS and OS. Continuous induction of OS is negatively correlated with survivability and longevity and positively correlated with ageing in animals. Thus, it can be predicted that continuous exposure of animals to acute or gradual rise in habitat temperature due to global warming may induce OS, reduced survivability and longevity in animals in general and poikilotherms in particular. A positive correlation between metabolism and temperature in general and altered O_2 consumption at elevated temperature in particular could also increase the risk of experiencing OS in homeotherms. Effects of global warming on longevity of animals through increased risk of protein misfolding and disease susceptibility due to OS as the cause or effects or both also cannot be ignored. Therefore, understanding the physiological impacts of global warming in relation to longevity of animals will become very crucial challenge to biologists of the present millennium.     

Cold stress increases reactive oxygen species formation via TRPA1 activation in A549 cells

Reactive oxygen species (ROS) are responsible for lung damage during inhalation of cold air. However, the mechanism of the ROS production induced by cold stress in the lung is still unclear. In this work, we measured the changes of ROS and the cytosolic Ca²⁺ concentration ([Ca²⁺]_c) in A549 cell. We observed that cold stress (from 20 to 5 °C) exposure of A549 cell resulted in an increase of ROS and [Ca²⁺]_c, which was completely attenuated by removing Ca²⁺ from medium. Further experiments showed that cold-sensing transient receptor potential subfamily member 1 (TRPA1) agonist (allyl isothiocyanate, AITC) increased the production of ROS and the level of [Ca²⁺]_c in A549 cell. Moreover, HC-030031, a TRPA1 selective antagonist, significantly inhibited the enhanced ROS and [Ca²⁺]_c induced by AITC or cold stimulation, respectively. Taken together, these data demonstrated that TRPA1 activation played an important role in the enhanced production of ROS induced by cold stress in A549 cell.     

光果甘草毛状根培养过程中对活性氧清除能力和总黄酮含量的变化

采用化学发光法, 分析了光果甘草(Glycyrrhiza glabra L.)毛状根培养过程中对3种活性氧(ROS: O-2(÷)、HO·和H-2O-2)清除能力的动态变化, 并测定了培养过程中总黄酮含量的动态变化. 实验结果表明, 毛状根在对数生长期(20～28 d)对3种ROS都有很强的清除能力,在生长停滞期(29～40 d)对HO·和H-2O-2的清除能力仍维持较高的水平,而对O-2(÷)的清除能力随培养时间的延长逐渐下降.总黄酮含量在对数生长期呈现增加的趋势,至31 d时达到最高含量(0.78%),随培养时间的延长含量逐渐降低.     

光果甘草与乌拉尔甘草清除活性氧能力的比较

采用化学发光法测定了光果甘草（Glycyrrhiza glabra L.）与乌拉尔甘草（G.uralensis Fisch.）中总黄酮和总皂甙对3种重要活性氧（O2^-、OH和H2O2）的清除能力,其中光果甘草对活性氧的清除能力超过乌拉尔甘草;黄酮类化合物对3种活性氧的清除能力强于皂甙类化合物。因此,若以清除活性氧为应用目标,应尽量使用光果甘草。     

Protective effect of exogenous polyamines on root tonoplast function against salt stress in barley seedlings

Salinity stress is one of the most serious factors limiting the productivity of agricultural crops. A possible survival strategy of plants under saline conditions is to sequester excess Na⁺ in the vacuole by vacuolar Na⁺/H⁺ antiport using a pH gradient generated by H⁺-ATPasc (EC 3.6.1.35) and H⁺-Pyrophosphatase (H⁺-PPase; EC 3.6.1.1) to maintain a higher K⁺/Na⁺ ratio in cytoplasm. The effect of exogenously applied polyamines (PAs) in stabilizing root tonoplast integrity and function against salt stress in the barley (Hordeum vulgare L.) seedlings was investigated. The NaCl-induced reductions in the contents of phospholipids and PAs in tonoplast vesicles isolated from barely seedling roots, as well as the activities of H⁺-ATPase, H⁺-PPase and vacuolar Na⁺/H⁺ antiport were all partially restored by the application of 0.5 mM putrescine and 0.5 mM spermidine, especially the former. The above results indicated that one of the mechanisms involved in attenuating salt injury in barley seedlings by exogenous PAs application was to maintain tonoplast integrity and function under saline conditions. Moreover, the possible mechanism involved in counteracting detrimental effects of salt on the barley seedlings by the application of exogenous PAs was discussed.     

Plasmodesmal widening accompanies the short-term increase in symplasmic phloem unloading in pea root tips under osmotic stress

Summary Root tips ofPisum sativum seedlings were exposed to 350 mM mannitol, which was shown to effect a transient but dramatic increase in phloem unloading, and investigated by electron microscopy. After chemical fixation and embedding, extremely thin sections of the root extension zone were examined. Outer, inner, and desmotubule diameters of 830 primary plasmodesmata in transverse walls of cortical cells were measured. Statistical analysis indicated that the majority of plasmodesmata had no neck constriction during osmoregulation. Compared to controls, a highly significant increase in mean plasmodesmata diameter was found, but the desmotubule diameter remained unchanged. Both loss of neck constriction and widening of the cytoplasmic sleeve indicate an increase in effective passage area of plasmodesmata. Spokes between plasma membrane and desmotubule were preserved. Continued exposure of the root tips to mannitol led to a return to control values for plasmodesmal diameters. In contrast to these responses, plasmolysis of cortical cells by 1,000 mM sucrose, diminishing phloem unloading, was accompanied by a reduction in those plasmodesmata classified as open. This is the first report showing a correlation between the ultrastructure of plasmodesmata and the rate of symplasmic transport. The role of the different plasmodesmal components in controlling the passage area of symplasmic transport is discussed.     

Peroxisomes and reactive oxygen species,a lasting challenge

Oxidases generating and enzymes scavenging H₂O₂ predestine peroxisomes (PO) to a pivotal organelle in oxygen metabolism. Catalase, the classical marker enzyme of PO, exhibits both catalatic and peroxidatic activity. The latter is responsible for the staining with 3,3′-diamino-benzidine, which greatly facilitated the visualization of the organelle and promoted further studies on PO. d-Amino acid oxidase catalyzes with strict stereospecificity the oxidative deamination of d-amino acids. The oxidase is significantly more active in the kidney than in liver and more in periportal than pericentral rat hepatocytes. Peroxisomes in these tissues differ in their enzyme activity and protein concentration not only in adjacent cells but even within the same one. Moreover, the enzyme appears preferentially concentrated in the central region of the peroxisomal matrix compartment. Urate oxidase, a cuproprotein catalyzing the oxidation of urate to allantoin, is confined to the peroxisomal core, yet is lacking in human PO. Recent experiments revealed that cores in rat hepatocytes appear in close association with the peroxisomal membrane releasing H₂O₂ generated by urate oxidase to the surrounding cytoplasma. Xanthine oxidase is exclusively located to cores, oxidizes xanthine thereby generating H₂O₂ and O₂ ^– radicals. The latter are converted to O₂ and H₂O₂ by CuZn superoxide dismutase, which has been shown recently to be a bona fide peroxisomal protein. Presented at the 50th Anniversary Symposium of the Society for Histochemistry, Interlaken, Switzerland, October 1-4, 2008.