Reactive Oxygen Species Tune Root Tropic Responses

The default growth pattern of primary roots of land plants is directed by gravity. However, roots possess the ability to sense and respond directionally to other chemical and physical stimuli, separately and in combination. Therefore, these root tropic responses must be antagonistic to gravitropism. The role of reactive oxygen species (ROS) in gravitropism of maize and Arabidopsis (Arabidopsis thaliana) roots has been previously described. However, which cellular signals underlie the integration of the different environmental stimuli, which lead to an appropriate root tropic response, is currently unknown. In gravity-responding roots, we observed, by applying the ROS-sensitive fluorescent dye dihydrorhodamine-123 and confocal microscopy, a transient asymmetric ROS distribution, higher at the concave side of the root. The asymmetry, detected at the distal elongation zone, was built in the first 2 h of the gravitropic response and dissipated after another 2 h. In contrast, hydrotropically responding roots show no transient asymmetric distribution of ROS. Decreasing ROS levels by applying the antioxidant ascorbate, or the ROS-generation inhibitor diphenylene iodonium attenuated gravitropism while enhancing hydrotropism. Arabidopsis mutants deficient in Ascorbate Peroxidase 1 showed attenuated hydrotropic root bending. Mutants of the root-expressed NADPH oxidase RBOH C, but not rbohD, showed enhanced hydrotropism and less ROS in their roots apices (tested in tissue extracts with Amplex Red). Finally, hydrostimulation prior to gravistimulation attenuated the gravistimulated asymmetric ROS and auxin signals that are required for gravity-directed curvature. We suggest that ROS, presumably H₂O₂, function in tuning root tropic responses by promoting gravitropism and negatively regulating hydrotropism.Plants evolved the ability to sense and respond to various environmental stimuli in an integrated fashion. Due to their sessile nature, they respond to directional stimuli such as light, gravity, touch, and moisture by directional organ growth (curvature), a phenomenon termed tropism. Experiments on coleoptiles conducted by Darwin in the 1880s revealed that in phototropism, the light stimulus is perceived by the tip, from which a signal is transmitted to the growing part (Darwin and Darwin, 1880). Darwin postulated that in a similar manner, the root tip perceives stimuli from the environment, including gravity and moisture, processes them, and directs the growth movement, acting like “the brain of one of the lower animals” (Darwin and Darwin, 1880). The transmitted signal in phototropism and gravitropism was later found to be a phytohormone, and its redistribution on opposite sides of the root or shoot was hypothesized to promote differential growth and bending of the organ (Went, 1926; Cholodny, 1927). Over the years, the phytohormone was characterized as indole-3-acetic acid (IAA, auxin; Kögl et al., 1934; Thimann, 1935), and the ‘Cholodny-Went’ theory was demonstrated for gravitropism and phototropism (Rashotte et al., 2000; Friml et al., 2002). In addition to auxin, second messengers such as Ca²⁺, pH oscillations, reactive oxygen species (ROS) and abscisic acid (ABA) were shown to play an essential role in gravitropism (Young and Evans, 1994; Fasano et al., 2001; Joo et al., 2001; Ponce et al., 2008). Auxin was shown to induce ROS accumulation during root gravitropism, where the gravitropic bending is ROS dependent (Joo et al., 2001; Peer et al., 2013).ROS such as superoxide and hydrogen peroxide were initially considered toxic byproducts of aerobic respiration but currently are known also for their essential role in myriad cellular and physiological processes in animals and plants (Mittler et al., 2011). ROS and antioxidants are essential components of plant cell growth (Foreman et al., 2003), cell cycle control, and shoot apical meristem maintenance (Schippers et al., 2016) and play a crucial role in protein modification and cellular redox homeostasis (Foyer and Noctor, 2005). ROS function as signal molecules by mediating both biotic- (Sagi and Fluhr, 2006; Miller et al., 2009) and abiotic- (Kwak et al., 2003; Sharma and Dietz, 2009) stress responses. Joo et al. (2001) reported a transient increase in intracellular ROS concentrations early in the gravitropic response, at the concave side of maize roots, where auxin concentrations are higher. Indeed, this asymmetric ROS distribution is required for gravitropic bending, since maize roots treated with antioxidants, which act as ROS scavengers, showed reduced gravitropic root bending (Joo et al., 2001). The link between auxin and ROS production was later shown to involve the activation of NADPH oxidase, a major membrane-bound ROS generator, via a PI3K-dependent pathway (Brightman et al., 1988; Joo et al., 2005; Peer et al., 2013). Peer et al. (2013) suggested that in gravitropism, ROS buffer auxin signaling by oxidizing the active auxin IAA to the nonactive and nontransported form, oxIAA.Gravitropic-oriented growth is the default growth program of the plant, with shoots growing upwards and roots downward. However, upon exposure to specific external stimuli, the plant overcomes its gravitropic growth program and bends toward or away from the source of the stimulus. For example, as roots respond to physical obstacles or water deficiency. The ability of roots to direct their growth toward environments of higher water potential was described by Darwin and even earlier and was later defined as hydrotropism (Von Sachs, 1887; Jaffe et al., 1985; Eapen et al., 2005).In Arabidopsis (Arabidopsis thaliana), wild-type seedlings respond to moisture gradients (hydrostimulation) by bending their primary roots toward higher water potential. Upon hydrostimulation, amyloplasts, the starch-containing plastids in root-cap columella cells, which function as part of the gravity sensing system, are degraded within hours and recover upon water replenishment (Takahashi et al., 2003; Ponce et al., 2008; Nakayama et al., 2012). Moreover, mutants with a reduced response to gravity (pgm1) and to auxin (axr1 and axr2) exhibit higher responsiveness to hydrostimulation, manifested as accelerated bending compared to wild-type roots (Takahashi et al., 2002, 2003). Recently, we have shown that hydrotropic root bending does not require auxin redistribution and is accelerated in the presence of auxin polar transport inhibitors and auxin-signaling antagonists (Shkolnik et al., 2016). These results reflect the competition, or interference, between root gravitropism and hydrotropism (Takahashi et al., 2009). However, which cellular signals participate in the integration of the different environmental stimuli that direct root tropic curvature is still poorly understood. Here we sought to assess the potential role of ROS in regulating hydrotropism and gravitropism in Arabidopsis roots.     

自噬发生中的ROS调节机制

活性氧是细胞代谢中产生的有很强反应活性的分子,易将邻近分子氧化,并参与细胞内多种信号转导途径,对相关生理过程进行调控．自噬是真核细胞通过溶酶体机制对自身组分进行降解再利用的过程,在细胞应激及疾病发生等过程中发挥重要作用．本文对活性氧和自噬相关调节进行分类介绍,根据新近研究进展,从活性氧参与的自噬性死亡、自噬性存活以及线粒体自噬3方面探讨了相关信号转导机制,对活性氧作为信号分子参与的自噬调控途径做一总结和介绍．     

活性氧的信号分子作用

活性氧 (ＲＯＳ)包括过氧化氢 (Ｈ2 Ｏ2 )、超氧阴离子 (Ｏ·-2 )、羟自由基 (·ＯＨ)等。过量的活性氧可引起细胞大分子的氧化损伤。另外 ,微量活性氧在某些生理现象的调控中也发挥重要的作用 ,特别是在细胞内信号转导方面。在配体与受体的相互作用及激动剂处理细胞的过程中 ,发现酶及转录因子的激活 ,基因的表达 ,细胞凋亡等过程的发生均与活性氧有一定关系。因此 ,活性氧被认为是一种新的第二信使。1 .酶的激活酶的活化是信号转导过程中的重要环节。最近几年的研究表明 ,某些酶的活化与ＲＯＳ参与有密切关系。当血小板源生长因子(ＰＤＧ…     

ROS 的信息分子功能

ROS在机体内主要由NADPH氧化酶系统产生,ROS作为信息分子对细胞功能如细胞生长,转化,凋亡,转录和衰老的调节及相关信息传递等方面的研究,在90年代后期有了明显的进展。并从细胞内环境的氧化还原状态变化和蛋白质的氧化修饰角度初步探讨了ROS参与信息传递的机理。     

The Role of Reactive Oxygen Species in Mitochondrial Permeability Transition

We have provided evidence that mitochondrial membrane permeability transition induced by inorganic phosphate, uncouplers or prooxidants such as t-butyl hydroperoxide and diamide is caused by a Ca²⁺-stimulated production of reactive oxygen species (ROS) by the respiratory chain, at the level of the coenzyme Q. The ROS attack to membrane protein thiols produces cross-linkage reactions, that may open membrane pores upon Ca²⁺ binding. Studies with submitochondrial particles have demonstrated that the binding of Ca²⁺ to these particles (possibly to cardiolipin) induces lipid lateral phase separation detected by electron paramagnetic resonance experiments exploying stearic acids spin labels. This condition leads to a disorganization of respiratory chain components, favoring ROS production and consequent protein and lipid oxidation.     

植物防御反应中活性氧的产生和作用

活性氧在植物抗病性中起着重要的作用。本文将对其在防御反应中的产生和作用进行简要的论述。     

活性氧在创伤愈合中作用的研究

：创伤愈合是一个复杂的生物学过程,包括出血与凝血、炎症渗出、血管和肉芽组织的形成、再上皮化、纤维化和瘢痕改建等,在这一系列的生物学活动过程中都需要能量支持;高等动物使用氧气作为终端氧化剂,通过对碳水化合物的氧化作用为愈合过程中的各种生命活动提供能量,但该过程却可以产生大量的活性氧,这些活性氧在创伤愈合的过程中扮演着重要的角色,在低浓度情况下可以促进伤口的愈合,而在高浓度时会抑制伤口愈合,而活性量浓度的过高过低都会影响创口的正常愈合过程。     

Extending Cassava Root Shelf Life via Reduction of Reactive Oxygen Species Production

One of the major constraints facing the large-scale production of cassava (Manihot esculenta) roots is the rapid postharvest physiological deterioration (PPD) that occurs within 72 h following harvest. One of the earliest recognized biochemical events during the initiation of PPD is a rapid burst of reactive oxygen species (ROS) accumulation. We have investigated the source of this oxidative burst to identify possible strategies to limit its extent and to extend cassava root shelf life. We provide evidence for a causal link between cyanogenesis and the onset of the oxidative burst that triggers PPD. By measuring ROS accumulation in transgenic low-cyanogen plants with and without cyanide complementation, we show that PPD is cyanide dependent, presumably resulting from a cyanide-dependent inhibition of respiration. To reduce cyanide-dependent ROS production in cassava root mitochondria, we generated transgenic plants expressing a codon-optimized Arabidopsis (Arabidopsis thaliana) mitochondrial alternative oxidase gene (AOX1A). Unlike cytochrome c oxidase, AOX is cyanide insensitive. Transgenic plants overexpressing AOX exhibited over a 10-fold reduction in ROS accumulation compared with wild-type plants. The reduction in ROS accumulation was associated with a delayed onset of PPD by 14 to 21 d after harvest of greenhouse-grown plants. The delay in PPD in transgenic plants was also observed under field conditions, but with a root biomass yield loss in the highest AOX-expressing lines. These data reveal a mechanism for PPD in cassava based on cyanide-induced oxidative stress as well as PPD control strategies involving inhibition of ROS production or its sequestration.     

植物乙烯生物合成过程中活性氧的作用

大量的研究结果表明,活性氧参与植物乙烯生物合成过程具有明显的普遍性,超氧阴离子自由基是参与乙烯生物合成过程的主要活性氧。近年来研究的焦点主要从乙烯生物合成的关键调控酶ACC合酶及ACC氧化酶的酶活性、酶动力学特性、酶蛋白空间结构、酶基因表达水平等方面来阐明活性氧调控植物乙烯生物合成的机制。最新的研究表明：植物在各种正常或应激的生长条件下首先诱导了活性氧产生水平的变化,活性氧在基因或蛋白质水平上影响ACC合酶和ACC氧化酶的活性水平,从而调节乙烯的生物合成。本文首次综述了活性氧影响植物乙烯生物合成过程的最新研究进展,并对活性氧在植物乙烯生物合成中具有诱导与抑制并存的“双重性”作用进行了探讨。     

The Role of Reactive Oxygen Species in the Antitumor Activity of Bleomycin

Calf thymus DNA was incubated with bleomycin and FeCl₃, in the presence of isolated rat liver microsomal NADH-cytochrome b₅ reductase, cytochrome b₅ and NADH which catalyze redox cycling of the bleomycin-Fe-complex. Furthermore, isolated rat liver nuclei were incubated with bleomycin, FeCl₃ and NADH, a system in which redox cycling of bleomycin-Fe leads to DNA damage. In both systems free bases from DNA were released. Furthermore, 8-hydroxy-guanine was also found in the supernatant. On the other hand, 8-hydroxy-deoxyguanosine was detected in DNA of cell nuclei indicating that hydroxylation of the guanine molecule occurred in intact DNA. The release of bases correlated with the release of malondialydehyde as well as with NADH and oxygen consumption. These results indicate that NADH-cytochrome b₅ reductase catalyzes redox cycling of the bleomycin-Fe-complex which results in the formation of reactive oxygen species which oxidize deoxyribose as well as bases of DNA. Both mechanisms may contribute to the cytotoxic and cytostatic effects of bleomycin observed in intact cells.     

Role of Reactive Oxygen Species in the Initiation of Plant Retrograde Signaling

The interaction between the nucleus and the different organelles is important in the physiology of the plant. Reactive oxygen species (ROS) are a by-product of the oxidation of organic molecules to obtain energy by the need to carry out the electron transfer between the different enzymatic complexes. However, they also have a role in the generation of what is known as retrograde signaling. This signal comes from the different organelles in which the oxidation of molecules or the electron transference is taking place such as mitochondria and chloroplasts. Furthermore, ROS can also induce the release of signals from the apoplast. It seems that these signals plays a role communicating to the nucleus the current status of the different parts of the plant cell to induce a changes in gene expression. In this review, the molecular mechanism of ROS retrograde signaling is described.

     

Diabetic Peripheral Neuropathy: Role of Reactive Oxygen and Nitrogen Species

The prevalence of diabetes has reached epidemic proportions. There are two forms of diabetes: type 1 diabetes mellitus is due to auto-immune-mediated destruction of pancreatic β-cells resulting in absolute insulin deficiency and type 2 diabetes mellitus is due to reduced insulin secretion and or insulin resistance. Both forms of diabetes are characterized by chronic hyperglycemia, leading to the development of diabetic peripheral neuropathy (DPN) and microvascular pathology. DPN is characterized by enhanced or reduced thermal, chemical, and mechanical pain sensitivities. In the long-term, DPN results in peripheral nerve damage and accounts for a substantial number of non-traumatic lower-limb amputations. This review will address the mechanisms, especially the role of reactive oxygen and nitrogen species in the development and progression of DPN.     

Springback in Root Gravitropism

Conditions under which a gravistimulus of Merit corn roots (Zea mays L.) is withdrawn result in a subsequent loss of gravitropic curvature, an effect which we refer to as `springback.' This loss of curvature begins within 1 to 10 minutes after removal of the gravistimulus. It occurs regardless of the presence or absence of the root cap. It is insensitive to inhibitors of auxin transport (2,3,5-triiodobenzoic acid, naphthylphthalmaic acid) or to added auxin (2,4-dichlorophenoxyacetic acid). Springback is prevented if a clinostat treatment is interjected to neutralize gravistimulation during germination, which suggests that the change in curvature is a response to a `memory' effect carried over from a prior gravistimulation.     

A Role for the TOC Complex in Arabidopsis Root Gravitropism

Arabidopsis (Arabidopsis thaliana) roots perceive gravity and reorient their growth accordingly. Starch-dense amyloplasts within the columella cells of the root cap are important for gravitropism, and starchless mutants such as pgm1 display an attenuated response to gravistimulation. The altered response to gravity1 (arg1) mutant is known to be involved with the early phases of gravity signal transduction. arg1 responds slowly to gravistimulation and is in a genetically distinct pathway from pgm1, as pgm1 mutants enhance the gravitropic defect of arg1. arg1 seeds were mutagenized with ethylmethane sulfonate to identify new mutants that enhance the gravitropic defect of arg1. Two modifier of arg1 mutants (mar1 and mar2) grow in random directions only when arg1 is present, do not affect phototropism, and respond like the wild type to application of phytohormones. Both have mutations affecting different components of the Translocon of Outer Membrane of Chloroplasts (TOC) complex. mar1 possesses a mutation in the TOC75-III gene; mar2 possesses a mutation in the TOC132 gene. Overexpression of TOC132 rescues the random growth phenotype of mar2 arg1 roots. Root cap amyloplasts in mar2 arg1 appear ultrastructurally normal. They saltate like the wild type and sediment at wild-type rates upon gravistimulation. These data point to a role for the plastidic TOC complex in gravity signal transduction within the statocytes.     

气孔运动中的活性氧信号

大量研究证明活性氧(ROS)在气孔运动中起信号分子的作用。保卫细胞中ROS的产生依赖于特定的酶, 其中NADPH氧化酶组分RBOH已得到深入研究, 并已证实其参与生物与非生物胁迫反应。植物激素包括脱落酸(ABA)、水杨酸(SA)、乙烯、生长素及细胞分裂素等, 它们均通过ROS的介导来调控气孔运动。生物胁迫(如毒性细菌和真菌)也会调控气孔运动。ROS参与这些调控过程。保卫细胞中存在多层次对ROS产生及其作用的调节, 抗氧化活性物质和ROS敏感蛋白(如蛋白激酶和磷酸酶)均可传递ROS信号并调节气孔运动。ROS对离子通道调节的证据也越来越多。保卫细胞由于可通过ROS整合复杂的信号途径, 已成为研究植物ROS信号转导过程的良好模式系统。     

气孔运动中的活性氧信号

大量研究证明活性氧(ROS)在气孔运动中起信号分子的作用。保卫细胞中ROS的产生依赖于特定的酶,其中NADPH氧化酶组分RBOH已得到深入研究,并已证实其参与生物与非生物胁迫反应。植物激素包括脱落酸(ABA)、水杨酸(SA)、乙烯、生长素及细胞分裂素等,它们均通过ROS的介导来调控气孔运动。生物胁迫(如毒性细菌和真菌)也会调控气孔运动。ROS参与这些调控过程。保卫细胞中存在多层次对ROS产生及其作用的调节,抗氧化活性物质和ROS敏感蛋白(如蛋白激酶和磷酸酶)均可传递ROS信号并调节气孔运动。ROS对离子通道调节的证据也越来越多。保卫细胞由于可通过ROS整合复杂的信号途径,已成为研究植物ROS信号转导过程的良好模式系统。     

The Role of Reactive Oxygen Species in Inflammatory Disease: Evaluation of Methodology

The production of reactive oxidants has been implicated in the pathology of a number of inflammatory conditions, including inflamed arthritic joints. Many assays for the detection of these oxidants in diseased states have been described, but there are a number of potential pitfalls in both experimental design and the interpretation of results obtained with these techniques. Here, we describe a number of commonly used assays to detect the production of reactive oxidants and critically discuss their usefulness and limitations. We focus on the role of xanthine oxidase in reactive oxidant production in inflammatory disease.     

植物中活性氧的检测方法

主要对超氧阴离子自由基(O2-·)、过氧化氢(H2O2)等活性氧的检测方法,包括化学发光法、分光光度法、荧光染色法,EPR波谱学方法、DAB组织染色法和电子显微技术检测法等进行了综述,并简单介绍了最近发展起来的一些新技术。