Barley leaf antioxidant system under photooxidative stress induced by Rose Bengal

The effects of photooxidative stress induced in green barley (Hordeum vulgare L.) leaves by xanthene dye Rose Bengal (RB) on the content of low-molecular antioxidants and the activity of antioxidant enzymes were studied. During illumination (24 h, 160 mol quanta/(m² s)) of the leaves preincubated in darkness on 10 and 100 M RB, ROS accumulated, and their level increased along with RB concentration and duration of illumination. Under these conditions, the content of reduced ascorbate and reduced glutathione (GSH) increased, the level of -and -tocopherol decreased, and the activity of ascorbate peroxidase, the enzyme participating in H₂O₂ degradation, increased. At the same time, the activity of catalase, also participating in H₂O₂ detoxification, decreased, which may be due to the enzyme inhibition during the photochemical stress. In the illuminated treated leaves, superoxide dismutase (SOD), the enzyme destroying the superoxide anion radicals, was activated. The cytosolic SOD isoform was the first to be activated and chloroplastic isoforms followed. It is supposed that photodynamic processes induced by RB in barley leaves are initiated in the cytosol. The activity of glutathione reductase, participating in glutathione oxidized form reduction, did not change at first and grew only after continuous illumination. Thus, the increase in the GSH level, which we have revealed on the initial stage of photooxidative stress development, was due to its synthesis de novo. In addition, under photooxidative stress induced by the sensitizer RB, the level of tocopherol reduced, whereas the amount of other low-molecular antioxidants increased. The exhaustion of the tocopherol pool, in its turn, could limit the resistance of barley leaves to the photooxidative stress.     

Antioxidative protection in a drought-resistant maize strain during leaf senescence

Leaves of 7- and 18-day-old plants of two maize strains, one resistant (LIZA) and one sensitive (LG11) to water stress, were floated in 1 m M paraquat and 1 m M H₂O₂ for 12 h in light and in darkness. The aim of this work was to analyse the effects of these substances on the activities of enzymes involved in the scavenging of active oxygen species during senescence. Three senescence parameters; chlorophyll loss, lipid peroxidation and conductivity; showed a general cell damage caused by both oxidative treatments and revealed a higher tolerance of LIZA than LG11 to paraquat and H₂O₂ both in light and in darkness. Activities of antioxidative enzymes increased by the effect of oxidative treatments in young and senescent leaves of the drought-resistant maize strain LIZA. These increases were about 3-to 6-fold in glutathione reductase. 3-to 4-fold in superoxide dismutase and 2-fold in ascorbate peroxidase activities. The possible correlation between water stress resistance. senescence and the potential of antioxidant enzymes was analysed.     

Identification of the Product of ndhA Gene as a Thylakoid Protein Synthesized in Response to Photooxidative Treatment

A 76 amino acid sequence of NDH-A (the protein encoded by plastidndhA gene) from barley (Hordeum vulgare L.) was expressed asa fusion protein with rß-galactosidase in E. coli.The corresponding antibody generated in rabbits was used toinvestigate localization, expression and synthesis in vitroof NDH-A. NDH-A was identified as a 35 kDa polypeptide localizedin thylakoid membrane. Western blots shows a large increasein NDH-A levels when barley leaves were incubated under photooxidativeconditions, which was more pronounced in mature-senescent leavesthan in young leaves. Immunoprecipitation of the [³⁵S]methioninelabelled proteins, synthesized in vitro by isolated chloroplasts,demonstrated the synthesis in chloroplasts of the NDH-A 35 kDapolypeptide when barley leaves had been incubated under photooxidativeconditions. The results indicate that ndh genes may be involvedin the protection of chloroplasts against photooxidative stress,particularly in mature-senescent leaves. (Received November 13, 1995; Accepted February 5, 1996)     

Senescence-associated mRNAs that may participate in signal transduction and protein trafficking

The differential display technique was used to generate cDNA probes in order to identify mRNAs that are up-regulated during senescence of Arabidopsis leaves. Three mRNAs were examined that had not previously been associated with senescence. The steady-state levels of these mRNAs are detectable in small amounts in mature green leaves, but increase considerably as chlorophyll levels begin to decline. This relationship to senescence occurs under natural circumstances as well as when senescence is accelerated by leaf detachment in the dark or by addition of 1-aminocyclopropane-1-carboxylic acid (ACC). Retardation of senescence by benzyladenine slows the increase of the mRNAs. One of these mRNAs appears to code for a protein (Sec 13) that may be involved in vesicle formation at the endoplasmic reticulum. Another mRNA codes for a protein with WD‐repeat motif whose function is as yet unidentified, and the third codes for a putative calcium-dependent protein kinase. A fourth cDNA has also been cloned by subtractive hybridization from senescing Arabidopsis leaves that encodes vacuolar-processing enzyme ( γ VPE). Incubation of detached leaves in darkness also caused an abrupt elevation in the steady-state levels of the γVPE , similar to that of the senescing attached leaves. The possible functions of the gene products and their involvement in cellular and biochemical processes during senescence are discussed.     

低温胁迫下嫁接对黄瓜叶片SOD和CAT基因表达与活性变化的影响

研究了低温胁迫下嫁接和自根黄瓜叶片Mn-SOD、Cu/Zn-SOD和CAT mRNA基因表达和酶活性变化及其与抗冷性的关系.结果表明:低温胁迫下,嫁接与自根黄瓜叶片Cu/Zn-SOD、Mn-SOD mRNA基因相对表达量变化分别与其Cu/Zn-SOD、Mn-SOD活性变化相吻合,而CATmRNA相对表达量变化与其CAT活性变化并不一致;嫁接黄瓜叶片Cu/Zn-SOD和Mn-SOD mRNA相对表达量及SOD、Cu/Zn-SOD和Mn-SOD活性均高于自根黄瓜,MDA含量和电解质渗漏率均低于自根黄瓜,嫁接黄瓜较高的SOD基因表达量调控的较高SOD活性是其抗冷性强于自根黄瓜的主要因素;嫁接黄瓜的功能叶CAT mRNA相对表达量略高于自根黄瓜,而幼叶CAT mRNA相对表达量低于后者,但两者CAT活性差异不大,说明低温胁迫对嫁接黄瓜叶片CAT mRNA相对表达量及CAT活性的影响不大.     

Superoxide dismutase and glutathione reductase overexpression in wheat protoplast: photooxidative stress tolerance and changes in cellular redox state

In previous works, we have established a correlation between antioxidant system response and tolerance to drought, osmotic stress and photooxidative stress of different wheat cultivars with contrasting drought tolerance. In the present work, a protocol to obtain and transform wheat protoplasts was established. Transgenic protoplasts with Manganese Superoxide Dismutase (Mn-SOD) (E.C.: 1.15.1.1) and Glutathione Reductase (GR) (E.C.: 1.6.4.2) overexpression in chloroplasts were obtained, and their responses to photooxidative stress were characterized. Protoplasts with Mn-SOD or GR overexpression, showed different responses and tolerance to photooxidative stress. Protoplasts with Mn-SOD overexpression showed lower levels of oxidative damage, higher level of endogenous hydrogen peroxide and a great induction of total SOD and GR activities during photooxidative treatments. In protoplasts with GR overexpression the oxidative damage provoked by the photooxidative treatment was similar to control protoplasts, the GSH content and GSH/GSH + GSSG ratio were higher than control and Mn-SOD transformed protoplast, and total SOD and GR activities were not induced. Our results suggest that the differential responses and tolerance to photooxidative stress given by Mn-SOD or GR overexpression, also depend on the effects of these enzyme activities over the cellular redox state balance, which modulate the responses to photooxidative stress.     

Effect of water stress on some oxidative enzymes and senescence in Vigna seedlings

Seedlings of Vigna catjang Endl. were subjected to water stress for 6, S and 10 days by withholding water to investigate the activities of some oxidative enzymes and the pattern of senescence in leaves of 17-day-old seedlings undergoing water stress. Increasing duration of stress produced a proportional increase in the activities of IAA-oxidase, AA-oxidase, peroxidase and glycolate oxidase but decreased catalase activity and the contents of both chlorophyll and protein, hastening senescence. Leaf water potential and relative water content were also lowered with incresing duration of stress. Permeability was increased in leaf tissue undergoing water stress for 8 days. Seed treatment with CaCl₂ (10⁻² and 10⁻¹⁴ M ) for 6 h improved the water status of leaves, decreased tissue permeability, activities of oxidative enzymes, decline of chlorophyll and protein contents and delayed senescence compared to untreated water stressed plants.     

Effects of light and regulators on senescence-related changes in soluble proteins in detached oat (Avena sativa L.) leaves

The rate of senescence and the two-dimensional pattern of soluble proteins from detached oat leaves senescing in either darkness or light were analyzed, and compared to those of leaves in which senescence was delayed by application of the cytokinin benzyladenine or enhanced through the action of abscisic acid.Senescence of detached leaves in light did not differ significantly from senescence in attached leaves on intact plants. In darkness, protein was lost at a higher rate than in light, but several individual proteins showed relative increases. Notably, proteins previously characterized as high-molecular-weight proteins and senescence-associated proteins (Klerk et al., 1992) increased. Changes observed during incubation in light or darkness appeared to be related to this condition rather than the rate or progress of senescence. Cytokinins delayed and abscisic acid accelerated the changes in protein pattern compared to water. Beside changes previously identified in leaves senescing on the plant, detached leaves show alterations that reflect their condition of incubation rather than their developmental progress.Abbreviations 2D-PAG two-dimensional polyacrylamide gel electrophoresis - ABA abscisic acid - BA N⁶-benzyladenine - BSA bovine serum albumin - EDTA ethylenediamine tetraacetic acid - IEF isoelectric focusing - Rubisco ribulosebisphosphate carboxylase/oxygenase - SDS sodium dodecyl sulfate - Tris tris (hydroxymethyl) aminomethane     

冬小麦旗叶旱促衰老过程中氧化伤害与抗氧化系统的尖

研究了土壤缓慢干旱胁迫下抗旱性不同的２个冬小麦品种旗叶老过程中氧化丰以及酶促与硕果发现,在抗旱性强的品种中,冬泪科叶片旱个衰老与膜脂过氧化程度之间并无直接的联系。超氧化物歧化酶（ＳＯＤ）活性在不同抗旱性品种中均呈现持志下降的趋势,但在生强的品种中下降幅度较小,过氧化氢酶（ＣＡＴ）活性在胁迫初期基本不变,至中后期明显下降,且在抗旱性弱的品种中下降幅度较大,抗坏血酸过氧化物酶（ＡＰ）和谷胱甘肽还原酶（     

棉花叶片衰老过程中激素和膜脂过氧化的关系

以陆地棉品种辽棉9号的去根幼苗为材料,对其进行暗诱导衰老培养.在培养液中分别加入6-BA、ABA、GSH、H2O2、CaCl2、A23187 和A23187 CaCl2,测定在不同培养条件下棉花去根幼苗叶片内源激素、SOD酶活性和MDA含量的变化.结果表明:棉花叶片衰老表现为细胞分裂素含量的下降和ABA含量的上升.6-BA、GSH和钙离子均延缓叶片的衰老,ABA和H2O2促进叶片的衰老.     

Retardation of Leaf Senescence by Triadimefon Treatment in Mung Bean Seedling

Treatment of triadimefon on detached leaves of mung bean (Phaseolus radiatus L. ) seedlings increased the levels of chlorophyll and soluble proteins. Declined activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate-peroxidase (AsA-POD) and contents of ascorbate (ASA) and glutathione (GSH) were observed during the senescence of detached young leaves. Triadimefon at concentration of 20 mg/L promoted the activities of POD, AsA-POD and levels of AsA and GSH, but had no effect on the activities of SOD and CAT. On the other hand, the levels of malondialdehyde (MDA) were increased and the increase of which was markedly negative correlated with the activities of POD, AsA-POD and with the contents of AsA and GSH during the senescence of leaves. MDA contents were decreased by triadimefon treatment. These resuits suggested that triadimefon retarded the senescence of leaves in mung bean seedlings in terms of enhancing the protective ability of plant tissues against membrane lipid peroxidation.     

夜间高温胁迫对水稻叶片光合机构的影响

The net photosynthetic rate (Pn), the apparent quantum yield (AQY), the photochemical efficiency of PSⅡ(Fv/Fm), the quantum yield of PSⅡ electron transport (Ф PSⅡ) and the coefficient of photochemical quenching (qp) were decreased in rice (Oryza sativa L.) leaves under high nocturnal temperature (42±1) ℃ stress, but the relative reduction state of PSⅡ (1-qp) was increased. With the increment of stress time, the chlorophyll content and the binding degree of chlorophyll-protein complex declined gradually, the O2(superoxide radical) production rate and H2O2 content in leaves elevated. The activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) increased for 2-3 days under high temperature stress yet decreased afterwards. The results showed that the photosynthetic efficiency declined in rice leaves under high nocturnal temperature stress with concurrent emergence of the symptom of oxidative damage. The gradual increase of the SOD, POD and CAT activities, as well as that of the ratio of Pr (photo-respiration rate) to Pr+Pn and non-photochemical quenching of chlorophyll fluorescence (NPQ) indicates that those mechanisms related to the changes of these parameters may play an important role in protecting rice leaves from oxidative damage under high nocturnal temperature stress.     

Antioxidative protection and proteolytic activity in tolerant and sensitive wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) varieties subjected to long-term field drought

Field drought studies were performed in order to assess oxidative stress, proteolytic activity and yield loss under natural stress conditions. Flag leaves of two drought-tolerant (Yantar and Zlatitsa) and two drought-sensitive (Miziya and Dobrudjanka) winter wheat varieties were analyzed. Stress intensity was assessed by relative electrolyte leakage and proline accumulation. Senescence progression was followed by loss of chlorophyll and protein. Lipid peroxidation, H₂O₂ content, activities of superoxide dismutase (SOD), catalase (CAT), and non-specific peroxidase (GPX) isoforms, as well as proteolytic activities were analyzed from heading throughout grain filling. Weakening of membrane integrity and oxidative damage to lipids were more pronounced in the sensitive varieties under field drought. The activities of Fe- and Cu/Zn SOD isoforms decreased in the controls, but remained high in drought-treated plants. The activities of MnSOD isoforms and CAT were enhanced towards grain filling, especially in the sensitive varieties under drought. GPX activities were rised under drought but progressively diminished. Accelerated senescence under field drought was linked to higher proteolytic activity with variety specific differences in the protease response, but without a clear correlation to drought resistance or sensitivity. Field drought led to higher oxidative stress more pronounced for drought sensitive varieties, especially during the grain filling period.