Abscisic acid cross-talking with hydrogen peroxide and osmolyte compounds may regulate the leaf rolling mechanism under drought

Leaf rolling observed in some crops such as maize, rice, wheat and sorghum is an indicator of decreased water status. Moderate leaf rolling not tightly or early increases the photosynthesis and grain yield of crop cultivars under environmental stresses. Moreover, the effects of exogenous abscisic acid (ABA) on stomatal conductance, water status and synthesis of osmotic compounds are a well-known issue in plants subjected to water deficit. However, it is not clear how the cross-talk of ABA with H₂O₂ and osmolyte compounds affects the leaf rolling mechanism. Regulation mechanism of leaf rolling by ABA has been first studied in maize seedlings under drought stress induced by polyethylene glycol 6000 (PEG 6000) in this study. ABA treatment under drought stress reduced hydrogen peroxide (H₂O₂) content and the degree of leaf rolling (%) while the treatment-induced ABA synthesis, osmolyte levels (proline, polyamine and total soluble sugars) and some antioxidant enzyme activities in comparison to the plants that were not treated with ABA. Furthermore, exogenous ABA up-regulated the expression levels of arginine decarboxylase (ADC) and pyrroline-5-carboxylate synthase (P5CS) genes and down-regulated polyamine oxidase (PAO), diamine oxidase (DAO) and proline dehydrogenase (ProDH) gene expressions. When endogenous ABA content was decreased by the treatment of fluoridone (FLU) that is an ABA inhibitor, leaf rolling degree (%), H₂O₂ content and antioxidant enzyme activities increased, but osmolyte levels, ADC and P5CS gene expressions decreased. Finally, the treatment of ABA to maize seedlings exposed to drought stress resulted in the stimulation of the antioxidant system, osmotic adjustment and reduction of leaf rolling. We concluded that ABA can be a signal compound cross-talking H₂O₂, proline and polyamines and thus involved in the leaf rolling mechanism by providing osmotic adjustment. The results of this study can be used to provide data for the molecular breeding of maize hybrids with high grain yield by means of moderately rolled leaves.     

Photosynthesis of <Emphasis Type="Italic">Rehmannia glutinosa</Emphasis> subjected to drought stress is enhanced by choline chloride through alleviating lipid peroxidation and increasing proline accumulation

Rehmannia glutinosa seedlings were pretreated with choline chloride (CC) in concentrations of 0, 0.7, 2.1 and 3.5 mM, and then subjected to drought and rewatering treatment to study the effects of CC on the generation of reactive oxygen species (O₂⁻, H₂O₂), lipid peroxidation, proline accumulation, water status and photosynthesis. The results showed that pretreatment with CC alleviated the inhibition of SOD and APX activity caused by drought stress, and therefore, the rate of O₂⁻ production and H₂O₂ concentration were reduced and lipid peroxidation decreased in pretreated plants. CC pretreatment also accelerated accumulation of proline, maintained higher Ψw and RWC, deferred leaf water loss during drought stress and retarded the drop in proline concentration after rewatering. Consequently, drought-induced decreases in F_m/F₀, F_v/F_m, Φ_PS2, q_P, and A and increase in q_NP were inhibited and the recovery of photosynthesis after rewatering was quicker in pretreated plants. Although differences in F_v/F_m, Φ_PS2 and q_P between treatments were not significant, there was a general trend that the effects of CC increased with the rise of its concentrations. The data suggested that 2.1 mM of CC be suitable for alleviating lipid peroxidation, promoting proline accumulation, retarding leaf water loss and improving photosynthesis of R. glutinosa seedlings under drought stress.     

不同BR施用方式诱导黄瓜幼苗对Ca(NO_3)_2胁迫抗性的研究

为探讨外源油菜素内酯(brassinosteroid,BR)诱导黄瓜幼苗对Ca(NO3)2胁迫抗性的效果,研究了3种外源BR施用方法(0.01mg·L-1 BR浸种、0.1mg·L-1 BR喷叶及其二者结合施用)对Ca(NO3)2胁迫(60mmol·L-1)下黄瓜幼苗生长、生理活动以及光合作用的影响。结果表明:(1)3种外源BR方法处理后,Ca(NO3)2胁迫下的黄瓜幼苗株高、茎粗、展开叶片数、叶面积、干重含水量均显著提高,同时其叶片游离脯氨酸和可溶性糖含量上升,过氧化物酶活性提高,而其丙二醛(MDA)含量趋于无Ca(NO3)2胁迫对照的水平;(2)外源BR处理还提高了Ca(NO3)2胁迫下黄瓜幼苗的净光合速率、蒸腾速率和气孔导度,却抑制了Ca(NO3)2胁迫下胞间CO2浓度的升高。研究认为,适宜浓度的外源BR浸种和喷叶处理均可有效增强黄瓜幼苗渗透调节能力,降低细胞膜质过氧化伤害程度,提高抗氧化酶活性和光合效率,从而表现出对Ca(NO3)2胁迫的抗性,并以操作简便、用量极低的0.01mg·L-1 BR浸种方法效果最佳。     

The efficiency of water use in water stressed plants is increased due to ABA induced stomatal closure

Gas exchange and abscisic acid content of Digitalis lanata EHRH. have been examined at different levels of plant water stress. Net photosynthesis, transpiration and conductance of attached leaves declined rapidly at first, then more slowly following the withholding of irrigation. The intercellular partial pressure of CO₂ decreased slightly. The concentration of 2-cis(S)ABA increased about eight-fold in the leaves of non-irrigated plants as compared with well-watered controls. A close linear correlation was found between the ABA content of the leaves and their conductance on a leaf area basis. In contrast, the plot of net assimilation versus ABA concentration was curvilinear, leading to an increased efficiency of water use during stress. After rewatering, photosynthesis reached control values earlier than transpiration, leaf conductance and ABA content. From these data it is concluded that transpiration through the stomata is directly controlled by the ABA content, whereas net photosynthesis is influenced additionally by other factors.Possible reasons for the responses of photosynthesis and water use efficiency to different stress and ABA levels are discussed.Abbreviations A net CO₂ assimilation - ABA abscisic acid - C_i intercellular CO₂ concentration - g stomatal conductance - T transpiration - WUE water use efficiency     

H_2S对干旱胁迫下白三叶幼苗抗氧化防御能力的影响

以‘拉丁诺’白三叶(Trifolium repens cv.‘Ladino’)为试验材料,研究外源H2S处理对PEG6 000(聚乙二醇)模拟干旱胁迫下白三叶叶片相对含水量(RWC)、膜脂过氧化、活性氧成分、抗氧化酶、抗坏血酸-谷胱甘肽循环代谢和非酶抗氧化物质的影响,以揭示H_2S调控白三叶抗旱性的生理机制。结果显示:(1)0.2 mmol/L的外源NaHS(H_2S供体)能显著提高干旱胁迫下白三叶的叶片相对含水量,维持显著较低的电解质渗透率(EL)和丙二醛(MDA)含量。(2)与直接干旱胁迫相比,干旱胁迫下外源添加NaHS处理的白三叶叶片内超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性显著增强,抗坏血酸-谷胱甘肽循环代谢中关键酶抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)、单脱水抗坏血酸还原酶(MDHAR)和谷胱甘肽还原酶(GR)活性及其抗氧化中间产物抗坏血酸(AsA)、谷胱甘肽(GSH)含量也显著提高。(3)叶片类黄酮、总酚和原花青素的含量在一定的胁迫时间范围内亦显著增加,并伴随着活性氧成分O_2~(-·)产生速率和H_2O_2水平降低。研究认为,外源H2S能通过促进干旱胁迫下白三叶体内的多重抗氧化防御能力来提高其幼苗的抗旱性。     

Hydrogen peroxide spraying alleviates drought stress in soybean plants

To ascertain the effect of exogenously applied hydrogen peroxide (H₂O₂) on drought stress, we examined whether the spraying of soybean leaves with H₂O₂ would alleviate the symptoms of drought stress. Pre-treatment by spraying leaves with H₂O₂ delayed foliar wilting caused by drought stress compared to leaves sprayed with distilled water (DW). Additionally, the relative water content of drought-stressed leaves pre-treated with H₂O₂ was higher than that of leaves pre-treated with DW. Therefore, we analyzed the effect of H₂O₂ spraying on photosynthetic parameters and on the biosynthesis of oligosaccharides related to water retention in leaves during drought stress. Under conditions of drought stress, the net photosynthetic rate and stomatal conductance of leaves pre-treated with H₂O₂ were higher than those of leaves pre-treated with DW. In contrast to DW spraying, H₂O₂ spraying immediately caused an increase in the mRNA levels of d-myo-inositol 3-phosphate synthase 2 (GmMIPS2) and galactinol synthase (GolS), which encode key enzymes for the biosynthesis of oligosaccharides known to help plants tolerate drought stress. In addition, the levels of myo-inositol and galactinol were higher in H₂O₂-treated leaves than in DW-treated leaves. These results indicated that H₂O₂ spraying enabled the soybean plant to avoid drought stress through the maintenance of leaf water content, and that this water retention was caused by the promotion of oligosaccharide biosynthesis rather than by rapid stomatal closure.     

Hydrogen peroxide production is an early event during bicarbonate induced stomatal closure in abaxial epidermis of <Emphasis Type="Italic">Arabidopsis</Emphasis>

The presence of 2 mM bicarbonate in the incubation medium induced stomatal closure in abaxial epidermis of Arabidopsis. Exposure to 2 mM bicarbonate elevated the levels of H₂O₂ in guard cells within 5 min, as indicated by the fluorescent probe, dichlorofluorescein diacetate (H₂DCF-DA). Bicarbonate-induced stomatal closure as well as H₂O₂ production were restricted by exogenous catalase or diphenylene iodonium (DPI, an inhibitor of NAD(P)H oxidase). The reduced sensitivity of stomata to bicarbonate and H₂O₂ production in homozygous atrbohD/F double mutant of Arabidopsis confirmed that NADP(H) oxidase is involved during bicarbonate induced ROS production in guard cells. The production of H₂O₂ was quicker and greater with ABA than that with bicarbonate. Such pattern of H₂O₂ production may be one of the reasons for ABA being more effective than bicarbonate, in promoting stomatal closure. Our results demonstrate that H₂O₂ is an essential secondary messenger during bicarbonate induced stomatal closure in Arabidopsis.     

Pre-treatment with H<Subscript>2</Subscript>O<Subscript>2</Subscript> induces salt tolerance in Barley seedlings

Barley seedlings were pre-treated with 1 and 5 μM H₂O₂ for 2 d and then supplied with water or 150 mM NaCl for 4 and 7 d. Exogenous H₂O₂ alone had no effect on the proline, malondialdehyde (MDA) and H₂O₂ contents, decreased catalase (CAT) activity and had no effect on peroxidase (POX) activity. Three new superoxide dismutase (SOD) isoenzymes appeared in the leaves as a result of 1 μM H₂O₂ treatment. NaCl enhanced CAT and POX activity. SOD activity and isoenzyme patterns were changed due to H₂O₂ pre-treatment, NaCl stress and leaf ageing. In pre-treated seedlings the rate of ¹⁴CO₂ fixation was higher and MDA, H₂O₂ and proline contents were lower in comparison to the seedlings subjected directly to NaCl stress. Cl⁻ content in the leaves 4 and 7 d after NaCl supply increased considerably, but less in pre-treated plants. It was suggested that H₂O₂ metabolism is involved as a signal in the processes of barley salt tolerance.     

Stomatal sensitivities to changes in leaf water potential, air humidity, CO2 concentration and light intensity, and the effect of abscisic acid on the sensitivities in six temperate deciduous tree species

To characterise the stomata of six temperate deciduous tree species, sets of stomatal sensitivities to all the most important environmental factors were measured. To compare the importance of abscisic acid (ABA) in the different stomatal responses, the effect of exogenous ABA on all the stomatal sensitivities was determined.Almost all the stomatal sensitivities: the sensitivity to a decrease in leaf water potential, air humidity, CO₂ concentration ([CO₂]) and light intensity, and to an increase in [CO₂] and light intensity were the highest in the slow-growing species, and the lowest in the fast-growing species. Drought increased the sensitivity to the environmental changes that induce a decrease in the stomatal conductance, and decreased the sensitivity to the changes that induce an increase in this conductance. The sensitivities of the slow-growers were most strongly affected by drought and ABA. Therefore the success of the slow-growers in their ecological niches can be based on the highly sensitive and strictly regulated responses of their stomata. The fast-growers had the highest sensitivity to an increase in leaf water potential and this sensitivity was sharply reduced by drought and ABA. Thus, the dominance of the trees in riparian areas can be based on the ability of their stomata to quickly reach high conductance in well-watered conditions and to efficiently decrease this rate during drought.Stomatal sensitivities to the hydraulic environmental factors (water potentials in plant and air) had higher values in well-watered trees and a more pronounced response to drought than the sensitivities to the photosynthetic environmental factors ([CO₂] and light intensity). Thus, the hydraulic factors most likely prevail over the photosynthetic factors in determining stomatal conductance in these species.In response to exogenous ABA, the rates of stomatal closure, following a decrease in air humidity and light intensity, and an increase in [CO₂], were accelerated. Stomatal opening following an increase in air humidity and light intensity and a decrease in [CO₂] was replaced by slow closing. The rate of stomatal opening following an increase in leaf water potential was reduced. As the sensitivities to changes in light were modified less by the ABA than the other stomatal sensitivities, the prediction of stomatal responses on the basis of the sensitivity to light alone should be excluded in stomatal models.     

Drought-induced H2O2 accumulation in subsidiary cells is involved in regulatory signaling of stomatal closure in maize leaves

Increasing H₂O₂ levels in guard cells in response to environmental stimuli are recently considered a general messenger involved in the signaling cascade for the induction of stomatal closure. But little is known as to whether subsidiary cells participate in the H₂O₂-mediated stomatal closure of grass plants. In the present study, 2-week-old seedlings of maize (Zea mays) were exposed to different degrees of soil water deficit for 3 weeks. The effects of soil water contents on leaf ABA and H₂O₂ levels and stomatal aperture were investigated using physiological, biochemical, and histochemical approaches. The results showed that even under well-watered conditions, significant amounts of H₂O₂ were observed in guard cells, whereas H₂O₂ concentrations in the subsidiary cells were negligible. Decreasing soil water contents led to a significant increase in leaf ABA levels associated with significantly enhanced O₂ ^? and H₂O₂ contents, consistent with reduced degrees of stomatal conductance and aperture. The significant increase in H₂O₂ appeared in both guard cells and subsidiary cells of the stomatal complex, and H₂O₂ levels increased with decreasing soil water contents. Drought-induced increase in the activity of antioxidative enzymes could not counteract the significant increase in H₂O₂ levels in guard cells and subsidiary cells. These results indicate that subsidiary cells participate in H₂O₂-mediated stomatal closure, and drought-induced H₂O₂ accumulation in subsidiary cells is involved in the signaling cascade regulating stomatal aperture of grass plants such as maize.     

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.     

Cross-talks between Ca<Superscript>2+</Superscript>/CaM and H<Subscript>2</Subscript>O<Subscript>2</Subscript> in abscisic acid-induced antioxidant defense in leaves of maize plants exposed to water stress

Here we examined whether Ca²⁺/Calmodulin (CaM) is involved in abscisic acid (ABA)-induced antioxidant defense and the possible relationship between CaM and H₂O₂ in ABA signaling in leaves of maize (Zea mays L.) plants exposed to water stress. An ABA-deficient mutant vp5 and its wild type were used for the experimentation. We found that water stress enhanced significantly the contents of CaM and H₂O₂, and the activities of chloroplastic and cytosolic superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR), and the gene expressions of the CaM1, cAPX, GR1 and SOD4 in leaves of wild-type maize. However, the increases mentioned above were almost arrested in vp5 plants and in the wild-type plants pretreated with ABA biosynthesis inhibitor tungstate (T), suggesting that ABA is required for water stress-induced H₂O₂ production, the enhancement of CaM content and antioxidant defense. Besides, we showed that the up-regulation of water stress-induced antioxidant defense was almost completely blocked by pretreatment with Ca²⁺ inhibitors, CaM antagonists and reactive oxygen (ROS) manipulators. Moreover, the analysis of time course of CaM and H₂O₂ production under water stress showed that the increase in CaM content preceded that of H₂O₂. These results suggested that Ca²⁺/CaM and H₂O₂ were involved in the ABA-induced antioxidant defense under water stress, and the increases of Ca²⁺/CaM contents triggered H₂O₂ production, which inversely affected the contents of CaM. Thus, a cross-talk between Ca²⁺/CaM and H₂O₂ may play a pivotal role in the ABA signaling.     

Photosynthetic rate,chlorophyll fluorescence parameters,and lipid peroxidation of maize leaves as affected by zinc deficiency

Pot trial in greenhouse was conducted using cumulic cinnamon soil from North China to study the effects of zinc deficiency on CO₂ exchange, chlorophyll fluorescence, the intensity of lipid peroxidation, and the activity of superoxide dismutase (SOD) in leaves of maize seedlings. Zn deficiency resulted in a reduction of net photosynthetic rate and stomatal conductance to H₂O. The maximum quantum efficiency of photosystem 2 (PS2) and the PS2 activity were depressed, while the pool size of the plastoquinone molecules was not affected by Zn deficiency. The content of super oxygen anion radical (O₂ ^·−) and the intensity of lipid peroxidation as assessed by malonyldialdehyde content in Zn-deficient leaves were higher than those in Zn-sufficient leaves. The activity of SOD increased with Zn application. The adverse influence of Zn-deficiency on the light stage of photosynthesis is probably one of possible reasons for the limitation of photosynthetic capacity in maize leaves.     

干旱胁迫对降香黄檀幼苗光合生理特性的影响

采用温室盆栽方法,设置对照(CK)、轻度(LS)、中度(MS)和重度(HS)干旱胁迫4个水分条件,研究不同水分条件对降香黄檀幼苗光合和生理特性的影响。结果表明:(1)随着干旱胁迫程度增加,降香黄檀幼苗叶片叶绿素总含量总体呈现出下降趋势。(2)降香黄檀幼苗叶片净光合速率、气孔导度、胞间CO2浓度和蒸腾速率随着干旱胁迫强度增加均呈现出先增加后降低趋势,且MS和HS处理下的气孔导度和胞间CO2浓度同时降低,此时幼苗光合能力的下降主要受气孔因素限制。(3)随着干旱胁迫强度的增加,降香黄檀幼苗叶片细胞膜相对透性、丙二醛含量、游离脯氨酸含量和POD活性均呈现出增加趋势,而同期SOD和CAT活性呈现出先升高后降低趋势。可见,降香黄檀幼苗在轻度干旱胁迫下可通过增加叶片保护酶活性来清除活性氧对其组织造成的伤害,但胁迫超过一定程度后保护酶活性下降,表明降香黄檀幼苗的耐旱能力有限。     

Effect of temperature on net CO2 assimilation and photosystem II quantum yield of electron transfer of French bean (Phaseolus vulgaris L.) leaves during drought stress

The effect of leaf temperature on stomatal conductance and net CO₂ uptake was studied on French bean (Phaseolus vulgaris L.) using either dehydrated attached leaves (25–40% water deficit) or cut leaves supplied with 10^–4 M abscisic acid (ABA) solution to the transpiration stream. Decreasing leaf temperature caused stomatal opening and increased net CO₂ uptake (which was close to zero at around 25° C) to a level identical to that of control leaves (without water deficit) at around 15° C. (i) The ABA effect on stomatal closure was modulated by temperature and, presumably, ABA is at least partly responsible for stomatal closure of french bean submitted to a drought stress. (ii) For leaf temperatures lower than 15° C, net CO₂ uptake was no longer limited by water deficit even on very dehydrated leaves. This shows that dehydrated leaves retain a substantial part of their photosynthetic capacity which can be revealed at normal CO₂ concentrations when stomata open at low temperature. In contrast to leaves fed with ABA, decreasing the O₂ concentration from 21% to 1% O₂ did not increase either the rate of net CO₂ uptake or the thermal optimum for photosynthesis of dehydrated leaves. The quantum yield of PSII electron flow (measured by F/F_m) was lower in 1% O₂ than in 21% O₂ for each leaf pretreatment given (non-dehydrated leaves, dehydrated leaves, and leaves fed with ABA) even within a temperature range in which leaf photosynthesis at normal CO₂ concentration was the same in these two O₂ concentrations. It is concluded that this probably indicates an heterogeneity of photosynthesis, since this difference in quantum yield disappears when using high CO₂ concentrations during measurements.Abbreviations and Symbols ABA abscisic acid - F_m maximum chlorophyll fluorescence - F difference between steady-state chlorophyll fluorescence and F_m - PPFD photosynthetic photon flux density We would like to thank Dr. J.-M. Briantais (Laboratoire d'écologie végétale, Orsay, France) for help during fluorescence measurements and Ms. J. Liebert for technical assistance.     

Effects of salicylic acid on the photosystem 2 of barley seedlings under osmotic stress

The effects of exogenous salicylic acid (SA) on photosystem 2 (PS 2) in barley (Hordeum vulgare L.) seedlings were investigated. SA pretreatment provided protection against subsequent osmotic stress. The highest protective effect of 0.25 mM SA was confirmed by determination of chlorophyll fluorescence, electrolyte leakage, malonyldialdehyde contents, PS 2 mRNAs and proteins. SA pretreatment increased reactive oxygen species (ROS), decreased net photosynthetic rate and stomatal conductance immediately, but prevented ROS accumulation during subsequent osmotic stress by activating antioxidant enzymes. Elimination of H₂O₂ during SA pretreatment inhibited almost all above mentioned SA effects. Therefore, SA pretreatment enhanced osmotic stress tolerance in barley seedlings mainly through ROS signals, rather than SA itself. The only SA-dependent and ROS-independent effect of exogenous SA on PS 2 was reduction of non-photochemical quenching.     

Growth response of barley and tomato to nitrogen stress and its control by abscisic acid,water relations and photosynthesis

Barley (Hordeum vulgare L.) and tomato Lycopersicon esculentum Mill.) were grown hydroponically and examined 2, 5, and 10 d after being deprived of nitrogen (N) supply. Leaf elongation rate declined in both species in response to N stress before there was any reduction in rate of dryweight accumulation. Changes in water transport to the shoot could not explain reduced leaf elongation in tomato because leaf water content and water potential were unaffected by N stress at the time leaf elongation began to decline. Tomato maintained its shoot water status in N-stressed plants, despite reduced water absorption per gram root, because the decline in root hydraulic conductance with N stress was matched by a decline in stomatal conductance. In barley the decline in leaf elongation coincided with a small (8%) decline in water content per unit area of young leaves; this decline occurred because root hydraulic conductance was reduced more strongly by N stress than was stomatal conductance. Nitrogen stress caused a rapid decline in tissue NO ₃ ^- pools and in NO ₃ ^- flux to the xylem, particularly in tomato which had smaller tissue NO ₃ ^- reserves. Even in barley, tissue NO ₃ ^- reserves were too small and were mobilized too slowly (60% in 2 d) to support maximal growth for more than a few hours. Organic N mobilized from old leaves provided an additional N source to support continued growth of N-stressed plants. Abscisic acid (ABA) levels increased in leaves of both species within 2 d in response to N stress. Addition of ABA to roots caused an increase in volume of xylem exudate but had no effect upon NO ₃ ^- flux to the xylem. After leaf-elongation rate had been reduced by N stress, photosynthesis declined in both barley and tomato. This decline was associated with increased leaf ABA content, reduced stomatal conductance and a decrease in organic N content. We suggest that N stress reduces growth by several mechanisms operating on different time scales: (1) increased leaf ABA content causing reduced cell-wall extensibility and leaf elongation and (2) a more gradual decline in photosynthesis caused by ABA-induced stomatal closure and by a decrease in leaf organic N.Abbreviation and symbols ABA abscisic acid - c_i leaf internal CO₂ concentration - L_p root hydraulic conductance     

Abscisic acid-induced apoplastic H<Subscript>2</Subscript>O<Subscript>2</Subscript> accumulation up-regulates the activities of chloroplastic and cytosolic antioxidant enzymes in maize leaves

The histochemical and cytochemical localization of abscisic acid (ABA)-induced H₂O₂ production in leaves of maize (Zea mays L.) plants were examined, using 3,3-diaminobenzidine (DAB) and CeCl₃ staining, respectively, and the relationship between ABA-induced H₂O₂ production and ABA-induced subcellular activities of antioxidant enzymes was studied. H₂O₂ generated in response to ABA treatment was detected within 0.5 h in major veins of the leaves and maximized at about 2–4 h. In mesophyll and bundle sheath cells, ABA-induced H₂O₂ accumulation was observed only in apoplast, and the greatest accumulation occurred in the walls of mesophyll cells facing large intercellular spaces. Meanwhile, ABA treatment led to a significant increase in the activities of the leaf chloroplastic and cytosolic antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR), and pretreatment with the NADPH oxidase inhibitor diphenyleneiodonium (DPI), the O ₂ ⁻ scavenger Tiron and the H₂O₂ scavenger dimethylthiourea (DMTU) almost completely arrested the increase in the activities of these antioxidant enzymes. Our results indicate that the accumulation of apoplastic H₂O₂ is involved in the induction of the chloroplastic and cytosolic antioxidant enzymes. Moreover, an oxidative stress induced by paraquat (PQ), which generates O ₂ ⁻ and then H₂O₂ in chloroplasts, also up-regulated the activities of the chloroplastic and cytosolic antioxidant enzymes, and the up-regulation was blocked by the pretreatment with Tiron and DMTU. These data suggest that H₂O₂ produced at a specific cellular site could coordinate the activities of antioxidant enzymes in different subcellular compartments.     

Implication of ABA and proline on cell membrane injury of water deficit stressed barley seedlings

The aim of this work was to examine the ability of ABA and proline to counteract the deleterious effect of water deficit stress on cell membrane injuries. Six-day-old seedlings of two barley genotypes (cv. Aramir, line R567) were treated with ABA (2·10⁻⁴ M) or proline (0.1 M) for 24 h, and then subjected to osmotic stress for 24h, by immersing their roots in polyethylene glycol (PEG 6000) solution of osmotic potential of −1.0 MPa and −1.5 MPa or by submerging the leaf pieces in PEG solution of osmotic potential of −1.6 MPa. Pretreatment of plants with ABA and proline caused an increase of free proline level in the leaves. Plants treated with ABA exhibited a lower membrane injury index under water stress conditions than those untreated even when no effect of this hormone on RWC in the leaves of stressed plants was observed. Pretreatment of plants with proline prevented to some extent membrane damage in leaves of the stressed seedlings, but only in the case when stress was imposed to roots. Improvement in water status of leaves was also observed in seedlings pretreatment with proline. The protective effect of both ABA and proline was more pronounced in line R567 that exhibited higher membrane injury under water deficit stress conditions.     

Electron transport chain of Saccharomyces cerevisiae mitochondria is inhibited by H2O2 at succinate-cytochrome c oxidoreductase level without lipid peroxidation involvement

The deleterious effects of H₂O₂ on the electron transport chain of yeast mitochondria and on mitochondrial lipid peroxidation were evaluated. Exposure to H₂O₂ resulted in inhibition of the oxygen consumption in the uncoupled and phosphorylating states to 69% and 65%, respectively. The effect of H₂O₂ on the respiratory rate was associated with an inhibition of succinate-ubiquinone and succinate-DCIP oxidoreductase activities. Inhibitory effect of H₂O₂ on respiratory complexes was almost completely recovered by β-mercaptoethanol treatment. H₂O₂ treatment resulted in full resistance to Q_O site inhibitor myxothiazol and thus it is suggested that the quinol oxidase site (Q_O) of complex III is the target for H₂O₂. H₂O₂ did not modify basal levels of lipid peroxidation in yeast mitochondria. However, H₂O₂ addition to rat brain and liver mitochondria induced an increase in lipid peroxidation. These results are discussed in terms of the known physiological differences between mammalian and yeast mitochondria.