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1.
Using Arabidopsis plants Col-0 and vtc2 transformed with a redox sensitive green fluorescent protein, (c-roGFP) and (m-roGFP), we investigated the effects of a progressive water stress and re-watering on the redox status of the cytosol and the mitochondria. Our results establish that water stress affects redox status differently in these two compartments, depending on phenotype and leaf age, furthermore we conclude that ascorbate plays a pivotal role in mediating redox status homeostasis and that Col-0 Arabidopsis subjected to water stress increase the synthesis of ascorbate suggesting that ascorbate may play a role in buffering changes in redox status in the mitochondria and the cytosol, with the presumed buffering capacity of ascorbate being more noticeable in young compared with mature leaves. Re-watering of water-stressed plants was paralleled by a return of both the redox status and ascorbate to the levels of well-watered plants. In contrast to the effects of water stress on ascorbate levels, there were no significant changes in the levels of glutathione, thereby suggesting that the regeneration and increase in ascorbate in water-stressed plants may occur by other processes in addition to the regeneration of ascorbate via the glutathione. Under water stress in vtc2 lines it was observed stronger differences in redox status in relation to leaf age, than due to water stress conditions compared with Col-0 plants. In the vtc2 an increase in DHA was observed in water-stressed plants. Furthermore, this work confirms the accuracy and sensitivity of the roGFP1 biosensor as a reporter for variations in water stress-associated changes in redox potentials.  相似文献   

2.
To explore the significance of the ascorbate–glutathione cycle under drought stress, the leaves of 2-year-old potted apple (Malus domestica Borkh.) plants were used to investigate the changes of each component of the ascorbate–glutathione cycle as well as the gene expression of dehydroascorbate reductase (DHAR, EC 1.8.5.1), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) under drought stress. The results showed that the malondialdehyde (MDA) and H2O2 concentrations in apple leaves increased during drought stress and began to decrease after re-watering. The contents of total ascorbate, reduced ascorbic acid (AsA), total glutathione and glutathione (GSH) were obviously upregulated in apple leaves when the soil water content was 40–45%. With further increase of the drought level, the contents of the antioxidants and especially redox state of AsA and GSH declined. However, levels of them increased again after re-watering. Moreover, drought stress induced significant increase of the activities of enzymes such as APX, scavenging H2O2, and also of monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), DHAR and GR used to regenerate AsA and GSH, especially when the soil water content was above 40–45%. During severe drought stress, activities of the enzymes were decreased and after re-watering increased again. Gene expression of cytoplasmic DHAR, cytoplasmic APX and cytoplasmic GR showed similar changes as the enzyme activities, respectively. The results suggest that the ascorbate–glutathione cycle is up-regulated in response to drought stress, but cannot be regulated at severe drought stress conditions.  相似文献   

3.
4.
Some plant-symbiotic strains of the genus Trichoderma colonize roots and induce profound changes in plant gene expression that lead to enhanced growth, especially under biotic and abiotic stresses. In this study, we tested the hypothesis that one of the protective mechanisms enhanced by T. harzianum T22 colonization is the antioxidant defense mechanism. Having established that strain T22 modulates the expression of the genes encoding antioxidant enzymes, the status of antioxidant defense of tomato seedlings in response to colonization by T22 and water deficit was investigated. Total ascorbate or glutathione levels were not affected by either stimuli, but under water deficit, antioxidant pools became more oxidized (lower ratios of reduced to oxidized forms), whereas colonized plants maintained redox state as high as or higher than unstressed and untreated plants. The enhanced redox state of colonized plants could be explained by their higher activity of ascorbate and glutathione-recycling enzymes, higher activity of superoxide dismutase, catalase, and ascorbate peroxidase, in both root and shoot throughout the experiment. Similar enzymes were induced in uncolonized plants in response to water-deficit stress but to a lower extent when compared with colonized plants. This orchestrated enhancement in activity of reactive oxygen species (ROS)-scavenging pathways in colonized plants in response to stress supports the hypothesis that enhanced resistance of colonized plants to water deficit is at least partly due to higher capacity to scavenge ROS and recycle oxidized ascorbate and glutathione, a mechanism that is expected to enhance tolerance to abiotic and biotic stresses.  相似文献   

5.
The hybrid Richter-110 (Vitis berlandieri x Vitis rupestris) (R-110) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to water withholding followed by re-watering. The goal was to analyze how stomatal conductance (g(s)) is regulated with respect to different physiological variables under water stress and recovery, as well as how water stress affects adjustments of water use efficiency (WUE) at the leaf level. Water stress induced a substantial stomatal closure and an increase in WUE, which persisted many days after re-watering. The g(s) during water stress was mainly related to the content of ABA in the xylem and partly related to plant hydraulic conductivity but not to leaf water potential. By contrast, low g(s) during re-watering did not correlate with ABA contents and was only related to a sustained decreased hydraulic conductivity. In addition to a complex physiological regulation of stomatal closure, g(s) and rate of transpiration (E) were strongly affected by leaf-to-air vapor pressure deficit (VPD) in a way dependent of the treatment. Interestingly, E increased with increasing VPD in control plants, but decreased with increasing VPD in severely stressed plants. All together, the fine stomatal regulation in R-110 resulted in very high WUE at the leaf level. This genotype is revealed to be very interesting for further studies on the physiological mechanisms leading to regulation of stomatal responsiveness and WUE in response to drought.  相似文献   

6.
Gas exchange, chlorophyll fluorescence and water potentials, together with ascorbate and glutathione concentrations, were studied during moderate and severe drought stress and in response to re-watering in Allocasuarina luehmannii seedlings. Moderate drought stress (MS) decreased stomatal conductance (gs) and net CO2 assimilation rates (A) to ∼40% and ∼60% of control values, respectively, and caused decreases in internal CO2 concentration (Ci) and maximum light use efficiency of light-acclimated photosystem II (PSII) centres (Fv'/Fm'). Severe drought stress (SS) decreased gs and A to ∼5% and ∼15% of the control values, respectively, and caused increases in Ci and PSII excitation pressure (1 − qP), as well as decreases in water potentials, effective quantum yield of PSII (ΦPSII), maximum efficiency of PSII (Fv/Fm) and Fv'/Fm'. Ascorbate and glutathione concentrations remained unaffected by drought treatments, but ascorbate became more oxidised under severe stress. MS seedlings recovered within 1 day (Ci, Fv'/Fm') to 1 week (A, gs) of re-watering. In comparison, SS seedlings had longer-lasting after-stress effects, with recovery of many variables (gs, water potentials, Fv/Fm, ΦPSII, Fv'/Fm') taking between 1 and 3 weeks from re-watering. We found no indication that interaction with antioxidants played a significant role in recovery. In conclusion, A. luehmannii seedlings appear to function normally under moderate drought, but do not seem to have particular metabolic tolerance mechanisms to endure severe drought, which may have implications for its persistence under climate change at the drier margins of its distribution.  相似文献   

7.
Water stress effects were studied on three cultivars ofEragrostis curvula. Leaf water potential, RWC, total plantleaf area, green dry weight mass percentage and CO2 gas-exchangeweremeasured during the onset of stress and after recovery. After 3 days of waterstress, RWC of cv Tanganyika plants was around 30–40% of controls,while RWC of cvs Ermelo and Consol was around 50–60% of controls.However midday and predawn water potentials were lower in cvs Tanganyka andErmelo than in cv Consol. After re-watering, RWC and water potentials recoveredonly in Consol plants. A strong decrease of leaf area was recorded in cvsErmeloand Consol during water stress (about 91–94% less than the leafarea of controls). Photosynthesis decreased as a function of the degree ofwaterstress severity in all cultivars. Also, light saturated photosynthesis,CO2 quantum yield and light at which saturated photosynthesisoccurred, were strongly reduced by water stress. Recovery of photosynthesis wasfound in cv Consol after five days re-watering. Cv Consol showed a betterconservation of water and higher resistance to water stress than the other twocvs.  相似文献   

8.
Transgenic tobacco plants expressing the ascorbate oxidase (AAO) gene in sense and antisense orientations, and an Arabidopsis mutant in which the T-DNA was inserted into a putative AAO gene, were used to examine the potential roles of AAO for salt-stress tolerance in plants. AAO activities in the transgenic tobacco plants expressing the gene in sense and antisense orientations were, respectively, about 16-fold and 0.2-fold of those in the wild type. Under normal growth conditions, no significant differences in phenotypes were observed, except for a delay in flowering time in the antisense plants. However, at high salinity, the percentage germination, photosynthetic activity, and seed yields were higher in antisense plants, with progressively lower levels in the wild type and the sense plants. The redox state of apoplastic ascorbate in sense plants was very low even under normal growth conditions. Upon salt stress, the redox state of symplastic and apoplastic ascorbate decreased among the three types of plants, but was lowest in the sense plants. The hydrogen peroxide contents in the symplastic and apoplastic spaces were higher in sense plants, progressively lower in the wild type, followed by the antisense plants. The Arabidopsis T-DNA inserted mutant exhibited very low ascorbate oxidase activity, and its phenotype was similar to that of antisense tobacco plants. These results suggest that the suppressed expression of apoplastic AAO under salt-stress conditions leads to a relatively low level of hydrogen peroxide accumulation and a high redox state of symplastic and apoplastic ascorbate which, in turn, permits a higher seed yield.  相似文献   

9.
以岩溶特有药用植物地枫皮为材料,研究土壤水分胁迫及复水条件下,其叶片光合参数、叶绿素荧光参数及光合色素含量的变化特性,进而探讨其对水分胁迫的生理生态适应性。结果表明:停止供水10 d,水分胁迫地枫皮叶片的P_n(净光合速率)、C_i(胞间CO_2浓度)、G_s(气孔导度)和L_s(气孔限制值)均下降,气孔限制是P_n降低的主要原因;停止供水15 d,水分胁迫地枫皮叶片的P_n日变化呈逐渐下降趋势,上午9:30以后全天的P_n值均接近零,非气孔限制成为P_n下降的主要因素;而对照地枫皮叶片的P_n日变化呈"双峰型",中午P_n下降的主要原因依然是气孔限制。水分胁迫下,地枫皮叶片叶绿素含量降低和Chl_(a/b)升高,减少了叶片对光能的捕获,减轻了光合机构遭受光氧化的破坏,而Car/Chl_(a+b)升高增强了光保护能力。水分胁迫下,地枫皮叶片的初始荧光(F_o)显著增大,最大荧光(F_m)、光系统Ⅱ(PSⅡ)潜在活性(F_v/F_o)和最大光化学效率(F_v/F_m)均显著降低,表明水分胁迫对地枫皮叶片的PSⅡ反应中心和电子传递造成了一定的破坏,从而使其PSⅡ的潜在活性和最大光化学效率降低。复水5 d后,地枫皮的上述生理生态参数均能恢复到对照水平,表明其复水后的生理修复能力很强。  相似文献   

10.
Ascorbate is an important antioxidant in plants and fulfills many functions related to plant defense, redox signaling and modulation of gene expression. We have analyzed the subcellular distribution of reduced and oxidized ascorbate in leaf cells of Arabidopsis thaliana and Nicotiana tabacum by high-resolution immuno electron microscopy. The accuracy and specificity of the applied method is supported by several observations. First, preadsorption of the ascorbate antisera with ascorbic acid or dehydroascorbic acid resulted in the reduction of the labeling to background levels. Second, the overall labeling density was reduced between 50 and 61% in the ascorbate-deficient Arabidopsis mutants vtc1-2 and vtc2-1, which correlated well with biochemical measurements. The highest ascorbate-specific labeling was detected in nuclei and the cytosol whereas the lowest levels were found in vacuoles. Intermediate labeling was observed in chloroplasts, mitochondria and peroxisomes. This method was used to determine the subcellular ascorbate distribution in leaf cells of plants exposed to high light intensity, a stress factor that is well known to cause an increase in cellular ascorbate concentration. High light intensities resulted in a strong increase in overall labeling density. Interestingly, the strongest compartment-specific increase was found in vacuoles (fourfold) and in plastids (twofold). Ascorbate-specific labeling was restricted to the matrix of mitochondria and to the stroma of chloroplasts in control plants but was also detected in the lumen of thylakoids after high light exposure. In summary, this study reveals an improved insight into the subcellular distribution of ascorbate in plants and the method can now be applied to determine compartment-specific changes in ascorbate in response to various stress situations.  相似文献   

11.
Wheat ( Triticum aestivum L.) seedlings of a drought-resistant cv. C306 were subjected to severe water deficit directly or through stress cycles of increasing intensity with intermittent recovery periods (drought acclimation). The antioxidant defense in terms of redox metabolites and enzymes in leaf cells, chloroplasts, and mitochondria was examined in relation to ROS-induced membrane damage. Drought-acclimated seedlings modulated growth by maintaining favorable turgor potential and RWC and were able to limit H2O2 accumulation and membrane damage as compared with non-acclimated plants during severe water stress conditions. This was due to systematic upregulation of H2O2-metabolizing enzymes especially ascorbate peroxidase (APX, EC 1.11.1.11) and by maintaining ascorbate–glutathione redox pool in acclimated plants. By contrast, failure in the induction of APX and ascorbate–glutathione cycle enzymes makes the chloroplast susceptible to oxidative stress in non-acclimated plants. Non-acclimated plants protected the leaf mitochondria from oxidative stress by upregulating superoxide dismutase (SOD, EC 1.15.1.1), APX, and glutathione reductase (GR, EC 1.6.4.2) activities. Rewatering led to rapid enhancement in all the antioxidant defense components in non-acclimated plants, which suggested that the excess levels of H2O2 during severe water stress conditions might have inhibited or downregulated the antioxidant enzymes. Hence, drought acclimation conferred enhanced oxidative stress tolerance by well-co-ordinated induction of antioxidant defense both at the chloroplast and at the mitochondrial level.  相似文献   

12.
We examined the influence of water stress (water deficit) induced by drought on the steady state levels of ascorbic acid (ASC), dehydroascorbate (DHA), and the ASC&rcolon;DHA redox status in leaflets of Glycine max (soybean) and leaves of Spinacia oleracea (spinach). Two soybean cultivars (cv. Essex and cv. Forrest) and one spinach cultivar (cv. Nordic) were grown in high-light growth chambers ( approximately 1000-1200 μmol m-2 s-1) or in the greenhouse during May, June, and July 1999. The cultivars were supplied with water until approximately 25-29 d postemergence, at which time one-half of the plants were not watered for a period of from 4.5 to 7.5 d; the other half of the plants were provided water daily and served as controls. On designated days, leaf water potential (PsiLeaf) was measured, and leaf disks of constant area were excised in the period between approximately 1230 and 1330 hours. Leaf disk samples were immediately frozen in liquid N2, samples were extracted, and ASC and DHA levels were measured and expressed as μmol per gram dry mass per time point. For the soybean cultivars, low PsiLeaf values ( approximately -3.00 to -3.95 MPa) were accompanied by slight decreases in ASC levels and slight increases in DHA levels per gram dry mass. In some cases, leaflet ASC levels of water-stressed soybeans were similar to controls or were even increased by as much as 1.2 times. In soybeans, the mole fraction of ASC remained at 93-99 mol% of the total ascorbate (ASC+DHA), indicating that most of the total ascorbate remained in the reduced form even at low water potential. In spinach plants subjected to water stress (-1.8 to -2.6 MPa), leaf ASC decreased as much as 38%, but the ASC remained at 96-99 mol% of the total ascorbate. It is concluded that during water stress, enzymes of the ascorbate-glutathione cycle in leaf mesophyll cells, as well as in the system that generates reductant to support DHA to ASC recycling, e.g., photosynthetic electron transport in chloroplasts, is able to remain active enough to maintain reduction of DHA to ASC.  相似文献   

13.
Photosynthesis, transpiration, stomatal conductance and chlorophyll fluorescence characteristics were examined in kidney bean plants, with developing gradually water stress for several days after watering and then permitted to recover by re-watering. The photosynthetic rate, transpiration rate, and stomatal conductance decreased rapidly by withholding water for 2 days. The Fv/Fm of chlorophyll fluorescence characteristics slightly decreased when the water was withheld for 7 days. After re-watering the rate of recovery of photosynthesis, transpiration, and stomatal conductance decreased gradually as the days without watering became longer. The differences existed in rates of recovery of photosynthesis, transpiration, and stomatal conductance following drought stress. Among the fractional recoveries the highest was photosynthesis, and the lowest was stomatal conductance. Photosynthesis rate following drought stress was rapidly recovered until 2 days after re-watering, then recovered slowly. The critical time for the recovery of photosynthesis was recognized. The results show clearly a close correlation between the leaf water potential and the recovery level and speed of photosynthesis, transpiration, and stomatal conductance.  相似文献   

14.
Exogenous salicylic acid (SA) can be used for chemical hardening to alleviate oxidative stress in plants exposed to salinity. The treatment of 5-week-old Arabidopsis thaliana plants with increasing doses of SA alters the ascorbate (ASC) and glutathione (GSH) pools, and modulates their redox status and the activity of several antioxidant enzymes, such as ascorbate peroxidase (APX) and glutathione reductase (GR). To investigate the role of GR in the maintenance of cytoplasmic redox homeostasis after hardening by SA, wild type (WT) and gr1 mutant plants, expressing the cytoplasmic redox-sensitive green fluorescent protein (c-roGFP1), were pre-treated with 10?7 and 10?5 M SA for 2 weeks and subsequently exposed to 100 mM NaCl. The redox status of the salt-stressed WT plants became more oxidized, which was prevented by pretreatment with 10?5 M SA. The gr1 mutants showed more positive redox potential than WT plants, which could be reversed by treatment with 10?5 M SA. In mutants, the increased GSH levels may have compensated for the deleterious effect of GR deficiency and stabilized the redox potential in plants exposed to salinity. The ASC regeneration in WT plants shifted from the GSH-dependent dehydroascorbate reductase (DHAR) reaction to the NAD(P)H-dependent monodehydroascorbate reductase (MDHAR) activity during chemical hardening, which contributed to the preservation of the GSH pool in plants under salt stress. Our results suggest that the maintenance of GSH levels and redox homeostasis by SA-mediated hardening play a major role in priming and defending against salt stress.  相似文献   

15.
Ascorbate (vitamin C) is a potent antioxidant protecting plants against oxidative damage imposed by environmental stresses such as ozone and drought. Dehydroascorbate reductase (DHAR; EC 1.8.5.1) is one of the two important enzymes functioning in the regeneration of ascorbate (AsA). To examine the protective role of DHAR against oxidative stress, we developed transgenic tobacco plants overexpressing cytosolic DHAR gene from Arabidopsis thaliana . Incorporation of the transgene in the genome of tobacco plants was confirmed by polymerase chain reaction and Southern blot analysis, and its expression was confirmed by Northern and Western blot analyses. These transgenic plants exhibited 2.3–3.1 folds higher DHAR activity and 1.9–2.1 folds higher level of reduced AsA compared with non-transformed control plants. The transgenic plants showed maintained redox status of AsA and exhibited an enhanced tolerance to ozone, drought, salt, and polyethylene glycol stresses in terms of higher net photosynthesis. In this study, we report for the first time that the elevation of AsA level by targeting DHAR overexpression in cytosol properly provides a significantly enhanced oxidative stress tolerance imposed by drought and salt.  相似文献   

16.
17.
马铃薯块茎膨大期不同程度干旱后复水的源库补偿效应   总被引:1,自引:0,他引:1  
旱后复水的补偿效应在多种作物的不同生育时期都存在,是植物抵抗逆境胁迫和伤害的重要自我调节机制,也是对有限水分高效利用的体现.本研究在马铃薯块茎膨大期进行两轮干旱后复水处理,明确马铃薯补偿效应产生的干旱胁迫阈值,并从源-库角度探索马铃薯旱后复水补偿效应产生的缘由.试验选取‘大西洋’马铃薯脱毒组培苗为材料,设置充分供水(W)、轻度干旱后复水(D1-W)、中度干旱后复水(D2-W)和重度干旱后复水(D3-W)4个水分处理并经过两个循环.结果表明:在经过两轮轻度干旱复水后,马铃薯产量表现出超补偿效应,水分利用效率和产量比充分供水分别提高了17.5%和6.3%;中度水分胁迫表现出近等量补偿效应,产量与充分供水差异不大,而水分利用效率提高了8.4%;而重度水分胁迫没有表现出产量补偿效应.不同程度的干旱胁迫均降低马铃薯叶片叶绿素含量、净光合速率、叶面积等源的大小和活性,而在复水后,轻度和中度胁迫出现了超补偿和补偿效应,增强了源的供应能力.同时,适度干旱后复水显著增强了块茎(库)中蔗糖-淀粉代谢途径关键酶的活性,提高了库活性,进而表现为块茎平均重量的增加.综上,马铃薯块茎膨大期适度的水分亏缺在复水后源-库均存在补偿和超补偿效应,以此来弥补干旱带来的损失,最终在产量上表现为补偿或者超补偿效应,并显著提高了水分利用效率.  相似文献   

18.
Spring-sown (vegetative) and autumn-sown (flowering) swards of Lolium perenne cvs Melle, Aurora and their hybrid growing in 0.9 m deep bins of soil in the glasshouse were compared, a) as drought progressed from June to August 1986, and b) after cutting, fertilising and re-watering.
During drought, vegetative plants produced more herbage, had initially higher leaf extension rates, had longer and wider leaves, maintained more tillers, had lower mortality, adjusted osmotic potential more effectively and had stomata less sensitive to stress than did flowering plants. On re-watering, previously stressed flowering plants regrew more slowly than the irrigated controls, whereas previously stressed vegetative plants regrew more rapidly than the controls.
Aurora was the most drought-resistant population, mainly because of low tiller death rates. The hybrid tended to have the lowest leaf water potentials and conductances and grew poorly during drought. Possible underlying physiological mechanisms are discussed.  相似文献   

19.
Since the higher redox potential of quinone molecules has been correlated with enhanced cellular deleterious effects, we studied the ability of the association of ascorbate with several quinones derivatives (having different redox potentials) to cause cell death in K562 human leukaemia cell line. The rationale is that the reduction of quinone by ascorbate should be dependent of the quinone half-redox potential thus determining if reactive oxygen species (ROS) are formed or not, leading ultimately to cell death or cell survival. Among different ROS that may be formed during redox cycling between ascorbate and the quinone, the use of different antioxidant compounds (mannitol, desferal, N-acetylcysteine, catalase and superoxide dismutase) led to support H2O2 as the main oxidizing agent. We observed that standard redox potentials, oxygen uptake, free ascorbyl radical formation and cell survival were linked. The oxidative stress induced by the mixture of ascorbate and the different quinones decreases cellular contents of ATP and GSH while caspase-3-like activity remains unchanged. Again, we observed that quinones having higher values of half-redox potential provoke a severe depletion of ATP and GSH when they were associated with ascorbate. Such a drop in ATP content may explain the lack of activation of caspase-3. In conclusion, our results indicate that the cytotoxicity of the association quinone/ascorbate on K562 cancer cells may be predicted on the basis of half-redox potentials of quinones.  相似文献   

20.
水分胁迫和复水对石灰岩地区柏木幼苗根系生长的影响   总被引:8,自引:0,他引:8  
刘锦春  钟章成 《生态学报》2009,29(12):6439-6445
利用适生植物自身的抗旱能力来进行石灰岩地区植被的恢复和重建是石灰岩生态研究中的重要问题.为了解石灰岩山地适生物种柏木对水分胁迫的适应机制,以柏木实生苗为材料,通过盆栽水分受控实验,研究了其根系生长特性及复水后的修复能力.结果表明:在水分胁迫程度不大、历时不长的情况下,水分胁迫有诱导根系下扎的趋势,复水后,根长在原有的基础上进一步伸长.随着胁迫的加剧和胁迫时间的延长,柏木幼苗的根长、根平均直径、根表面积和根体积等根系大小参数均呈降低趋势,以采取小根系来增强竞争力.复水后,除重度干旱外,其余处理组根系大小参数都能恢复到对照水平,补偿作用与胁迫程度有关.一定程度的水分胁迫对柏木幼苗的茎叶生长没有产生影响,而对根的抑制作用明显,根冠比降低,但胁迫程度加剧和历时延长后,柏木幼苗的根冠比有增大的趋势.柏木幼苗在水分胁迫较轻时把较多的碳水化合物分配到茎叶中,而胁迫严重时把较多的碳水化合物分配到根部.复水促进根、冠干物质的积累,但不同的胁迫程度和胁迫时间促进作用不同.  相似文献   

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