Inducible and constitutive mechanisms of salt stress resistance in <Emphasis Type="Italic">Geum urbanum</Emphasis> L.

The avens (Geum urbanum L.) seedlings were grown for 6 weeks until the expansion of five to six leaves and then exposed to salinity shock (300 mM NaCl in the nutrient medium) or to a gradual (within 4 days) increase in NaCl concentration from 100 to 400 mM. The dynamics of stress-dependent accumulation of Na⁺, Cl^?, proline, and polyamines in leaves and roots was measured, together with activities of antioxidant enzymes, namely, superoxide dismutase (SOD) and guaiacol-dependent peroxidase occurring in soluble, ionically bound, and covalently bound forms. It is shown that avens plants can adapt to gradual salinization by mobilizing stressinducible protective mechanisms (accumulation of proline and spermine) and by activating constitutive enzyme systems (SOD and peroxidase).     

Homeostasis of polyamines and antioxidant systems in roots and leaves of <Emphasis Type="Italic">Plantago major</Emphasis> under salt stress

Functioning of the antioxidant system in roots and leaves of Plantago major L. in water culture at the stage of 5–6 genuine leaves of the plants subjected to NaCl (100 mM) action for 96 h was investigated. This plant exhibited a pronounced organ specificity of antioxidant defense system functioning. The roots were characterized by high constitutive activities of superoxide dismutase and three forms of peroxidase, and a lower catalase activity. Constitutive level of polyamines in roots was higher than in leaves. In both leaves and roots during first 24 h, the polyamine content declined but spermidine remained to be a predominant polyamine. The analysis of differential expression of the genes encoding enzymes of polyamine biosynthesis demonstrated certain differences in these plant organs. The changes in expression of genes MET1, SPMS1, and SPMS2 were observed in roots, whereas in leaves expression of MET1, SAMDC1, SPDS1, and SPMS1 was altered. These changes are possibly one of the mechanisms responsible for the regulation of polyamine endogenous level under salinity. In contrast to leaves, in roots, the oxidative degradation of spermidine by polyamine oxidase can take part in the regulation of endogenous spermidine level. Taken together, these findings allowed us to conclude that, unlike leaves, the roots of P. major under salinity conditions possessed a higher activity of the antioxidant system protecting plants from injurious action of oxidative stress, thereby providing survival of this plant species under stress conditions. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 3, pp. 359–368. This text was submitted by the authors in English.     

Role of antioxidant systems in wild plant adaptation to salt stress

Wild plants differing in the strategies of adaptation to salinity were grown for six weeks in the phytotron and then subjected to salt stress (100 mM NaCl, 24 h). The activities of principal antioxidant enzymes and the accumulation of sodium ions and proline were studied. Independently of the level of constitutive salt tolerance, plants of all species tested accumulated sodium ions under salinity conditions but differed in their capability of stress-dependent proline accumulation and superoxide dismutase (SOD) and guaiacol-dependent peroxidase activities. Proline-accumulating species were found among both halophytes (Artemisia lerchiana and Thellungiella halophila) and glycophytes (Plantago major and Mycelis muralis). The high activities of ionically-bound and covalently bound peroxidases were characteristic of Th. halophila plants. High constitutive and stress-induced SOD activities were, as a rule, characteristic of glycophytes with the low constitutive proline level: Geum urbanum and Thalictrum aquilegifolium. Thus, a negative correlation was found between proline content and SOD activity in wild species tested; it was especially bright in the halophyte Th. halophila and glycophyte G. urbanum. An extremely high constitutive and stress-induced levels of proline and peroxidase activity in Th. halophila maybe compensate SOD low activity in this plant, and this contributed substantially into its salt resistance. Thus, monitoring of stress-dependent activities of some antioxidant enzymes and proline accumulation in wild plant species allowed a supposition of reciprocal interrelations between SOD activity and proline accumulation. It was also established that the high SOD activity is not obligatory trait of species salt tolerance. Moreover, plants with the high activity of peroxidase and active proline accumulation could acclimate to salts stress (100 mM NaCl, 24 h) independently of SOD activity.     

Antioxidant systems in the leaf apoplast compartment of Pinus pinaster Ait. and Pinus radiata D. Don. Plants exposed to SO2

The activities of guaiacol peroxidase (GuPOD), ascorbate peroxidase (ASAp), superoxide dismutase (SOD) and ascorbate/glutathione cycle (AGC) enzymes, together with ascorbate (ASC) and glutathione contents, were determined in apoplastic-fluid and cell-wall fractions of needles of Pinus pinaster Ait. and Pinus radiata D. Don. exposed for up to 6 months to SO₂ (0.01 ppm or 0.30 ppm) in fumigation chambers. AGC enzyme activities (monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase) were in all cases undetectable, as was glutathione content. In needles of P. pinaster plants exposed to SO₂, ascorbate content and all enzyme activities considered (except AGC enzymes) increased. The increases were most marked in response to the higher SO₂ concentration. In needles of P. radiata, similar but less marked responses were observed. These findings suggest a) that enzyme activities and ascorbate contents increase in order to deal with the reactive oxygen intermediates produced during long-term contamination with SO₂, and b) that P. pinaster has more effective defences against contamination of this type than P. radiata.     

Effects of salinity on the contents of polyamines and some other compounds in sunflower plants differing in salt tolerance

The effects of salt stress (50, 100, and 150 mM NaCl) on the levels of free, bound, and total polyamines were studied in the leaf tissues of salt-tolerant (Coban) and salt-sensitive (Sanbro) cultivars of sunflower (Helianthus annuus L.) plants grown for 15 or 25 days under salinity. The amounts of free, acid-soluble bound, and total spermine increased in leaf tissues of sunflower plants subjected to salt stress while the levels of other polyamines decreased or no significant changes occurred. The increase in some PA titers suggests their potential role in overcoming the adverse effect of salinity stress. The salt sensitivity of the sunflower plants was associated with the excessive accumulation of total polyamines in the leaf tissues of salt-sensitive cultivar (Sanbro) under saline condition. The content of other compounds such as proline, protein, and ions (Na⁺, K⁺, Cl^-, K⁺/Na⁺) in leaf tissue changed depending on salt concentration and the cultivars used.From Fiziologiya Rastenii, Vol. 52, No. 1, 2005, pp. 36–42.Original English Text Copyright © 2005 by Mutlu, Bozcuk.This article was presented by the authors in English.     

Cytokinin-induced activity of antioxidant enzymes in transgenic Pssu-ipt tobacco during plant ontogeny

Cytokinin (CK) content and activities of several antioxidant enzymes were examined during plant ontogeny with the aim to elucidate their role in delayed senescence of transgenic Pssu-ipt tobacco. Control Nicotiana tabacum L. (cv. Petit Havana SR1) and transgenic tobacco with the ipt gene under the control of the promoter of small subunit of Rubisco (Pssu-ipt) were both grown either as grafts on control rootstocks or as rooted plants. Both control plant types showed a decline in total content of CKs with proceeding plant senescence. Contrary to this both transgenic plant types exhibited at least ten times higher content of CKs than controls and a significant increase of CK contents throughout the ontogeny with maximal values in the later stages of plant development. Significantly higher portion of O-glucosides was found in both transgenic plant types compared to control ones. In transgenic plants, zeatin and zeatin riboside were predominant type of CKs. Generally, Pssu-ipt tobacco exhibited elevated activities of antioxidant enzymes compared to control tobacco particularly in the later stages of plant development. While in control tobacco activity of glutathione reductase (GR) and superoxide dismutase (SOD) showed increasing activity up to the onset of flowering and then gradually decreased, in both transgenic types GR increased and SOD activity showed only small change throughout the plant ontogeny. Ascorbate peroxidase (APOD) was stimulated in both transgenic types. The manifold enhancement of syringaldazine and guaiacol peroxidase activities was observed in transgenic grafts throughout plant ontogeny in contrast to control and transgenic rooted plants, where the increase was found only in the late stages. Electron microscopic examination showed higher number of crystallic cores in peroxisomes and abnormal interactions among organelles in transgenic tobacco in comparison with control plant. The overproduction of cytokinins resulted in the stimulation of activities of AOE throughout the plant ontogeny of transgenic Pssu-ipt tobacco.     

Modulation of expression of SOD isoenzymes in mud crab (Scylla serrata): effects of inhibitors,salinity and season

Presence of several isoenzymes of superoxide dismutase (SOD) were demonstrated in tissues (abdominal muscle: 7 number, hepatopancreas: 13 number and gills: 7 number) of mud crabs (Scylla serrata) by employing specific staining of the enzyme in native-PAGE. SOD isoenzymes in tissues of mud crab were found to be thermolabile. The intensity of a major SOD band in tissues of crabs was reduced by the treatment of H₂O₂ or chloroform:ethanol. KCN treatment resulted in splitting of that major SOD band into two or more distinct bands. SDS treatment resulted in disruption of SOD bands. A sex-specific SOD isoenzyme band of higher molecular weight was observed in gills and muscle in winter and summer seasons, respectively. The observed different SOD isoenzyme pattern in tissues at altered salinities and seasons suggests separate tissue-specific antioxidant adaptation strategies of crabs against abiotic factors.     

Growth, photosynthesis and respiration in Plantago coronopus as affected by salinity

Plantago coronopus was grown in a non-saline culture solution and in a culture solution containing 50 m M NaCl. The rates of dry matter accumulation in both roots and shoots were not affected by 50 m M NaCl. Photosynthesis, expressed per shoot, was also the same in both environments. Neither the rate of shoot respiration nor that of root respiration was affected by salinity. In both environments the alternative respiratory pathway contributed to the same extent in root respiration. The activity of the alternative pathway decreased with increasing age. Since the respiratory activities were the same in plants grown under both saline and non-saline conditions and since the alternative respiratory pathway was also equally active in roots under both environmental conditions, it is concluded that respiratory costs involved in growth in 50 m M NaCl are negligible in terms of the plant's total energy costings.     

Thellungiella halophila, a salt-tolerant relative of Arabidopsis thaliana, has specific root ion-channel features supporting K+/Na+ homeostasis under salinity stress

Thellungiella halophila is a salt-tolerant relative of Arabidopsis thaliana with high genetic and morphological similarity. In a saline environment, T. halophila accumulates less sodium and retains more potassium than A. thaliana. Detailed electrophysiological comparison of ion currents in roots of both species showed that, unlike A. thaliana, T. halophila exhibits high potassium/sodium selectivity of the instantaneous current. This current differs in its pharmacological profile from the current through inward- and outward-rectifying K(+) channels insofar as it is insensitive to Cs(+) and TEA(+), but resembles voltage-independent channels of glycophytes as it is inhibited by external Ca(2+). Addition of Cs(+) and TEA(+) to the growth medium confirmed the key role of the instantaneous current in whole-plant sodium accumulation. A negative shift in the reversal potential of the instantaneous current under high-salt conditions was essential for decreasing sodium influx to twofold lower than the corresponding value in A. thaliana. The lower overall sodium permeability of the T. halophila root plasma membrane resulted in a smaller membrane depolarization during salt exposure, thus allowing the cells to maintain their driving force for potassium uptake. Our data provide quantitative evidence that specific features of ion channels lead to superior sodium/potassium homeostasis in a halophyte compared with a closely related glycophyte.     

Stress Tolerance Parameters in Different Genotypes of Soybean

Free proline content, superoxide-dismutase activity, and lipid peroxidation were measured in sixteen Yugoslav and introduced genotypes of soybean. More tolerant genotypes with higher free proline content and high superoxide-dismutase activity, and low lipid peroxidation were chosen. The selected genotypes could be used in field production, as well as in breeding.     

Morphological and biochemical changes in maize under drought and salinity stresses in a semi-arid environment

Abstract

This study investigated some morphological and biochemical responses of maize to drought and salinity in open field in Tunisia with the aim of gaining insights into tolerance mechanisms. After seedling emergence, five treatments were applied until maturity: optimal water supply (control, 100% of maximum evapotranspiration - ETM); irrigation at 70% ETM (moderate drought) and at 35% ETM (severe drought); optimal supply of water containing 3?g NaCl L^?1 (moderate salinity) and 6?g NaCl L^?1 (severe salinity). Here, we demonstrated that extreme drought and salinity severely decreased the leaf area (?74% and ?55%, respectively) and the above-ground biomass (?35% and ?31%, respectively) at silking stage, indicating that the photosynthetic leaf apparatus is highly sensitive and that drought has a greater effect than salinity. Grain yield losses were also exacerbated under extreme stress conditions, viz. severe drought (?85% versus controls) and severe salinity (?73%), while productivity under moderate salinity approximated that of moderate drought, possibly due to increases in leaf chlorophyll and carotenoid content and K/Na ratio. The leaf area and its relative water content were positively correlated with grain yield under both salinity and drought stresses, and may therefore be used as markers for effective screening of maize genotypes for better stress tolerance.     

Cadmium effects on growth and antioxidant enzymes activities in Miscanthus sinensis

Plants of Miscanthus sinensis (cv. Giganteus) were grown in hydroponics for three months in nutrient solution with 0, 2.2, 4.4 and 6.6 μM CdNO₃. Growth parameters, catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and superoxide dismutase (SOD) activities were analysed in leaves and roots collected after 1-and 3-month exposure. Dry biomass of all miscanthus organs was affected by Cd concentration both after 1-and 3-month exposure. No visible symptoms of Cd toxicity were observed in shoots and rhizomes of plants grown in presence of Cd. In contrast, roots became shorter and thicker and the whole root system more dense and compact already after one month of treatment with 6.6 μM Cd. The lower Cd concentration increased the enzymes activities after 3 months in leaves and only after 1-month in roots, while a decrease in activity was observed at higher Cd concentrations.     

The effect of salinity on lipid composition and on activity of Ca2+- and Mg2+-stimulated ATPases in salt-sensitive and salt-tolerant Plantago species

Lipid composition of the roots and the shoot of the salt-sensitive Plantago media L., the salt-tolerant P. maritima L. and the less salt-tolerant P. coronopus L. was followed under saline conditions. In the roots of P. media the level of phospho-, galacto- and sulpholipids decreased strongly with increased NaCl concentration, indicating decreased control of permeability of the root cell membranes. In the roots of the two salt-tolerant species the level of most lipid classes was maintained or even raised up to 75 mM NaCl, and a decrease was noted only at higher NaCl concentrations. In P. maritima, a species from relatively nutrient-rich habitats, decreased lipid levels in the roots and shoot were observed with increasing salinity in combination with a low nutrient availability. The Ca²⁺- and Mg²⁺-stimulated ATPase activities of the microsomal fraction of the roots of P. maritima was decreased at a salinity level in excess of 150 mM, while in P. coronopus they were decreased at all NaCl levels tested. The obtained results are discussed as part of the adaptation of the species to salinity.     

Heat shock responses of bean plants: involvement of free radicals, antioxidants and free radical/active oxygen scavenging systems

In non-acclimated bean plants heat shock induced oxidative damage (increase of free radical concentration and drop of bound thiols, indicating aggregation of proteins) which was regulated by the enhanced activities of peroxidase and superoxide dismutase, as well as by the accumulation of polyphenols and especially of polyamines. In the plants acclimated to high temperature no oxidative damage occurred following heat shock. This revised version was published online in July 2006 with corrections to the Cover Date.     

盐胁迫对盐芥生长及硝酸还原酶活性的影响

盐胁迫处理导致盐芥植株鲜重、干重、含水量、肉质化程度和根冠比都下降;根中有机物含量上升,而无机物含量下降,叶的变化与根的相反;渗透调节能力、Na 含量和根系活力上升;硝酸还原酶活性显著增加;超氧阴离子(O2-)含量先降低后升高.表面扫描电镜图像显示:盐芥叶片表面没有盐腺或盐囊泡,所以它不是泌盐盐生植物.盐芥生长状况、Na 含量和Na X-ray微区分析结果表明:盐芥也不是拒盐盐生植物,而很可能是稀盐盐生植物.     

Interactions between osmoregulation and the alternative respiratory pathway in Plantago coronopus as affected by salinity

Plantago coronopus L., a species from the coastal zone, was grown in culture solution with and without 50 mM NaCl. In addition it was transferred from a non-saline solution to a solution containing 50 mM NaCl. Short term effects of NaCl on growth and various aspects of energy metabolism, including photosynthesis, shoot dark respiration, root respiration and the contribution of the SHAM-sensitive alternative pathway to root respiration were investigated. The concentrations of soluble and insoluble non-structural carbohydrates and of sorbitol a compatible osmotic solute in Plantago, in both shoots and roots were also determined. Growth of shoots and roots was largely unaffected by addition of 50 mM NaCl. Net photosynthesis, shoot dark respiration and the concentration of non-structural carbohydrates in both shoots and roots were also unaffected by salinity. The rate of root respiration immediately decreased upon addition of 50 mM NaCl. This decrease was almost exclusively attributed to a decreased activity of the SHAM-sensitive alternative pathway. The concentration of sorbitol in the roots increased quickly after addition of 50 mM NaCl, whilst the increase in sorbitol concentration in the shoots started later. The time course of the increase of sorbitol concentration was similar to that of the decrease in activity of the alternative pathway. During the first 12 h after exposure to 50 mM NaCl, the amount of carbohydrates which was saved in respiration, due to the decreased activity of the alternative pathway, was the same as that used for sorbitol synthesis in the roots. It is concluded that the activity of the alternative pathway decreased due to increased utilization of carbohydrates for sorbitol synthesis, according to a proposed ‘energy overflow model’. After 24 h, the sorbitol concentration in the cytoplasm of the root cells of plants transferred to a saline solution reached a level that was sufficient to compensate for 50 mM NaCl, assuming a cytoplasmic volume of ca. 10% of the total cell volume. The sorbitol concentration in roots of plants grown in a saline environment for several weeks was lower than that in roots of plants transferred to a saline environment for c. 24 h. It is suggested that sorbitol accumulated in roots of Plantago coronopus as an immediate reaction upon salinity, whilst other adaptations may occur thereafter.     

Is proline involved in stomata regulation of Commelina communis plants recovering from salinity stress?

Plants of Commelina communis L. were grown in culture solution to which NaCl was added for 48 h. The solutions were then replaced with normal medium, so that the plants could recover from the stress. The water potential increased almost to that of the controls during 4 h of recovery, but stomatal resistance stayed high. Cytokinin treatment of leaf discs failed to enhance recovery of stomatal aperture, although it enhanced stomatal recovery of identically treated epidermal tissue. Proline levels in leaves correlated well with stomatal resistance. Incubation of epidermal tissue in D-proline inhibited stomatal opening. NaCl and benzyladenine interacted with the effect of proline, and the effect of abscisic acid and was additive to that of proline.     

Proline and glycine betaine accumulation in two succulent halophytes under natural and experimental conditions

Proline (Pro) and glycine betaine (GB) contents were determined in two Mediterranean halophytes, Plantago crassifolia and Inula crithmoides, to assess their possible role in salt tolerance of both taxa. Plant material was collected in a littoral salt marsh under different environmental conditions, and from plants subjected to salt treatments in a growth chamber. Relative growth inhibition by NaCl indicated that I. crithmoides is more salt-tolerant than P. crassifolia, in agreement with the distribution of the two species in nature. Field and laboratory data confirmed GB as the major osmolyte responsible for osmotic adjustment in I. crithmoides, but with only a minor role – if any – as “osmoprotectant” in the salt tolerance of P. crassifolia. Under natural conditions, Pro contents were very low in both taxa, but increased to levels high enough to contribute significantly to osmotic balance when plants were artificially treated with 450–600 mM NaCl – higher salt concentrations than those they would normally encounter in their natural habitats. These data suggest that halophytes possess built-in mechanisms, such as accumulation of additional osmolytes, to rapidly adapt to increasing salinity levels in their natural ecosystems; for example, those expected to be caused by climate change in salt marshes in the Mediterranean region.     

Genetically modified cyanobacteriumNostoc muscorum overproducing proline in response to salinity and osmotic stresses

In the parentNostoc muscorum an active proline oxidase enzyme is required to assimilate exogenous proline as a fixed nitrogen source. Cyanobacterial mutants, resistant to growth inhibitory action of proline analogue L-azetidine-2-carboxylate (Ac-R), were deficient in proline oxidase activity, and were over-accumulators of proline. Proline over-accumulation, resulting either from mutational acquisition of the Ac-R phenotype, or from salinity-induced uptake of exogenous proline, confirmed enhanced salinity/osmotic tolerance in the mutant strain. The nitrogenase activity and photosynthetic O₂ evolution of the parent were sensitive to both salinity as well as osmotic stresses than of Ac-R mutant strain. In addition, the mutation to Ac-resistant phenotype showed no alteration in salinity inducible potassium transport system in the cyanobacterium.     

Effect of lead exposure on liver lipid peroxidative and antioxidant defense systems of protein-undernourished rats

The effect of the liver mitogen, lead nitrate [Pb(NO₃)₂], on protein-undernutrition-induced increased lipid peroxidation and reduced antioxidants levels was investigated in rats. Animals were divided into four groups: A, B, C, and D of five animals each. Animals in groups C and D were placed on a low-protein diet (5% casein) and animals in groups A and B were maintained on a normal diet (16% casein) for 14 wk and fed ad libitum. Animals in groups B and D were each given a single intravenous injection of Pb(NO₃)₂ (100 μmol/kg body weight) 72 h before sacrifice. The results confirm that protein undernutrition (PU) induced an increase in lipid peroxidation with concomitant reductions in catalase (CAT) activity, glutathione (GSH) level, and superoxide dismutase (SOD) activity. Lead (Pb) treatment, however, provoked increased lipid peroxidation, CAT activity, and GSH level but resulted in reduced SOD activity in both normal and PU-rats. These results suggest that Pb exacerbates liver lipid peroxidation in PU rats and suggests the involvement of free radicals in the pathogenesis of Pb poisoning. In addition, the results show that Pb affects well-fed and PU rats in similar ways but that the CAT activity of PU rats is more sensitive to the effect of Pb than that of normal rats.