Improved Tolerance of Three Saudi Pearl Millet Cultivars (<i>Pennisetum spicatum</i>) to Salt Stress by Mycorrhiza

Seeds of three Saudi pearl millet cultivars (Pennisetum spicatum) from three regions (Madinah, Khulais and Jaizan) were inoculated with arbuscular mycorrhizal fungus Glomus mosseae obtained from the Agriculture Research Center of Giza, Egypt to enhance their salt tolerance. Five different NaCl concentrations (0, 30, 60, 90, and 120 mM) were used for treating cultivars with and without mycorrhiza. Growth rates, chlorophyll content, chlorophyll fluorescence (Fv/Fm), proline content and gas exchange were measured to determine the effect of salinity on these cultivars. The results indicated that compared to cultivars without mycorrhiza, all cultivars with mycorrhiza had enhanced growth and physiological parameters including shoot and root length, area and number of leaves, fresh and dry weights of shoots and roots, chlorophyll contents and gas exchanges at 0 and 30 mM of salinity. In addition, the measurements of the different growth rates showed higher growth performance of the cultivars from Madinah and Khulais than the cultivar from Jaizan. However, all cultivars with and without mycorrhiza showed significant reductions in growth rates, chlorophyll contents and gas exchanges at a salinity of 60 mM than those grown at 0 and 30 mM. Moreover, the values of Fv/Fm were significantly reduced in all cultivars with and without mycorrhiza grown at 60 mM than in those grown at 0 mM and 30 mM. Proline contents indicated a progressive increase with the elevation of NaCl concentration stress. The proline contents in the mycorrhiza-inoculated cultivars were significantly higher than those in the non-inoculated cultivars. On the other hand, all cultivars with and without mycorrhiza underwent senescence within four weeks of growth at salinity concentrations of 90 mM and 120 mM. Therefore, relatively low salinity must be maintained to achieve high growth rates and gas exchanges of these inoculated cultivars.     

Anatomical changes induced by increasing NaCl salinity in three fodder shrubs, <Emphasis Type="Italic">Nitraria retusa</Emphasis>, <Emphasis Type="Italic">Atriplex halimus</Emphasis> and <Emphasis Type="Italic">Medicago arborea</Emphasis>

Nitraria retusa and Atriplex halimus (xero-halophytes) plants were grown in the range 0–800 mM NaCl while Medicago arborea (glycophyte) in 0–300 mM NaCl. Plants were harvested after 120 days of salt-treatment. The present study was designed to study the effect of salinity on root, stem and leaf anatomy, water relationship, and plant growth in greenhouse conditions. Salinity induced anatomical changes in the roots, stems and leaves. The cuticle and epidermis of N. retusa and A. halimus stems were unaffected by salinity. However, root anatomical parameters (root cross section area, cortex thickness and stele to root area ratio), and stem anatomical parameters (stem cross section area and cortex area) were promoted at 100–200 mM NaCl. Indicating that low to moderate salinity had a stimulating effect on root and stem growth of these xero-halophytic species. At higher salinities, root and stem structures were altered significantly, and their percentages of reduction were higher in A. halimus than in N. retusa whereas, in M. arborea, they were strongly altered as salinity rose. NaCl (100–300 mM) reduced leaf water content by 21.2–56.2% and specific leaf area by 51–88.1%, while increased leaf anatomical parameters in M. arborea (e.g. increased thickness of upper and lower epidermis, palisade and spongy mesophyll, entire lamina, and increased palisade to spongy mesophyll ratio). Similar results were evidenced in A. halimus leaves with salinity exceeding 100 mM NaCl. Leaves of N. retusa were thinner in salt-stressed plants while epidermis thickness and water content was unaffected by salinity. The size of xylem vessel was unchanged under salinity in the leaf’s main vein of the three species while we have increased number in M. arborea leaf main vein in the range of 200–300 mM NaCl. A longer distance between leaf vascular bundle, a reduced size and increased number of xylem vessel especially in stem than in root vascular system was evidenced in M. arborea treated plants and only at (400–800 mM) in the xero-halophytic species. The effects of NaCl toxicity on leaf, stem and root ultrastructure are discussed in relation to the degree of salt resistance of these three species. Our results suggest that both N. retusa and A. halimus show high tolerance to salinity while M. arborea was considered as a salt tolerant species.     

Differential Responses of <i>NHX1</i> and <i>SOS1</i> Gene Expressions to Salinity in two <i>Miscanthus sinensis</i> Anderss. Accessions with Different Salt Tolerance

The lignocellulosic crop Miscanthus spp. has been identified as a good candidate for biomass production. The responses of Miscanthus sinensis Anderss. to salinity were studied to satisfy the needs for high yields in marginal areas and to avoid competition with food production. The results indicated that the relative advantages of the tolerant accession over the sensitive one under saline conditions were associated with restricted Na⁺ accumulation in shoots. Seedlings of two accessions (salt-tolerant ‘JM0119’ and salt-sensitive ‘JM0099’) were subjected to 0 (control), 100, 200, and 300 mM NaCl stress to better understand the salt-induced biochemical responses of genes involved in Na⁺ accumulation in M. sinensis. The adaptation responses of genes encoding for Na⁺ /H⁺ antiporters, NHX1 and SOS1 to NaCl stress were examined in JM0119 and JM0099.The cDNA sequences of genes examined were highly conserved among the relatives of M. sinensis based on the sequencing on approximate 600 bp-long cDNA fragments obtained from degenerate PCR. These salt-induced variations of gene expression investigated by quantitative real-time PCR provided evidences for insights of the molecular mechanisms of salt tolerance in M. sinensis. The expression of NHX1 was up-regulated by salt stress in JM0119 shoot and root tissues. However, it was hardly affected in JM0099 shoot tissue except for a significant increase at the 100 mM salt treatment, and it was salt-suppressed in the JM0099 root tissue. In the root tissue, the expression of SOS1 was induced by the high salt treatment in JM0119 but repressed by all salt treatments in JM0099. Thus, the remarkably higher expression of NHX1 and SOS1 were associated with the resistance to Na⁺ toxicity by regulation of the Na⁺ influx, efflux, and sequestration under different salt conditions.     

Effect of salinity on osmotic adjustment characteristics of <Emphasis Type="Italic">Kandelia candel</Emphasis>

To elucidate the osmotic adjustment characteristics of mangrove plants, inorganic ion and organic solute contents of intermediate leaves were investigated in 3-month-old Kandelia candel (L.) Druce seedlings during 45 days of NaCl treatments (0, 200, and 500 mM NaCl). The contents of Na⁺, Cl⁻, total free amino acids, proline, total soluble sugars, pinitol and mannitol increased to different degree by salinity, whereas, K⁺ content decreased by salinity compared with control. NaCl treatment induced an increase of inorganic ion contribution while a decrease of organic solute contribution. It was concluded that accumulating a large amount of inorganic ions was used as the main osmotic adjustment mechanism under salinity treatment. However, accumulation of organic osmolytes might be considered to play much more important role in osmoregulation under severe salinity (500 mM NaCl) than under moderate salinity (200 mM NaCl), thus the damage caused by high toxic ions (Na⁺ and Cl⁻) concentration in K. candel leaves could be avoided.     

Photosystem II photochemistry and physiological parameters of three fodder shrubs, <Emphasis Type="Italic">Nitraria retusa</Emphasis>, <Emphasis Type="Italic">Atriplex halimus</Emphasis> and <Emphasis Type="Italic">Medicago arborea</Emphasis> under salt stress

Nitraria retusa and Atriplex halimus (xero-halophytes) plants were grown in the range 0–800 mM NaCl while Medicago arborea (glycophyte) in 0–300 mM NaCl. Salt stress caused a marked decrease in osmotic potential and a significant accumulation of Na⁺ and Cl⁻ in leaves of both species. Moderate salinity had a stimulating effect on growth rate, net CO₂ assimilation, transpiration and stomatal conductance for the xero-halophytic species. At higher salinities, these physiological parameters decreased significantly, and their percentages of reduction were higher in A. halimus than in N. retusa whereas, in M. arborea they decreased linearly with salinity. Nitraria retusa PSII photochemistry and carotenoid content were unaffected by salinity, but a reduction in chlorophyll content was observed at 800 mM NaCl. Similar results were found in A. halimus, but with a decrease in the efficiency of PSII (F′v/F′m) occurred at 800 mM. Conversely, in M. arborea plants we observed a significant reduction in pigment concentrations and chlorophyll fluorescence parameters. The marked toxic effect of Na⁺ and/or Cl⁻ observed in M. arborea indicates that salt damage effect could be attributed to ions’ toxicity, and that the reduction in photosynthesis is most probably due to damages in the photosynthetic apparatus rather than factors affecting stomatal closure. For the two halophyte species, it appears that there is occurrence of co-limitation of photosynthesis by stomatal and non-stomatal factors. Our results suggest that both N. retusa and A. halimus show high tolerance to both high salinity and photoinhibition while M. arborea was considered as a slightly salt tolerant species.     

Contractile roots are the most sensitive organ in <Emphasis Type="Italic">Crocus sativus</Emphasis> to salt stress

Crocus sativus corms were grown in Perlite and watered by half-strength modified Hoagland nutrient solution containing 0, 50, 100, 150, 200 mM NaCl. Growth parameters and contents of proteins, proline, polyphenols, minerals and saccharides were studied in fibrous roots, contractile roots, corms and leaves. All plants remained alive and did not display any sign of foliar damage even at 200 mM NaCl. However, the salinity decreased growth, relative water content and increased contents of proline and Na⁺ in all organs. Total protein content was increased in corms and contractile roots but decreased in fibrous roots. Changes in protein pattern were also observed. Polyphenol content was increased by salinity in all organs except the leaves. As salinity increased, content of soluble saccharides decreased except in the contractile roots.     

Differences in efficient metabolite management and nutrient metabolic regulation between wild and cultivated barley grown at high salinity

Physiological and biochemical responses of Hordeum maritimum and H. vulgare to salt stress were studied over a 60‐h period. Growth at increasing salinity levels (0, 100, 200 and 300 mM NaCl) was assessed in hydroponic culture. H. maritimum was shown to be a true halophyte via its typical behaviour at high salinity. Shoot growth of cultivated barley was gradually reduced with increasing salinity, whereas that of wild barley was enhanced at 100 and 200 mm NaCl then slightly reduced at 300 mM NaCl. The higher salt tolerance of H. maritimum as compared to H. vulgare was due to its higher capacity to maintain cell turgor under severe salinity. Furthermore, H. maritimum exhibited fine regulation of Na⁺ transport from roots to shoots and, unlike H. vulgare, it accumulated less Na⁺ in shoots than in roots. In addition, H. maritimum can accumulate more Na⁺ than K⁺ in both roots and shoots without the appearance of toxicity symptoms, indicating that Na⁺ was well compartmentalized within cells and substituted K⁺ in osmotic adjustment. The higher degree of salt tolerance of H. maritimum is further demonstrated by its economic strategy: at moderate salt treatment (100 mm NaCl), it used inorganic solutes (such as Na⁺) for osmotic adjustment and kept organic solutes and a large part of the K⁺ for metabolic activities. Indeed, K⁺ use efficiency in H. maritimum was about twofold that in H. vulgare; the former started to use organic solutes as osmotica only at high salinity (200 and 300 mm NaCl). These results suggest that the differences in salt tolerance between H. maritimum and H. vulgare are partly due to (i) differences in control of Na⁺ transport from roots to shoots, and (ii) H. maritimum uses Na⁺ as an osmoticum instead of K⁺ and organic solutes. These factors are differently reflected in growth.     

<i>In vitro</i> Evaluation of Seed Germination in Twelve Alfalfa Cultivars under Salt Stress

Alfalfa (Medicago sativa L.), when exposed to abiotic stress such as salinity, suffers significant losses in yield and productivity. The present study evaluated the salinity tolerance of 12 alfalfa cultivars in vitro using five concentrations of sodium chloride (NaCl), ranging from 0 to 250 mmol L⁻¹ . The results obtained in the current study revealed that the Saudi cultivars, Kasimi and Hassawi, and the German cultivar (Berlin) had the highest salinity tolerance in terms of germination percentage (GP), corrected germination rate index (CGRI), days to reach 50% germination (GT50), and ability to form cotyledonary and true leaves. Under mmol L⁻¹ NaCl, the Saudi cultivar Kasimi cultivar showed GP, CGRI, and GT50 of 55.20%, 123.15, and 3.77 days, respectively. Similarly, the German cultivar (Berlin) showed GP, CGRI, and GT50 of 50.06%, 86.61, and 5.17 days, respectively. These findings might reveal a pivotal aspect in salt tolerance in alfalfa. Our results will help to select salt-tolerant alfalfa cultivars that could thrive in arid and semi-arid areas with salinity problems.     

Photosynthetic Behaviour and Mineral Nutrition of <i>Tamarix gallica</i> Cultivated Under Aluminum and NaCl Combined Stress

The lack of knowledge of plant tolerance and differential response to aluminum (Al) encouraged many researchers, in the last decade, to elucidate Al toxicity and tolerance mechanisms. The current study reported the impact of Al, a toxic element with negative effects on plant growth and development, in halophytic plant Tamarix gallica. Plants were subjected to different Al concentrations (0, 200, 500 and 800 μM) with or without NaCl (200 mM) supplementation. Growth, photosynthesis and mineral content were assessed. Al stress had a significant decrease on shoots’ biomass production between 19 to 41%, and a little variation on chlorophyll content and photosynthetic efficiency (Fo, Fm, Fv fluorescence’s and Fv/Fm). Furthermore, the Al-treatments did not affect significantly the content of potassium, calcium, and magnesium in different plant parts, whereas NaCl addition to the medium induced a decrease in these elements’ concentrations. Our results have shown that T. gallica is able to accumulate the high levels of Al in shoots and roots, 6288 μg.g^-1 DW and 7834 μg.g^-1 DW respectively. It is considered as a hyperaccumulator plant of Al. In addition, Na⁺ contents in shoots and roots exceed 23000 μg.g^-1 DW. Therefore, T. gallica presents a high tolerance at the same time to Al and NaCl phytotoxicity, so it is interesting to use in phytoremediation programs.     

Carbohydrate and Proline Contents in Leaves, Roots, and Apices of Salt-Tolerant and Salt-Sensitive Wheat Cultivars1

Intra-specific variations in nonstructural carbohydrates and free proline were determined in leaves, apices, roots, and maturing seeds of two salt-tolerant cultivars (CR and Kharchia-65) and one salt-sensitive cv. Ghods of spring wheat (Triticum aestivum L.) grown in sand culture at various levels of salinity (0, 100, 200, and 300 mM NaCl and CaCl₂ at 5 : 1 molar ratio) under controlled environmental conditions. The levels of leaf, apex, and root ethanol-soluble carbohydrates, fructans, starch, and proline increased in line with elevating level of salinity in all three cultivars under investigation. The contents of proline, soluble and insoluble carbohydrates in the apex increased to levels exceeding those in the leaves and roots. Soluble carbohydrate content of salt-sensitive cv. Ghods was higher in the leaves, apices, and roots and lower in the maturing seeds than in the other cultivars at all levels of salinity except at 300 mM. The results show considerable variation in the amount of soluble, insoluble sugars, and proline among plant tissues and wheat genotypes in response to salinity. Higher soluble carbohydrates and fructan in leaves, roots and maturing seeds of stressed plants indicate that their accumulation may help plant to tolerate salinity. Salt-sensitive cv. Ghods accumulated less soluble sugars in the maturing seeds and higher soluble sugars in the apices, which might be used as an indicator in screening wheat genotypes for salinity tolerance.     

Biochemical responses of Hyacinth bean (<Emphasis Type="Italic">Lablab purpureus)</Emphasis> to salinity stress

Effect of salinity on Hyacinth bean, Lablab purpureus (HA-4 cultivar) was evaluated in 10-day old seedlings with 100–500 mM NaCl over 72 h of exposure. The stress reduced dry and fresh weight, leaf surface area, root and shoot length, total chlorophyll, and RWC. Oxidative stress markers, H₂O₂, glutathione, TBARS, proline, ascorbic acid, total phenols, and total soluble sugar contents were significantly elevated. Salinity enhanced antioxidant enzymes, POX, and GR activities and reduced that of CAT in concentration and time dependent manner in leaves. Antioxidant enzymes in roots showed inverse relationship with concentration and time of exposure. Metabolic enzyme β-amylase activity increased in both leaves and roots. Acid phosphatase decreased in leaves and elevated in roots. Intensity of constitutive isozymes correlated with in vitro levels under stress, but the protein band patterns differed from controls. Lablab showed reasonable tolerance up to 300 mM NaCl, but leaves and roots differed in their response.     

Effects of Different Arbuscular Mycorrhizal Fungi on Physiology of <i>Viola prionantha</i> under Salt Stress

Arbuscular mycorrhizal (AM) fungi distribute widely in natural habits and play a variety of ecological functions. In order to test the physiological response to salt stress mediated by different AM fungi, Viola prionantha was selected as the host, the dominant AM fungus in the rhizosphere of V. philippica growing in Songnen saline-alkali grassland, Rhizophagus irregularis, and their mixtures were used as inoculants, and NaCl stress was applied after the roots were colonized. The results showed that V. philippica could be colonized by AM fungi in the field and the colonization rate ranged from 73.33% to 96.67%, and Claroideoglomus etunicatum was identified as the dominant AM fungi species in the rhizosphere of V. philippica by morphology combined with sequencing for AM fungal AML1/AML2 target. Inoculation with both the species resulted in the formation of mycorrhizal symbiosis (the colonization rate was more than 70%) and AM fungi significantly enhanced plants’ tolerance to salt stress of varying magnitude. Higher activity of antioxidant enzymes and augmented levels of proline and other osmoregulators were observed in AM plants. The content of MDA in CK was higher than that in the inoculations with the stress of 100, 200, and 250 mM. All indices except soluble protein content and MDA content were significantly correlated with AM fungal colonization indices. The analysis for different AM fungal effects showed that the mixtures and R. irregularis worked even better than C. etunicatum. These results will provide theoretical support for the exploration and screening of salt-tolerant AM fungi species and also for the application of AM-ornamental plants in saline-alkali urban greening.     

Exogenous Glycine Betaine Reduces Drought Damage by Mediating Osmotic Adjustment and Enhancing Antioxidant Defense in <i>Phoebe hunanensis</i>

Drought stress negatively impacts growth and physiological processes in plants. The foliar application of glycine betaine (GB) is an effective and low-cost approach to improve the drought tolerance of trees. This study examined the effect of exogenously applied GB on the cell membrane permeability, osmotic adjustment, and antioxidant enzyme activities of Phoebe hunanensis Hand.-Mazz under drought stress. Two levels (0 and 800 mL) of water irrigation were tested under different applied GB concentrations (0, 50, 100, and 200 mM). Drought stress decreased the relative water content by 58.5% while increased the electric conductivity, malondialdehyde, proline, soluble proteins, soluble sugars, and antioxidant enzyme activities (superoxide dismutase, catalase, peroxidase) by up to 62.9%, 42.4%, 87.0%, 19.1%, 60.5%, 68.3%, 71.7%, and 83.8%, respectively, on the 25^th day. The foliar application of GB, especially at 100 mM, increased the relative water content of P. hunanensis leaves under drought stress. The concentration of GB from 50 to 100 mM effectively alleviated the improvement of cell membrane permeability and inhibited the accumulation of membrane lipid peroxidation products. Under drought stress, the concentrations of proline, soluble proteins, and soluble sugars in the leaves of P. hunanensis increased as the applied GB concentration was increased and the water stress time was prolonged. Exogenously applied GB decreased oxidative stress and improved antioxidant enzyme activities as compared with treatments without GB application. Furthermore, the physiological and biochemical indexes of P. hunanensis showed a certain dose effect on exogenous GB concentration. These results suggest that GB helps maintain the drought tolerance of P. hunanensis.     

Contribution of inorganic cations and organic compounds to osmotic adjustment in root cultures of two <Emphasis Type="Italic">Centaurium</Emphasis> species differing in tolerance to salt stress

The effect of reduced availability of sugars on growth and essential metabolic processes in roots, resulting from decreased photosynthesis under salinity, was excluded by establishing a non-photosynthetic model-system in this study: root cultures of Centaurium maritimum (L.) Fritch and Centaurium spicatum (L.) Fritch. The contribution of inorganic cations and organic compounds (e.g. carbohydrates and amino acids) to the osmotic adjustment (OA) in roots during short-term exposure to various salt concentrations (0, 50, 100 or 200 mM NaCl) was emphasized. Observed morphological and histological changes in roots were species specific, and were dependent on salinity level. Although C. spicatum appears to be more tolerant to salt stress, both species employed similar strategies in response to elevated salinity to different extents, and displayed effective OA mechanisms. Under low and moderate salinity, inorganic cations were the major contributors to OA in roots of both species, followed by soluble sugars, while the relative contribution of proline (Pro) and free amino acids was insignificant. Osmotic adjustment under severe stress appears to be mediated by increased accumulation of organic compounds. The analysis of the intraspecies variability in salt response of C. spicatum and C. maritimum roots enabled the identification of some organic compounds which could be used as potential biochemical markers in screening for salt tolerance, including Pro in C. spicatum, and trehalose and polyols in C. maritimum.     

Germination and Photosynthetic Responses to Salinity and Alkalinity in <i>Avicenna marina</i> Propagules

Avicenna marina (Forssk.) Vierh is a halophytic mangrove. The reproductive unit is green and has photosynthetic propagules. Mangroves are naturally exposed to fluctuations in some abiotic factors at the soil surface, including salinity and alkalinity. The objective of this study was to determine the effects of two salts including NaCl and NaHCO₃ on germination processes and discuss the relationships between cotyledon photosynthesis and embryo axis growth in A. marina propagules. These propagules came from Al Birk, located on the shoreline of the Saudi Red Sea. The results showed that the studied salts did not affect neither the final germination percentage nor the embryo axis growth. However, rooting and root growth were delayed by both salts at 300 mM and were strongly inhibited by 600 mM NaHCO₃. Both NaCl and NaHCO₃ reduced the photosynthetic activity. These two salts did not affect the other photosynthetic parameters, including stomatal conductance, net transpiration, and intercellular CO₂. Thereafter, the reduction in net photosynthesis was not related to any limitation of stomatal conductance. The early germination phase was independent of cotyledon photosynthesis, whereas rooting and root growth may be limited by reduced photosynthesis under NaCl and NaHCO₃.     

Effects of Barium Stress in <i>Brassica juncea</i> and <i>Cakile maritima</i>: The Indicator Role of Some Antioxidant Enzymes and Secondary Metabolites

Soil contamination by toxic trace metal elements, like barium (Ba), may stimulate various undesirable changes in the metabolic activity of plants. The plant responses are fast and with, direct or indirect, generation of reactive oxygen species (ROS). To cope with the stress imposed by the ROS production, plants developed a dual cellular system composed of enzymatic and non-enzymatic players that convert ROS, and their by-products, into stable nontoxic molecules. To assess the Ba stress response of two Brassicaceae species (Brassica juncea, a glycophyte, and Cakile maritime, a halophyte), plants were exposure to different Ba concentrations (0, 100, 200, 300 and 500 μM). The plants response was evaluated through their morphology and development, the determination of plant leaves antioxidant enzymatic activities and by the production of plants secondary metabolites. Results indicated that the two Brassicaceae species have the ability to survive in an environment containing Ba (even at 500 μM). The biomass production of C. maritima was slightly affected whereas an increase in biomass B. juncea was noticed. The stress imposed by Ba activated the antioxidant defense system in the two species, noticed by the changes in the leaves activity of catalase (CAT), ascorbate peroxidase (APX) and guaicol peroxidase (GPX), and of the secondary metabolites, through the production of total phenols and flavonoids. The enzymatic response was not similar within the two plant species: CAT and APX seem to have a more important role against the oxidative stress in C. maritima while in B. juncea is GPX. Overall, total phenols and flavonoids production was more significant in the plants aerial part than in the roots, of the both species. Although the two Brassicaceae species response was different, in both plants catalytic and non-catalytic transformation of ROS occurs, and both were able to overcome the Ba toxicity and prevent the cell damage.     

Regulation of gene expression governing proline metabolism in <Emphasis Type="Italic">Thellungiella salsuginea</Emphasis> by NaCl and paraquat

Differential expression of the proline metabolism genes in Thellungiella salsuginea (Pall) E. Schulz was investigated under salinity (100 and 300 mM NaCl), upon the effect of paraquat (0.1 μM), and at their joint action. It was shown that, depending on the intensity of stress factor, expression of the P5CS1 gene was induced in the leaves (at 100 mM NaCl) or roots (at 300 mM NaCl). When the plants on control medium were treated with paraquat, the proline content changed only in the leaves. Time course of proline content in the leaves complied with the dynamic of P5CS1 gene expression, while expression of PDH gene essentially did not change. When the plants, which experienced salt stress, were treated with paraquat, the content of proline and the P5CS1 mRNA level increased even more. The obtained results suggest a complicated nature of signaling between the organs of the halophyte Th. salsuginea causing expression of the proline biosynthesis genes in the leaves and roots under the effect of salinity, paraquat, or upon their joint action. The proline catabolism in these plants was maintained essentially unchanged, which is probably related to the participation of proline and/or the products of its degradation in the pathways of other metabolite biosynthesis. We suggested that proline took part in ROS scavenging process and proline level was under strong control in Th. salsuginea.     

Effect of salt stress on growth parameters, enzymatic antioxidant system, and lipid peroxidation in wild chicory (Cichorium intybus L.)

Efficient utilization of saline land for food cultivation can increase agricultural productivity and rural income. To obtain information on the salt tolerance/susceptibility of wild chicory (Cichorium intybus L.), the influence of salinity (0–260 mM NaCl) on chicory seed germination and that of two salinity levels of irrigation water (100 and 200 mM NaCl) on plant growth, antioxidative enzyme activity, and accumulation of proline and malondialdehyde (MDA) were investigated. The trials were performed outdoors, in pots placed under a protective glass covering, for two consecutive years. Seeds showed a high capacity to germinate in saline conditions. The use of 100 mM NaCl solution resulted in 81 % germination, whereas seed germinability decreased below 40 % using salt concentrations above 200 mM NaCl. Wild chicory showed tolerance to medium salinity (100 mM NaCl), whereas a drastic reduction in biomass was observed when 200 mM NaCl solution was used for irrigation. MDA, present in higher amounts in leaves than in roots, decreased in both tissues under increasing salinity. Proline content increased remarkably with the level of salt stress, more so in roots than in leaves. In salt stress conditions, the activity of antioxidant enzymes (APX, CAT, POD, SOD) was enhanced. The electrophoretic patterns of the studied enzymes showed that the salinity of irrigation water affected only the intensity of bands, but did not activate new isoforms. Our results suggest that wild chicory is able to grow in soil with moderate salinity by activating antioxidative responses both in roots and leaves.     

Expression of a carbonic anhydrase gene is induced by environmental stresses in Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Expression of the gene (OsCA1) coding for carbonic anhydrase (CA) in leaves and roots of rice was induced by environmental stresses from salts (NaCl, NaHCO₃ and Na₂CO₃), and osmotic stress (10%, w/v, PEG 6000). CA activity of rice seedlings more than doubled under some of these stresses. Transgenic Arabidopsis over-expressing OsCA1 had a greater salt tolerance at the seedling stage than wild-type plants in 1/2 MS medium with 5 mM NaHCO₃, 50 mM NaCl, on 100 mM NaCl. Thus CA expression responds to environmental stresses and is related to stress tolerance in rice.     

Salinity stress response of non-transformed and AtCKX transgenic centaury (Centaurium erythraea Rafn.) shoots and roots grown in vitro

Common centaury (Centaurium erythraea Rafn.) is a plant species that can inhabit saline soils. It is known as a plant with high spontaneous regeneration potential in vitro. In the present work we evaluated shoots and roots salinity tolerance of non-transformed and three AtCKX transgenic centaury lines to graded NaCl concentrations (0, 50, 100, 150, 200 mM) in vitro. Overexpression of AtCKX genes in transgenic centaury plants resulted in an altered cytokinins (CKs) profile leading to a decline of bioactive CK levels and, at the same time, increased contents of storage CK forms, inactive CK forms and/or CK nucleotides. Significant increment of fresh shoot weight was obtained in shoots of non-transformed and AtCKX1 transgenic line only on medium supplemented with 50 mM NaCl. However two analysed AtCKX2 transgenic lines reduced shoot growth at all NaCl concentrations. In general, centaury roots showed higher tolerance to salinity than shoots. Non-transformed and AtCKX1 transgenic lines tolerated up to 100 mM NaCl without change in frequency of regeneration and number of regenerated plants. Roots of two analysed AtCKX2 transgenic lines showed different regeneration potential under salt stress. Regeneration of transgenic AtCKX2-26 shoots even at 200 mM NaCl was recorded. Salinity stress response of centaury shoots and roots was also evaluated at biochemical level. Free proline, malondialdehyde and hydrogen peroxide content as well as antioxidative enzymes activities were investigated in shoots and roots after 1, 2, 4 and 8 weeks. In general, adition of NaCl in culture medium elevated all biochemical parameters in centaury shoots and in roots. Considering that all analysed AtCKX transgenic centaury lines showed altered salt tolerance to graded NaCl concentrations in vitro it can be assumed that CKs might be involved in plant defence to salt stress conditions.