The effects of NaCl on growth,water relations,osmolytes and ion content in <Emphasis Type="Italic">Kochia prostrata</Emphasis>

The effects of NaCl (0, 50, 100, 150 and 200 mM) on growth, water relations, glycinebetaine, free proline, ion contents, stomata number and size of Kochia prostrata (L.) Schard were determined. Shoot and root fresh and dry matter, root and shoot length, relative growth rate, net assimilation rate, relative water content, water use efficiency, soluble sugars and glycinebetaine contents were not changed at low NaCl concentrations, but they were significantly decreased at 200 mM NaCl. The K⁺, Mg²⁺ and Ca²⁺ contents, water potential, chlorophyll a+b and carotenoides contents, and stomata number and size were reduced already at low concentrations of NaCl. In contrast, the Na⁺, Cl^– and proline contents increased several times with increasing NaCl concentration. Kochia prostrata is a salt tolerant species, the optimal growth of this plant occurred up to 150 mM NaCl. The mechanisms of salt tolerance in the plant may be balance among ion accumulation and production of glycinebetaine, proline, soluble sugars for maintenance of pressure potential.     

Identification of two phenotypes of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> under <Emphasis Type="Italic">in vitro</Emphasis> salt stress conditions

This study describes two phenotypes of Arabidopsis thaliana (ecotype Columbia) developed in vitro under salt stress (75 mM NaCl). The phenotypes 01 and 02 appeared visibly distinguishable by rosette morphology and competence to produce flowers. Phenotype 01, sensible to salt stress, accumulated high quantities of Na⁺, showed a slight reduction in dry mass, and high protein and chlorophyll contents. Moreover, its anatomy exhibited some xeromorphic traits. Phenotype 02, clearly salt tolerant, showed a morphology similar to control plants, displaying typical phyllotactic rosette and flowering stalk production. Accumulation of Na⁺, protein and chlorophyll contents were close to control plants. Reversion experiments on NaCl free MS medium, showed a partially recovered phenotype 01. A threshold salt stress concentration that permits the simultaneous development of two phenotypes, was found.     

A new <Emphasis Type="Italic">Em</Emphasis>-like protein from <Emphasis Type="Italic">Lactuca sativa</Emphasis>, <Emphasis Type="Italic">LsEm1</Emphasis>, enhances drought and salt stress tolerance in <Emphasis Type="Italic">Escherichia coli</Emphasis> and rice

Late embryogenesis abundant (LEA) proteins are closely related to abiotic stress tolerance of plants. In the present study, we identified a novel Em-like gene from lettuce, termed LsEm1, which could be classified into group 1 LEA proteins, and shared high homology with Cynara cardunculus Em protein. The LsEm1 protein contained three different 20-mer conserved elements (C-element, N-element, and M-element) in the C-termini, N-termini, and middle-region, respectively. The LsEm1 mRNAs were accumulated in all examined tissues during the flowering and mature stages, with a little accumulation in the roots and leaves during the seedling stage. Furthermore, the LsEm1 gene was also expressed in response to salt, dehydration, abscisic acid (ABA), and cold stresses in young seedlings. The LsEm1 protein could effectively reduce damage to the lactate dehydrogenase (LDH) and protect LDH activity under desiccation and salt treatments. The Escherichia coli cells overexpressing the LsEm1 gene showed a growth advantage over the control under drought and salt stresses. Moreover, LsEm1-overexpressing rice seeds were relatively sensitive to exogenously applied ABA, suggesting that the LsEm1 gene might depend on an ABA signaling pathway in response to environmental stresses. The transgenic rice plants overexpressing the LsEm1 gene showed higher tolerance to drought and salt stresses than did wild-type (WT) plants on the basis of the germination performances, higher survival rates, higher chlorophyll content, more accumulation of soluble sugar, lower relative electrolyte leakage, and higher superoxide dismutase activity under stress conditions. The LsEm1-overexpressing rice lines also showed less yield loss compared with WT rice under stress conditions. Furthermore, the LsEm1 gene had a positive effect on the expression of the OsCDPK9, OsCDPK13, OsCDPK15, OsCDPK25, and rab21 (rab16a) genes in transgenic rice under drought and salt stress conditions, implying that overexpression of these genes may be involved in the enhanced drought and salt tolerance of transgenic rice. Thus, this work paves the way for improvement in tolerance of crops by genetic engineering breeding.     

Hydrogen peroxide-induced salt tolerance in the <Emphasis Type="Italic">Arabidopsis</Emphasis> salicylate-deficient transformants <Emphasis Type="Italic">NahG</Emphasis>

The effect of hydrogen peroxide treatment on the salt tolerance of wild-type Arabidopsis thaliana L. plants (Col-0) and plants transformed with the bacterial salicylate hydroxylase gene (NahG) was studied. The base tolerance to salt stress caused by 200 mM of NaCl in solution culture was higher in plants with the NahG genotype in comparison with the wild-type plants. Growth inhibition was observed for wild-type plants under the action of exogenous hydrogen peroxide, which was not observed for the NahG transformants; salt tolerance increased in the both types of plants after treatment, which was assessed based on the growth indicators and the ability to preserve the chlorophyll pool following NaCl treatment. The content of endogenous Н2О2 in the leaves of wild-type plants increased significantly following exogenous hydrogen peroxide treatment and salt stress, while it practically did not change in the leaves of the NahG genotype. The SOD activity increased in both genotypes after treatment with exogenous hydrogen peroxide, and remained at an elevated level after salt stress in comparison with the nontreated plants. Furthermore, the catalase activity increased in leaves of the salicylate-deficient genotype but not in the Col-0 genotype. The guaiacol peroxidase activity increased in plants of both genotypes under the action of hydrogen peroxide and salt stress, with the NahG plants demonstrating a higher degree of increase. The Н₂О₂ treatment facilitated the increase of the proline content in leaves of the plants of both genotypes under conditions of salt stress. It was concluded that there were hydrogen peroxide signal transduction pathways in Arabidopsis plants that were salicylic acid independent and that the antioxidant system functioned more effectively in salicylate-deficient Arabidopsis plants.     

A <Emphasis Type="Italic">Vitis vinifera</Emphasis> xanthine dehydrogenase gene, <Emphasis Type="Italic">VvXDH</Emphasis>, enhances salinity tolerance in transgenic <Emphasis Type="Italic">Arabidopsis</Emphasis>

Xanthine dehydrogenase (EC1.1.1.204; XDH) plays an important role in purine catabolism that catalyzes the oxidative hydroxylation of hypoxanthine to xanthine and of xanthine to uric acid. Long attributed to its role in recycling and remobilization of nitrogen, recently, XDH is implicated in plant stress responses and acclimation, such research efforts, however, have thus far been restricted to Arabidopsis XDH-knockdown/knockout studies. This study, using an ectopic overexpression approach, is expected to provide novel findings. In this study, a XDH gene from Vitis vinifera, named VvXDH, was synthesized and overexpressed in Arabidopsis, the transgenic Arabidopsis showed enhanced salt tolerance. The VvXDH gene was investigated and the results demonstrated the explicit role of VvXDH in conferring salt stress by increasing allantoin accumulation and activating ABA signaling pathway, enhancing ROS scavenging in transgenic Arabidopsis. In addition, the water loss and chlorophyll content loss were reduced in transgenic plants; the transgenic plants showed higher proline level and lower MDA content than that of wild-type Arabidopsis, respectively. In conclusion, the VvXDH gene has the potential to be applied in increasing allantoin accumulation and enhancing the tolerance to abiotic stresses in Arabidopsis and other plants.     

Mutation induced by ethylmethanesulphonate (EMS), in vitro screening for salt tolerance and plant regeneration of sweet potato (<Emphasis Type="Italic">Ipomoea</Emphasis><Emphasis Type="Italic">batatas</Emphasis> L.)

Salt tolerant cultivars of sweet potato (Ipomoea batatas L.) can be obtained from induced mutation. The objective of the present study was to induce mutation for salt tolerance using ethylmethanesulphonate (EMS) in calli of sweet potato, followed by cell line selection and subsequent plant regeneration. Calli initiated from leaf explants were treated with 0.5% EMS for 0, 1, 1.5, 2, 2.5 and 3 h, followed by rinsing with sterile distilled water for four times. Preliminary experiments showed that 200 mM NaCl could be used as selection pressure. Salt tolerant calli were sub-cultured on medium supplemented with 200 mM NaCl for selection of mutant cell lines and this process repeated 5 times (20 days each). The selected calli were transferred onto somatic embryo formation medium, which was Murashige and Skoog (MS) medium supplemented with 4 mg l⁻¹ abscisic acid (ABA), 10 mg l⁻¹ gibberellic acid (GA). After 15 days, somatic embryos were transferred onto MS medium supplemented with 0.05 mg l⁻¹ ABA, 0.2 mg l⁻¹ zeatin (ZT) for regeneration. Plants designated as ML1, ML2 and ML3 were regenerated from the somatic embryos formed by calli treated with 0.5% EMS for 2 and 2.5 h. After propagation, salt tolerance of these mutants was investigated. Data suggested the mutants were more salt tolerant than control plants.     

Growth and antioxidant production of <Emphasis Type="Italic">Spirulina</Emphasis> in different NaCl concentrations

Objective

To evaluate the quantity of Spirulina cultured in seawater, salt-tolerant strains were screened out and their growth and antioxidant accumulation were studied in different salt concentrations

Results

Salt tolerance of five Spirulina strains were investigated with modified Zarrouk medium (with 200–800 mM NaCl). All strains grew well with 400 mM NaCl; their growth rates were almost same as in the control medium. Spirulina strains FACHB-843 (SP843) and FACHB-972 (SP972) had the highest salt tolerance their growth rates in 600 mM NaCl were nearly same as the control. Both strains produced more carotene, phycocyanin, polysaccharides, proline and betaine in 400–600 mM NaCl than the control. Salt stress also induced them to produce higher activities of superoxide dismutase and peroxidase. Total antioxidant capacities of SP843 and SP972 peaked at 600 and 400 mM NaCl, respectively.

Conclusion

Spirulina strains cultured with seawater accumulate more bioactive substances and will have a higher nutritive value.

     

NaCl induced morpho-biochemical and anatomical changes in mulberry (<Emphasis Type="Italic">Morus</Emphasis> spp.)

Mulberry is an economically important tree, used for feeding the silkworm Bombyx mori L. Effect of different levels of NaCl on growth and development of mulberry has been studied using five mulberry genotypes selected on the basis of their performance under in vitro salinity. The study while endorsing the efficacy of in vitro screening of axillary buds of mulberry for salt tolerance, showed genotypic variability in its response to salinity. Salinity reduced growth and development of all genotypes. However, the putative tolerant genotypes showed better performance than the putative susceptible genotypes. Under low salinity (<0.5% NaCl) salt tolerant genotypes showed an increase in chlorophyll and protein concentrations, while in susceptible genotypes both were reduced by 3–58% at 0.5% NaCl and 50–64% at 1.00% NaCl. Leaf thickness increased by 16% at 1.00% NaCl in C776 and reduced by 1.0% in Mandalaya. The increase in chlorophyll concentration and leaf thickness under high salinity can be considered as preliminary selection parameters for salt tolerance in mulberry. The study confirmed the efficacy of in vitro method for screening of large number of genotypes for salt tolerance in mulberry.     

Effects of nickel toxicity on morphological and physiological aspects of osmoregulation in <Emphasis Type="Italic">Typha domingensis</Emphasis> (<Emphasis Type="Italic">Typhaceae</Emphasis>) populations

The present work examined morphological and physiological aspects of osmoregulation in some differently adapted populations of Typha domingensis. Six populations of T. domingensis were grown in waterlogged conditions under four levels of nickel (0, 50, 100 and 1500 mg/kg of Ni). The Ni tolerance was explored on the basis of different characteristics of shoots. Along with the increasing level of Ni stress, all populations showed an increment of proline and glycinebetaine content, low photosynthetic and transpiration rates and decreased chlorophyll content. Analyzed morphological parameters revealed possible adaptations to deal with Ni stress. The less tolerant Sahianwala population produced more aerenchyma, less accumulation of osmolytes and reduced shoot fresh weight under Ni stress than the tolerant ones. Our results suggested that T. domingensis has the capacity to survive under Ni stress and could be used for the absorption of toxic metal ions from industrially polluted water.     

<Emphasis Type="Italic">Dunaliella salina</Emphasis> exopolysaccharides: a promising biostimulant for salt stress tolerance in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>)

Microalgal exopolysaccharides represent a potential sustainable alternative for the enhancement and protection of agricultural crops including management of both biotic and abiotic stress. In the present study, we investigated the potential of Dunaliella salina exopolysaccharides (PS) to attenuate the effect of salt stress on growth of Solanum lycopersicum, which was grown under different salinity levels (3 and 6 g L^?1 NaCl). The effects of PS treatment on plant growth, osmoprotectant molecules, protein content, and antioxidant enzymes activities of tomato plants under salt stress were analyzed. A metabolomics study showed that the exopolysaccharides released by D. salina contained sulfated moiety along with carbohydrates and uronic acids. The application of sulfated exopolysaccharides on tomato plants alleviated the salt stress and mitigated the decrease in length and dry weight of the plant’s shoot and root systems, as well as that of potassium (K⁺), and K⁺/Na⁺ ratio. Furthermore, the increase in proline, phenolic compounds, Na⁺, and antioxidant enzymes (CAT, POD, SOD) activities caused by salt stress were attenuated after the exopolysaccharide treatment. GC-MS metabolomics analysis showed that PS treatment allowed the activation and/or inhibition of various metabolic pathways involved in the plant’s tolerance to stress such as jasmonic acid-dependent pathways. This study shows the potential of microalgal exopolysaccharides for enhancing tomato tolerance to salt stress and highlights the possibility of their use as plant growth biostimulants under harsh environmental conditions.     

Effects of paclobutrazol on cultivars of Chinese bayberry (<Emphasis Type="Italic">Myrica rubra</Emphasis>) under salinity stress

Salt stress is one of the most critical factors hindering the growth and development of plants. Paclobutrazol (PBZ) is widely used to minimize this problem in agriculture because it can induce salt stress tolerance in plants. This study investigated the effects of PBZ on salt tolerance of seedlings from two Chinese bayberry cultivars (i.e., Wangdao and Shenhong). Plants were treated with three salt concentrations (0, 0.2, and 0.4 % NaCl) and two PBZ concentrations (0 and 2.0 μmol L^–1). Application of PBZ increased a relative water content, proline content, chlorophyll (a+b) content, and antioxidant enzyme activities in both cultivars, resulting in a better acclimation to salt stress and an increase in dry matter production. We concluded that PBZ ameliorated the negative effects of salt stress in Chinese bayberry seedlings.