Comparison of ultrastructural and physiological changes of potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) plantlets subjected to salt and modeling drought stresses

Salinity and drought are two severe abiotic stresses that affect plant growth and decrease food production worldwide. Compared to the field plants, test-tube plantlets could be more direct and fast to investigate the mechanism of stress adaptation. In the present study, the ultrastructural and physiological differences of potato (Solanum tuberosum L. c.v. “Longshu No. 3”) plantlets in response to the gradient saline (0, 25, 50, 100, and 200 mM NaCl) and the modeling drought stresses with polyethylene glycol (PEG) at the concentrations of 0, 2, 4, 6, and 8 % were analyzed. The results show that the severe salt (200 mM NaCl) and the modeling drought stresses (8 % PEG) inhibited the plantlet growth. There are considerable differences in their ultrastructural alteration under salt and modeling drought adaptation: PEG caused the increase in the number of stacked chloroplast, plastoglobuli, and starch; NaCl induced the decrease in the number of chloroplast and plastoglobuli. Moreover, plantlet has higher free proline content, less malondialdehyde (MDA) content, and higher activities of catalase (CAT) and superoxide dismutase (SOD) under the gradient NaCl treatments than the gradient PEG treatments. The results of this study will provide theoretical and practical insights into characterizing the ultrastructural and physiological differences of plants adapting to various stressful environments.     

Large-scale de novo transcriptome analysis reveals specific gene expression and novel simple sequence repeats markers in salinized roots of the euhalophyte <Emphasis Type="Italic">Salicornia europaea</Emphasis>

Glycophytic plants suffer from severe stress and injury when roots are exposed to high salinity in the rhizosphere. In contrast, the euhalophyte Salicornia europaea grows well at 200 mM NaCl and can withstand up to 1000 mM NaCl in the root zone. Analysis of gene expression profiles and the underlying molecular mechanisms responsible for this tolerance have been largely overlooked. Using the Illumina sequencing platform and the short-reads assembly programme Trinity, we generated a total of 40 and 39 million clean reads and further 140,086 and 122,728 unigenes from the 200 mM NaCl and 0 mM NaCl treated tissues of S. europaea roots, respectively. All unigenes in this study were functionally annotated within context of the COG, GO and KEGG pathways. Unigene functional annotation analysis allowed us to identify hundreds of ion transporters related to homeostasis and osmotic adaptation as well as a variety of proteins related to cation, amino acid, lipid and sugar transport. We found significant enrichment in response to stress including the functional categories of “antioxidant activity”, “catalytic activity” and “response to stimuli”. These findings represent for a useful resource for the scientific community working on salt tolerance mechanisms. Conversely, a total of 8639 EST-SSRs from 131,594 unigenes were identified and 4539 non-redundant SSRs primers pairs were developed. These data provide a good foundation for future studies on molecular adaptation mechanisms of euhalophytes roots under saline environments and will likely facilitate the identification of critical salt tolerance traits to be transferred in economically important crops.     

The modulation of various physiochemical changes in <Emphasis Type="Italic">Bruguiera cylindrica</Emphasis> (L.) Blume affected by high concentrations of NaCl

Bruguiera cylindrica is a major mangrove species in the tropical mangrove ecosystems and it grows in a wide range of salinities without any special features for the excretion of excess salt. Therefore, the adaptation of this mangrove to salinity could be at the physiological and biochemical level. The 3-month-old healthy plantlets of B. cylindrica, raised from propagules were treated with 0 mM, 400 mM, 500 mM and 600 mM NaCl for 20 days under hydroponic culture conditions provided with full strength Hoagland medium. The modulation of various physiochemical changes in B. cylindrica, such as chlorophyll a fluorescence, total chlorophyll content, dry weight, fresh weight and water content, Na⁺ accumulation, oxidation and antioxidation (enzymatic and non-enzymatic) features were studied. Total chlorophyll content showed very minute decrease at 500 mM and 600 mM NaCl treatment for 20 days and the water content percentage was decreased both in leaf and root tissues with increasing concentration. A significant increase of Na⁺ content of plants from 84.505 mM/plant dry weight in the absence of NaCl to 543.38 mM/plant dry weight in plants treated with 600 mM NaCl was recorded. The malondialdehyde and the metabolites content associated with stress tolerance (amino acid, total phenols and proline) showed an increasing pattern with increasing NaCl concentration as compared to the control in both leaf and root tissues but the increase recorded in plantlets subjected to 500 mM was much less, indicating the tolerance potential of this species towards 500 mM NaCl. The significant decrease of sugar content was found only in 600 mM NaCl on 20 days of treatment, showing that the process of sugar synthesis was negatively affected but the same process remains less affected at 500 mM NaCl. A slight reduction in ascorbate and glutathione content and very less increase in carotenoid content were observed at 500 mM and 600 mM NaCl stress. Antioxidant enzymes (APX, GPX, SOD and CAT) showed an enhanced activity in all the treatments and the increased activity was more significant in 600 mM treated plants. The result establishes that B. cylindrica tolerates high NaCl concentration, to the extent of 500 mM NaCl without any major inhibition on photosynthesis and metabolite accumulation. Understanding the modulation of various physiological and biochemical changes of B. cylindrica at high levels of NaCl will help us to know the physiochemical basis of tolerance strategy of this species towards high NaCl.     

Morpho-physiological and antioxidant response to NaCl-induced stress in in vitro shoots of pomegranate (<Emphasis Type="Italic">Punica granatum</Emphasis> L.)

The physiological and antioxidant response to salinity was studied in pomegranate (Punica granatum L.) by exposing in vitro growing shoots of the Italian variety Profeta Partanna to 125 or 250 mM NaCl for 10 and 20 days. 250 mM NaCl significantly reduced shoot length, leaf area and water content of the shoots, regardless the length of the salt treatment,with respect to the control and to the 125 mM NaCl treatment. After 20 days the shoots treated with 250 mM NaCl also showed a significant reduction in relative growth rate (RGR) together with marked necroses and abscission of the oldest leaves. Salt treatments significantly decreased the contents of chlorophylls and carotenoids in both exposure times, depending on NaCl concentration. Proline, total phenolic compounds and ellagic acid did not increase or even decrease with the salt treatments. The levels of lipid peroxidation decreased, ascorbate peroxidase (APX) activity significantly increased in both treatment times and concentrations, while guaiacol peroxidase (G-POD) activity significantly increased in shoots treated with 250 mM NaCl for 20 days suggesting the rapid involvement of APX in controlling the oxidative stress in this species, even at low salt concentrations, and a delayed complementary role of G-POD.     

In situ histone landscape of nephrogenesis

In the developing kidney, self-renewing progenitors respond to inductive signaling from the adjacent branching ureteric bud by undergoing mesenchyme-to-epithelium transition. Nascent nephrons subsequently undergo elongation, segmentation, and differentiation into a mature renal epithelium with diverse functions. Epigenetic mechanisms have been implicated in impacting cell fate decisions during nephrogenesis; however, the chromatin landscape of nephron progenitors and daughter differentiating cells are largely unknown. Here, we examined the spatiotemporal expression patterns of histone H3 methylation and histone methyltransferases in E15.5 mouse kidneys. Kidney sections were probed with antibodies against histone modifications, enzymes, and markers of progenitors and differentiation. The results revealed that: (1) nephron progenitor cells exhibit a broad histone methylation signature that comprises both “active” and “repressive” marks (H3K4me3/K9me3/K27me3/R2me2/R17me2); (2) nascent nephrons retain high H3K4me3 but show downregulation of H3K9/K27me3 and; (3) maturing epithelial tubules acquire high levels of H3K79me2/3. Consistent with respective histone marks, the H3K4 methyltransferase, Ash2l, is expressed in progenitors and nascent nephrons, whereas the H3K9/K27 methyltransferases, G9a/Ezh2, are more enriched in progenitors than nascent nephrons. We conclude that combinatorial histone signatures correlate with cell fate decisions during nephrogenesis.     

Silicon increases salt tolerance by influencing the two-phase growth response to salinity in wheat (Triticum aestivum L.)

Salt usually stresses plants in two ways, osmotic stress and ion toxicity. Plant responds to salinity in two distinct phases through time. It is known that silicon (Si) could alleviate salt stress by decreasing the Na⁺ accumulated in the leaf. In order to determine the function of Si in the two-phase growth response (osmotic and ion toxicity) to salinity, we selected the wheat cultivar “Changwu 134” out of 10 wheat cultivars, and confirmed that it responds to salinity in two distinct phases through time. The fresh weight, leaf area, and leaf Na⁺ concentration were measured during 31 days of 120 mM NaCl supplemented with 1 mM Si treatment. The results revealed that the growth of plants under salinity conditions both with and without Si application were in accordance with the two-phase growth model. Si alleviated the salt stress in the both two-phase growth, but the alleviative effects were more pronounced in the osmotic stress phase than ion toxicity phase. These results clearly showed that Si can enhance plant salt tolerance by alleviating the salt-induced osmotic stress.     

In vitro olive (Olea europaea L.) cvs Frantoio and Moraiolo microshoot tolerance to NaCl

Abstract

In vitro culture of rooted and unrooted olive microshoots, established from seed lines of free-pollinated “Frantoio” and “Moraiolo” cultivars, were evaluated for NaCl tolerance. The aim was to use growth and physiological parameters in order to identify salt-adapted genotypes. Leaf tissue elemental distribution of Na, Cl and K was also investigated in unrooted plantlets by cryo-scanning electron microscopy and energy-dispersive X-ray microanalysis. Both in unrooted and rooted plantlets, increased concentrations of NaCl reduced shoot growth, whereas plant survival was not affected. However, no significant interactions between line and NaCl concentration were found. Elemental distribution showed that Moraiolo J and Frantoio Z accumulated more Na and Cl inside leaves, and that these elements followed a tissue-dependent pattern. Rooting capacity was reduced at the higher levels of NaCl. Significant interactions between seed line and salt treatment were found. Seed lines showed different abilities to develop roots at different salt levels. In particular, Frantoio Z showed a significant and different behaviour relative to the other seed lines at 50 mM NaCl with regard to both length and dry weight of roots. The results obtained suggest that rooting parameters are the most useful tools in the evaluation and screening of salt-tolerant olive genotypes through in vitro shoot culture.     

Drought and cadmium may be as effective as salinity in conferring subsequent salt stress tolerance in Cakile maritima

Plants are often exposed to a combination of stresses, which can occur simultaneously or at different times throughout their life. In this study, the effects of salinity, drought and cadmium pre-treatments were evaluated on the subsequent response of Cakile maritima, a halophytic species, to various levels of salinity (from 100 to 800 mM NaCl) after a recovery time of 2 weeks. Studies were performed in two sets of experiments in a glasshouse under short and long photoperiod (November and July, respectively). In both experiments and in contrast to control plants (not exposed to any previous stress), plants previously exposed to drought, salt or cadmium stress showed lower levels of hydrogen peroxide and malondialdehyde, an indicator of lipid peroxidation, upon salt treatment, particularly at high NaCl concentrations. Oxidative stress alleviation was not only observed at 800 mM NaCl under short photoperiod, but also at 600 and 800 mM NaCl under long photoperiod in terms of reduced salt-induced increases in hydrogen peroxide and malondialdehyde levels in plants previously exposed to drought, salt or cadmium stress. Previous exposure of plants to all stresses additionally caused decreased levels of jasmonic acid, which might be associated with a lower oxidative stress, differences being observed again at 800 mM NaCl only under short photoperiod and at 600 and 800 mM NaCl under long photoperiod. In conclusion, a relatively long-term stress memory was found in C. maritima pre-exposed to salinity, drought or cadmium, which resulted in a lower oxidative stress when subsequently exposed to salinity. The positive effects of drought and cadmium were of similar magnitude to those provided by salt pre-exposure, which indicated an effective cross-tolerance response in this species.     

Allocation pattern,ion partitioning,and chlorophyll a fluorescence in Arundo donax L. in responses to salinity stress

Biometric and physiological analyses of salt stress responses were performed in two time-course experiments on giant reed (Arundo donax L). Experiment I evaluated biomass production in plants exposed to 128, 256, 512 mM NaCl for 84 days. For Experiment II, plants grown under 256 mM NaCl were further assessed for chlorophyll a fluorescence, ionic partitioning, and proline content at 14 and 49 days after treatment (DAT). Biomass allocation was affected with all the concentrations of NaCl used from 28 DAT onward. Proline biosynthesis in leaves was more stimulated than that in roots after salt stress. Photosynthetic efficiency of photosystem II (PSII) was not affected by salt stress up to 42 DAT, while 49 DAT plants exhibited a significant reduction of both potential (Φ_PSII) and maximal (F_v/F_m) PSII quantum yield. A. donax resulted a moderately sensitive species in response to 256 and 512 mM NaCl, concentrations that are however higher than that commonly found in most marginal lands (such as 128 mM or lower), where the biomass yield is appreciable, especially in short-term cultivation (56 DAT here). Altogether, this study indicates that A. donax can be considered as a promising and valuable energy crop for exploiting the Mediterranean marginal land.     

Effects of salinity and nitric oxide donor sodium nitroprusside (SNP) on development and salt secretion of salt glands of Limonium bicolor

NO, as a signaling molecule, is involved in abiotic stresses. Limonium bicolor seedlings were treated with 200 mM NaCl combined with 0.05 mM SNP for 20 days to study the effects of NO on development and salt-secretion rates of salt glands. It was shown that the total number of salt glands on adaxial surfaces under condition of 200 mM NaCl containing 0.05 mM SNP treatment increased significantly compared with that under 200 mM NaCl treatment. Na⁺ secretion rate per leaf under 200 mM NaCl containing 0.05 mM SNP was significantly higher than that under 200 mM NaCl without SNP. However, there was no significant difference in salt-secretion rate of individual salt glands between 200 mM NaCl containing 0.05 mM SNP treatment and 200 mM NaCl treatment. Although there was no significant difference in salt-secretion rate of individual glands, Na⁺ concentration in the leaves treated with 200 mM NaCl solution containing SNP was significantly lower than that treated with 200 mM NaCl solution. Treatment with 200 mM NaCl solution containing SNP caused a remarkable increase in Na⁺ concentration in salt glands. Obviously, the efficiency of the secretion process per gland was enhanced by adding SNP to NaCl. The results showed NO may enhance the salt secretion by inducing more dermatogen cells to develop into salt glands and by enhancing the efficiency of the secretion process per gland.     

Physiological and proteomic analysis of salinity tolerance of the halotolerant cyanobacterium <Emphasis Type="Italic">Anabaena</Emphasis> sp

The halotolerant cyanobacterium Anabaena sp was grown under NaCl concentration of 0, 170 and 515 mM and physiological and proteomic analysis was performed. At 515 mM NaCl the cyanobacterium showed reduced photosynthetic activities and significant increase in soluble sugar content, proline and SOD activity. On the other hand Anabaena sp grown at 170 mM NaCl showed optimal growth, photosynthetic activities and comparatively low soluble sugar content, proline accumulation and SOD activity. The intracellular Na⁺ content of the cells increased both at 170 and 515 mM NaCl. In contrast, the K⁺ content of the cyanobacterium Anabaena sp remained stable in response to growth at identical concentration of NaCl. While cells grown at 170 mM NaCl showed highest intracellular K⁺/Na⁺ ratio, salinity level of 515 mM NaCl resulted in reduced ratio of K⁺/Na⁺. Proteomic analysis revealed 50 salt-responsive proteins in the cyanobacterium Anabaena sp under salt treatment compared with control. Ten protein spots were subjected to MALDI-TOF–MS/MS analysis and the identified proteins are involved in photosynthesis, protein folding, cell organization and energy metabolism. Differential expression of proteins related to photosynthesis, energy metabolism was observed in Anabaena sp grown at 170 mM NaCl. At 170 mM NaCl increased expression of photosynthesis related proteins and effective osmotic adjustment through increased antioxidant enzymes and modulation of intracellular ions contributed to better salinity tolerance and optimal growth. On the contrary, increased intracellular Na⁺ content coupled with down regulation of photosynthetic and energy related proteins resulted in reduced growth at 515 mM NaCl. Therefore reduced growth at 515 mM NaCl could be due to accumulation of Na⁺ ions and requirement to maintain higher organic osmolytes and antioxidants which is energy intensive. The results thus show that the basis of salt tolerance is different when the halotolerant cyanobacterium Anabaena sp is grown under low and high salinity levels.     

Enhanced tolerance to salinity following cellular acclimation to increasing NaCl levels in Medicago truncatula

Salinity is a major abiotic stress that limits plant productivity. Plants respond to salinity by switching on a coordinated set of physiological and molecular responses that can result in acclimation. Medicago truncatula is an important model legume species, thus understanding salt stress responses and acclimation in this species is of both fundamental and applied interest. The aim of this work was to test whether acclimation could enhance NaCl tolerance in calli of M. truncatula. A new protocol is described incorporating multi-step up acclimation over 0–350 mM exogenous NaCl. By the end of the experiment, calli were tolerant to 150 mM and competent for embryogenesis at 100 mM NaCl. Positive and negative linear relationships between Na⁺ and K⁺ uptake and exogenous NaCl concentration intercepted at 160 mM suggesting a Na⁺/K⁺ homeostasis. Proline level peaked at 100/150 mM whilst highest osmolarity and lowest water content occurred at 250/350 mM NaCl. The concentration of water soluble sugars was positively related to 0–250 mM NaCl whilst callus growth and embryogenesis occurred regardless of endoreduplication. Expression of genes linked to growth (WEE1), in vitro embryogenesis (SERK), salt tolerance (SOS1), proline synthesis (P5CS) and ploidy level (CCS52 and WEE1) peaked at 100/150 mM NaCl. Hence, these genes and various physiological traits except sugar levels, served as useful markers of NaCl tolerance. To our knowledge, this is the first report of a multi-step acclimation conferring tolerance to 150 mM NaCl in leaf-derived calli of M. truncatula.     

Paclobutrazol mitigates salt stress in <Emphasis Type="Italic">indica</Emphasis> rice seedlings by enhancing glutathione metabolism and glyoxalase system

Stress-induced methylglyoxal (MG) functions as a toxic molecule, inhibiting plant physiological processes such as photosynthesis and antioxidant defense systems. In the present study, an attempt was made to investigate the MG detoxification through glutathione metabolism in indica rice [Oryza sativa L. ssp. indica cv. Pathumthani 1] under salt stress by exogenous foliar application of paclobutrazol (PBZ). Fourteen-day-old rice seedlings were pretreated with 15 mg L^?1 PBZ foliar spray. After 7 days, rice seedlings were subsequently exposed to 0 (control) or 150 mM NaCl (salt stress) for 12 days. Prolonged salt stress enhanced the production of MG molecules and the oxidation of proteins, leading to decreased activity of glyoxalase enzymes, glyoxalase I (Gly I) and glyoxalase II (Gly II). Consequently, the decreased glyoxalase activities were also associated with a decline in reduced glutathione (GSH) content and glutathione reductase (GR) activity. PBZ pretreatment of rice seedlings under salt stress significantly lowered MG production and protein oxidation, and increased the activities of both Gly I and Gly II. PBZ also increased GSH content and GR activity along with the up-regulation of glyoxalase enzymes, under salt stress. In summary, salinity induced a high level of MG and the associated oxidative damage, while PBZ application reduced the MG toxicity by up-regulating glyoxalase and glutathione defense system in rice seedlings.     

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.     

Accumulation of gamma-aminobutyric acid in the halotolerant cyanobacterium <Emphasis Type="Italic">Aphanothece halophytica</Emphasis> under salt and acid stress

γ-Aminobutyric acid (GABA) is known as an inhibitory neurotransmitter in human, while in plants, GABA is an intermediate for amino acid metabolism and also is accumulated in response to a wide range of environmental stress. In the present study, GABA accumulation in Aphanothece halophytica was increased 2-fold in mid-log phase cells grown under salt stress (2.0 M NaCl). When mid-log phase cells were subjected to changes in NaCl concentrations and pH for 4 h, the highest GABA accumulation was observed in cells adapted in medium that contained 2.0 M NaCl and that was adjusted to pH 4.0, respectively. The increase of GABA accumulation was accompanied by an increased glutamate decarboxylase activity. Addition of glutamate to growth medium stimulated GABA accumulation under acid stress but had no effect under salt stress. However, the highest GABA accumulation was detected in cells exposed to both high salt and acid stresses combined with the 5 mM glutamate supplementation with an approximately 3-fold increase as compared to the control. The unicellular A. halophytica showed a similarly high content of GABA to that of a filamentous Arthrospira platensis suggesting the possibility of genetic manipulation of the genes of A. halophytica involved in GABA synthesis to increase GABA yield.     

Plant growth promoting rhizobacteria alleviate salinity induced negative effects on growth, oil content and physiological status in Mentha arvensis

This study was aimed to investigate the effect of inoculation on three salt-tolerant, plant-growth-promoting rhizobacteria (PGPR) STR2 (Bacillus pumilus), STR8 (Halomonas desiderata) and STR36 (Exiguobacterium oxidotolerans), for their growth-promoting potential and efficacy in augmenting salt tolerance in Mentha arvensis, an essential oil-bearing crop and natural source of l-menthol, under varying levels of NaCl stress (0, 100, 300 and 500 mM) imposed through irrigating water. Increase in the levels of salt concentration led to a decrease in the growth, fresh weight, leaf–stem ratio, oil content and yield. However, the negative effects of salinity were observed to be convalesced in the PGPR inoculated plants. At salinity levels of 100 and 300 mM NaCl, H. desiderata inoculated plants recorded the highest herb yield, whereas at 500 mM NaCl, the plants inoculated with E. oxidotolerans yielded maximum herb. The oil content in non-inoculated, salt-stressed plants was observed to be 0.46, 0.42 and 0.35 % at 100, 300 and 500 mM NaCl, respectively, whereas the plants inoculated with H. desiderata recorded the oil content of 0.71 and 0.60 and 0.48 % at similar levels of NaCl stress, respectively. The halotolerant PGPR minimized the deleterious effects of salt toxicity producing at par or higher yields at lower and medium salinity levels (100, 300 mM NaCl) than the un-inoculated non-salt-stressed plants through improved foliar nutrient uptake and enhanced antioxidant machinery. Based on the results of the experiments reported herein, the use of salt-tolerant, plant-growth-promoting bacteria may provide an effective means of facilitating M. arvensis growth in salt-stressed environments.     

Glucose-6-phosphate dehydrogenase acts as a regulator of cell redox balance in rice suspension cells under salt stress

The reduced coenzyme nicotinamide-adenine dinucleotide phosphate (NADPH) is an important molecule in cellular redox balance. Glucose-6-phosphate dehydrogenase (G6PDH) is a key enzyme in the pentose phosphate pathway, the most important NADPH-generating pathway. In this study, roles of G6PDH in maintaining cell redox balance in rice suspension cells under salt stress were investigated. Results showed that the G6PDH activity decreased in the presence of 80 mM NaCl on day 2. Application of exogenous glucose stimulated the activity of G6PDH and NADPH oxidase under salt stress. Exogenous glucose also increased the ion leakage, thiobarbituric acid reactive substances and hydrogen peroxide (H₂O₂) contents in the presence of 80 mM NaCl on day 2, implying that the reduction of the G6PDH activity was necessary to avoid serious damage caused by salt stress. The NAPDH/NADP⁺ ratio increased on day 2 but decreased on day 4 under 80 mM NaCl plus glucose treatment. Diphenyleneiodonium, an NADPH oxidase inhibitor, decreased the H₂O₂ content under 80 mM NaCl treatment on day 2. These results imply that the H₂O₂ accumulation induced by glucose treatment under salt stress on day 2 was related to the NADPH oxidase. Western-blot analysis showed that the G6PDH expression was slightly induced by glucose and was obviously blocked by DPI on day 2 under salt stress. In conclusion, G6PDH plays a key role in maintaining the cell redox balance in rice suspension cells under salt stress. The coordination of G6PDH and NADPH oxidase is required in maintaining cell redox balance in salt tolerance.     

Histone methyltransferase G9a and H3K9 dimethylation inhibit the self-renewal of glioma cancer stem cells

Epigenetic modification is crucial to keep the self-renewal and the “stemness” states of stem cells, not letting them to differentiate. The actual roles of Histone 3 Lysine 9 dimethylation (H3K9me2) and its methyltransferase G9a in this process are still unclear, especially in cancer stem cells. In our study, we found an interesting observation that most CD133-positive cells were H3K9me2 negative, both in glioma tissues and in cultured cells, although most cancer cells were detected to be H3K9me2 immunopositive. This implied that the G9a-dependent H3K9me2 was one of the crucial barriers of cancer stem cell self-renewal. To test the hypothesis, we examined the loss-of-function and gain-of-function of G9a. We found that bix01294, the selective inhibitor of G9a, can stimulate the sphere formation rate of glioma cancer stem cells, together with increasing Sox2 and CD133 expressions. The increase of CD133-active stem cells was confirmed by flow cytometry. On the other aspect, overexpression of G9a increased the H3K9me2 and decreased the sphere formation rate as well as the CD133 and Sox2 expressions. Since H3K9me2 modification is the major repressive switch, we predict that the repressive H3K9me2 modification may happen at the CD133 promoter regions. By chromatin precipitation assay, we confirmed that the CD133 and Sox2 promoter regions were modified by the H3K9me2. Therefore, we concluded that the G9a-dependent H3K9me2 repression on CD133 and Sox2 was one of the main switches of the self-renewal in glioma cancer stem cells.     

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.