Growth and soluble amino acid composition inPenicillium corylophilum andHalobacterium halobium grown under salt stress

Penicillium corylophilum can grow in a medium containing NaCl up to 3 mol/L, whereasHalobacterium halobium can grow at 5 mol/L NaCl. At 1 and 3 mol/L NaCl the number of spores decreased with constrictions in the conidiophore and damage in phialides and metulae while there were no morphological changes in bacterial cells grown in the presence of NaCl. Addition of some amino acids or sugar alcohol to the growth medium enhanced the growth of both organisms in the presence of NaCl. Total protein in the fungal cells grown at 1 mol/L NaCl increased by 50% more than in the control cells but it decreased in the bacterial cells with increasing NaCl concentration in the growth medium. Total saccharides and lipids increased in both organisms with increasing NaCl in the growth medium. The endogenous amino acid pool increased in fungal and bacterial cells in the presence of NaCl. Proline was the major amino acid in the fungal cells at 1 mol/L NaCl, representing 41.3% of the total identified amino acids at this concentration, being 8.4 times higher than in the control cells. Glutamate and aspartate showed the highest amount in bacterial cells at 3 mol/L NaCl. Isoleucine showed the highest increase at 3 mol/L NaCl, being 54 times higher than in the control cells.     

Salt concentration-dependency of vitellogenin processing by cathepsin D in Xenopus laevis

The mechanism by which cathepsin D produces only limited proteolysis of vitellogenins (VTG) was studied in Xenopus oocytes. We first examined mature oocytes for the existence of cathepsin D; immunoblot and biochemical analyses revealed the existence of a 43kDa enzyme protein and its proteolytic activity in oocytes during and after the vitellogenesis. By determining the proteolytic activity of the fractions after subcellular fractionation of oocytes, we confirmed that cathepsin D is preserved in the yolk plasma of mature yolk platelets. The reaction of VTG with cathepsin D was examined in vitro at pH 5.6 as a function of NaCl concentrations. Lipovitellins generated from the VTG were preserved for several days at 37°C in the presence of the enzyme if the NaCl concentration was 0.15 mol/L or lower. The amount of lipovitellins decreased with increased molarity of the salt and at 0.5 mol/L NaCl they were rapidly degraded. The precipitates, growing in the reaction tube with 0.15 mol/L NaCl, included all constituents of yolk proteins and were ultrastructurally shown to have crystal structures perforated by empty cavities. No precipitates appeared at 0.5 mol/L NaCl. The results indicate that the limitation on proteolysis of the VTG by cathepsin D is due to the insolubility of yolk proteins at physiological salt concentrations, which explains why yolk can be stored stably in the presence of acid hydrolases over a long period.     

盐胁迫对石蒜叶片形态结构和生理指标的影响

以石蒜为试材,测定了0.1、0.5、1.0mol/L NaCl胁迫下叶片的超氧化物歧化酶（SOD）和过氧化物酶（POD）的活性,可溶性蛋白、不溶性蛋白以及脯氨酸、丙二醛含量等生理指标的动态变化。用石蜡切片法对不同浓度NaCl胁迫后的叶片结构特征作比较,进行其抗盐性分析。结果表明：随NaCl浓度增加,对石蒜形态结构的影响表现在其叶片长度的变化及叶细胞的受损程度等方面;随NaCl浓度的增加和处理时间的延长,SOD和POD活性,不溶性蛋白、脯氨酸含量均呈先升后降趋势;可溶性蛋白呈先降后升趋势;丙二醛含量呈整体上升趋势。石蒜叶片各生理指标的变化通常在NaCl处理后的第7周和第8周为转折点。石蒜对低浓度的NaCl胁迫具有一定的耐受性。     

Effects of trace elements on the telomere lengths of hepatocytes L-02 and hepatoma cells SMMC-7721

The effects of selenium, zinc, iron, chromium, and lead on telomere lengths of human cells have not been investigated. This article adopted flow cytometry and fluorescence in situ hybridization to investigate the impact of different elements on cellular apoptosis and telomere lengths of human hepatocytes L-02 and hepatoma cells SMMC-7721. Results showed that these trace elements under the following dosages did not have remarkable effect on cellular apoptosis. However, sodium selenite at doses of 0.5 and 2.5 μmol/L significantly extended the telomere length of hepatocytes L-02; 0.5 μmol/L lead acetate remarkably shortened the telomere length of L-02 cells; 80 μmol/L zinc sulfate, 20 μmol/L ferric chloride, and 200 μmol/L chromic chloride only had slight impact on the telomere length, respectively. Regarding hepatoma cells SMMC-7721, sodium seleite at 0.5 and 2.5 μmol/L had little impact on the telomere length; 80 μmol/L zinc sulfate significantly accelerated the loss of telomere length, whereas 20 μmol/L ferric chloride, 200 μmol/L chromic chloride, and 0.5 μmol/L lead acetate remarkably extended the telomere lengths, respectively. The results revealed differential effects of each trace element on the life-span of human hepatocytes and hepatoma cell lines, which suggested further research on somatic hepatocytes and hepatoma in vivo.     

Long-term hyposaline and hypersaline stresses produce distinct antioxidant responses in the marine alga Dunaliella tertiolecta

Tolerance to salinity stress in higher plants correlates to levels of antioxidant enzymes and/or substrates. Do hyperosmotic and hypoosmotic stress induce antioxidant responses in salt tolerant algae, and if so, are these responses the same for both excess and minimal salinity? To answer these questions, cultures of the marine alga Dunaliella tertiolecta (Chlorophyta) were grown in seven salinities covering a 60-fold range from 0.05 to 3.0 mol/L NaCl. Long-term effects of salinity on growth and antioxidant parameters were determined. Growth rates were reduced at the salinity extremes (0.05 mol/L NaCl and 3 mol/L NaCl) indicating the cultures were stressed. The levels of six antioxidant enzymes and three antioxidant substrates were quantified at these growth salinities. Compared to growth at optimum salinities (i.e. 0.2-0.5 mol/L NaCl), high salinities produced a 260% increase in monodehydroascorbate reductase, a doubling of ascorbate peroxidase activity and a three-fold increase in the rate of dark respiration. Cells acclimated to low growth salinities (hyposaline stress, i.e. < 0.2 mol/L NaCl) showed major increases in glutathione and alpha-tocopherol coupled with decreases in Fv/Fm ratios and in total and reduced ascorbate compared to moderate and high external salinities. Cell volumes remained unchanged, except at the lowest salinity where they doubled. Catalase, superoxide dismutase, dehydroascorbate reductase and glutathione reductase activities were not altered by extreme salinities. The involvement of oxidative stress at both salinity extremes is implied by the alterations in antioxidant enzymes and substrates, but the specific changes are very different between hypo and hypersaline stresses.     

Resistance of fully imbibed tomato seeds to very high salinities

Abstract Seeds of Lyeopersicon esculentum cv. VF36 (a salt-sensitive cultivar), L. esculentum var. Edkawi (which is fairly salt-resistant), and a wild relative, L. cheesmanii, were exposed to various concentrations of NaCl, up to 460 mol m^?3, either directly or following imbibition in non-saline nutrient solution. After 10 d exposure to salt, they were transferred to non-saline solution. All taxa showed some germination at the lowest salinity tested, 92 mol m^?3 NaCl, but virtually no germination occurred at 184 mol m^?3 NaCl or higher salinities. Within 2 d after removal of the salt stress, however, the seeds of L. esculentum reached control levels of germination, even if they had already been on the verge of germination when the stress was imposed. The seeds of L. cheesmanii were less resistant to NaCl. The physiological basis for the resistance of VF36 seeds is discussed.     

Properties of Scottish isolates of cocksfoot mild mosaic virus and their comparison with others

Two isolates of cocksfoot mild mosaic virus obtained from cocksfoot (Dactylis glomerata) in Scotland differed in symptomatology, and apparently in host range, from isolates obtained in Germany and Wales. They were serologically more closely related to a Dutch isolate from cocksfoot, and to a Scottish isolate from timothy (Phleum pratense), than was the German isolate from cocksfoot. The Scottish isolate from timothy was somewhat more virulent than, but serologically closely related to a Welsh isolate from timothy. Particles of Scottish isolates from cocksfoot and timothy were best preserved for electron microscopy by fixation with osmium tetroxide. In 1.0 m KCl or 0.01 m ethylene diamine tetraacetate they were stable at pH 5.2–5.3 but unstable above pH 7; they were disrupted by 0.5% sodium dodecyl sulphate. The particles contained major and minor RNA components of mol. wt c. 1.5. 10⁶(RNA-1) and 0.5. 10⁶(RNA-2) respectively, together with polydisperse RNA of intermediate mol. wt and protein of mol. wt c. 27 000. In CsCl gradients, major and minor nucleoprotein components of density 1.39 and 1.38 g/ml respectively were distinguished. The less dense particles contained a larger proportion of intermediate-sized RNA molecules and of RNA- 2 , and a smaller proportion of RNA- 1 , than did the denser particles. Particles seem to contain either RNA-1 or various combinations of smaller RNA molecules. Despite the differences in antigenic constitution, symptomatology and particle stability between virus isolates obtained from cocksfoot and timothy in different countries, these isolates seem sufficiently similar to be considered one virus.     

Effect of chloride and glutamate ions on in vitro protein synthesis by the moderate halophile Vibrio costicola.

Vibrio costicola grown in the presence of different NaCl concentrations contains cell-associated Na+ and K+ ions whose sum is equal to or greater than the external Na+ concentration. In the presence of 0.5 M NaCl, virtually no in vitro protein is synthesized in extracts of cells grown in 1.0 M NaCl. However, we report here that active in vitro protein synthesis occurred in 0.6 M or higher concentrations of Na2SO4, sodium formate, sodium acetate, sodium aspartate, or sodium glutamate, whereas 0.6 M NaF, NaCl, or NaBr completely inhibited protein synthesis as measured by polyuridylic acid-directed incorporation of [14C]phenylalanine. Sodium glutamate, sodium aspartate, and betaine (0.3 M) counteracted the inhibitory action of 0.6 M NaCl. The cell-associated Cl- concentration was 0.22 mol/kg in cells grown in 1.0 M NaCl. Of this, the free intracellular Cl- concentration was only 0.02 mol/kg. Cells contained 0.11 mol of glutamate per kg and small concentrations of other amino acids. All of the negative counterions for cell-associated Na+ and K+ have not yet been determined. In vitro protein synthesis by Escherichia coli was inhibited by sodium glutamate. Hybridization experiments with ribosomes and the soluble (S-100) fractions from extracts of E. coli and V. costicola showed that the glutamate-sensitive fraction was found in the soluble, not the ribosomal, part of the system. The phenylalanyl-tRNA synthetase of V. costicola was not inhibited by 0.5 M or higher concentrations of NaCl; it was slightly more sensitive to high concentrations of sodium glutamate. Therefore, this enzyme was not responsible for the salt response of the V. costicola in vitro protein-synthesizing system.     

Influence of sodium chloride on the beta-glucuronidase activity of Clostridium perfringens and Escherichia coli

While the beta-glucuronidase activity of intact cells of Clostridium perfringens was higher in 0.95% sodium chloride (NaCl) than that in 0, 0.1 or 0.5%, that of Escherichia coli was higher in 0.1% NaCl than that in 0, 0.5 or 0.95% NaCl in 0.1 mol l-1 KH2PO4. However, the enzyme activity of both species of intact cells was higher in buffer containing 16 mEq sodium, 134 mEq potassium and 16 mEq chloride per litre than in that containing 146 mEq sodium, 13 mEq potassium and 146 mEq chloride. These findings suggest that bacterial cells are affected by the presence of NaCl and that the effect of NaCl on the activity of bacterial beta-glucuronidase may differ by location in the large intestine.     

苜蓿中华根瘤菌与耐盐有关的DNA片段的克隆

以耐盐的苜蓿中华根瘤菌(\%Sinorhizobium meliloti) \%042B为材料,制备其总DNA,经过限制性内切酶\%Eco\%RⅠ的部分酶解,利用电洗脱方法回收15～25kb大小的DNA片段。以碱法制备载体质粒pLAFRⅠ,用\%Eco\%RⅠ将其切成线状,然后用T\-4DNA连接酶将回收片段与线状载体连接,利用包装蛋白进行包装后,感染大肠杆菌(Escherichia coli)S17\|1,构建了042B的基因文库。以固体亚硝基胍作为诱变剂处理出发菌株,在05mol/LNaCl的条件下,从2000个菌落中筛选得到12株042B的盐敏感突变株,以其中稳定的盐敏突变株GZ17为受体菌,利用两亲本杂交将含有042B的DNA片段的pLAFRⅠ重组质粒转移到GZ17中,在含有四环素和05mol/LNaCl的基本培养基上筛选出能够耐盐的阳性克隆,获得了与耐盐有关的7kb长的DNA片段。对该片段进行亚克隆,最终获得了4kb与耐盐有关的片段。     

A new isolate of Hydrogenobacter,an obligately chemolithoautotrophic,thermophilic, halophilic and aerobic hydrogen-oxidizing bacterium from seaside saline hot spring

An obligately chemolithoautotrophic and aerobic hydrogen-oxidizing bacterium was isolated from a seaside saline hot spring in Izu Peninsula, Japan. The isolate was a Gram-negative, non-motile, non-spore-forming rod cell measuring 0.3 to 0.5 by 1.0 to 2.5 m. The optimal temperature for growth was around 70°C, and no growth was observed at 40°C or 80°C. Elemental sulfur or thiosulfate could be an alternative to molecular hydrogen as the sole energy source. The DNA base composition of the isolate was 46.0 mol% G+C. 2-Methylthio-3-VI,VII-tetrahydromultiprenyl⁷-1,4-naphthoquinone (methionaquinone) was the major component of the quinone system. C_18:0, C_18:1 and C_20:1 were the major components of the cellular fatty acids. These properties clearly indicate that the isolate belongs to genus Hydrogenobacter, but differed from H. thermophilus in some respects. Specifically, the isolate was a halophile which grew optimally at around 0.3–0.5 M NaCl, while H. thermophilus could not grow at such NaCl concentration levels. A new species name H. halophilus is proposed for this new halophilic isolate.     

Regulation of cellular glutathione peroxidase by different forms and concentrations of selenium in primary cultured bovine hepatocytes

The expression and activity of cellular glutathione peroxidase (GPx1) are regulated by selenium (Se). Generally speaking, organic forms of Se have less toxicity and greater bioavailability compared with inorganic forms. In this study, the effects of different forms and concentrations of Se on the regulation of mRNA level and activity of GPx1 in bovine hepatocytes were evaluated, and the optimal doses of different forms of Se that supported the full expression of GPx1 were determined. Primary cultured bovine hepatocyte monolayers derived from neonatal male Holstein calves (aged 1–2 days) were incubated for 24 h with 0 (control), 0.5, 1, 1.5, 2, 3, 4 or 5 μmol/L of Se from dl-selenomethionine (Se-Met), sodium selenite (Na₂SeO₃) or Kappa-selenocarrageenan (Se-Car). Compared with controls, a significantly lower level of release of lactic dehydrogenase (LDH) was observed at 0.5–5 μmol/L of Se-Met, 0.5–1 μmol/L of Na₂SeO₃ and 0.5 μmol/L of Se-Car, but significantly higher LDH release was observed at 2–5 μmol/L of Na₂SeO₃ and 3–5 μmol/L of Se-Car, and the response occurred in a dose-dependent manner. The intracellular content of reduced glutathione in all hepatocytes treated with Se was significantly lower than that of controls. Significant increases in GPx1 mRNA were obtained in all hepatocytes treated with Se, with maximal effects at 3 μmol/L of Se-Met, 1.5 μmol/L of Na₂SeO₃ and 2 μmol/L of Se-Car, respectively. Furthermore, 3 μmol/L of Se from Se-Met resulted in peak levels of GPx1 mRNA. After reaching a maximal level, higher Se supplementation led to a reduction of GPx1 mRNA. The activity of GPx1 showed similar patterns but of lower magnitude. We conclude that (a) the regulation of mRNA level and activity of GPx1 in primary cultured bovine hepatocytes by different forms of Se varies and (b) the optimal doses of Se to support the full expression of GPx1 in bovine hepatocytes when supplied as Se-Met, Na₂SeO₃ and Se-Car are 3, 1.5 and 2 μmol/L, respectively.     

Effects of sodium chloride on biodegradation of crude oils by two species ofAeromonas

Summary The biodegradation of five weathered crude oils by two species ofAeromonas, (B59-4 and E. BOB) was investigated in varying concentrations of sodium chloride. A minimal salts medium whose NaCl concentration increased serially by 0.5% w/v up to 1.5% w/v was used to investigate the growth of these strains in glucose, and their biodegradation of the crude oils. The latter was also investigated in fresh and aged sea water. Strain B59-4 was more potent than E. BOB in the degradation of all five crude oils and at all four levels of salt concentration tested. The amount of oil degraded by each strain increased initially to a maximum level at 0.5% w/v NaCl, but thereafter decreased with increasing salt concentration, and the patterns were similar to those of aged and fresh sea water, respectively. The Forties and Nigerian crude oils with lower specific gravity, were more readily degraded than the Libyan and Venezuelan with higher specific gravity. The growth of the two strains ofAeromonas in glucose and their biodegradation of crude oils was optimal at 0.5% w/v NaCl, and thereafter decreased with increasing salt concentration of the basal medium.     

Statistical optimization of lovastatin production by Omphalotus olearius (DC.) singer in submerged fermentation

In this study, culture conditions were optimized to improve lovastatin production by Omphalotus olearius, isolate OBCC 2002, using statistical experimental designs. The Plackett–Burman design was used to select important variables affecting lovastatin production. Accordingly, glucose, peptone, and agitation speed were determined as the variables that have influence on lovastatin production. In a further experiment, these variables were optimized with a Box–Behnken design and applied in a submerged process; this resulted in 12.51 mg/L lovastatin production on a medium containing glucose (10 g/L), peptone (5 g/L), thiamine (1 mg/L), and NaCl (0.4 g/L) under static conditions. This level of lovastatin production is eight times higher than that produced under unoptimized media and growth conditions by Omphalotus olearius. To the best of our knowledge, this is the first attempt to optimize submerged fermentation process for lovastatin production by Omphalotus olearius.     

龙珠果茎段离体培养和组培苗耐盐性分析

为建立龙珠果(Passiflora foetida)的快繁再生体系,以实生苗茎段为外植体,研究了植物生长调节剂对丛生芽诱导、壮苗生根的影响,同时对组培苗的耐盐性进行研究.结果表明,MS+6-BA 0.5 mg/L+NAA 0.05 mg/L培养基有利于诱导丛生芽并促进芽的生长;MS+6-BA 3.0 mg/L+NAA ...     

Effect of 5-Aminolevulinic Acid on Development and Salt Tolerance of Potato (Solanum tuberosum L.) Microtubers in vitro

The effects of 5-aminolevulinic acid (ALA), a key precursor in the biosynthesis of porphyrins such as chlorophyll and heme, on development and salt tolerance of microtubers of two potato (Solanum tuberosum L.) cultivars Jingshi-2 and Zihuabai were examined under in vitro conditions. ALA at 0.3–3 mg/l promoted microtuber formation by increasing the average number, diameter, and fresh weight of microtubers especially under 0.5% NaCl stress conditions, but further increase in ALA concentration resulted in a reduction of microtuber yield irrespective of NaCl stress. Under 1.0% NaCl stress conditions, microtuberization was seriously repressed and could not be restored by the addition of ALA. The accumulation of malondialdehyde in the microtubers treated with 30 mg/l ALA increased by 22% compared to the controls (no salinity), while only a 7% increase was observed when the microtubers were exposed to 0.5% NaCl, indicating that ALA functions as a protectant against oxidative damages of membranes. Under 0.5% NaCl stress conditions, the highest activities of peroxidase and polyphenoloxidase were detected in microtubers treated with ALA at 0.3 and 3 mg/l, being by 73% and by 28% greater than those in the untreated controls, respectively. These results demonstrate that ALA at lower concentrations of 0.3–3 mg/l promotes development and growth of potato microtubers in vitro and enhances protective functions against oxidative stresses, but ALA at 30 mg/l and higher concentrations seems to induce oxidative damage probably through formation and accumulation of photooxidative porphyrins.     

Effect of NaCl salinity on growth and mineral partitioning in Quercus robur L., a rhythmically growing species

 Sodium salt sensitivity of common oak (Quercus robur L.) was evaluated in hydroponic culture using INRA-Morizet solution. Addition of NaCl to the nutrient solution reduced only length and weight of roots and first flush stems. In contrast, the second flush was properly expanded even in the presence of 40 mM of NaCl in culture medium. Both leaf number and leaf area were not affected by increasing salt concentration in medium culture while this increase induced significant leaf damage especially in first flush leaves. Stem starch storage was reduced only at 40 mM NaCl treatment. Common oak seedlings seemed to be able to better compartmentalize sodium than chloride when the NaCl concentration increased in the medium culture. Chloride presented a lower uptake than sodium. Sodium was preferentially accumulated in roots and this accumulation occurred at the expense of potassium uptake. The decrease of ATP content in leaves of common oak seedlings submitted to NaCl treatments could indicate that it was used for sodium exclusion out of the leaves, especially in second flush leaves. Relationships between growth responses, starch and mineral element distribution in common oak seedlings will be discussed. Received: 20 November 1997 / Accepted: 3 March 1998     

Alleviation of salt-induced stress on seed emergence using soil additives in a greenhouse

Use of sodium chloride (NaCl) as a deicing salt results in high concentrations of ions in roadside soils, which decreases seedling emergence in these areas. Greenhouse experiments performed in soil culture tested the efficacy of three soil fertilizers, gypsum (CaSO₄), potash (potassium chloride, KCl) and potassium nitrate (KNO₃), in alleviating NaCl stress on seedling emergence of three grass species exhibiting a range of salt tolerance, Poa pratensis (Kentucky bluegrass), Bouteloua gracilis (blue grama), and Puccinellia distans (alkali grass). Two-factor factorial designs were utilized for each species-fertilizer combination. Treatments of 5000 mg/L (0.086 M) NaCl with or without fertilizer, in concentrations that were equal to 0.5, 1, and 2 times the molar equivalent of 5000 mg/L NaCl were applied biweekly. Salt stress on Poa pratensis emergence was alleviated by all fertilizers with CaSO₄ having the greatest effect in alleviating NaCl stress and potash and potassium nitrate alleviating stress at lower treatment levels. Emergence of Bouteloua gracilis and Puccinellia distans was in most cases negatively effected by soil amendments.     

Distribution of saccharides and salts on amphoteric ion-exchange resin

An amphoteric ion-exchange resin hardly shrank in 550 and 300 g/L glucose and sodium chloride solutions, respectively; however, the bed packed with a cation-exchange resin shrank considerably. From the distribution coefficients of some saccharides, the swelling pressure of the amphoteric ion-exchange resin was estimated to be 2.0 MPa at 25 °C. The distribution coefficients of glucose, galactose, fructose, and mannose were independent of their concentration and were about 0.621. On the other hand, the apparent distribution coefficients of NaF, NaCl, NaBr, NaI, LiCl, KCl, and CsCl largely depended on concentration. A model for the distribution of salts on the amphoteric resin was proposed, assuming an interaction between the anion of the salt and the positively charged fixed ions with binding constant B. The B values of the chloride salts were nearly the same (1.69–2.94 L/mol), while the values of the sodium salts were largely different depending on the anion.     

盐胁迫对蓝蓟种子萌发及早期幼苗的影响

研究了不同浓度盐(NaCl)处理对蓝蓟种子发芽势、发芽率及幼苗生长的影响。结果表明:(1)不同盐胁迫处理对蓝蓟种子的萌发具有显著影响。随着盐处理浓度的增加,发芽率呈下降趋势,蓝蓟在无盐环境(蒸馏水)和低盐环境中(0.05 mol/L)长势最好、发芽率最高、发芽速度最快。(2)将不同盐溶液处理5 d的未萌发种子转移到蒸馏水后,蓝蓟种子的萌发能力均可恢复,原来较高盐浓度(0.3 mol/L～0.5 mol/L)下的种子在恢复后,其萌发恢复率均在75%以上,发芽率基本上随着原浓度的升高呈上升的趋势。研究表明,NaCl处理对蓟种子没有造成伤害,其抑制萌发是通过渗透效应而不是离子毒害。