Phosphorus contamination in polyethylene glycol

Concentrations of Fe, Mn, Cu, Zn, Ca, Mg, K, and P were examined in untreated and ion exchange resin-treated solutions of polyethylene glycol, molecular weight 3000 to 3700, polyethylene glycol (PEG 4000). Relatively high levels of P were found in untreated PEF-4000 solutions. The concentration of contaminating P in solutions prepared from untreated PEG 4000, even at high water potentials (−1 to −3 bars), was greater than what is usually found in soil solution. Occurrence of significant amounts of P in untreated PEG could introduce problems in experiments where ³²P and PEG are used together and where phosphate interactions may occur.     

Optimization of conditions for acid protease partitioning and purification in aqueous two-phase systems using response surface methodology

Aspergillopepsin I, an acid protease, was purified using an aqueous two-phase system that comprised various combinations of polyethylene glycol (PEG), NaH₂PO₄ and NaCl. Partition of the enzyme depended upon the molecular mass of the PEG and the presence of NaCl. With PEG 1500, 4000 and 6000, the partition coefficients were increased by 1,500-, 1,800- and 560-fold compared to values without NaCl. The presence of NaCl (8.75%, w/w) increased purification by 3.8, 9.5 and 2.8 times into these respective PEGs. The optimal aqueous two-phase system for acid protease purification was developed using response surface methodology. This system contained 17.3% of PEG 4000 (w/w), 15% NaH₂PO₄ (w/w) and 8.75% NaCl (w/w) and provided the best partition coefficient (Ke > 1,100) and yield over 99% in the same phase. The optimal ATPS purification factor of acid protease was over 5.     

The osmotic potential of polyethylene glycol 6000

Osmotic potential (ψ_s) of aqueous solutions of polyethylene glycol 6000 (PEG-6000) was curvilinearly related to concentration. At given concentrations, ψ_s increased linearly with temperature. The effects of concentration and temperature on ψ_s of PEG-6000 solutions differ from those for most salts and sugars and apparently are related to structural changes in the PEG polymer. Measurements of ψ_s with thermocouple psychrometers are more negative than those with a vapor pressure osmometer, with the psychrometer probably giving the more nearly correct ψ_s for bulk solutions. An empirical equation permits calculation of ψ_s from known concentrations of PEG-6000 over a temperature range of 15 to 35 C. Viscometery and gravimetric analysis are convenient methods by which the concentrations of PEG-6000 solutions may be measured.     

The effect of H2O2 and abscisic acid (ABA) interaction on β-amylase activity under osmotic stress during grain development in barley

The effects of exogenous abscisic acid (ABA) and polyethylene glycol (PEG 6000) treatments on grain H₂O₂, ABA and β-amylase activity were studied during grain development in the spike culture experiments with variety Triumph and its ABA-insensitive mutant TL43 as the plant materials. The results showed that during grain development the two genotypes were similar in the pattern of ABA concentration change, but differed greatly in the pattern of H₂O₂ concentration and β-amylase activity changes. The β-amylase activity was positively correlated with H₂O₂ concentration, negatively correlated with ABA concentration, and it is mainly closely associated with continued high levels of ABA with respect to H₂O₂. Water stress (PEG treatment) induced β-amylase was associated with H₂O₂ concentration but not with ABA concentration. Exogenous application of H₂O₂ and Ascorbic acid (AsA) increased β-amylase activity in Triumph but reduced that of TL43. However, the endogenous H₂O₂ concentration in grains was always consistent with β-amylase activity. A novel model was hypothesized from the current results to illustrate the relationship between H₂O₂, ABA and β-amylase synthesis for the barley exposed to abiotic stresses.     

Acetylsalicylic acid ameliorates negative effects of NaCl or osmotic stress in Solanum stoloniferum in vitro

The role of acetylsalicylic acid (0, 1 and 10 μM) pre-treatment in amelioration of salt and osmotic stress in a wild species of potato (Solanum stoloniferum) was investigated. We compared the effects of iso-osmotic concentrations of polyethylene glycol 6000 (15 %) and NaCl (80 mM) on the physiological responses of this species in explants grown in the liquid Murashige and Skoog medium. Both salt and drought reduced shoot growth parameters, photosynthetic pigment contents and increased lipid peroxidation, electrolyte leakage, H₂O₂ content and lipoxygenase activity. The effect of NaCl was more severe than that of polyethylene glycol. Salinity also increased Na⁺ content and decreased K⁺ content and K⁺/Na⁺ ratio. Under both stresses, the activities of superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase, catalase and glutathione reductase enzymes were increased. Acetylsalicylic acid pre-treatment alleviated the adverse effects of both stresses on all parameters measured.     

Polyethylene Oxide (PEO) and Polyethylene Glycol (PEG) Polymer Sieving Matrix for RNA Capillary Electrophoresis

The selection of sieving polymer for RNA fragments separation by capillary electrophoresis is imperative. We investigated the separation of RNA fragments ranged from 100 to 10,000 nt in polyethylene glycol (PEG) and polyethylene oxide (PEO) solutions with different molecular weight and different concentration. We found that the separation performance of the small RNA fragments (<1000 nt) was improved with the increase of polymer concentration, whereas the separation performance for the large ones (>4000 nt) deteriorated in PEG/PEO solutions when the concentration was above 1.0%/0.6%, respectively. By double logarithmic plot of mobility and RNA fragment size, we revealed three migration regimes for RNA in PEG (300-500k) and PEO (4,000k). Moreover, we calculated the smallest resolvable nucleotide length (N_min) from the resolution length analysis.     

2种玉米幼苗耐旱性生理机制研究

以白种皮(白玉米)和黄种皮(黄玉米)2个玉米栽培品种为材料,在水培条件下进行聚乙二醇(PEG-6000)模拟干旱胁迫处理,分析玉米叶片抗旱性相关生理特性和质膜H+-ATP酶活性的变化,探讨2种玉米幼苗耐旱性生理机制。结果表明:(1)在2%、5%、10%PEG-6000处理条件下,随处理浓度和时间的增加,2种玉米幼苗植株失水率上升,叶片蒸腾速率降低,气孔传导率下降;在所有相同处理条件下,白玉米植株失水率明显小于黄玉米,而叶片蒸腾速率和气孔传导率下降幅度明显大于黄玉米,即白玉米的耐旱性比黄玉米强。(2)在相同浓度PEG-6000处理下,白玉米叶片可溶性蛋白、可溶性糖含量、游离脯氨酸含量均高于黄玉米,它在干旱胁迫下的渗透调节能力强于黄玉米。(3)在抗氧化酶体系中,随着PEG-6000胁迫浓度的升高,2种玉米叶片CAT活性呈下降趋势,但白玉米CAT活性在2%和5%PEG-6000胁迫下均显著高于黄玉米,其叶片中H2O2含量显著低于黄玉米。(4)随着PEG-6000胁迫浓度的升高,白玉米叶片质膜H+-ATPase磷酸化水平及其与14-3-3蛋白的结合受到的抑制作用比黄玉米强,白玉米叶片质膜H+-ATPase活性比黄玉米叶片低,叶片气孔开度小于黄玉米,叶片蒸腾速率和气孔传导率均低于黄玉米,这可能是白玉米耐旱性强于黄玉米的一个重要机制。     

Priming responses of leek (Allium porrum L.) seeds to different dissolved oxygen levels in the osmoticum

Leek seeds (Allium porrum L.) cv. Winterreuzen were treated for 5 days in stirred bioreactors containing a -1.0 MPa polyethylene glycol (PEG 6000) solution. The level of dissolved oxygen was controlled from zero to approximately two and a half times that of saturation with respect to atmospheric air by bubbling through 0₂/N₂ gas blends from 0% O₂ to 50% O₂. The treatment with 0% O₂ (i.e. bubbling with N₂) gave no reduction in germination time compared to untreated seeds after allowing for the time for water imbibition. As the proportion of oxygen was increased, a rapid reduction was observed until a concentration equivalent to air was used. Further increases to the maximum, produced only a marginal further improvement. The reduction in germination times for seeds that had been cabinet dried was longer for all treatments due to the time required for re-imbibition of water but the general response to priming was the same as with undried seeds. Only dried seeds treated with air or higher oxygen concentrations showed enhanced synchronicity of germination times. The percentage germination of seeds from all treatments was the same, including the treatment with nitrogen gas. These tests help to explain the recently reported advantages of using enriched air for seed priming and are one of the necessary protocols for ensuring satisfactory bulk priming.     

A simple procedure to overcome polyethelene glycol toxicity on whole plants

A procedure is described that can be used to minimize toxic effects of polyethylene glycol (PEG) to plants. The procedure is based on recycling nutrient solutions containing PEG-6000 through two plant cultures. Tomato plants grown in −0.3 megapascals PEG solutions used after two growth cycles exhibited minimal toxic effects. Long-term responses like dry matter production and chlorophyll content as well as short-term responses like CO₂ fixation rates and leaf conductance were severely inhibited by fresh PEG-6000 and only slightly reduced by recycled PEG-6000. Complete osmotic adjustment was obtained with tomatoes grown in recycled but not in fresh PEG solutions.     

A method for the preparation of heavy-atom derivatives of yeast cytochrome c peroxidase.

This Letter describes a strategy for preparation of heavy-atom derivatives which has succeeded with yeast cytochrome c peroxidase and which may be useful for other crystalline proteins. Transfer of the crystals from methylpentanediol solutions, in which they were grown, to solutions of polyethylene glycol (M_r 6000) maintains the crystal structure so that crystals tolerate heavy-atom concentrations at least ten times larger than could be attained in methylpentanediol mother liquors.     

Extraction of protease from Calotropis procera latex by polyethylene glycol–salts biphasic system

The protease from the latex of Calotropis procera was isolated by an aqueous two-phase system (ATPS). The systems consist of polyethylene glycol (PEG 4000, 6000 and 8000) at concentrations of 9, 12 and 15% (w/w) with salts (Na-citrate, MgSO₄, K₂HPO₄, and (NH₄)₂SO₄) at concentrations of 11, 14 and 17% (w/w) were investigated. The highest protease recovery was found in the PEG-rich phase of the system, comprising of 12% PEG 4000–17% MgSO₄. For optimization of the system to obtain the higher yield of protease, the system pH (4, 7 and 10) or NaCl addition (2, 4 and 6%, w/w) was studied. At acidic (pH 4.0) and alkaline (9.0) conditions of the systems the reduction of K_E and protease recovery was clearly observed compared to that of the neutral pH (7.0). The addition of NaCl up to a final concentration of 6% (w/w) significantly increased the yield to 107% of the control. Molecular weight distribution and activity staining showed that the isolated protease had the molecular weight of ∼38 kDa. However, the isolated protease had no activity under reducing condition (βME). Under cathodic electrophoresis, protease from C. procera showed the same protein pattern to purified papain.     

Hydrogen peroxide pretreatment induces osmotic stress tolerance by influencing osmolyte and abscisic acid levels in maize leaves

Hydrogen peroxide (H₂O₂) functions as a signal molecule in plants under abiotic and biotic stresses. Leaves of detached maize (Zea mays L.) seedlings were used to study the function of H₂O₂ pretreatment in osmotic stress resistance. Low H₂O₂ concentration (10 mM) which did not cause a visual symptom of water deficit (leaf rolling) was applied to the seedlings. Exogenous H₂O₂ alone increased leaf water potential, endogenous H₂O₂ content, abscisic acid (ABA) concentration, and metabolite levels including soluble sugars, proline, and polyamines while it decreased lipid peroxidation and stomatal conductance. Osmotic stress induced by polyethylene glycol (PEG 6000) decreased leaf water potential and stomatal conductance but enhanced lipid peroxidation, endogenous H₂O₂ content, the metabolite levels, and ABA content. H₂O₂ pretreatment also induced the metabolite accumulation and improved water status, stomatal conductance, lipid peroxidation, ABA, and H₂O₂ levels under osmotic stress. These results indicated that H₂O₂ pretreatment may alleviate water loss and induce osmotic stress resistance by increasing the levels of soluble sugars, proline, and polyamines thus ABA and H₂O₂ production slightly decrease in maize seedlings under osmotic stress.     

Bisulfite interacts with binding sites of the auxin-transport inhibitor N-1-naphthylphthalamic acid

The affinity of the auxin-transport inhibitor N-1-naphthylphthalamic acid (NPA) for membrane particles as well as for solubilized binding sites from Cucurbita pepo L. hypocotyls was reduced by low concentrations of bisulfite (half-maximal inhibition at 2·10^-3–3·10^-3 M). Two membrane fractions obtained by sedimentation aided with polyethylene glycol showed differential sensitivity to bisulfite. Other oxidizing or reducing substances tested at 1 mM had no effect, except for N-ethylmaleimide (80% inhibition) and iodine (complete inhibition), both of which reduced the number of binding sites but not their affinity. Addition of bisulfite to either the isoalloxane ring of flavoproteins or to pyridoxal phosphate or quinones is proposed as a possible mechanism of action. Sulfur dioxide, at concentrations measured in polluted air, can lead to bisulfite concentrations in plant tissue sufficient to interfere with NPA-binding sites and hence with auxin transport.Abbreviations DTE dithioerythritol - DTT dithiothreitol - IC₅₀ concentration of half-maximal inhibition - NAA 1-naphthylacetic acid - NEM N-ethylmaleimide - NPA N-1-naphthylphthalamic acid - PEG polyethylene glycol, 6000 molecular weight     

Aerenchyma formation and associated oxygen movement in seminal and nodal roots of wheat

Abstract The present paper describes the effects of growth of roots of wheat (Triticum aestivum cv. Gamenya) in hypoxic nutrient solutions on acrenchyma formation and O₂ movement from shoots to roots. Two types of roots were investigated: (1) seminal roots of 4–7-d-old seedlings, and (2) seminal and nodal roots of 10–28-d-old plants. Gas-filled porosity of seminal and nodal roots increased from 3 to 12% and from 5–7 to 11–15%, respectively, when the roots emerged in stagnant or N₂-flushed solutions (0.003 mol m ^?3 O₂) compared with growth in continuously acrated solutions (0.26 mol m ^?3 O₂). However, neither root type increased in porosity when they were longer than 100–200 mm at the start of the exposure to these stagnant or N₂-flushed treatments. A vernier microscope and cylindrical platinum-electrode were used to examine the relationship between root extension and transport of O₂ from shoots to roots via the gas spaces. Measurements were made when the roots were in an anoxic medium and were dependent solely on O₂ supplied from the shoots. For seminal roots of 5–7-d-old seedlings raised in stagnant solutions (90–100 mm), internal O₂ transport was sufficient to support a rate of root elongation in the O₂-free medium of between 0.03 and 0.17 mm h^?1. When the O₂ pressure around the shoots was increased from 20 to 100 kPa O₂, the O₂ concentrations at the walls of the expanding zone (2–7 mm from the tip) of these roots increased from 0.006 mol m^?3 to between 0.04 and 0.26 mol m^?3, and the rate of root extension increased five-fold. Oxygen transport to roots grown continuously in acrated solutions was considerably less than for roots raised in stagnant solutions; this difference was greater for seminal than for nodal roots. When the acrated seminal roots were longer than 100 mm and transferred to an O₂-free root medium, O₂ concentration became zero at the root tip causing elongation to cease. After 24 h of anoxia, none of these roots were able to resume elongation following a return to acrated solutions.     

The effect of reactive oxygen species on ethylene production induced by osmotic stress in etiolated mungbean seedling

After 10 h osmotic stress in 25% polyethylene glycol (PEG6000) solution (–1.8 MPa) at 25 °C in darkness, the etiolated mungbean seedlings were transferred to pure water for recovery. The ethylene release rate and the level of reactive oxygen species (ROS), including superoxide radical (O₂^–) and hydrogen peroxide (H₂O₂), were investigated during the recovery process. The results showed that ethylene production rate and amount of ROS increased dramatically after osmotic stress, and a close correlation was observed between ethylene release rate and concentrations of ROS. Inhibitors of ethylene biosynthesis, aminoethoxyvinylglycine (AVG) or aminooxyacetic acid (AOA), could reduce the ethylene release rate, but had no significant influence to the content of O₂^– and H₂O₂. As well as, silver thiosulfate (STS), an inhibitor of ethylene action, exhibited no obvious effect to the concentration of ROS, showing stress-inducible ethylene was not the cause for the increase of stress-inducible ROS. On the other hand, exogenous generator of superoxide radical (methylviologen, MV, or sodium dithionite, Na₂S₂O₄) could enhance the ethylene production evidently, which could be inhibited by exogenous scavenger of superoxide radical (superoxide dismutase, SOD, or 1, 4-diazabicyclo (2,2,2) octane, DABCO). However, either exogenous H₂O₂ or catalase (CAT) had no significant influence on ethylene production. The results suggested that it was superoxide radical but not H₂O₂which was involved directly in osmotic stress-inducible ethylene biosynthesis. The dual-role of superoxide radical on stress ethylene biosynthesis was also discussed.     

Effect of PEG-6000 imposed drought stress on RNA content,relative water content (RWC), and chlorophyll content in peanut leaves and roots

Drought, one of the environmental stresses, plays crucial role in reduction in plant production on majority of agricultural fields of world, In order to evaluate drought stress on RNA content Relative water content (RWC), and chlorophyll content, Water deficit was induced by Polyethylene glycol (PEG) in peanut (Arachis hypogaea), accession number ICGV 91114. In this current study we evaluate RNA content and Relative water content (RWC) both in leaves and roots and chlorophyll content in leaf. The present study was undertaken with the aim to investigate the effect of water deficit imposed by PEG-6000, 40 old day seedlings were treated with varying concentrations of polyethylene glycol-6000 (PEG-6000; w/v-5%, 10%, 15% & 20%) for 24 h. The results showed that RNA content and Relative water content (RWC) content was significantly reduced in both leaves and roots with increased concentration of PEG, In leaves, a concentration dependent decline in chlorophyll content with increasing concentration of polyethylene glycol-6000 (PEG-6000). Reduction in chlorophyll ‘a’ level was to a greater extent than the chlorophyll ‘b’. Thus, this attributes can be used as screening tool for drought tolerance in peanut.     

Crystallization of pig mitochondrial aspartate aminotransferase by seeding with crystals of the chicken mitochondrial isoenzyme

Pig mitochondrial aspartate aminotransferase has been crystallized from polyethylene glycol solutions (M_r = 4000) with the aid of small seed crystals of the chicken mitochondrial isoenzyme. The “hanging drop” vapour diffusion technique was used. The unit cells of the pig and chicken mitochondrial isoenzymes are roughly isomorphous. Diffraction data have been collected to a resolution of 2.8 Å.     

Protection of cellulose synthesis in detached cotton fibers by polyethylene glycol

Detachment of the cotton fiber cell from the ovule results in loss of over 90% of the in vivo capacity for synthesis of [¹⁴C]cellulose from [¹⁴C]glucose. However, over 50% of the capacity for cellulose synthesis in the detached fiber population is protected when polyethylene glycol 4000 is present during detachment and incubation. Radioautography shows that approximately full capacity is restored in about half the fibers, whereas the other half of the population are incapable of cellulose synthesis from supplied glucose. The rate of cellulose synthesis in such fibers has a pH optimum of 6 and the optimum polyethylene glycol 4000 concentration is 0.06 molal (−9 bars). Cellulose synthesis in such detached fibers is synergistically stimulated by Ca²⁺ and Mg²⁺ and inhibited by K⁺.     

A comparison of different chemicals for osmotic treatment of vegetable seed

Samples of carrot, celery, leek and onion seed were treated before sowing by imbibition in osmotic solutions of polyethylene glycol 6000 (PEG), glycerol and potassium dihydrogen orthophosphate (KH₂PO₄). The solutions were sufficiently concentrated to prevent germination during treatment, and within each species, the amount of water taken up by the seeds during treatment did not vary greatly between solutions. All treatments increased the rates of seed germination and seedling emergence compared to untreated controls, but glycerol-treated seeds germinated and emerged significantly more slowly than did PEG- and KH₂PO₄-treated seeds. The effects of PEG and glycerol treatments on percentage germination and seedling emergence were small, but KH₂PO₄ treatment tended to reduce percentage germination and emergence, most markedly in leek and one cultivar of celery. It is concluded that PEG treatment gives the most consistently beneficial effects for the range of species tested.     

Steady-State, Hemoglobin-Facilitated O2 Transport in Human Erythrocytes

We measured the rate of oxygen transport through thin (165 µ) films of packed erythrocytes (Hb concentration = 30 g/100 ml). Under optimal conditions steady-state O₂ diffusion was nearly three times that found when the hemoglobin was prevented from acting as a carrier molecule by carbon monoxide binding or high oxygen back pressure. After each experiment we measured hemolysis and found that it averaged less than 1%. Hemolysis could not account for the facilitation, thus proving that facilitated transport of O₂ by hemoglobin can occur in red blood cells. The rate of facilitated transport was identical for Hb solutions of equal concentration to the cells. We interpret this to mean that under the conditions of our experiments the red cell membrane offers no detectable diffusion resistance to O₂ and that the mobility of Hb in intact red cells is the same as in concentrated Hb solution.