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1.
Partitioning of 15 proteins in polyethylene glycol (PEG)–sodium sulfate aqueous two-phase systems (ATPS) formed by PEG of two different molecular weights, PEG-600 and PEG-8000 in the presence of different buffers at pH 7.4 was studied. The effect of two salt additives (NaCl and NaSCN) on the protein partition behavior was examined. The salt effects on protein partitioning were analyzed by using the Collander solvent regression relationship between the proteins partition coefficients in ATPS with and without salt additives. The results obtained show that the concentration of buffer as well as the presence and concentration of salt additives affects the protein partition behavior. Analysis of ATPS in terms of the differences between the relative hydrophobicity and electrostatic properties of the phases does not explain the protein partition behavior. The differences between protein partitioning in PEG-600–salt and PEG-8000–salt ATPS cannot be explained by the protein size or polymer excluded volume effect. It is suggested that the protein–ion and protein–solvent interactions in the phases of ATPS are primarily important for protein partitioning.  相似文献   

2.
The present study was aimed to evaluate the effect of different seed priming methods to enhance the sodium chloride (NaCl) and polyethylene glycol-8000 (PEG-8000) stress tolerance in Indian mustard (Brassica juncea L.). Seeds subjected to different priming treatments such as water (hydro-priming), calcium chloride (CaCl2) (chemo-priming), and abscisic acid (ABA) (hormonal-priming) showed increased rate of germination as compared to non-primed seeds. The primed and non-primed seeds were grown for 15 days and then the seedlings were independently subjected to iso-osmotic salt (150 mM NaCl) or PEG-8000 (20%) stress. The different biochemical responses were studied 10 days after treatment. Under NaCl and PEG stress, the dry weight and total chlorophyll content were higher in primed sets as compared to non-primed treatment which was also evident by the phenotype of the seedlings. In general, the higher activities of superoxide dismutase and glutathione reductase resulted in lower oxidative damage, in terms of malondialdehyde content, under NaCl and PEG stress in hydro-primed set as compared to non-primed, ABA-, and CaCl2-primed treatments. Besides, the level of total phenolics and accumulation of osmolytes such as free proline, glycine betaine, and total soluble sugars was also lower in hydro-primed set as compared to other primed and non-primed treatments. The study thus suggests the use of hydro-priming as a simple and cost-effective strategy to alleviate the NaCl and PEG induced stress in B. juncea.  相似文献   

3.
Immunoadsorbents were modified with monomethoxy-polyethylene glycol (PEG; average molecular weights of 5000 (PEG-5000) and 1900 (PEG-1900)) activated with cyanuric acid (activated PEG) by four different methods. In the two methods, anti-BSA antibodies were modified with activated PEG with and without protection of antigen binding sites with BSA and then were coupled to CNBr-activated Sepharose 4B. In the other two methods, Immunoadsorbents, which were prepared by coupling anti-BSA antibodies to CNBr-activated Sepharose 4B, were modified with activated PEG with and without the protection. The effects of PEG modification by these four methods on the binding ratio (the ratio of the numbers of moles of antigen adsorbed to the numbers of moles of binding sites of antibody coupled), the antigen binding property and the resistance to proteolytic digestion of immunoadsorbents were studied. The decrease in the binding ratio by the modification with activated PEG was small enough to use modified immunoadsorbents for industrial purification processes. The resistance to proteolytic digestion of immunoadsorbents was improved by modification with activated PEG. The modification without protection of antigen binding sites gave higher resistance to proteolytic digestion than that with protection, while the former caused larger decrease in the binding ratio of modification. The immunoadsorbents modified with activated PEG-5000 showed higher resistance to proteolytic digestion than those modified with activated PEG-1900.  相似文献   

4.

Hyperhydricity is a physiological disorder during plant tissue culture that seriously affects regeneration and micropropagation. In this study, Dendrobium officinale plantlets were cultured on solid Murashige and Skoog (MS) medium supplemented with plant growth regulators and various concentrations of sucrose, agar, and polyethylene glycol (PEG)-6000 to explore the effect of osmotic stress on hyperhydricity. The results show that low concentrations of sucrose or agar, as well as PEG-6000 at various concentrations, significantly increase the hyperhydric rate of D. officinale, whereas high concentrations of sucrose or agar did not. Furthermore, high concentrations of PEG-6000 significantly increase total water content, free-water content, relative electrical conductivity, and peroxidase (POD) activity of D. officinale plantlets, whereas they significantly decrease bound-water content, proline content, soluble protein content, soluble sugar content, and superoxide dismutase (SOD) activity. These results indicate that PEG-6000 disrupts the antioxidant system and water metabolism in D. officinale plantlets, as well as increases cell membrane permeability, which might be the key factors for the occurrence of hyperhydricity in this species.

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5.
To promote the economic feasibility of Nannochloropsis oculata, efficacy of using polyethylene glycol (PEG) to increase microalgal growth and lipid accumulation was investigated. We first examined the effects of PEG concentrations on microalgal growth using 0–5 % (w/v) PEG-6000, and followed by exploring the effects of PEG molecular weights (400, 600, 2,000, 4,000, 6,000, and 20,000) on microalgal growth, size, as well as on yields of biomass, total lipids, and eicosapentaenoic acid. In addition, the capacity of PEG to reduce the effect of oxygen inhibition on microalgal growth was also investigated to evaluate its adaptability for use in large-scale and closed setting. Our results showed that PEG-induced osmotic stress (Π) in the range of 2.465–2.472 MPa can raise microalgal growth. The PEG with higher molecular weight exhibited greater efficacy of growth promotion but less lipid content under equal concentration. In this study, 0.5 % (w/v) PEG-20000 (Π = 2.466 MPa) remarkably enhanced microalgal growth without interference of intracellular lipid productivity and cellular size, yielding >50 % (w/w) increases in biomass, total lipid, and eicosapentaenoic acid amounts after 7 days that provided the optimal condition for microalgal cultivation. These positive effects possibly resulted from the moderate enhancement of osmotic stress in the medium and stronger chaotrope-like behavior from higher molecular weight PEG. With further verification that 0.5 % (w/v) PEG-20000 enabled to reduce the effect of oxygen inhibition on microalgal growth, the PEG-20000-mediated cultivation offers a feasible means for mass culture of N. oculata in closed setting.  相似文献   

6.
Seedlings of Cyamopsis tetragonoloba were grown on Petri dishes either in water or water plus 3 % PEG-6000 to induce water stress. The senescing cotyledons experiencing the stress exhibited loss in contents of leaf proteins and chlorophyll (Chl) and a decline in oxygen evolution. The effect of PEG treatment was more pronounced at moderate (MI) than low (LI) irradiance. The stress-induced loss in the activity of superoxide dismutase and increase in the thylakoid lipid peroxidation accompanied a change in the physical status of the bilayer membrane as demonstrated by an enhancement of room temperature Chl a fluorescence polarization and decrease in energy transfer efficiency in pigment assembly. This resulted in a sustained decrease in photosystem 2 activity blocking channels of energy utilization. The absorbed quanta, thus unutilized, were excess even at MI, leading to photoinhibitory response.  相似文献   

7.
In the present study, we evaluated the protective effect of nitric oxide(NO) against senescence of rice leaves enhanced by water deficit. Dehydration(DH), polyethylene glycol (PEG) and sorbitol (ST) treatments were used toinducewater deficit. Senescence of rice leaves was determined by the decrease ofprotein content. NO donors[N-tert-butyl--phenylnitrone (PBN), sodiumnitroprusside (SNP), 3-morpholinosydnonimine (SIN-1), and ascorbic acid +NaNO2] were effective in inhibiting senescence of dehydrated andPEG-treated rice leaves, but had no effect on senescence of ST-treated riceleaves. PEG or DH resulted in a marked increase in malondialdehyde (MDA)contentand decrease in superoxide dismutase (SOD) activity, but ST had no effect onMDAcontent and SOD activity. Treatment with NO donors caused a reduction of PEG-and DH-induced increase in MDA content and decrease in SOD activity. Theprotective effect of NO donors on promotion of senescence, increase in lipidperoxidation and decrease in SOD activity induced by PEG and DH was reversed by2-(4-carboxy-2-phenyl)-4,4,5,5- tetra-methylinmidazoline-1-oxyl-3-oxide, a NOspecific scavenger, suggesting that the protective effect of NO donors isattributed to NO released. The inhibition of PEG- and DH- enhanced senescenceofrice leaves by NO is most likely mediated through increasing SOD activity anddecrease in lipid peroxidation.  相似文献   

8.
吴敏怡  李霞  何亚飞  张琛  严婷 《植物研究》2017,37(3):402-415
为了研究高表达转玉米C4-磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因水稻(PC)的耐旱性机制,本研究以PC和未转基因野生型原种kitaake为材料,分别在光照和黑暗预处理24 h,其中光照处理中光强为600 μmol·m-2·s-1,预处理后稻苗再在15%聚乙二醇-6000模拟干旱胁迫下,同时使用药理学的方法,通过加入脱落酸和己糖激酶的专一性抑制剂100 μmol·L-1去甲二氢愈创木酸和10 mmol·L-1葡萄糖胺,观察两种水稻4~5叶期稻苗耐旱表现。结果发现,与WT水稻相比,在PEG-6000处理后,经过光预处理的PC水稻叶片相对含水量下降较少,始终比WT的高,而且丙二醛含量则较少,脯氨酸诱导增加,表现耐旱;而经过暗预处理后PC植株显著降低这个优势,表明光预处理有利于PC耐旱性的表现;黑暗预处理均显著下调了2供试材料植株叶片中可溶性糖的含量,而对可溶性蛋白的含量影响不显著;而光预处理后PC水稻叶片内可溶性糖含量比WT增加,而在黑暗预处理则PC的显著低于WT的,其中葡萄糖胺对光预处理下PC的可溶性糖含量的下调作用最显著;暗预处理逆转或消除了NO,H2O2和钙离子含量变化趋势,这些变化与暗预处理减少了两材料叶片蔗糖和葡萄糖含量变化同步;光暗预处理对两材料的PEPC酶活性的差异影响不大,表明外源玉米C4-PEPC在水稻中是组成型表达。可见PC叶片可部分通过糖组分,参与内源糖介导ABA和HXK信号途径,缓解干旱处理对叶片的伤害,稳定光合能力。  相似文献   

9.
Summary -Amylase production with Bacillus amyloliquefaciens was investigated in two different aqueous two-phase systems and in polyethylene glycol (PEG) 600 solutions of different concentrations. The cells did not partition totally to the bottom phases of the aqueous two-phase systems, and the enzyme production was repressed in both systems as well as in PEG 600 solutions. Concomitantly, the cultivation time was prolonged, indicating an increased maintenance metabolism. The surface properties of cells grown in 200 g/kg PEG 600 were investigated by phase partitioning and compared to the surface properties of Bacillus subtilis, which under these conditions showed increased -amylase production. The cells of B. amyloliquefaciens partitioned to the top phase in a PEG-dextran system, whereas the cells of B. subtilis partitioned to the bottom phase. The results are discussed in relation to water activity, oxygen transfer rate and PEG-induced changes of the surface properties of the cells. The possible role of PEG as an uncoupler of the proton motive force at high concentrations is also discussed.  相似文献   

10.
The effect of colonization of tissue-cultured strawberry ( Fragaria × ananassa Duch. cv. Kent) plantlets in vitro by the arbuscular mycorrhizal fungus (AMF) Glomus intraradices on plantlet response to poly(ethylene glycol) (PEG)-8000-induced water stress was investigated. The plantlets were inoculated axenically and co-cultured with the AMF for 4 wk, then transferred to 15% PEG-8000 solutions for 4, 8 and 12 h. Relative water content, water potential, osmotic potential, leaf conductance for water vapour diffusion and photosynthetic efficiency as estimated by chlorophyll a fluorescence were all affected by the PEG treatment and its duration but not by the presence of the intraradical phase of the AMF. However, distinct differences in PEG-induced changes in amino acid content were observed between nonmycorrhizal and mycorrhizal plantlets. In the latter, the treatment with PEG caused a substantial decrease in asparagine levels in leaves that was accompanied by a marked increase in asparagine concentration in roots. The opposite was observed in nonmycorrhizal plantlets. Furthermore, concentrations of aspartic acid, serine, threonine, amino- N -butyric acid, alanine and starch increased in roots of mycorrhizal and decreased in nonmycorrhizal plantlets. Our results suggest the presence of a mobile pool of asparagine that can be translocated from leaves to roots or vice versa in response to PEG-induced water stress, depending on the mycorrhizal status of the plantlets. These opposite patterns suggest different strategies of mycorrhizal and nonmycorrhizal plantlets to water stress, which seem to involve different adjustments in nitrogen and carbon metabolism.  相似文献   

11.
The effect of polyethylene glycol (PEG-6000), abscisic acid (ABA), 2-chloroethylphosphonic acid (ethephon) and 1-aminocyclopropane-1-carboxylic acid (ACC) on the germination of Amaranthus caudatus L. seeds was examined. Both PEG-6000 and ABA inhibited the rate and percentage of seed germination. ABA potentiated the effect of PEG. Ethephon was highly effective in reversing the inhibitory effect of PEG and ABA or combinations of both. ACC relieved inhibition by ABA and the combined effect of ABA and PEG. Aminoethoxyvinylglycine (AVG) increased the inhibition of seed germination caused by ABA. The inhibition of seed germination by ABA seems to be related more or less to ethylene biosynthesis or is associated with a change of tissue sensitivity to ethylene. The possibility of ethylene control of water uptake by seeds is also considered.  相似文献   

12.
Biotechnology industry has recently been demanding nanoparticulate products (20-200 nm) such as viruses, plasmids, virus-like particles and drug delivery assemblies. These products are mainly used as gene delivery systems in gene therapy protocols. During the process development for the manufacture of these products, it is crucial to optimize the recovery and purification steps. Unfortunately, the high value of some bio-nanoparticles complicates the optimization studies. The solvent extraction method with aqueous two-phase systems (ATPS) has been used to successfully recover bioproducts on a large scale. In this study, the potential miniaturization of ATPS is presented. The partition behavior of pure bovine serum albumin (BSA) in PEG-800-phosphate and bacteriophage T4 in PEG 8000-phosphate and PEG 600-sulphate systems were studied at three different scales (10 g, 2 g and 300 microl). The results obtained showed that the volume ratio (V(R)) for BSA (V(R)=1.0) was comparable to the blank systems at the scales studied. Additionally, the partition coefficient (K) was also similar (K=0.05) with more than 82% of BSA concentrated in the bottom phase. Same system was challenged with bacteriophage T4 showing a V(R)=1.0 and K greater than 5 with the infective particles concentrated in the top phase. The bacteriophage T4 was concentrated in opposite phase in the PEG-600-sulfate system with a consistent V(R)=0.8 and K<0.2 for the scales analyzed. The partition behavior the bacteriophage T4 was comparable to that reported previously for adenoviral vectors in same system at 15 ml scale. The results obtained demonstrated that the miniaturization of ATPS is feasible and reproducible for the two models selected. This provides significant information about the miniaturization process of such ATPS for their potential generic applications in the recovery of different bio-nanoparticle products.  相似文献   

13.
Extraction in a polyethylene glycol (PEG)–phosphate aqueous two-phase system was considered as a primary step in purification of the acetohydroxy acid synthase III large catalytic subunit from an E. coli extract. Extraction optimization was achieved by varying the system parameters. Two systems with the following weight compositions were chosen for purification: PEG-2000 (16%)–phosphate (6%) and PEG-4000 (14%)–phosphate (5.5%)–KCl (8%), both at pH 7.0 and 1 mg total protein per 1 g system. Significant purification was achieved by a single extraction step with 70% recovery of the enzyme. After an additional ion-exchange chromatography step, pure enzyme was obtained in a 50% overall yield.  相似文献   

14.
The rate of synthesis of RNA was studied under salt and water stresses created by the addition of four different salts and polyethylene glycol-6000 (PEG-6000) in excised embryo axis of mung bean usingl4C-uracil. Iso-osmotio concentrations of salts and PEG resulted in differential effect on RNA synthesis. PEG inhibited, whereas salt stimulated incorporation of labelled uracil into RNA. Ionic differences on RNA synthesis were very clear as synthesis of RNA was more affected with sulphate than chloride and with sodium than potassium ions.  相似文献   

15.
马文静  魏小红  宿梅飞  骆巧娟  赵颖 《生态学报》2019,39(21):8068-8077
以紫花苜蓿(Medicago sativa)为材料,采用盆栽试验方法,用聚乙二醇(PEG-6000)作为渗透介质模拟干旱胁迫,外源喷施NO供体硝普钠,NO清除剂(carboxy-PTIO,cPTIO),对紫花苜蓿幼苗叶片、根系中非结构性碳水化合物含量及相关酶活性的变化进行研究,探讨NO对紫花苜蓿耐旱机制的作用。结果表明:外源NO促进了紫花苜蓿叶片中淀粉的分解、根系中淀粉的积累,提高叶片及根系中可溶性糖(蔗糖、果糖和葡萄糖)含量,降低了渗透势,促进细胞吸水,缓解干旱造成的损伤。此外,外源NO能提高干旱胁迫下紫花苜蓿叶片中蔗糖合成酶(SS)、酸性转化酶(AI)和中性转化酶(NI)活性,降低了蔗糖磷酸合成酶(SPS)的活性,提高根系中SS、SPS和转化酶活性,使蔗糖的合成与分解处于高水平的动态平衡,增强了紫花苜蓿的抗旱性。而NO清除剂cPTIO则会不同程度的抑制紫花苜蓿幼苗中非结构性碳水化合物(NSC)及其相关酶活性。因此,NO可以通过调控NSC的代谢响应干旱胁迫,缓解干旱胁迫造成的不利影响,在紫花苜蓿的抗旱中扮演着重要的角色。  相似文献   

16.
Three strains of Pseudomonas oleovorans, a well known poly(hydroxyalkanoate) (PHA) producer, were tested for the ability to control PHA molar mass and end group structure by addition of poly(ethylene glycol) (PEG) to the fermentation medium. Each strain of P. oleovorans - NRRL B-14682 (B-14682), NRRL B-14683 (B-14683), and NRRL B-778 (B-778) - synthesized a different type of PHA from oleic acid when cultured under identical growth conditions. Strain B-14682 produced poly(3-hydroxybutyrate) (PHB), while B-14683 synthesized a medium-chain-length PHA ( mcl-PHA) with a repeat unit composition ranging from C4 to C14 and some mono-unsaturation in the C14 alkyl side chains. Strain B-778 synthesized a mixture of PHB (95 mol%) and mcl-PHA (5 mol%). The addition of 0.5% (v/v) PEG (M(n) =200 g/mol, PEG-200) to the fermentation broth of strains B-14682 and B-778 resulted in chain termination through esterification at the carboxyl terminus of the PHB with PEG chain segments, thus reducing the molar mass by 54% and 23%, respectively. The molar mass of the mcl-PHA produced by strains B-14683 and B-778 also showed a 34% and 47% reduction in the presence of PEG-200, respectively, but no evidence of esterification was present. PEG-400 (M(n) =400 g/mol) had a reduced effect on PHA molar mass. In fact, the molar masses of the mcl-PHA derived from strain B-14683 and both the PHB and mcl-PHA from B-778 were unchanged by PEG-400. In contrast, the PHB produced by B-14682 showed a 35% reduction in molar mass in the presence of PEG-400.  相似文献   

17.
Azotobacter chroococcum MAL-201 (MTCC 3853), a free-living nitrogen-fixing bacterium accumulates poly(3-hydroxybutyric acid) [PHB, 69% of cell dry weight (CDW)] when grown on glucose and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [PHBV with 19.2 mol% 3HV] when grown on glucose and valerate. Use of ethylene glycol (EG) and/or polyethylene glycols (PEGs) of low molecular weight as sole carbon source were detrimental to A. chroococcum growth and polymer yields. PEG-200, however, in the presence of glucose was incorporated into the polyhydroxyalkanoate (PHA) polymer. Addition of PEG-200 (150 mM) to culture medium during mid-log phase growth favored increased incorporation of EG units (12.48 mol%) into the PHB polymer. In two-step culture experiments, where valerate and PEG simultaneously were used in fresh medium, EG was incorporated most effectively in the absence of glucose, leading to the formation of a copolymer containing 18.05 mol% 3HV and 14.78 mol% EG. The physico-mechanical properties of PEG-containing copolymer (PHBV–PEG) were compared with those of the PHB homopolymer and the PHBV copolymer. The PHBV–PEG copolymer appeared to have less crystallinity and greater flexibility than the short-chain-length (SCL) PHA polymers.  相似文献   

18.
巴东木莲原生种群天然更新差,林下鲜见更新幼苗或幼树.为探究其种子萌发和芽苗在生长期对干旱胁迫的耐受性,该文以巴东木莲种子为材料,采用不同质量浓度的聚乙二醇(PEG-6000)模拟干旱胁迫,分析干旱胁迫对其种子萌发、芽苗生长及其相关的生理生化指标的影响.结果表明:(1)不同质量浓度的PEG溶液对巴东木莲种子萌发和芽苗生长...  相似文献   

19.
周玲  王乃江  张丽楠 《西北植物学报》2012,32(11):2293-2298
在种子发芽箱中,以清水为对照,用5%、10%、15%、20%、25%、30%和35%共7个浓度聚乙二醇(PEG-6000)溶液浇灌栽培基质,观察文冠果种子的萌发和幼苗生长情况,以及幼苗叶片脯氨酸、丙二醛的含量和细胞膜相对透性的变化,探讨文冠果种子萌发对土壤水分的要求及其幼苗忍耐干旱的能力。结果显示:(1)在PEG浓度5%~35%范围内,文冠果种子发芽率、成苗率、幼苗根长和细胞膜透性均随着胁迫强度的增加呈明显的下降趋势;细胞膜透性随着胁迫强度的增加先升高后降低,但脯氨酸、丙二醛含量和幼苗根数量则呈现逐渐上升趋势。(2)5%的PEG浓度能够促进文冠果种子萌发,提高成苗率和根数量,降低幼苗死亡率;5%~10%的PEG能够明显促进苗高生长,但对根长影响不大;当PEG浓度高于15%后,种子萌发受到抑制,成苗率明显降低。(3)当PEG浓度高于25%时幼苗死亡率急剧上升,幼苗脯氨酸含量、丙二醛含量和细胞膜透性显著增加,细胞膜结构受到严重伤害。研究表明,低浓度PEG处理有利于文冠果种子萌发和幼苗生长,但过高浓度却对文冠果种子萌发和幼苗生长具有一定的抑制作用;文冠果种子萌发及其幼苗可忍受5%~25%的PEG渗透胁迫,即文冠果种子萌发和幼苗生长可忍受土壤水势为-7.94~-11.05MPa的干旱胁迫。  相似文献   

20.
The partitioning behavior of the pertechnetate anion was studied in aqueous biphasic systems (ABS) formed from (NH4)2SO4 and four types of polymers – poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), Pluronic (a PEG/PPG block copolymer), and polyvinylpyrrolidone (PVP). Phase diagrams are reported for five (NH4)2SO4-polymer ABS systems including the polymers PEG-2000, PEG-3400, PEG-12 000, Pluronic-L64 (average molecular mass ≈ 2900), and PVP-K15 (average Mr≈10 000). Distribution ratios for the TcO4 anion in each of these ABS were investigated as a function of increasing salt concentration. In addition, the water-insoluble polymer PPG-2000 was studied. Pertechnetate partitions nearly quantitatively to the polymer-rich phase in each ABS, however, distribution ratios of near one were found for the PPG system. The relative ordering of the distribution ratios is PPGPVP-2000PEG-3400>PEG-12 000, exhibiting the expected increase in phase incompatibility with increasing polymer Mr. Investigation of pertechnetate partitioning in two additional ABS based on K3PO4 and NaOH with Pluronic-L64 revealed trends similar to those reported for PEG-2000; the distribution ratio (D) values increase in the order NaOH<(NH4)2SO43PO4. Despite the higher distribution ratios from Pluronic-L64 at lower concentrations than found for PEG, the limited useable range of salt concentrations available may limit the practical utilization of this polymer in ABS separations.  相似文献   

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