Visualization of single and aggregated hulless oat (Avena nuda L.) (1-->3),(1-->4)-beta-D-glucan molecules by atomic force microscopy and confocal scanning laser microscopy

Surfactants were used to disperse oat beta-glucan. Atomic force microscopy (AFM) images of the resulting samples revealed a distribution of extended chainlike molecules and allowed, for the first time, direct visualization of single oat beta-glucan molecules with cross-sectional heights of about 0.44 nm. The number-average contour length (L(n)) and root-mean-square end-to-end distance ((R(ee)2)(1/2)) measured from the AFM images were 938 and 912 nm, respectively. The calculated persistence length (L(p)) was 526 nm. The weight-average molecular weight (M(w)) calculated from single beta-glucan molecules was 4.43 x 10(5). Samples without surfactant showed a strong tendency to form aggregates. The sample concentration, reserving time, and calcofluor as well as freezing could affect the formation of aggregates. These aggregates were visualized by both AFM and confocal scanning laser microscopy. The shape of the aggregates changed from small dots with diameters of approximately 20-50 nm to microfibrils over 3 microm long with the increasing of the concentration of oat beta-glucan from 10 to 100 microg/mL. The particle size distribution obtained by a laser particle size analyzer was 926 nm, which confirmed the size of oat beta-glucan molecules obtained from AFM images.     

碱解还原扩散法测定缓释复合肥氮素释放特性研究

以自制的新型非包膜有机无机缓释复合肥料（简称缓释复合肥,Slow-release compound fertilizer,SRCF）为供试材料,采用连续碱解还原扩散法研究氮素养分缓释过程。试验结果表明,1）以硼酸为吸收液时,NaOH浓度在0～0.1 mol/L范围内, SRCF在40 d时的氮素累积扩散率随碱的浓度增加而提高,且累积扩散率显著高于较高浓度碱液（0.5～2 mol/L NaOH）的处理效果;2）同一NaOH浓度下,0.1 mol/L H2SO4作吸收液时,SRCF的累积扩散率高于0.1 mol/L H3BO3作吸收液的累积扩散率;3）0.05和0.01 mol/L NaOH浓度下,SRCF的氮素扩散特征与土壤肥包培养下的氮素矿化过程具有较好的相似性,能充分指示有机无机缓释复合肥氮素形态的多样性。连续还原扩散法测定有机无机缓释复合肥氮素扩散释放的适宜条件为:0.05～0.1 mol/L NaOH为扩散介质,0.1 mol/L H2SO4为吸收介质,25℃恒温培养（40 d）,在不同时段滴定吸收介质。     

Production of extracellular water-insoluble polysaccharide from Pseudomonas sp

Curdlan is a microbial polysaccharide composed exclusively of β-(1,3)-linked glucose residues. Until now only bacteria belonging to the Alcaligenes and Agrobacterium species have been reported to produce Curdlan. In this study, a bacterium capable of producing extracellular Curdlan, identified as Pseudomonas sp. on the basis of 16S rDNA gene sequencing, was isolated from soil samples. From the HPLC, permethylation linkage analysis, (13)C NMR, and FT-IR analytical data, the polysaccharide consisted exclusively of glucose; the most prominent sugar was 1,3-linked glucose, and most glycosidic bonds joining these sugar residues were of the β-type. This also supported that the exopolysaccharide produced by Pseudomonas sp. was actually Curdlan. In addition, the Pseudomonas sp. was studied for the production of Curdlan by conventional "one-factor-at-a-time technique" and response surface methodology (RSM). It was observed that glucose and yeast extract were the most suitable carbon source and nitrogen source for Curdlan production, respectively. By using RSM, Curdlan production was increased significantly by 188%, from 1.25 to 2.35 g/L, when the strain was cultivated in the optimal condition developed by RSM, and the highest Curdlan production rate of 0.81 g/(L h) was obtained. To the best of the authors' knowledge, this is the first report on Curdlan production by Pseudomonas sp.     

铵态氮营养下水稻根系分泌氢离子与细胞膜电位及质子泵的关系

为了研究水稻在铵态氮营养下分泌氢离子的机理,采用不同浓度的铵态氮（0.1～1.0 mol/L）处理水稻幼苗根系,4h后用1 mol/L NaOH滴定培养液,计算氢离子的分泌量;同时,将水稻根系用多聚糖PEG-DEXTRAN两相系统分离出细胞膜囊体,并测定细胞膜H+-ATPase的水解活性和质子泵活性。另外,利用毛细管微电极测定水稻根细胞在上述不同铵浓度下膜电位的变化,以阐明水稻根系吸收铵态氮后分泌氢离子与细胞膜电位及细胞膜质子泵之间的关系。结果表明,随着培养液中铵离子浓度的升高,根系分泌氢离子的量随之增加;分离细胞膜后,离体细胞膜囊体H+-ATPase的水解活性和质子泵活性也相应增强。原位测定细胞膜电位时,膜电位去极化程度亦随NH4+浓度的升高而升高;氢离子分泌量与细胞膜电位、细胞膜H+-ATPase水解活性及质子泵活性之间的均具有一定的相关性。说明根系在NH4+-N营养下分泌氢离子是由于细胞膜上H+-ATPase主动泵出氢离子造成的,这与根系吸收NH4+后引起细胞膜去极化,需要通过提高质子泵活性来维持膜电位有关。     

~(60)Coγ射线诱变选育热凝胶多糖高产菌株的研究

利用60 Coγ射线对产生热凝胶多糖 (Curdlan)出发菌株GM 2 4的菌体细胞与原生质体分别进行辐照处理 ,发现60 Coγ射线对GM 2 4原生质体的诱变效应明显优于对其菌体细胞的作用。对最终诱变筛选获得的 1株高产稳定的Curdlan生产突变株A81研究表明 :与GM 2 4相比 ,其Curdlan产量提高了 5 0 4%,发酵周期缩短了 1 8%,发酵终止后底物残糖由 1 5 3g L降至7 4g L。糖的转化率从 38 7%提高至 5 8 2 %。这说明在Curdlan生产菌株的筛选中60 Coγ辐照是一种有效的方法     

Dynamic interfacial rheology as a tool for the characterization of whey protein isolates gelation at the oil-water interface.

Heat-induced interfacial aggregation of a whey protein isolate (WPI), previously adsorbed at the oil-water interface, was studied by interfacial dynamic characteristics coupled with microscopic observation and image analysis of the drop after heat treatment. The experiments were carried out at temperatures ranging from 20 to 80 degrees C with different thermal regimes. During the heating period, competition exists between the effect of temperature on the film fluidity and the increase in mechanical properties associated with the interfacial gelation process. During the isothermal treatment, the surface dilational modulus, E, increases, and the phase angle, delta, decreases with time to a plateau value. The frequency dependence of E and delta is characteristic of viscoelastic films with increasing delta and decreasing E at lower frequencies. The effects of heat treatment depend on the conditions at which the gelation process takes place. Microscopic observation of gelled films gives complementary information on the effect of heat treatment on WPI adsorbed films.     

The maize primary cell wall microfibril: a new model derived from direct visualization

Understanding the molecular architecture of the plant cell wall is critical to reducing the biomass recalcitrance problem, which currently impedes economic bioconversion processing. The parenchyma cell walls from field senesced, maize stem pith have been directly visualized without extraction processes using high-resolution atomic force microscopy (AFM). By imaging the cell wall inner surfaces from different cells and different faces of the same cell, we were able to map the native primary cell wall ultrastructures. Depending on the thickness of non-cellulosic deposition, the parallel-microfibrils appear in various morphologies ranging from clearly defined to completely embedded in the wall matrixes forming cell wall lamella. Macrofibrils were found to exist only on the uppermost layer of the native primary cell wall and appeared to be bundles of elementary fibrils. This novel observation led us to a new hypothesis for the cell wall fibrillar network and biosynthesis processes. Put concisely, a number of elementary fibrils are synthesized at one locus, that of the cellulose synthase complex (CelS), and coalesce into much larger macrofibrils. These macrofibrils eventually split at the ends to form parallel microfibrils with deposition of other cell wall components (i.e. hemicelluloses, pectin, etc.) also evident. On the basis of these AFM surface measurements and current supportive evidence from cell wall biophysics, biosynthesis, and genomics, we propose a new molecular model consisting of a 36-glucan-chain elementary fibril, in which the 36-glucan chains form both crystalline and subcrystalline structures. We also propose a modified model of CelS based on recently reported experimental evidence from plant cell wall biosynthesis.     

Rheological Studies on the Reaction of Zein with Polyethylenemaleic Anhydride

There continues to be interest in developing solvent‐resistant articles from biobased renewable materials to successfully compete with petrochemical products. It was previously shown that reaction of zein with polyethylenemaleic anhydride (PEMA) provides articles that are solvent resistant. The gelation kinetics for the reaction of PEMA with zein was investigated rheologically to better understand this chemistry. The reaction of the nucleophilic groups on zein with the anhydrides on PEMA is the main cause for the gelation reaction. The gelation time was defined as being the point when the elastic modulus (G′) and viscous modulus (G″) cross. In this work, the rate of reaction, in terms of time to gelation, was studied in N,N‐dimethylformamide solution for which the amount of PEMA, the reaction temperature, and the overall reaction concentration were varied. Exponential relationships were found between the gelation time and % PEMA, temperature, and % solids, as well as between elastic modulus with either % PEMA or % solids. The concentration of PEMA had the largest impact on gelation time, for which going from 2.5% PEMA to 6% PEMA reduced the gelation time from 63,114 to 1,576 s. The temperature dependence of this gelation reaction was well described by an Arrhenius plot with an apparent activation energy of 50.5 kJ/mol.     

Waste water from citrus processing as a source of hesperidin by concentration on styrene-divinylbenzene resin

This paper describes a procedure for recovering hesperidin from the waste water of orange juice processing, namely, yellow water, by concentration of diluted extracts on styrene-divinylbenzene resin. Turbid raw material flowing out from centrifuges of essential oil separation contains considerable amount of hesperidin ( approximately 1 g/L) mainly associated with solid particles. Yellow water was treated with calcium hydroxide until pH 12 to solubilize hesperidin, filtered, neutralized at pH 6, and loaded on resin up to saturation. Desorption with 10% ethanol aqueous solutions at different NaOH concentrations (0.23-0.92 M) assured high concentration of hesperidin in selected fractions (10-78 g/L), from which it precipitated in high yield and purity immediately after acidification at pH 5. Best results were obtained using 0.46 M NaOH as eluent: 71.5% of the adsorbed hesperidin was desorbed in 300 mL, with an overall 64% yield of isolated product at 95.4% purity (HPLC). These experiments can constitute a useful starting point for an industrial application.     

Identification of (poly)phenolic compounds in concord grape juice and their metabolites in human plasma and urine after juice consumption

Analysis of Concord grape juice by HPLC with ESI-MS(n), PDA, and fluorescence detection resulted in the identification and quantification of 60 flavonoids and related phenolic compounds, which were present at an overall concentration of 1508 ± 31 μmol/L. A total of 25 anthocyanins were detected, which were mono- and di-O-glucosides, O-acetylglucosides, O-p-coumaroyl-O-diglucosides, and O-p-coumaroylglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. The anthocyanins represented 46% of the total phenolic content of the juice (680 μmol/L). Tartaric esters of hydroxycinnamic acids, namely, trans-caftaric and trans-coutaric acids, and to a lesser extent trans-fertaric acid accounted for 29% of the phenolic content, with a total concentration of 444 μmol/L, of which 85% comprised trans-caftaric acid. Free hydroxycinnamic acids were also quantified but contributed to <1% of the total phenolic content (8.4 μmol/L). The other groups of polyphenolic compounds present in the juice, accounting for 24% of the total, comprised monomeric and oligomeric units of (epi)catechin and (epi)gallocatechin (248 μmol/L), flavonols (76 μmol/L), gallic acid (51 μmol/L), and trans-resveratrol (1.5 μmol/L). The bioavailability of the (poly)phenolic compounds in 350 mL of juice was investigated following acute intake by healthy volunteers. Plasma and urine were collected over 0-24 h and analyzed for parent compounds and metabolites. In total, 41 compounds, principally metabolites, were identified.     

Controlled proteolysis and the properties of milk gels

Milk gels induced by partial proteolysis of the kappa-casein followed by acidification were studied, and their gelation behavior was compared to that of milk gels induced by simultaneous acidification and renneting, using dynamic rheology. There were generally two stages (at pH values below and above 5.0) in the gelation of the milk whose kappa-casein had been partially proteolyzed and acidified. The onset of gelation was at higher pH as the degree of kappa-casein proteolysis increased. The development of G' immediately after the onset of gelation was faster in the milk gels induced by simultaneous acidification and renneting, because of the continuing kappa-casein proteolysis. Preheat treatment caused the onset of gelation to occur at higher pH than for unheated milk. However, the maximum tan delta during gelation always occurred at the same pH (for a given concentration of acidulant), and its value and position were independent of the extent of renneting and whether the milks had been heat treated. The results are discussed in terms of the interactions between casein micelles occurring during gelation.     

红壤中铝的形态

以改进的连续分级提取方法,用１ｍｏｌ／Ｌ,ＫＣｌ,０．２ｍｏｌ／Ｌ,ＨＣｌ,０．１ｍｏｌ／Ｌ Ｎａ４Ｐ２Ｏ７（ｐＨ８．５）,ＤＣＢ溶液,０．３３ｍｏｌ／Ｌ,柠檬酸钠和０．５ｍｏｌ／Ｌ ＮａＯＨ为提取剂,把红壤中可提取的非晶态铝区分为交换态铝,肿附态无机羟基铝,有机配合态铝,氧化铁结合态铝,层间铝和非晶态铝硅酸盐。     

有机酸根与铝氧化物表面吸附磷的解吸

研究了有机酸根离子与合成铝氧化物表面吸附磷解吸的相互关系。结果表明:（1）有有机酸时比无有机酸时吸附的磷具有更高的解吸率,无草酸且加磷pH为4时,0.01molL-1KCI对磷的解吸率最低,草酸与磷共存且pH为6时磷的解吸率最高;（2）高浓度有机酸可解吸低浓度有机酸难解吸的磷,草酸难解吸的磷可为等浓度的柠檬酸极解吸;（3）不同浓度草酸和柠檬酸对铝-草酸复合物吸附磷的解吸率比对Al（OH)x的低,而对铝-柠檬酸复合物吸附磷的解吸率则比对Al（OH）x的高;（4）柠檬酸对铝-磷复合物中磷的解吸量随柠檬酸浓度升高而增大。这些结果证明,有机酸对铝氧化物吸附磷的解吸机理包括配位交换和溶解,有机酸可促进磷的解吸,提高磷的有效性。     

魔芋辐照接枝丙烯酸及其产物吸水性能的研究

研究了魔芋-丙烯酸辐照接枝共聚条件及产物的吸水性能,结果表明接枝最佳配方为:魔芋粉∶丙烯酸∶NaOH(9.5mol/L)∶水=3g∶21ml∶17.8ml∶131ml。丙烯酸与魔芋粉配比为7∶1(体积质量比);丙烯酸中和度55%;辐照后胶体最佳烘干温度30℃,该温度下40~80目颗粒吸水倍数较大,常温120min时达最大吸水866.5倍。温度对产物吸水和胶体保水有一定影响,以60℃以下为宜。pH对产物吸水倍数影响明显,但不及离子浓度对产物吸水倍数的影响大。     

栝楼ISSR－PCR反应体系的建立和优化

用改良SDS法提取栝楼（Trichosanthes kirilowii Maxim）叶片DNA为模板,通过单因素试验研究模板、引物、Tat／DNA聚合酶和dNTPs等4因素各7个水平对栝楼ISSR-PCR扩增结果的影响,并通过正交试验对各因子的浓度进行优化,首次建立了栝楼ISSR-PCR分析的最适反应体系：20μL的反应体系中,1×buffer,模板含量为50ng,引物的浓度为0．5μmol／L,TaqDNA聚合酶的用量为0．8U,dNTPs的浓度为200μmol／L。随机选取10条引物验证体系稳定性,并对190条ISSR引物初筛,得到条带清晰、多态性好的引物126条。经验证,本实验所建立的栝楼ISSR-PCR反应体系具有稳定可靠、可重复性好、多态性较强等特点,能够较好的应用于栝楼的ISSR分子生物学分析。     

Study on the supramolecular interaction of thiabendazole and beta-cyclodextrin by spectrophotometry and its analytical application

The beta-cyclodextrin-thiabendazole (beta-CD-TBZ) inclusion complex was synthesized and its structure characterized by (1)H NMR and IR. The mechanism of the supramolecular interaction of TBZ and beta-CD has been studied and discussed by spectrophotometry. The results showed that the phenyl ring of TBZ was included in the beta-CD cavity to form a 1:1 host-guest complex with an apparent formation constant of 1.60 x 10(3) mol(-1).L. On the basis of the enhancement of the absorbance of TBZ produced through complex formation, a spectrophotometric method for the determination of TBZ in bulk aqueous solution in the presence of beta-CD was developed. The linear relationship between the absorbance and TBZ concentration was obtained in the range of 8.86 x 10(-7)-1.45 x 10(-5) mol/L. The detection limit was 2.71 x 10(-7)mol/L, and the relative standard deviation was 0.86%. The interference of 48 coexisting substances was slight. The proposed method has been successfully applied to the determination of TBZ in fruits with recoveries of 96-103%.     

Limited enzymatic treatment of skim milk using chymosin affects the micelle/serum distribution of the heat-induced whey protein/kappa-casein aggregates

The effects of heat treatment and limited kappa-casein hydrolysis on the micelle/serum distribution of the heat-induced whey protein/kappa-casein aggregates were investigated as a possible explanation for the gelation properties of combined rennet and acid gels. Reconstituted skim milk was submitted to combinations of 0-67% hydrolysis of the kappa-casein at 5 degrees C and heat treatment at 90 degrees C for 10 min. The protein composition of the ultracentrifugal fractions was obtained by reverse-phase high-performance liquid chromatography (RP-HPLC). The aggregates contained in each phase were isolated by size-exclusion chromatography and analyzed by RP-HPLC and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Upon heating only, 20-30% of the total kappa-casein dissociated, while 20-30% of the total whey protein attached to the micelles. When heated milk was renneted, little changes were observed in the distribution and composition of the aggregates. Conversely, the heat treatment of partially renneted milk induced the formation of essentially micelle-bound aggregates. The results were discussed in terms of the preferred interaction between hydrophobic para-kappa-casein and denatured whey proteins.     

A role for pectin in the control of cell expansion

Uptake of nutrients and water depends on the growth of roots through elongation of individual cells near the root tip. Many of the numerous components of Type I primary cell walls, those of dicotyledons and monocotyledons other than grasses (Poaceae), have been determined, and many hypotheses have been proposed for the control of cell expansion. This important aspect of plant growth still needs elucidation, however. A model is proposed in which pectin, which occurs as a calcium (Ca) pectate gel between the load-bearing cellulose microfibrils and xyloglucan (XG) chains, controls the rate at which cells expand. It is considered that the increasing tension generated by the expanding cell is transmitted to interlocked XG chains and cellulose microfibrils. The resulting deformation of the embedded Ca pectate gel elicits the excretion of protons from the cytoplasm, possibly via compounds such as cell wall-associated kinases, that weakens the Ca pectate gel, permitting slippage of XG molecules through the action of expansin. Further slippage is prevented by deformation of the pectic gel, proton diffusion, and the transfer of residual tension to adjacent XG chains. Evidence for this model is based on the effects of pH, Ca, and aluminum (Al) on root elongation and on the reactions of these cations with Ca pectate. This model allows for genetic selection of plants and adaptation of individual plants to root environmental conditions.     

Bowman-Cole石灰性土壤有机磷分组法的改进

本文研究了Bowman-Cole土壤有机磷分组法应用于石灰性土壤存在的若干问题,并提出了改进法。改进法的四组土壤有机磷(P0)的分组为:在氯仿预处理土样后用0.5molL-1NaHCO3浸提活性P0;然后,先用0.05molL-1NaOH、后用1molL-1H2SO4处理残余土样浸提中度活性P0,0.05molL-1NaOH提取的无机磷和1molL-1H2SO4提取的有机磷属于中度活性P0;在氯仿和0.5molL-1NaHCO3处理后,用0.05molL-1NaOH浸提稳定性P0,把浸提液的酸度调至pH3.00时,上清液中的有机磷为中度稳定性P0,而沉淀中的有机磷为高稳性P0。结果表明,改进法能更好地区分具有不同矿化率的有机磷化合物,例如,核酸、甘油磷酸、植酸钙、肌醇三磷酸铁、植酸铁等。改进法增加了超声波处理,使震荡时间也明显缩短。因此,石灰性土壤有机磷分组的改进法比Bowman-Cole法更理想。     

Remediation of a Diesel Contaminated,Sandy-Loam Soil Using Low Concentrated Surfactant Solutions (5 pp)

Background, Aims and Scope   Surfactant enhanced ex-situ soil washing can be used to remediate diesel contaminated soils. Surfactants enhance the diesel removal from soils by two processes: mobilization and solubilization. Mobilization occurs at surfactant concentrations below the critical micelle concentration (CMC), while solubilization occurs at surfactant concentrations beyond the CMC of the surfactant. In this paper, the leaching of diesel from a self contaminated (10 000 mg diesel/kg) sandy-loam soil was studied using low concentrated non-ionic surfactant solutions of Tergitol NP-10 (10-6 to 10-3 mol/L). Methods   The surface tension of the supernatant solutions of soil suspensions in non-ionic surfactant (Tergitol NP-10) solutions (10-6 to 10-3 mol/L) was measured to determine the total surfactant concentration (surfactant sorbed onto the soil + surfactant in solution) at which micelles are present in the water phase of the soil suspension, or 'effective critical micelle concentration' (ECMC), and to calculate the sub-CMC sorption isotherm of Tergitol NP-10 onto the soil. The diesel removal was measured by soil leaching experiments with Tergitol solutions ranging from 10-6 to 10-3 mol/L. Results and Discussion   The sub-CMC sorption isotherm of Tergitol onto the soil could be described by the Freundlich equation. Even at very low surfactant concentrations (10-6 mol/L) the surfactant enhanced the diesel removal from the soil. Up to the surfactants' ECMC the diesel removal increased in a linear way with an increasing surfactant concentration in the wash water. A significant part (20%) of the diesel oil was removed in the presence of surfactant, but in absence of micelles. Beyond the surfactants' ECMC, the increase in diesel removal efficiency with increasing surfactant concentration was less pronounced. When the added surfactant concentration was increased to 10-3 mol/L, which corresponds to an equilibrium surfactant concentration in the supernatant solution (6.77.10-4 mol/L) above the CMC, the diesel oil removal was increased up to 50%. At this surfactant concentration emulsion formation was observed. Conclusion   Surfactant aided leaching of diesel from a self-contaminated, sandy-loam soil with surfactant solutions below their ECMC was able to remove in one step 20% of diesel from the soil by the mobilization mechanism. Succesive treatments or continuous leaching with surfactant solutions below their ECMC was not studied but is likely to reduce the diesel contamination further and at the same time avoid problems with emulsion formation.