Significance of LMW-GS and HMW-GS for Dough Extensibility: «Addition» versus «Incorporation» Protocols

Low molecular weight (LMW-GS) and high molecular weight glutenin subunits (HMW-GS) were added to a base flour using both «addition» and «incorporation» protocols. «Incorporation» of glutenin subunits into the glutenin network of the base flour was performed by partial (reversible) reduction and subsequent reoxidation of the glutenin network in the presence of the added glutenin subunits whereas, in the «addition» protocol, glutenin subunits were added without reduction/oxidation. The effects of both «addition» and «incorporation» of alkylated and unalkylated LMW-GS and HMW-GS on dough extension parameters maximum resistance (MR) and extensibility (EX) were compared and thoroughly discussed. HMW-GS and LMW-GS had totally different effects on dough extensibility. «Addition» of LMW-GS significantly decreased both MR and EX whereas HMW-GS caused a significant increase in MR. «Incorporation» of LMW-GS caused a decrease in MR whereas HMW-GS clearly increased MR. The similarity in effects obtained with «addition» and «incorporation» of glutenin subunits indicated that, even with «addition», glutenin subunits can be partially incorporated into the glutenin network in the presence of oxygen. Alkylated and unalkylated glutenin subunits had different effects. This was probably caused by the effect of free sulphydryl groups in unalkylated subunits (possibility of SS/SH exchanges and/or incorporation) and/or the effect induced by introduction of alkylated derived substituents. A protocol for «incorporation with excess KIO₃» was developed to exclude the possible effect of a lowering of the available oxidant concentration by oxidation of free sulphydryl groups in glutenin subunits. However, the use of high levels of oxidant in the «incorporation with excess KIO₃» protocol seems to overrule the effects of added glutenin subunits or may force glutenin subunits to incorporate differently from what can be observed under gentle oxidation conditions. Therefore, «incorporation with excess KIO₃» is not suitable for studying the effects of incorporation of glutenin subunits on dough extensibility.     

四川小麦品种高、低分子量麦谷蛋白基因和1B/1R易位的分子标记鉴定

为给四川小麦品质育种提供参考信息,利用7个HMW-GS、17个LMW-GS和1个1B/1R易位的特异分子标记,对105份2000年后育成的四川小麦品种进行上述基因检测。结果表明:(1)针对HMWGS,在Glu-A1位点,含Ax2*的品种有2份,频率为1.9%;在Glu-B1位点,含Bx7、Bx20、Bx17、By8和By9的品种分别有73、26、4、45和30份,频率分别为69.5%、24.8%、3.8%、42.9%和28.6%,未检测到含Bx7OE的品种;在Glu-D1位点,含Dx5的品种有65份,频率为61.9%。(2)针对LMW-GS,在Glu-A3位点,含Glu-A3a、Glu-A3b、Glu-A3c、Glu-A3d和Glu-A3f的品种分别有2、2、63、29和9份,频率分别为1.9%、1.9%、60.0%、27.6%和8.6%,未检测到含Glu-A3e和Glu-A3g的品种;在Glu-B3位点,含Glu-B3b、Glu-B3d、Glu-B3f、GluB3g和Glu-B3i的品种分别有18、10、1、75和1份,频率分别为17.1%、9.5%、1.0%、71.4%和1.0%,未检测到含Glu-B3a、Glu-B3c、Glu-B3e和Glu-B3h的品种。(3)含1B/1R易位的品种有36份,频率为34.3%。(4)组合6种和5种以上优质基因的品种分别有2份(频率为3.8%)和15份(频率为14.3%)。可利用这些品种作为亲本,在四川小麦品质育种中逐步导入优质基因Ax2*、Bx7、By8、Dx5、Glu-A3d、Glu-B3d、Glu-A3b和Glu-B3b,并淘汰1B/1R易位,优化四川小麦面筋优质基因组成。     

Molecular Weight-Dependent Immunostimulative Activity of Low Molecular Weight Chitosan via Regulating NF-κB and AP-1 Signaling Pathways in RAW264.7 Macrophages

Chitosan and its derivatives such as low molecular weight chitosans (LMWCs) have been found to possess many important biological properties, such as antioxidant and antitumor effects. In our previous study, LMWCs were found to elicit a strong immunomodulatory response in macrophages dependent on molecular weight. Herein we further investigated the molecular weight-dependent immunostimulative activity of LMWCs and elucidated its mechanism of action on RAW264.7 macrophages. LMWCs (3 kDa and 50 kDa of molecular weight) could significantly enhance the mRNA expression levels of COX-2, IL-10 and MCP-1 in a molecular weight and concentration-dependent manner. The results suggested that LMWCs elicited a significant immunomodulatory response, which was dependent on the dose and the molecular weight. Regarding the possible molecular mechanism of action, LMWCs promoted the expression of the genes of key molecules in NF-κB and AP-1 pathways, including IKKβ, TRAF6 and JNK1, and induced the phosphorylation of protein IKBα in RAW264.7 macrophage. Moreover, LMWCs increased nuclear translocation of p65 and activation of activator protein-1 (AP-1, C-Jun and C-Fos) in a molecular weight-dependent manner. Taken together, our findings suggested that LMWCs exert immunostimulative activity via activation of NF-κB and AP-1 pathways in RAW264.7 macrophages in a molecular weight-dependent manner and that 3 kDa LMWC shows great potential as a novel agent for the treatment of immune suppression diseases and in future vaccines.     

普通小麦F1代籽粒高分子量谷蛋白亚基的遗传表现

为给小麦品质改良提供依据，采用SDS-PAGE方法,以来自国内外的6个小麦品种（系）为材料，分析了15个杂交组合F1代高分子量谷蛋白亚基（HMW-GS）的遗传表现。结果表明：①HMW-GS在F1代的遗传具有剂量效应，即部分正反交组合F1代籽粒中同一亚基带的颜色深浅不同，来自母本的亚基带比来自父本的亚基带颜色深,说明来自母本的亚基比来自父本的亚基含量高;②2个国内与国内品种间的杂交组合及2个国外与国外品种间的杂交组合，其F1代HMW-GS均表现为完全共显性，11个国内和国外品种间的杂交组合，有4个组合的F1代出现了HMW-GS的不完全表达，缺失率占11个组合总数的36％。     

HMW-GS分子标记多重PCR体系的建立及新疆春小麦的检测

为给新疆优质春小麦品种选育提供参考依据,利用小麦优质高分子量谷蛋白亚基(HMW-GS)基因的特异性标记,基于17份已知HMW-GS组成的品种(系),构建了3套多重PCR,体系Ⅰ可用于同时检测AxNull和Dx5基因,体系Ⅱ可同时检测Ax2*和By8基因,体系Ⅲ可同时检测Bx14和Dx5基因;用3套多重PCR体系分别检测...     

圆锥小麦（Triticum turgidumL．）贮藏蛋白的遗传多样性研究

为发掘可供利用的优异基因资源,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和酸性聚丙烯酰胺凝胶电泳（APAGE）对来自17个国家和地区的95份圆锥小麦的贮藏蛋白进行了分析。结果表明,圆锥小麦贮藏蛋白位点存在丰富的遗传多样性。供试材料中共检测到17种高分子量麦谷蛋白亚基和34种亚基组合类型,其中亚基1和7＋8分别为Glu-A1和Glu-B1位点优势亚基,也检测到优质亚基14＋15和17＋18。亚基组合1／7＋8和null／7＋8虽为优势组合,但所占频率低;醇溶蛋白检测共分离出29条多态性带纹,其中每份材料可分离出5～20条,平均12条,平均遗传相似系数为0．471。供试材料在0．402水平上可分为五类,其聚类结果与材料来源地并不完全一致。试验结果表明,中国圆锥小麦地方品种在高分子量麦谷蛋白亚基表现上具有独特性。     

Isolation and Characterisation of Single Chain Amylose

Five amylose preparations of different origins (cassava, potato, smooth seeded pea, wheat and maize), obtained from native starch granules by thymol complexation, were ultracentrifuged as the final decontamination step to remove a high molecular weight population contaminating the amylose solutions. The efficiency of this ultracentrifugation procedure was assessed by dynamic light scattering (DLS): decreases in apparent hydrodynamic radii,R?_H, from 46·1–72·6 nm before ultracentrifugation to 16·1–29·3 nm after were observed. Amylose solutions were then characterised by size exclusion chromatography coupled on-line to multi-angle laser light scattering (SEC–MALLS). Under these conditionsM?_wof cassava, potato, smooth seeded pea, wheat and maize amyloses were, respectively, 1·05×10⁶, 7×10⁵, 6·2×10⁵, 5·1×10⁵and 3·4×10⁵g/mol. Using a specific optimisation algorithm, experimental molecular weight distributions (MWD) were fitted by two mathematical models of ‘Most Probable’ distribution and ‘log-normal’ distribution. The best fit was obtained for the second model, but fittedM?_wwere higher than experimentalM?_w. When a ‘Most Probable’ model was used, the fittedM?_wwere consistent with experimentalM?_wbut with a lower quality of fit. Exponentscin the power lawR_G=KM^cwere between 0·6 and 0·7, indicating an extended linear random coil in the range of MW analysed (3×10⁵–9×10⁶g/mol). This procedure was also applied to the characterisation of different commercial amylose products.     

小麦低分子量谷蛋白亚基品质效应的研究

为了在小麦品质遗传改良中更好地利用低分子量麦谷蛋白亚基(LMW-GS)的品质效应,以35个小麦品种为材料,对17种LMW-GS的品质效应进行了研究.结果表明,在 Glu-A3位点,对面筋数量的正向效应为A3c、A3b、A3d、A3e＞A3a,对面筋强度的正向效应为A3c＞A3b、A3d＞A3a、A3e.在Glu-B3位点,对面筋数量的正向效应为B3i、B3d＞B3f＞B3e、B3h＞B3g＞B3j,对面筋强度的正向效应为B3d＞B3h＞B3j、B3e＞B3i、B3f＞B3g.在Glu-D3位点,对面筋数量的正向效应为D3e＞D3c＞D3d＞D3b、D3a,对面筋强度的正向效应为D3a＞D3c、D3e＞D3b、D3d.综合各个亚基的品质效应,本研究提出了包含17种小麦LMW-GS的评分体系,品种品质类型与LMW-GS的对应分析结果验证了LMW-GS品质效应的分析结果.研究结果还表明,不同位点优异LMW-GS的品质效应具有累加作用.     

Immunostimulative Activity of Low Molecular Weight Chitosans in RAW264.7 Macrophages

Chitosan and its derivatives such as low molecular weight chitosans (LMWCs) have been reported to exert many biological activities, such as antioxidant and antitumor effects. However, complex and molecular weight dependent effects of chitosan remain controversial and the mechanisms that mediate these complex effects are still poorly defined. This study was carried out to investigate the immunostimulative effect of different molecular weight chitosan in RAW264.7 macrophages. Our data suggested that two LMWCs (molecular weight of 3 kDa and 50 kDa) both possessed immunostimulative activity, which was dependent on dose and, at the higher doses, also on the molecular weight. LMWCs could significantly enhance the the pinocytic activity, and induce the production of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), interferon-γ (IFN-γ), nitric oxide (NO) and inducible nitric oxide synthase (iNOS) in a molecular weight and concentration-dependent manner. LMWCs were further showed to promote the expression of the genes including iNOS, TNF-α. Taken together, our findings suggested that LMWCs elicited significantly immunomodulatory response through up-regulating mRNA expression of proinflammatory cytokines and activated RAW264.7 macrophage in a molecular weight- and concentration-dependent manner.     

人工合成六倍体小麦的高分子量谷蛋白亚基组成分析

利用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳技术(SDS-PAGE)对103份人工合成六倍体小麦(2n=6x=42,AABBDD)的高分子量谷蛋白亚基组成进行了分析。结果发现,所分析材料具有丰富的等位变异。其中,在Glu-A1住点上共有5种变异类型,在Glu-B1和Glu-D1位点上均出现了12种等住变异类型;有些材料在Glu-D1位．占、上只表达1Dy类型亚基,而在1Dx亚基位置上出现空位形式;同时还发现了一些新的亚基组合形式和一未知新亚基。这些人工合成六倍体小麦材料中丰富的高分子量谷蛋白亚基变异,可能对小麦品质改良具有重要的应用价值。     

小麦 Glu A1位点1亚基和Null亚基间品质效应的差异

为了解小麦Glu-A1位点1亚基和Null亚基间的品质效应差异,用郑麦9405/郑麦98//周麦16配制杂交组合,利用系谱法辅助生化标记连续多代选择后得到纯合的1亚基和Null亚基的近等基因系,并对近等基因系1亚基和Null亚基间的面筋特性和粉质特性进行了测定和分析.结果表明,对于面筋指数、形成时间、稳定时间和粉质质量指数,1亚基的贡献明显大于Null亚基,1亚基比Null亚基的面筋指数、形成时间、稳定时间和粉质质量指数分别提高9.79％、68.73％、25.89％和13.99％,成组数据的t测验显示差异达到极显著水平(P值分别为0.007、0.000、0.002和0.006),亚基效应为1亚基＞Null亚基;对于湿面筋含量、干面筋含量和吸水率,1亚基和Null亚基的贡献无明显差别,成组数据t检验显示差异不显著(0.978、0.654和0.206).     

甘肃小麦品种(系)HMW-GS遗传变异分析

为了了解甘肃省小麦品种HMW-GS的遗传变异和组成，为甘肃优质专用小麦品种筛选和品种改良提供依据,采用SDS-PAGE法，分析了254份甘肃省小麦材料(育成品种(系)和农家品种)Glu-1位点的HMW-GS变异，共检测到22种HMW-GS变异，Glu-A1位点3种，Glu-B1位点11种，Glu-D1位点8种。其中110个育成品种(系)的15种HMW-GS有27种亚基组合类型。Glu-A1位点有2种亚基，47．3％的品种该位点具有优质亚基；Glu-B1位点有7种亚基，76．4％的品种具优质亚基；Glu-D1位点有6种亚基，32．7％的品种具优质亚基。14．5％的品种(n：15)Glu-1位点具优质亚基。144份农家品种的18种HMW-GS共有29种亚基组合形式，Glu-A1位点有3种亚基,Glu-B1位点有9种亚基，Glu-D1位点有6种亚基，无优质亚基组合。     

小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达

为了给小麦优质亚基研究奠定基础,将高分子量谷蛋白亚基1Dx5基因的核苷酸序列与载体pRSET A重组,将构建好的质粒转化大肠杆菌BL21(DE3)pLysS菌株,通过IPTG使其得到了成功表达.大肠杆菌中表达的1Dx5亚基在SDS-PAGE上与小麦品种钱尼中的1Dx5亚基具有相似的迁移率.用Western blot技术成功检测到了目的基因产物.通过改变IPTG浓度、诱导时间、培养基成分及初始菌液浓度研究了最适表达条件.结果表明,最适表达条件为:LB培养基,菌液初始浓度(OD600)0.4～0.6,IPTG浓度0.4mmol/L,诱导时间为3～5 h.     

Effect of high molecular weight glutenins and rye translocations on soft wheat flour cookie quality

The influence of high molecular weight glutenin subunits (HMW-GS) on wheat breadmaking quality has been extensively studied but the effect of different Glu-1 alleles on cookie quality is still poorly understood. This study was conducted to analyze the effect of HMW-GS composition and wheat-rye translocations on physicochemical flour properties and cookie quality of soft wheat flours. Alleles encoded at Glu-A1, Glu-B1 and Glu-D1 locus had a significant effect over physicochemical flour properties and solvent retention capacity (SRC) profile. The null allele for Glu-A1 locus presented the highest cookie factor observed (CF = 7.10), whereas 1BL/1RS and 1AL/1RS rye translocations had a negative influence on CF. The three cultivars that showed the highest CF (19, 44 and 47) had the following combination: Glu-A1 = null, Glu-B1 = 7 + 8, Glu-D1 = 2 + 12 and no secalins. Two prediction equations were developed to estimate soft wheat CF: one using the HMW-GS composition and the other using physicochemical flour parameters, where SRCsuc, SRC carb, water-soluble pentosans, damaged starch and protein turned out to be better CF predictors. This data suggests that grain protein allelic composition and physicochemical flour properties can be useful tools in breeding programs to select soft wheat of good cookie making quality.     

High frequency of abnormal high molecular weight glutenin alleles in Chinese wheat landraces of the Yangtze-River region

A total of 485 common landraces of bread wheat were collected from the Yangtze-River region of China. Their high molecular weight glutenin subunit (HMW-GS) composition was analyzed by Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry (MALDI-TOF-MS). Among all landraces tested, 453 were homogeneous for HMW-GS, 32 were heterogeneous, and 37 contained abnormal subunits. A total of 22 alleles were detected, including 3 at Glu-A1, 13 at Glu-B1 and 6 at Glu-D1, respectively. Higher variations occurred at the Glu-B1 locus compared with Glu-A1 and Glu-D1. Glu-A1c (74.0%), Glu-B1b (40.4%), Glu-D1a (84.9%) appeared to be the most frequent alleles at Glu-A1, Glu-B1 and Glu-D1, respectively. Two alleles ("null" and 1) at the Glu-A1 locus, three allele compositions (7 + 8, 7OE + 8, 7 + 9) at the Glu-B1 locus, and two (2 + 12 and 5 + 10) at the Glu-D1 locus appeared to be the common types in the 485 landraces. Sixteen new alleles represented by abnormal subunits were identified at the Glu-B1 and the Glu-D1 locus.     

中国小麦代表性地方品种高分子量谷蛋白亚基组成分析

为了明确中国小麦地方品种的遗传多样性,并为小麦品质改良提供基础材料,利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,分析了源自中国22个省市地区的200份代表性地方品种的高分子量麦谷蛋白亚基组成.结果表明,Glu-1位点共检测到35种亚基组合,null/7 8/2 12组合的频率最高,为59%,其它亚基组合的频率均低于10%;Glu-A1、Glu-B1和Glu-D1位点上的等位变异分别为3、12和9种,三位点中的优势亚基依次为null、7 8和2 12,其频率分别达86.5%、73.0%和85.0%.另外在Glu-B1位点上还检测到6 8 19和6 9、7、22等稀有亚基.本研究所选200份代表性地方品种的HMW-GS组成类型虽然丰富,但与品质相关的优质亚基频率较低.亚基组合得分在8分以上的品种为19个;得分在9分以上的品种有4个,它们是草鞋边(ZM011396)、中81Ⅲ-6218(ZM013120)、高8(ZM0094z8)和京红7号(ZM008935).     

Effects of mechanical strain on the electric conductivity of multiwalled carbon nanotube (MWCNT)/polyurethane (PU) composites

Conducting polymers have been under development for more than thirty years as replacements for metals in various applications, such as fuel cells, solar cells, actuators, etc. In this study, we investigate conducting polymer composites and attempt to fabricate composite polyurethane/multiwalled carbon nanotubes. The multiwalled carbon nanotubes (MWCNTs) were acid-treated to add functional groups such as -OH or -COOH so they could then be chemically bonded to diisocyanate to form a urethane linkage. Because they have fewer impurities and reduced surface roughness (as confirmed by TEM micrographs), acid-treated MWCNTs can be better dispersed in a polyurethane (PU) matrix than untreated MWCNTs, and acid-treated MWCNTs exhibit better adhesion with the PU matrix, as well. In addition, the conductance test of MWCNT/PU films as a function of elongation showed that the conductance of the acid-treated MWCNT/PU increased up to a certain % elongation, while that of the untreated MWCNT/PU decreased monotonically with % elongation.     

Low Molecular Weight Chitosan–Insulin Polyelectrolyte Complex: Characterization and Stability Studies

The aim of the work reported herein was to investigate the effect of various low molecular weight chitosans (LMWCs) on the stability of insulin using USP HPLC methods. Insulin was found to be stable in a polyelectrolyte complex (PEC) consisting of insulin and LMWC in the presence of a Tris-buffer at pH 6.5. In the presence of LMWC, the stability of insulin increased with decreasing molecular weight of LMWC; 13 kDa LMWC was the most efficient molecular weight for enhancing the physical and chemical stability of insulin. Solubilization of insulin-LMWC polyelectrolyte complex (I-LMWC PEC) in a reverse micelle (RM) system, administered to diabetic rats, results in an oral delivery system for insulin with acceptable bioactivity.