57份陕西新育成小麦品种（系）HMW-GS组成及品质分析

为探究陕西关中地区小麦HMW-GS亚基与品质性状间的关系,采用SDS-PAGE法对57份陕西关中地区小麦品种（系）HMW-GS亚基组成及相关品质性状进行了分析。结果表明,供试品种（系）中共检测出7种HMW-GS亚基类型和8种HMW-GS亚基组合;Glu-A1位点上有3种亚基类型,分别为1、2^*和Null,以1亚基为主（78.95%）;Glu-B1位点上检测到7+8（61.40%）与7+9（38.60%）两个类型;Glu-D1位点上检测到5+10（70.18%）和2+12（29.82%）两个类型。3个HMW-GS基因位点编码亚基共组成8种亚基组合,品质得分6～10分,其中1/7+8/5+10组合品质得分10分,出现频率最高。就HMW-GS不同位点对品质性状效应进行分析发现,Glu-D1位点对b^*值、形成时间、稳定时间、弱化度和粉质质量指数的影响达到极显著水平（P＜0.01）;对面团流变学特性的影响,Glu-D1>Glu-B1。不同类型亚基对小麦品质的效应存在差异,7+8亚基对蛋白质含量、湿面筋含量和容重具有正效应,7+9和5+10亚基对形成时间和稳定时间的影响显著高于其他亚基（P＜0.05）;携带1/7+8/5+10亚基组合小麦的蛋白质、湿面筋含量和容重最高;携带1/7+9/5+10亚基组合具有较高面粉L^*值和面团流变学特性指标值。     

影响小麦面团强度的贮藏蛋白基因表达研究

小麦品质主要由籽粒贮藏蛋白的含量和类型决定,高分子量麦谷蛋白亚基(HMW-GSs)组合是决定加工品质的主要因素。为了解HMW-GSs组合完全一致、蛋白含量相似的两个品种面团稳定时间相差近一倍的原因(郑麦158:高面团强度;郑麦369:低面团强度),通过转录组测序比较了籽粒发育中期3个时间点(花后14、21和28 d)的贮藏蛋白基因表达差异,分析了差异表达基因编码蛋白对面粉质量的贡献以及面粉的巯基含量差异等。结果表明,在3个时间点,两个品种间的HMW-GSs基因表达均无显著差异;郑麦158较郑麦369共有24个贮藏蛋白编码基因有显著表达差异,其中上调表达基因12个(23次),下调表达基因12个(23次)。12个显著上调表达基因中,包括9个燕麦类似蛋白基因(18次)、2个γ-醇溶蛋白基因(4次)和1个α-醇溶蛋白基因;在12个下调表达基因中,包括11个α-或α/β-醇溶蛋白基因(21次)和1个燕麦类似蛋白基因(2次)。两品种比较,郑麦158面粉的硫元素含量较低,但自由巯基和二硫键含量较高。基于蛋白二硫键预测和面粉蛋白质量评价模型分析结果,差异表达基因编码的燕麦类似蛋白和γ-醇溶蛋白对面团强...     

50份哈萨克斯坦小麦高分子量谷蛋白亚基解析

为发掘优良的小麦种质资源,利用SDS-PAGE技术分析了50份哈萨克斯坦小麦种质高分子量谷蛋白亚基组成及其品质得分。结果表明,50份供试材料中共检测到8种亚基类型,其中在 Glu-A1位点有2种亚基类型（Null,1）, Glu-B1位点有2种亚基类型（7＋8,7＋9）,在 Glu-D1位点有4种亚基类型（5＋10, 2＋10,2＋12,4＋12）。利用PCR分子标记对所得到的高分子量谷蛋白亚基进行验证,结果一致。在50份哈萨克斯坦小麦中,优质亚基1、7＋8、5＋10的出现频率较高,分别为34%、22%、56%。亚基组合Null/7＋9/5＋10在所有亚基组合类型中出现频率最高,为28%,品质得分为7分,而组合1/7＋8/5＋10出现频率仅为8%,品质得分为10分;供试材料品质得分大多为7、8分,平均为7.10分。出现频率较高的优质亚基可以作为改善现有品种亚基组成状况的亲本材料。     

小麦品质性状与蛋白组份含量关系的研究

通过对黑龙江省大面积主栽小麦品种不同品质类型进行蛋白组份含量以及麦谷蛋白亚基组合的份析表明 ,小麦品质性状与麦谷蛋白 /醇溶蛋白比值显著相关 ,随麦谷蛋白含量的增加 ,面筋、沉降值、稳定时间都有明显增大 ,醇溶蛋白高于麦谷蛋白含量 ,其面团筋性弱、稳定时间短。     

利用多重PCR技术鉴定小麦背景中的1BL·1RS易位和Glu D1d基因

高分子量谷蛋白亚基（HMWGS）对小麦面粉加工品质有促进作用,尤其是GluD1d 基因编码的1Dx5+1Dy10亚基能增加面团的筋度和弹性。小麦背景中的1BL·1RS易位对小麦面粉加工品质有显著的负面影响。因此,在小麦品质育种中如何判定小麦背景中是否含有1BL·1RS易位和HMWGS的GluD1d基因具有重要意义。本研究利用3对分别检测1BL·1RS易位、GluB3和GluD1位点的共显性特异标记,结合SDSPAGE鉴定,对16份已知遗传背景和GluD1x等位基因材料及38株（周麦18×烟农19）F₂群体进行了分析,探索出适合同时鉴定小麦背景中1BL·1RS易位和GluD1d基因的多重PCR技术实验体系,并采用该体系对国内外352份小麦品种（系）进行了鉴定。结果表明,该体系是同时鉴定小麦背景中1BL·1RS易位和GluD1d基因的一种非常有效、简便可行的实验方法,可在标记辅助选择（MAS）育种中应用。     

97份安徽省种植小麦品种HMW-GS组成及品质分析

为探究安徽小麦品种高分子量麦谷蛋白亚基（HMW-GS）组成及品质性状,对97份安徽省主要种植的小麦品种进行了HMW-GS组成及品质性状分析。结果表明,供试材料中共检测出10种HMW-GS亚基类型和19种亚基组合,在 Glu-A1位点检测出3种亚基类型,以1亚基（57.73%）和Null亚基 （40.21%）为主;在 Glu-B1位点共检测出3种亚基类型,以7+8亚基（52.58%）为主;在 Glu-D1位点共鉴定出4种亚基类型,优质亚基5+10占比最大,为45.36%;供试品种亚基组合品质得分在4~10分之间,10分的亚基组合1/7+8/5+10和1/17+18/5+10共19份材料（19.59%）,8分以下的共44份材料（45.36%）。对供试材料HMW-GS与品质性状进行相关分析发现, Glu-1的三个位点与硬度指数、溶剂保持力和吸水率相关达显著或极显著水平,对溶剂保持力影响表现为 Glu-A1> Glu-B1> Glu-D1;不同亚基组合对品质性状的影响存在差异,亚基组合为1/17+18/5+10的品种具有最高的硬度指数、4种溶剂保持力、吸水率、稳定时间和粉质质量指数,具有最低的L^*和W,该类品种有烟农19、泰农19、山农17和糯小麦1012;亚基组合为Null/7+9/4+12的品种具有最低的硬度指数、4种溶剂保持力、吸水率、稳定时间和粉质质量指数,具有最高的弱化度、L^*和W,该类品种有荃麦725、皖麦52和未来0818。     

小麦高分子量麦谷蛋白亚基近等基因系及其应用研究进展

介绍了影响小麦烘烤品质的遗传因素和环境因素,根据影响小麦烘烤品质的种因素对小麦高分子量麦谷蛋白亚基与小麦烘烤品质的各种相关性研究方法进行了分析,介绍了小麦高分子量麦谷蛋白亚基近等基因系在小麦烘烤品质研究中的作用。     

不同HMW麦谷蛋白亚基类型小麦品种（系）的沉降值及其与面筋质和量的关系

小麦沉降值是衡量小麦面筋质和量的综合指标ＨＭＷ（高分子量）麦谷蛋白亚基类型与小麦加工品质密切相关,本研究通过对具有不同ＨＭＷ麦谷蛋白亚基类型小麦品种（系）的ＳＤＳ和Ｚｅｌｅｙ沉降值的测定,分析其与湿面筋含量和面团稳定时间的相关关系,结果表明,具有５＋１０亚基的品种,品种间品质具有高度的不稳定性,ＳＤＳ沉降值与小麦面筋质量关系密切,可作为小麦品质育种低世代材料的品质筛选指标,能更确切地反映的品种水平     

陕西小麦黄色素含量基因 TaZds-A1和 TaZds-D1的组成与分布

面粉或面制品色泽是小麦重要的品质评价指标,ζ-胡萝卜素脱氢酶(ζ-carotene desaturase, ZDS)活性基因是控制小麦籽粒或面粉中黄色素(Yellow pigment, YP)含量的重要基因。为给陕西小麦品质改良提供参考依据,利用2A和2D染色体上的 YP2A-1和 YP2D-1标记,对陕西194份小麦品种(系)分别进行检测,分析 TaZds-A1和 TaZds-D1两位点等位变异的组成和分布特点。结果表明,在陕西小麦中, TaZds-A1位点存在 TaZds-A1a和 TaZds-A1b两种等位变异类型,分别占43.3%和56.7%;而在 TaZds-D1位点,全为 TaZds-D1a等位变异类型。陕西小麦两位点存在 TaZds-A1a/ TaZds-D1a和 TaZds-A1b/ TaZds-D1a两种等位变异的组合类型,在不同地区小麦中的分布不同：在陕西渭北旱塬和关中地区小麦中,两种组合类型的频率相当,约各占一半;在陕南地区小麦中,高YP含量的 TaZds-A1b/ TaZds-D1a组合类型占三分之二。ZDS活性基因的选择将成为以后陕西小麦色泽品质遗传改良研究中的一个重要部分。     

小麦品质性状与蛋白组份含量关系的研究

通过对黑龙江省大面积主栽小麦品种不同品质类型进行蛋白组份进行含量以及麦谷蛋白 基组合的份析表明,小麦品质性状与麦谷蛋白／醇溶蛋白比值显著相关,随麦谷蛋白含量的增加,面筋、沉降值、稳定时间都有明显增大,醇溶蛋白高于麦谷蛋白含量,其面团筋性弱,稳定时间短。     

Quality-related Epitopes of High Mr Subunits of Wheat Glutenin

Three hundred and eighty four immobilised overlapping nonapeptides, corresponding to the full amino acid sequences of three high M_r subunits of glutenin from bread wheat (Triticum aestivum) grain, were used to determine the linear epitopes recognised by four monoclonal antibodies. These antibodies were selected on the basis of significant and positive correlations between their binding to wheat flour extracts in a two-site ('sandwich') enzyme immunoassay and rheological measures of dough strength, an important aspect of bread wheat quality. The antibodies did not bind to a single, specific sequence but bound a series of related peptides in each high M_r glutenin subunit examined. The sequences recognised were not identical for the four antibodies, but in each case were in the central repeating domain of the high M_r glutenin subunits, and usually comprised regions that overlapped the degenerate repeat nonamer and hexamer sequences. High M_r glutenin subunits that have been associated with greater dough strength, such as the D-genome allelic products 1Dx5 and 1Dy10, displayed an increased number of the epitope sequences. The location of the epitopes in sequences of overlapping β-turns in the repetitive region supports the hypothesis that dough elasticity arises partly from β-turn-forming secondary structure in the repeat regions of the M_r glutenin subunits. Additional β-turn within high M_r subunits may extend their structure to allow increased interaction between the glutenin subunits and with the other proteins of the gluten complex, thus improving dough strength.     

小麦高分子量麦谷蛋白基因沉默研究进展

高分子量麦谷蛋白是小麦种子贮藏蛋白的重要组成部分,其数量和质量对小麦面筋含量及面粉烘烤品质有直接影响.因此,小麦高分子量麦谷蛋白基因的表达与沉默备受关注.本文主要介绍了高分子量麦谷蛋白基因沉默材料的发现及其沉默机理的研究现状,认为小麦高分子量麦谷蛋白基因沉默包括两种情况即在一个基因座位上所有编码基因均不表达和只有一个编码基因不表达.第一种情况可能是染色体部分缺失的结果,有时也被认为是由于调控元件发生了突变;而基因突变即DNA调控序列和/或编码区的微小缺失及其他变化则可能是第二种情况发生的机制.本文还简要介绍了离子束生物技术、转基因技术、化学诱变、组织培养等可以用于创制基因沉默材料的技术.     

Thermal stability of wheat gluten protein: its effect on dough properties and noodle texture

There is a need to develop more sensitive and reliable tests to help breeders select wheat lines of appropriate quality. Gluten thermostability, measured by the viscoelasticity of heated gluten, was assessed for its usefulness in evaluating quality of wheats in breeding programs. Two sets of wheat samples were used: Set I consisting of 20 cultivars and/or breeders' lines (BL), with diverse dough strengths and allelic variations of high M_r glutenin subunits coded at the Glu-A1, Glu-B1 and Glu-D1 loci (N=20) and Set II consisting of 16 near isogenic BL of F7 generation that had been in a quality selection program for three years. Thermostability of the isolated wet gluten was determined by measuring its viscoelastic properties, and was related to noodle texture, flour protein content, protein composition, dough physical properties and other quality predicting tests.Viscoelasticity of heat-treated gluten, isolated with 2% NaCl solution, significantly correlated with most of the tests used to measure dough and/or gluten strength and Chinese white salted noodle texture. The rate of thermal denaturation of proteins depends on M_r and packing density. High ratios of monomeric proteins such as gliadins and low M_r glutenin subunits to high M_r glutenin subunits increase the thermostability of the gluten. The measurement of viscoelasticity of heat-denatured gluten can be a useful test to determine gluten quality. Our study showed that gluten viscoelasticity and most of the tests related to dough and/or gluten strength are independent of allelic variations of the high molecular weight glutenin subunits. This test has been developed for predicting white salted noodle quality.     

Incorporation of high-molecular-weight glutenin subunits into doughs using 2 gram mixograph and extensigraphs

To study the contributions of high-molecular-weight glutenin subunits (HMW-GS) to the gluten macropolymer and dough properties, wheat HMW-GS (x- and y-types) are synthesized in a bacterial expression system. These subunits are then purified and used to supplement dough mixing and extensigraph experiments through dough partial reduction and reoxidation to allow these exogenously added HMW-GS to incorporate into gluten polymers. Detailed results are given for seven mixing and two extension parameters. HMW-GS synthesized in bacteria behaved similarly under these conditions to the same HMW-GS extracted from wheat flour. These experiments initially focused on the HMW-GS of the D-genome of hexaploid wheat encoded at the Glu-D1 locus; e.g. the Dx2, Dx5, Dy10, and Dy12 subunits. Experiments used five different flours and results are shown to be consistent when normalized to results from Dx5. The incorporation of Dx-type subunits into the gluten disulfide bonded network has greater effects on dough parameters than incorporation of Dy-type subunits. When Glu-D1 x- and y-type subunits are incorporated together, there are synergistic effects greater than those with either subunit type alone. This synergistic effect was greatest with approximately equal amounts of Dx- and Dy-type subunits - implying a 1:1 stoichiometric relationship.     

Mixing properties and dough functionality of transgenic lines of a commercial wheat cultivar expressing the 1Ax1, 1Dx5 and 1Dy10 HMW glutenin subunit genes

In this work we report the effects of the HMW-GS 1Ax1, 1Dx5 and 1Dy10 on the breadmaking quality of the bread wheat cultivar Anza that contains the HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8, and is null for the Glu-A1 locus. This allows the characterization of individual subunits 1Dx5 and 1Dy10 in the absence of subunit 1Dx5, and the interactions between these subunits and subunits 1Dx2 and 1Dy12 to be determined. Three transgenic lines termed T580, T581 and T590, containing, respectively, the HMW-GS 1Ax1, 1Dx5 and 1Dy10 were characterized over 3 years using a range of widely-used grain and dough testing methods. The transgenic subunits 1Ax1, 1Dx5 and 1Dy10 accounted for 25.2%, 20.3% and 17.9%, respectively, of the total HMW-GS in the three transgenic lines. Although lines T581 and T590 expressed similar levels of subunits 1Dx5 and 1Dy10 they had different effects on other aspects of protein composition, including changes in the ratios of glutenin/gliadin, of HMW/LMW-GS, the 1Dx2/1Dy12, the x-type/y-type HMW-GS and the proportions of high molecular mass glutenin polymers. In contrast, lines transformed to express subunits 1Ax1 and 1Dx5 showed similar changes in protein composition, with higher protein contents and decreased ratios of glutenin/gliadin and 1Dx2/1Dy12. In addition, both transgenic lines showed similar increases in the ratio of x-type/y-type subunits compared to the control line. The transgenic lines were analysed using Farinograph, Mixograph and Alveograph. This confirmed that the expression of all three subunits resulted in increased dough strength (and hence breadmaking quality) of the cultivar Anza. A beneficial effect of subunit 1Dx5 has not been reported previously, transgenic wheat lines expressing this subunit giving overstrong dough unsuitable for breadmaking. However, the expression of subunit 1Dy10 had a greater effect on breadmaking quality than subunits 1Ax1 and 1Dx5. The Farinograph parameters such as dough stability and peak time were increased by 9.2-fold and 2.4-fold, respectively, in line T590 (expressing 1Dy10) with respect to the control line. Similarly, the Mixograph mixing time was increased by four-fold and the resistance breakdown decreased by two-fold in line T590 compared with the control line. The Alveograph W value was also increased by 2.7-fold in line T590 compared to the control line. These transgenic lines are of value for studying the contribution of specific HMW-GS to wheat flour functional properties.     

Development of an Aegilops longissima substitution line with improved bread-making quality

This study focuses on the effect of Aegilops longissima on wheat bread making quality. Chromosome 1S^l disomic addition line of Ae. longissima (DAL1S^l) had significantly higher dough strength, grain hardness, mixographic peak height, band width, and unextractable polymeric protein content compared with wheat. DAL1S^l also had additional glutenin and gliadin proteins contributed by Ae. longissima. The larger size of 1S^l coded HMW-GSs sequenced from DAL1S^l and their phylogenetic similarity to the D-genome-coded subunits were suspected to be one of the major reasons for the increased dough strength of DAL1S^l. To transfer the chromosome 1S^l genes responsible for the good bread-making quality to wheat, we generated a chromosome-specific disomic substitution line [DSL1S^l(1A)] by crossing DAL1S^l with nulli 1A tetra 1B genetic stock and further selection. Grain quality analysis revealed significantly lower grain hardness and significantly higher dough strength, farinograph development time, stability time, gluten index, bread loaf volume, and bread quality score in DSL1S^l(1A), compared with wheat. However, the increased bread loaf volume and quality were not proportional to the relatively higher increases in dough strength and gluten index, indicating importance of other traits influencing bread making quality. The presence of a minor hardness locus on chromosome 1A is speculated.     

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.     

The low-molecular-weight glutenin subunits of wheat gluten

Low-molecular-weight glutenin subunits (LMW-GS) are polymeric protein components of wheat endosperm and like all seed storage proteins, are digested to provide nutrients for the embryo during seed germination and seedling growth. Due to their structural characteristics, they exhibit features important for the technological properties of wheat flour. Their ability to form inter-molecular disulphide bonds with each other and/or with high-molecular-weight glutenin subunits (HMW-GS), is important for the formation of the glutenin polymers, which are among the biggest macromolecules present in nature, and determine the processing properties of wheat dough. Explanation of the structural basis for these correlations continues to intrigue researchers and, while earlier emphasis had been on HMW-GS, considerable attention is now being focused on the LMW-GS.LMW-GS are a highly polymorphic protein complex, including proteins with gliadin-type sequences. Difficulty in separating single components, arising from the complexity of the group, has limited the characterisation of the individual proteins and the establishment of clear-cut relationships with quality parameters.Here we review results concerning different aspects of LMW-GS, including their structural characteristics, genetic control, and relationships with quality parameters. In addition, we emphasise the distinction between the components with sequences unique to the LMW-GS fraction and those behaving like glutenin subunits (incorporated into polymers), but with sequences corresponding to gliadins.     

Distribution and function of LMW glutenins,HMW glutenins,and gliadins in wheat doughs analyzed with ‘in situ’ detection and quantitative imaging techniques

The different gluten subunits, gliadins, LMW glutenins, and HMW glutenins have been reported to play different key roles in different type of wheat products. This paper studied the interaction between gliadin, LMW and HMW glutenins in soft, hard and durum semolina flour doughs during different stages of mixing. In order to see how do the gluten subunits (gliadin, LMW glutenin and HMW glutenin) redistribute during mixing, dough samples were taken at maximum strength and 10 min after maximum strength. The doughs have been mixed with the same level of added water (55%), therefore they all have different strengths values due to their changes in proteins content. Oscillatory rheological measurements were performed on the doughs. It has been found that HMW glutenins are relatively immobile because of their less molecular mobility and do no redistribute themselves especially at high strength for doughs such as hard wheat flour. LMW glutenins and gliadins on the other hand redistribute themselves at even at high dough strengths forming a more stable network. In weaker doughs such as soft wheat, the breakdown of the three proteins subunits is responsible for the decay in dough strength. We have also visualized how the greater amount of LMW glutenins in semolina is in constant interaction with HMW glutenins and gliadins allowing the dough to maintain a stable strength for an extended mixing time. Finally, we have found the ‘in situ’ detection and quantitative analysis techniques to be more sensitive to the changes occurring in the gluten network of the dough than the oscillatory rheological analysis.