山东省普通小麦醇溶蛋白Gli-1和Gli-2位点等位基因的遗传变异

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE)，分析了山东省种植面积较大的37个小麦品种醇溶蛋白Gli-1和Gli-2位点等位基因的组成特点。结果表明，山东小麦在醇溶蛋白Gli-1 (Gli-A1、Gli-B1和Gli-D1) 和Gli-2 (Gli-A2、Gli-B2和Gli-D2) 位点存在多样性，共鉴定出58个等位基因，出现频率较高的有6个，分别为Gli-A1a (48.6 %)、Gli-B1l (35.1%)、Gli-D1k (35.1%)、Gli-A2b (35.1％)、Gli-B2g (35.1％)和Gli-D2a (29.7％)，其中Gli-B1l出现频率较高，表明1BL/1RS易位系在山东小麦中存在比较普遍。醇溶蛋白6个主要位点的遗传变异系数较高，平均为0.7930，变幅为0.7297～0.8269，其中Gli-D2位点遗传多样性最高，Gli-A1最低。对具有优质醇溶蛋白等位基因Gli-B1b或Gli-A2b的品种进行了高分子量谷蛋白亚基的组成分析,表明烟农15、烟优361、山农98-1和山农93-52同时含有优质谷蛋白5＋10亚基，在小麦育种中可利用这些优质亚基基因。     

乌克兰普通小麦品种储藏蛋白分析

为了更好的利用乌克兰小麦品种资源,并了解引进品种的品质,采用SDS-PAGE和A-PAGE技术,对从乌克兰引进小麦材料的高分子量麦谷蛋白亚基(HMW-GS)和醇溶蛋白亚基的组成进行分析。结果表明,在16个普通小麦品种中,由Glu-1位点控制的高分子量亚基组合类型共有7种,最常见的是（1,7+8,5+10）占37.5%,其次是（1,2+12,6+8）和（1,7+9,5+10）,各占18.75%,其中Glu-A1位点有3种等位变异,以1亚基为主（75%）;Glu-B1位点有3种等位变异,以7+8为主（43.75%）;Glu-D1位点有3种等位变异,以5+10为主（68.75%）。醇溶蛋白方面,从供试材料的6个位点中,共鉴定了33个不同的醇溶蛋白等位基因,41条迁移率不同的醇溶蛋白带纹,其中Gli-A1,Gli-B1和Gli-D1分别有6,5,5个等位基因;Gli-A2,Gli-B2和Gli-D2各有6,5,6个等位基因,优质亚基Gli-B1b出现频率较高（43.75%）,这些材料有可能会成为比较有价值的品质改良中间材料。     

小麦资源胚乳蛋白Glu-1、Glu-3、Gli-1基因位点变异特点

141个普通小麦品种及农家种中，由Glu-1位点控制的高分子量谷蛋白亚基共27种图谱，最常见的图谱是（N，7+8，2+12）占22%和（N，7+9，2+12）占19.9%，Glu-A1、Glu-B1、Glu-D1位点控制的均为正效应亚基，其图谱（1，7+8，5+10），（1，14+15，5+10），（1，13+16，5+10），（1，17+18，5+10），（2*，7+8，5+10），（2*，13+16，5+10）占13.4%; 由Glu-3位点控制的低分子量谷蛋白亚基共48种以上的图谱，最常见的图谱是（a, j, c）, Glu-A3位点存在6个以上等位基因，新发现的占5.7%, Glu-B3位点存在10个以上等位基因，新发现的占2.8%, Glu-D3位点存在3个等位基因；由Gli-1位点控制的醇溶蛋白共81种以上图谱，Gli-1A1位点存在7个以上等位基因，新发现的占7.1%, Gli-B1位点存在12个以上等位基因，新发现的等位基因占3.5%, Gli-D1位点存在10个等位基因，Gli-B1位点的l为1B/1R易位系，占总数的33.6%; 由Gli-1位点控制的醇溶蛋白和由Glu-3位点控制的低分子量谷蛋白亚基基因变异远比由G1u-1位点控制的高分子量谷蛋白亚基复杂和丰富。     

小偃系列小麦品种的HMW-GS和醇溶蛋白分析

为探讨小偃系列小麦品种的品质相关性状的遗传基础,为小麦品质改良和品种选育提供参考。采用SDS-PAGE和A-PAGE技术对14个小偃系列小麦品种的高分子量谷蛋白亚基（HMW-GS）和醇溶蛋白进行了研究。结果表明：14份小偃系列小麦品种共出现了7种亚基和5种亚基组合类型。Glu-A1位点,1亚基是主要亚基,其频率达85．7％;G1u-B1位点,7＋9亚基是主要亚基。其频率为50％;Glu-D1位点,2＋12是主要亚基,其频率高达92．9％。1,7＋9,2＋12是主要的亚基组合类型,其频率为35．7％;小偃系列小麦品种整体品质得分较高,为7．21。14个小偃系列小麦品种共检测到19条醇溶蛋白带型。平均13．6条;其遗传距离（GD）在0．23-0．59之间,当遗传距离为0．50时,14个品种可聚为4个大类,来源相同的品种,遗传相似性大。可以聚在一起。小偃系列小麦品种的高分子量谷蛋白亚基和醇溶蛋白的遗传变异较小。遗传基础较狭窄。     

河南省主推小麦品种籽粒储藏蛋白质遗传多样性分析

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对河南省主推小麦品种进行了醇溶蛋白位点特异性检测和高分子谷蛋白亚基(HMWGS)组成分析。结果表明:41份小麦品种(系),共分离出31条不同迁移率的谱带,绝大多数品种在α、β、γ和ω4个区中存在着较大差异,其中α区共有6条带,β区共有10条带,γ区共有8条带,ω区共有7条带;供试品种在遗传相似系数(GS值)为0.68的水平下可明显聚为6类。45份河南小麦品种(系)共出现了11种亚基和15种亚基组合类型,在Glu-A 1位点,主要亚基是1亚基,其频率达75.6%;在Glu-B 1位点,主要以7+8和7+9为主,分别占33.3%和55.6%;Glu-D 1位点,5+10、2+12和5+12均较多,分别占33.3%、35.6%和28.9%。频率较高的组合形式有1、7+9、5+12和1、7+9、5+10,其频率依次为17.8%和15.6%。说明河南小麦品种高分子量谷蛋白亚基和醇溶蛋白的遗传变异丰富,遗传基础较为广泛。     

小麦远缘杂交后代节燕98-2分离类型贮藏蛋白的研究

采用A-PAGE和SDS-PAGE聚丙烯酰胺凝胶电泳方法,对远缘组合分离出来的节燕98-2类型入选11个遗传性基本稳定的具有高产、多抗的小麦株系的醇溶蛋白和高分子量谷蛋白亚基进行了分析.结果表明,在A-PAGE电泳分析中,11个供试株系具有11种不同的醇溶蛋白带型.在SDS-FAGE电泳分析中,出现了7种不同的高分子量谷蛋白亚基(HMW-GS)及6种亚基组合类型,优质亚基及亚基组合所占的比例较少,品质评分偏低,其变幅为5～8分,平均为6.36分.但在所分析的材料中,出现了一个少见的特殊亚基:2 10 12.并研究了这些HMW-GS和组合频率及特点.11个株系中7个具有45 10优质亚基和2个具有2*亚基,它们可供小麦优质育种利用.研究表明,通过远缘杂交能够选育出具有高产、抗病和优质的小麦新材料.     

小麦籽粒蛋白质组分含量及其加工品质的关系

应用反相高效液相色谱(RP-HPLC)法,对12个小麦品种籽粒的清蛋白＋球蛋白、醇溶蛋白和谷蛋白,高分子量谷蛋白亚基(HMW-GS)、低分子量谷蛋白亚基(LMW-GS)进行了分离量化,并根据谷蛋白含量、贮藏蛋白含量及面团稳定时间3个指标对其聚类分析。结果表明,不同小麦品种蛋白质各组分含量存在差异,其中贮藏蛋白的含量是决定蛋白质总含量的主要因素。HMW-GS含量、LMW-GS含量、谷蛋白总含量均与面团形成时间、稳定时间及沉降值呈极显著正相关;HMW-GS含量与LMW-GS含量的比值(HMW/LMW)与面团形成时间和稳定时间呈极显著正相关;醇溶蛋白含量与谷蛋白含量的比值(Gli/Glu)与面团稳定时间呈显著负相关,醇溶蛋白含量与HMW-GS含量的比值(Gli/HMW-GS)与面团形成时间和稳定时间均呈极显著负相关。籽粒中具有较高的贮藏蛋白含量、HMW-GS含量、LMW-GS含量和HMW/LMW及较低的Gli/Glu有利于提高强筋小麦的加工品质。     

小麦籽粒蛋白质组分含量及其加工品质的关系

应用反相高效液相色谱(RP-HPLC)法,对12个小麦品种籽粒的清蛋白＋球蛋白、醇溶蛋白和谷蛋白,高分子量谷蛋白亚基(HMW-GS)、低分子量谷蛋白亚基(LMW-GS)进行了分离量化,并根据谷蛋白含量、贮藏蛋白含量及面团稳定时间3个指标对其聚类分析。结果表明,不同小麦品种蛋白质各组分含量存在差异,其中贮藏蛋白的含量是决定蛋白质总含量的主要因素。HMW-GS含量、LMW-GS含量、谷蛋白总含量均与面团形成时间、稳定时间及沉降值呈极显著正相关;HMW-GS含量与LMW-GS含量的比值(HMW/LMW)与面团形成时间和稳定时间呈极显著正相关;醇溶蛋白含量与谷蛋白含量的比值(Gli/Glu)与面团稳定时间呈显著负相关,醇溶蛋白含量与HMW-GS含量的比值(Gli/HMW-GS)与面团形成时间和稳定时间均呈极显著负相关。籽粒中具有较高的贮藏蛋白含量、HMW-GS含量、LMW-GS含量和HMW/LMW及较低的Gli/Glu有利于提高强筋小麦的加工品质。     

普通小麦及近缘粗山羊草α-醇溶蛋白基因的克降、定位与进化分析

通过特异PCR引物设计,从普通小麦品种(豫麦34和烟农19)和粗山羊草(T9、T197、T48、T176和T17)中扩增、克隆了7个新的α-醇溶蛋白基因,分别命名为Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176和Gli-T17,基因序列长度为846~891 bp,编码282~297个氨基酸残基,都具有α-醇溶蛋白的典型结构特点。其中Gli-YM34和Gli-YN19基因推导的醇溶蛋白都含有一个额外的半胱氨酸残基,可能对面筋品质有正向作用。根据α-醇溶蛋白氨基酸序列所具有的4种T细胞抗原表位和多聚谷氨酰胺重复区的平均长度以及中国春缺体四体分析,将来自普通小麦品种的Gli-YM34和Gli-YN19基因定位在6D染色体上的Gli-D2位点,而且Gli-YM34和Gli-YN19与来自粗山羊草的α-醇溶蛋白基因具有很高的序列相似性,进一步证明粗山羊草是普通小麦D基因组的供体。在克隆的4个典型α-醇溶蛋白基因中检测到21个SNP和1个9 bp的缺失。系统进化分析表明,α-醇溶蛋白基因与低分子量谷蛋白亚基基因关系较近,在大约43.69百万年时分化,与ω-醇溶蛋白和HMW-GS基因亲缘关系较远,它们的分化时间大约为79.39百万年。     

普通小麦及近缘粗山羊草α-醇溶蛋白基因的克隆、定位与进化分析

通过特异PCR引物设计,从普通小麦品种(豫麦34和烟农19)和粗山羊草(T9、T197、T48、T176和T17)中扩增、克隆了7个新的α-醇溶蛋白基因,分别命名为Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176和Gli-T17,基因序列长度为846～891bp,编码282～297个氨基酸残基,都具有α-醇溶蛋白的典型结构特点。其中Gli-YM34和Gli-YN19基因推导的醇溶蛋白都含有一个额外的半胱氨酸残基,可能对面筋品质有正向作用。根据α-醇溶蛋白氨基酸序列所具有的4种T细胞抗原表位和多聚谷氨酰胺重复区的平均长度以及中国春缺体四体分析,将来自普通小麦品种的Gli-YM34和Gli-YN19基因定位在6D染色体上的Gli-D2位点,而且Gli-YM34和Gli-YN19与来自粗山羊草的α-醇溶蛋白基因具有很高的序列相似性,进一步证明粗山羊草是普通小麦D基因组的供体。在克隆的4个典型α-醇溶蛋白基因中检测到21个SNP和1个9bp的缺失。系统进化分析表明,α-醇溶蛋白基因与低分子量谷蛋白亚基基因关系较近,在大约43.69百万年时分化,与ω-醇溶蛋白和HMW-GS基因亲缘关系较远,它们的分化时间大约为79.39百万年。     

Gliadin allele composition of Yugoslav winter wheat cultivars

Summary The complete gliadin allele composition of 57 Yugoslav common winter wheat cultivars was studied. Large differences were found in gliadin genotypes among cultivars bred at different Yugoslav breeding centres as well as between early and recent wheats bred in the Novi Sad breeding centre. Yugoslav wheats have limited variation in gliadin alleles, of which Gli-A1a, Gli-B1b, Gli-B1l, Gli-D1b, Gli-A2e, Gli-B2b, Gli-D2a are shown to be the most frequent. Examples of favoured alleles to new cultivars are described. 40% of the studied wheat cultivars have natural biotypes differing in gliadin allele composition. The frequency of cultivars with the biotypes differs greatly among breeding centres.     

The high and low molecular weight glutenin subunits and ω-gliadin composition of bread and durum wheats commonly grown in Portugal

A collection of 63 bread wheats (Triticum aestivum L.) and 21 durum wheats (Triticum durum Desf.) commonly grown in Portugal since 1982 were characterized for the composition of wheat storage proteins (WSP), high molecular weight glutenin subunits (HMW-GS), low molecular weight glutenin subunits (LMW-GS) and ω-gliadins. The composition of HMW-GS, LMW-GS and &-gliadins, encoded at loci Glu-1, Glu-3 and Gli-1, respectively, was revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. WSP allelic compositions of bread and durum wheat patterns were given. In the bread wheats, a total of 24, 24 and 18 patterns were observed for HMW-GS, LMW-GS and ω-gliadins, respectively. Forty-two different alleles were identified for the nine loci studied, Glu-A1 (3), Glu-B1 (7), Glu-D1 (4), Glu-A3 (5), Glu-B1 (7), Glu-D3 (2), Gli-A1 (2), Gli-B1 (8) and Gli-D1 (4). In the case of durum wheats, 19 alleles were identified: one allele at Glu-A1, two at Glu-B3, Glu-B2 and Gli-A1, three at Glu-B1, four at Glu-A3 and five at Gli-B1. For HMW-GS, LMW-GS and ω-gliadins, three, six and six different patterns were revealed, respectively. This study represents the first attempt to discriminate the bread and durum wheat varieties commonly grown in Portugal by the allelic variation of storage proteins. The database is useful for varietal identification and for plant breeders who seek to devise effective programmes aimed at improving wheat quality.     

Allelic variation at the gliadin coding loci of improved Ethiopian durum wheat varieties

The objective of this study was to determine gliadin allele compositions of 20 improved Ethiopian durum wheat varieties using acid-polyacrylamide gel electrophoresis (A-PAGE). Each block of co-dominantly inherited polypeptides encoded by gliadin loci were identified and their genetic diversities were estimated using statistical analyses. A total of 30 electrophoretic blocks were identified at five major gliadin loci. In addition, four novel gliadin blocks were identified. Gli-B1 and Gli-A2 loci had higher numbers of gliadin alleles (nine and ten, respectively) compared to other loci. Alleles Gli-A1c on chromosome 1A, Gli-B1c on chromosome 1B, Gli-A2a, and Gli-A2o on chromosome 6A, and Gli-B2h on chromosome 6B had maximal frequencies in their corresponding loci. Varieties were classified into three main clusters and one singleton based on genetic distances of detected gliadin alleles. These results indicate that Ethiopian durum wheat varieties are genetically diverse with unique allele compositions at gliadin-coding loci.     

Seed storage protein diversity in triticale varieties commonly grown in Portugal

A collection of 14 triticale (X Triticosecale Wittmack) varieties commonly grown in Portugal were analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to describe allelic diversity in the storage proteins encoded at the Glu-1 (Glu-A1, Glu-B1 and Glu-R1), Gli-1 (Gli-A1 and Gli-B1), Glu-3 (Glu-A3 and Glu-B3), Glu-B2 and Gli-R2 loci. Several alleles were identified: 10 at the loci encoding for high molecular weight(HMW)subunits (seven for glutenin and three for secalins), eight for low molecular weight glutenin subunits, four for γ-gliadins and three for 75K γ-secalins. These results showed that triticale varieties grown in Portugal exhibit great genetic diversity. Knowledge of the diversity of these storage proteins, which are the major gluten components, will greatly increase our understanding of the quality differences that might exist between triticale varieties.     

Cumulative and interaction effects of prolamin allelic variation and of 1BL/1RS translocation on flour quality in bread wheat

The allelic variation of prolamin loci was studied in three F₂ progenies from three crosses between the 1BL/1RS cultivar Triana and Yécora Rojo, Pavón and Florence Aurora, cultivars without the translocation. According to the 1:2:1 theoretical proportions observed in the allelic variants of the Glu-B3/Gli-B1 loci of the parent without the translocation, the inheritance as a block of the rye chromosome arm was confirmed. A group of F₃-F₄ recombinant lines, developed from these crosses was evaluated using the SDS-sedimentation test and the mixograph and alveograph tests. The presence of the 1BL/1RS translocation was not associated with significantly lower grain protein content values or with the optimum mixing time in the mixograph of the genotypes. The effect of the 1BL/1RS translocation on most of the quality parameters was highly dependent on the genetic pool. Significant increases in gluten strength and better mixing properties associated with the presence of some alleles of the Glu-A1, Glu-A3/ Gli-A1 and Gli-D2 loci were detected. The additivity and the interaction of prolamin gene effects with the rye translocation in the 1BL/1RS lines and its possible use in plant breeding are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Allelic variation at the storage protein loci of 55 US-grown white wheats

Fifty soft white and hard white wheat cultivars (Triticum aestivum L.), and five club wheat cultivars (T. compactum L.) were partially characterized in terms of their storage protein compositions, i.e. gliadins, and high molecular weight and low molecular weight glutenin subunits (HMW-GS and LMW-GS, respectively). At the Glu-1 loci, HMW-GS composition 1,7 + 9,2+ 12 was found to be predominant, being expressed in 11 cultivars out of 55. The most common alleles at the loci coding for gliadins and LMW-GS were found to be Gli-A1/Glu-A3a (43.6%), Gli-B1/Glu-B3b (36.4%), Gli-D1a/Glu-D3a (38.1%) and Gli-Dli/Glu-D3a (21.8%). Two-dimensional fractionation (acid-poly-acrylamide gel electrophoresis (A-PAGE) × sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)) of reduced and alkylated glutenins revealed that the number and the relative mobility of LMW-GS polypeptides were different from those reported for the corresponding Glu-3 alleles of hard-bread wheat cultivars. This result could account for the different technological properties of soft white wheats compared with hard-bread wheat cultivars, owing to the major impact of LMW-GS on dough quality.     

Genetic diversity of wheat storage proteins and bread wheat quality

To understand the genetic and biochemical basis of the bread makingquality of wheat varieties, a large experiment was carried out with a set of162 hexaploid bread wheat varieties registered in the French or EuropeanWheat Catalogue. This material was used to analyse their allelic compositionat the twelve main storage protein loci. A large genetic and biochemicaldiversity of the gluten proteins was found. Several gliadin encoding lociexhibited the highest allelic diversity whereas the lowest diversity was foundfor Glu-A1 and Glu-D3 loci encoding some high molecularweight glutenin subunits (HMW-GS) and LMW-GS respectively. Thevarieties were grown in three experimental locations in France. Qualityevaluation was carried out from material harvested in each location usingseven technological tests: grain protein content (Prot), grain hardness(GH), Zeleny sedimentation test (Zel), Pelshenke test (Pel), water solublepentosans (relative viscosity: Vr ), mixograph test (giving 11 parameters)and the alveograph test (dough strength W, tenacity P , extensibility L,swelling G, ratio P/L and the elasticity index Ie). Genetic and locationeffects as well as broad-sense heritability of each of the 22 technologicalparameters were calculated. GH, corresponding to the major Ha gene, Pel,and MtxW (mixograph parameter) had the highest heritability coefficients,alveograph parameters like W, P, the relative viscosity Vr and severalmixograph parameters had medium heritability coefficients whereas Protand L had the lowest. Variance analysis (using GLM procedure) allowed theeffect of the allelic diversity of the storage proteins, on the geneticvariations of each quality parameters, to be estimated. Glu-1 and Glu-3 loci had significant additive effects in the genetic variations of manyparameters. Gliadin alleles encoded at Gli-1 and Gli-2 were alsofound to play significant effect on several quality parameters. The majorpart of the phenotypic variation of the different quality parameters like Zel,Pel, W or mixograph peak time MPT was explained with the GH and allelesencoded at Glu-1 and Glu-3. Allelic variants encoded at Glu3and Gli-2 had similar contribution to the phenotypic variations ofquality parameters and accounted for 4% up to 21% each.     

Identification of Rye Chromosome 1R Translocations and Substitutions in Hexaploid Wheats Using Storage Proteins as Genetic Markers

Grain protein compositions of 106 advanced generation backcross lines from crosses involving ‘Amigo’ (1AL.1RS), ‘Aurora’, ‘Kavkaz’, ‘Skorospelka-35’ and ‘Sunbird’ (all 1BL.1RS) and ‘Gabo’ 1DL.1RS parents and 152 cultivars with unknown pedigree were analysed by one-dimensional SDS-PAGE. Eighty seven backcross lines and 16 cultivars carried one or other of these translocations, 2 cultivars had a 1R (1B) substitution, whereas 5 backcross lines were found to be heterogeneous for the 1BL.1RS translocation. The translocation lines were easily identified by the presence of secalins (Sec-1) controlled by rye chromosome arm IRS and a simultaneous loss of the gliadin (Gli-1) and/or triticin (Tri-1) protein bands controlled by the replaced wheat chromosome arm (1AS, 1BS or 1DS). Certain gliadins, showing no allelic variation among the genotypes analysed, were identified as markers for chromosome arms 1AS (M_r= 34 kd) and IBS (M_r= 42,33 kd). The whole chromosome substitutions 1R (1B) were recognized by scoring for the presence of Sec-1 and HMW secalin bands, Sec-3 (controlled by rye chromosome arm 1RL) and the absence of Gli-B1 and HMW glutenin subunits, Glu-B1 (controlled by wheat chromosome arm 1BL). The results have shown that protein electrophoresis provides a rapid and reliable technique for screening genotypes for these translocations and substitutions in a breeding programme.     

High genetic diversity of Spanish common wheats as judged from gliadin alleles

Gliadin alleles were identified in 100 common wheat cultivars registered and/or grown in Spain during the last 40 years. A very high level of genetic polymorphism was found: in total, 103 allelic variants including one null‐allele were found at the six major Gli loci in the Spanish wheats studied. An average genetic diversity for these six loci was found to be higher (H=0.844) than in any group of wheat cultivars studied previously. Spanish wheats bred in Spain demonstrated even higher genetic diversity (H=0.868), probably because of the occurrence in this group of some landraces (local varieties) assumed to be strongly differentiated to fit local environments. The high level of genetic diversity of wheats grown in Spain was maintained by the introduction of distantly related wheat germplasm from different sources, especially from Italy and CIMMYT. A slight decrease of genetic diversity in recently registered cultivars might be caused by the excessive introduction of French wheats. Thirteen new alleles found in Spanish wheats were catalogued, including Gli‐D2w which encodes the first Gli‐D2‐controlledγ‐gliadin to be found.