建国以来山东省小麦品种及其亲本Glu-1位点的亚基组成和多样性分析

探讨建国以来山东省小麦品种Glu-1的亚基特点及其多样性,并有针对性地筛选含有优质亚基的亲本。用SDS-PAGE法对山东省建国以来推广的主要品种及其亲本的HMW-GS组成和多样性进行了分析。山东省小麦品种及骨干亲本在Glu-A1位点主要为两种等位变异类型“Null”和“1”;Glu-B1有“7+8”、“7+9”、“14+15”、“17+18”、“6+8”5个等位变异类型,但主要以“7+8”和“7+9”为主;Glu-D1有“2+12”、“2+10”、“5+10”、“4+10”、“4+12”、“2+11”6个等位变异类型,以“2+12”为主。在3个位点中,Glu-B1位点的多样性最丰富,其次为Glu-D1位点,Glu-A1位点的多样性最差。Glu-1的多态性指数从50年代的高点开始下降,在60年代达到最低点,之后缓慢上升,在90年代达到最高点,当前表现为下降的趋势。山东省小麦品种及其亲本缺少优质亚基组合,这为山东省小麦品种的改良提供了方向。     

陇东地区部分冬小麦品种(系)HMW-GS的组成与遗传变异分析

本实验通过SDS-PAGE技术,对陇东地区部分冬小麦品种(系)的高分子量谷蛋白亚基组成及亚基组成频率进行了分析.结果表明陇东地区冬小麦品种(系)存在16种亚基组合类型,以N,7+8,2+12亚基含量丰富,其出现频率占品种(系)总数的36.50%,而含5+10亚基的亚基组合类型只有3种,仅占品种(系)数量的9.62%.Glu-1位点遗传多样性单一,Glu-A 1位点仅有2种等位变异形式,N亚基频率最高(71.15%);Glu-B 1位点有4种等位基因变异形式,7+8亚基频率最高(59.62%);Glu-D 1位点上有6种等位基因变异形式,2+12亚基频率最高(61.54%).通过对地方品种、育成品种(系)和引进品种HMW-GS组成比较发现,地方品种亚基的组成类型呈现出组成单一,优质亚基少的特点;而外引品种和育成品种亚基类型较为丰富,且包含一定数量的优质亚基.     

黄淮麦区南片小麦Glu-3位点等位变异分析

小麦的加工品质与低分子量谷蛋白亚基(LMW-GS)组成息息相关。为了尽快改良黄淮麦区小麦加工品质,应用STS分子标记与SDS-PAGE相结合的方法对小麦低分子量谷蛋白亚基Glu-A3、Glu-B3与Glu-D3位点的等位基因变异类型进行检测。结果表明:黄淮麦区南片356份小麦品种(系)中共检测到15种Glu-3位点等位变异,Glu-A3位点含Glu-A3a、Glu-A3b、Glu-A3c和Glu-A3d共4种等位变异,其分布频率分别为12.1%,13.5%,41.0%,33.4%;GluB3位点含Glu-B3a、Glu-B3b、Glu-B3d、Glu-B3f、Glu-B3g、Glu-B3h、Glu-B3i和Glu-B3j共8种等位变异,其分布频率分别为8.71%,8.99%,23.0%,5.90%,7.30%,3.65%,0.28%,42.1%;Glu-D3位点含Glu-D3a、Glu-D3b和Glu-D3c共3种等位变异,其分布频率分别为40.2%,29.8%,30.0%。等位变异组合Glu-A3c/Glu-B3j/Glu-D3a分布频率最高(10.7%)。通过优质亚基的转育,多个优质亚基的聚合,将有助于改良我国小麦的品质。     

Glu-1位点等位变异及表达量对麦谷蛋白聚合体粒度分布的影响

选用我国春播麦区23份(试验I)和北部冬麦区21份(试验II)品种(系),研究了Glu-1位点等位变异及其亚基表达量对谷蛋白聚合体粒度分布的影响。结果表明,Glu-1位点等位变异及其亚基表达量显著影响谷蛋白聚合体的粒度分布,且影响程度受蛋白质含量,尤其是高分子量谷蛋白总量水平的影响。在高分子量谷蛋白总量较低时(试验I),Glu-B1和Glu-D1位点对不溶性谷蛋白大聚体含量(UPP)及其占聚合体蛋白总量的百分比(%UPP)的加性效应都达1%显著水平;Glu-B1和Glu-D1位点单个亚基对两者的贡献分别为7^OE+8^* >7+9 >17+18 >7+8和5+10 >2+12,具有5+10亚基组合的%UPP显著高于具有2+12的亚基组合。高分子量谷蛋白的亚基表达量与UPP含量呈高度正相关,相关系数为0.79~0.93(P < 0.01)。而在高分子量谷蛋白总量较高时(试验II),仅Glu-D1位点对%UPP的加性效应达5%显著水平,5+10亚基对%UPP的贡献显著高于2+12和4+12;亚基组合间的聚合体粒度分布无显著差异。高分子量谷蛋白的亚基表达量与UPP含量的相关系数为0.42~0.86(P < 0.05或0.01)。结合高分子量谷蛋白表达量和优质亚基进行选择,能有效提高不溶性谷蛋白大聚体的含量和相对比例,有利于面筋强度和加工品质的进一步提高。     

麦谷蛋白亚基Clu-1和Glu-3位点基因等位变异对小麦品质特性的影响

甲单向一步SDS-PAGE方法分析表明亲本品种Suneca和Cook在麦谷蛋白亚基的5个位点(Glu-B1,Glu-D1,Glu-A3,Glu-B3和Glu-D3)均含不同等位基因。本研究重点对Suneca×Cook的F_4代群体中在麦谷蛋白亚基位点均为纯合基因的60个系的出粉率(FY),面粉蛋白质含量(FP)及和面时间(PTM)进行了分析,以研究麦谷蛋白各亚基位点等位基因变异及位点间互作对小麦品质特性的影响。结果表明,不同基因型间出粉率无显著差异,Glu-D1位点等位基因d和a对FP的效应存在显著差异,Glu-Dld基因(编码5 10亚基)的正效应显著高于Glu-Dla基因(编码2 12亚基);Glu-D1、Glu-A3和Glu-B3位点上基因的等位变异对PTM有显著和极显著影响,含Glu-Dld、Glu-A3b和Glu-B3b基因的系分别比含Glu-Dla,Glu-A3d和Glu-B3h基因的系有较长的和面时间;Glu-B1位点上等位变异i和u以及Glu-D3位点等位基因b和e分别对PTM无明显影响。在这种遗传背景下,麦谷蛋白亚基位点对PTM的效应大小依次排列为Glu-D1>Glu-B3>Glu-A3>GIu-B1=Glu-D3。Glu-1位点和Glu-3位点间对和面特性的影响存在累加效应和互作效应。     

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。     

利用HMW-GS全缺失突变体快速构建Glu-1位点近等渗入系

小麦高分子量麦谷蛋白亚基(highmolecular weight glutenin subunit, HMW-GS)由Glu-A1、Glu-B1、Glu-D1位点中含有的复等位基因编码,评价和优化Glu-1位点组合是认识与改良HMW-GS表达与功能的重要途径。本研究创制了以小偃81为背景的HMW-GS基因完全缺失突变体DLGlu1。将DLGlu1与加拿大优质强筋小麦品种Glenlea杂交,结合后代幼胚培养与分子标记辅助选择技术,在BC₃F₃种子中快速鉴定出来自Glenlea的Glu-A1a、Glu-B1al和Glu-D1d位点不同组合的7种渗入系材料,可进一步发展成一套完整的Glu-1位点有差异的近等渗入系。本研究表明,DLGlu1可用于Glu-1位点近等渗入系的快速创制,对Glu-1位点功能研究和改良具有重要价值。     

小麦-黑麦远缘杂交后代高分子麦谷蛋白亚基变异分析

采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE) 方法, 分析了小麦-黑麦远缘杂交后代的66个株系的高分子量谷蛋白亚基(HMW-GS) 组成.结果表明:(1)在29个母本为MY 11的株系中,有6个株系的Glu-1等位基因相对于母本MY 11发生了变异,1 B/1 R易位系中Glu-1位点发生变异的频率为66.67%,非1 B/1 R易位系中Glu-1位点发生变异的频率为8.69%.在36个母本为A 42912株系中,3个株系的Glu-1等位基因发生变异且这3个株系皆为1 B/1 R易位系,1 B/1 R易位系中Glu-1位点发生变异的频率为60.0%,非1 B/1 R易位系Glu-1位点没有变异;(2)对于母本为MY 11的株系,Glu-1三个位点上等位基因的变异均只有2种,即Glu-A 1 a(86.20%)、Glu-A 1 c(13.79%)、Glu-B 1 b(89.65%)、Glu-B 1 c(10.34%)、Glu-D 1 a(3.45%)、Glu-D 1 d(96.55%).对于母本为A 42912的株系,仅在Glu-B 1位点发生变异,即Glu-B 1 b(91.66%)、Glu-B 1 c(8.33%);(3)供试株系共出现6种高分子谷蛋白亚基组合,即(1,7 8,5 10)、(Null,7 8,5 10)、(Null,7 9,5 10)、(Null,7 8,2 12)、(1,7 8,5 10)、(1,7 9,5 10),两种母本不同的株系类型均是母本型HMW-GS组合(1,7 8,5 10)占绝对优势.本文分析了1 B/1 R易位株系具有高频率HMW-GS变异的原因,并就这些材料在小麦优质育种中的利用策略作了探讨.     

黄淮麦区小麦新品系高分子量谷蛋白亚基的组成及结构变化分析

为了解高分子量谷蛋白亚基(HMW-GS)组成对小麦品质产生的影响,利用SDS-PAGE技术对黄淮麦区221份小麦新品系的HMW-GS组成进行鉴定,对其系谱来源进行分析,并比较近10年来黄淮麦区小麦品种(系)的HMW-GS结构变化。结果表明,黄淮麦区小麦新品系的杂交亲本来源较为单一,具有周8425B和豫麦2号血统的材料高达155个,占到全部供试材料的70.1%,以郑麦9023等小偃系列和济麦20等鲁麦系列为亲本的材料分别有34,40个,占全部供试材料的15.4%和18.1%。221份供试材料中共出现13种亚基及亚基组合,其中在Glu-A1位点,亚基1出现频率最高,占供试材料的58.8%,自2008年以来,亚基1的出现频率高于Null类型;在Glu-B1位点上,共出现7种亚基变异类型,出现频率较高的亚基组合是7+9和7+8,分别占供试材料的57.5%和30.3%,自2007年以来,7+9亚基组合类型的出现频率最高,优质强筋的17+18和13+16亚基组合的出现频率相对较低;在Glu-D1位点上,强筋亚基组合5+10占供试材料的15.4%,自2007年以来,5+10和2+12亚基组合的出现频率整体呈平行的趋势。本试验还检测出普通小麦品种中很少出现的5+12亚基组合,占供试材料的9.0%。     

四川藏区小麦地方品种高分子量谷蛋白亚基组成分析

利用SDS-PAGE分析检测了36份四川藏区小麦地方品种的高分子量谷蛋白亚基组成,从中可检测到7种亚基组合类型.其中,Null、7+8、2+12为四川藏区小麦的优势亚基组合,其频率高达58.33%.在Glu-A 1、Glu-B 1和Glu.D 1位点中,分别有3,5和3种等位变异类型,各位点出现频率最高的亚基分别是Null、7+8和2+12,频率分别为86.11%、63.89%和94.44%.在WL25的Glu-B 1位点发现了一个未知的y型亚基,其迁移率比By 8慢,与Bx7以亚基组合形式同时出现.在wL 16的Glu-D 1位点,Dx亚基沉默,而仅表达了Dy 12亚基.参试材料的面包品质评分为4-8分,无评分达9分以上的品种.     

Genetic diversity of European spelt wheat (Triticum aestivum ssp. spelta L. em. Thell.) revealed by glutenin subunit variations at the Glu-1 and Glu-3 loci

Summary High and low molecular weight glutenin subunit (HMW-GS and LMW-GS) compositions of 270 European spelts, 15 Iranian spelts and 25 bread wheat cultivars were analyzed by one- and two-dimensional gel electrophoresis. The results revealed a total of 22 HMW-GS alleles (4 at Glu-A1, 11 at Glu-B1 and 7 at Glu-D1) and 32 allele combinations among the three Glu-1 loci. Two major genotypes of HMW-GS: 1, 13+16, 2+12 and 1, 6.1+22.1, 2+12 were found to occur in Central European spelt wheat cultivars and landraces at higher frequencies of 35 and 28%, respectively. The Glu-B1 locus displayed the greatest variation and genetic diversity index (H) was 0.69 whereas Glu-A1 and Glu-D1 showed H index values of 0.26 and 0.19, respectively. The dendrogram constructed by HMW and LMW glutenin subunit bands revealed that European spelts form a separated cluster from common wheat suggesting that spelt and common wheat form distinct groups. In addition, all 15 Iranian spelt land variety accessions differed from European spelts and possessed similar HMW-GS alleles to common wheat. Because of a wider polymorphism Central European spelt wheats are an important genetic reserviour for improving common wheat quality. Both authors contributed equally to this work     

Allelic variation in high-molecular-weight glutenin subunit Loci of Glu-1 in Japanese common wheats

Seed storage proteins of 131 Japanese Norin wheat (Triticum aestivum) varieties were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis to determine allelic make-up in varieties at each of three loci that control high-molecular-weight (HMW) glutenin subunits. Three alleles were identified at the Glu-A1 locus, six at the Glu-B1 locus and five at the Glu-D1 locus. Twenty-four different, major glutenin HMW subunits were identified and each contained three to five subunits and seventeen different glutenin subunit patterns were observed for 19 subunits in the 131 Japanese Norin varieties. Fourteen alleles were identified by comparison of subunit mobility with that previously found in hexaploid wheat. Japanese Norin varieties showed a specific pattern of allelic variation in glutenin HMW subunits, different from that of Chinese and other country common wheats in allelic frequency at Glu-1 loci. This revised version was published online in July 2006 with corrections to the Cover Date.     

小麦重组自交系群体HMW-GS的检测与分析

利用重组自交系群体--RILL-8群体的131个系为材料,检测和分析了其高分子量麦谷蛋白亚基及亚基组合.结果表明,RIL-8群体Glu-A1、Glu-B1、Glu-D1位点编码的亚基分别为1、N,7 9、7 8和5 10、2 12,主要存在7种亚基组合类型.不同亚基及亚基组合类型在相同位点上仅存在1对等位基因差异,可以用其进行相同位点不同亚基及亚基组合对品质性状效应值的估算.     

Biochemical and molecular characterisation of Glu-1 loci in Argentinean wheat cultivars

The high molecular weight glutenin subunit (HMW-GS) composition of acollection of 107 Argentinean bread wheat cultivars was analysed bysodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE).Allelic variation at the Glu-1 loci was identified and its frequencycalculated. Eleven alleles were detected, three encoded at the Glu-A1locus, six at the Glu-B1 locus and two at the Glu-D1 locus. Alow frequency of the null allele at the Glu-A1 locus and a highfrequency of subunits 5+10 at the Glu-D1 locus were observed.Reversed phase-high performance liquid chromatography (RP-HPLC)analysis was used to further characterise HMW-GS. Two different types ofBx subunit 8 (named subunits 8 and 8) were detected, the latterhaving shorter elution time. Subunit 8 was not identifiable bySDS-PAGE. According to quantification by RP-HPLC analysis, two groupsof subunit 7 were observed. One group, with a relatively high proportionof subunit 7 (approximately 39% of the total amount of HMW-GS)appeared in cultivars with allele 7+8 at the Glu-B1 locus; asecond group of subunit 7 (around 24% of the total amount ofHMW-GS), was found in alleles 7+8, 7+8 and 7+9. Restrictionfragment length polymorphisms (RFLP) analyses of HMW-GS genes werealso carried out after digestion of genomic DNA with HindIII andTaqI restriction enzymes. The relationship between DNA fragment sizeand glutenin subunits, as estimated by electrophoretic mobility inSDS-PAGE, was also examined. The restriction enzyme TaqIdemonstrated to be a useful tool to detect homozygous plants in selectionprograms against the Glu-A1 null allele.     

CIMMYT引入小麦材料HMW-GS组成研究

利用SDS-PAGE技术对385份CIMMYT种质进行高分子量麦谷蛋白亚基组成分析。结果表明:Glu-A1、Glu-B1、Glu-D1位点上变异类型丰富,共存在11亚基与24种组合类型,优质亚基出现的频率非常高,Glu-A1位点上的1亚基的频率达到了33.5%,Glu-B1优质亚基7+8出现的频率11.2%,Glu-D1位点优质亚基5+10出现的频率达到81.3%。评分在7分以上的材料达37.8%,23个材料评分在9分以上,高分子量麦谷蛋白亚基的平均得分7.08分。通过在这些材料中选取评分较高的与贵州小麦进行杂交,可有效提高贵州小麦品种的产量与品质。     

Multiplex-PCR typing of high molecular weight glutenin alleles in wheat

In Australian commercial cultivars, each high molecular weight glutenin (Glu-1) homoeologous locus consists of one of two predominant alleles: Glu-A1a (subunit Ax1) or Glu-A1b (subunit Ax2*) at the GluA1 locus, Glu-B1b (Bx7 and By8 subunits) or Glu-B1i (Bx17 and By18 subunits) at the Glu-B1 locus, and Glu-D1d (Dx5 and Dy10 subunits) or Glu-D1a (Dx2 and Dy12 subunits) at the Glu-D1 locus. PCR-based assays have been developed in this study to discriminate between these common alleles at each locus. Primers specific for the Glu-A1 Ax2* gene give a single fragment of 1319 bp only in the presence of this gene. Primers targeting the Glu-B1 locus resulted in a co-dominant marker for which the Bx7 genotype produced two fragments (630 bp and 766 bp) and the Bx17 genotype a single fragment (669 bp). The third pair of primers was specific for the Dx5 gene and resulted in a single band of 478 bp. A multiplexed PCR assay was established which permitted the discrimination of the major HMW glutenins in a single PCR reaction and agarose gel assay. As the HMW glutenin composition of a wheat line is extremely important in determining the functional properties of wheat gluten, these markers are useful for the purposes of marker-assisted breeding. These markers may also be useful for the purpose of DNA-based identification of wheat varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

南疆冬小麦品种(系)高分子量谷蛋白亚基组成分析

采用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳技术，系统分析了新疆的56份冬小麦品种及品系的高分子量谷蛋白亚基的组成与分布。在3个位点上一共检测到15种不同的亚基类型。在Glu—Al位点上有三种等位变异类型“0”，“1”，“2”；在Glu—B1上有“7”，“7 8”，“7 9”，“22．7 9”，“14 15”，“14 15．9”．“7．17 18”七个等位变异类型，但主要以“7 8”和“7 9”为主(分别占48．2％和32．1％)；在Glu—D1位点上有“2 12”，“5 1l”，“5 10”，“2 1l”，“5 12”五个等位变异类型，其中以“2 12”(75％)和“5 10”(16．1％)为主，另外，本试验还研究了这些高分子量谷蛋白亚基及其组合形式出现的频率和特点。     

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.