Allelic variation and composition of HMW-GS in advanced lines derived from d-genome synthetic hexaploid / bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

The objective of this study was to identify allelic variations at Glu-1 loci of wheat (Triticum aestivum L.) advanced lines derived from hybridization of bread wheat and synthetic hexaploid wheats (2n = 6x = 42; AABBDD). Locally adapted wheat genotypes were crossed with synthetic hexaploid wheats. From the 134 different cross combinations made, 202 F₈ advanced lines were selected and their HMW-GS composition was studied using SDS-PAGE. In total, 24 allelic variants and 68 HMW-GS combinations were observed at Glu-A1, Glu-B1, and Glu-D1 loci. In bread wheat, the Glu-D1 locus is usually characterized by subunits 1Dx2+1Dy12 and 1Dx5+1Dy10 with the latter having a stronger effect on bread-making quality. The subunit 1Dx5+1Dy10 was predominantly observed in these advanced lines. The inferior subunit 1Dx2+1Dy12, predominant in adapted wheat germplasm showed a comparative low frequency in the derived advanced breeding lines. Its successful replacement is due to the other better allelic variants at the Glu-D1 locus inherited in these synthetic hexaploid wheats from Aegilops tauschii (2n = 2x = 14; DD).     

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.     

小麦优质谷蛋白亚基分子标记多重PCR体系的建立与应用

Ax1/Ax2*、Dx5和过量表达的Bx7亚基(Bx7^OE)被认为是对小麦品质有正向效应的优质亚基。根据以上亚基特异性分子标记建立相应的多重PCR体系,经品种和群体检验,证明利用该体系鉴定亚基的结果稳定可靠、成本较低。利用该多重PCR技术对89份西藏小麦育成和推广品种(系)的优质亚基频率进行鉴定,结果表明, Dx5和Ax1/Ax2*亚基的频率均为12.4%,没有检测到Bx7^OE,同时携带两个优质亚基的材料频率为10.1%,该麦区品质育种必须加强优质亚基的引入。针对优质亚基Ax1/Ax2*、Dx5和Bx7^OE的多重PCR体系,为小麦品质育种亲本评价和通过杂交方法聚合优质亚基基因提供了一种实用可靠的标记辅助选择技术。     

高分子量麦谷蛋白亚基HPCE高效分离及图谱鉴定

高分子量麦谷蛋白亚基(HMW-GS)是决定小麦加工品质的重要因素。高效毛细管电泳技术(HPCE)以其用样少、速度快、精确度高等特点被越来越多地应用到分离鉴定中来。目前小麦HMW-GS的HPCE研究尚处于起步阶段, 对标准鉴定图谱的研究甚少,且已有分离体系在连续多次试验中的分离速度及分辨率仍有提升空间。本研究通过SDS-PAGE结合分子标记的方法鉴定了不同小麦品种的HMW-GS, 以“中国春”小麦为标样确立了HPCE高效分离体系, 体系条件为75 mmol L^-1 IDA+0.05% HPMC+15% ACN, pH 2.5, 电泳参数为毛细管内径25 μm, PDA检测波长200 nm, 分离电压20 kV, 运行温度30°C。通过混合进样方式对不同类型亚基进行HPCE分析, 建立了18个亚基的标准图谱, 亚基迁移顺序为1Dy12→ 1Dy10→ 1By9→ 1By8→ 1By18→ 1By16→ 1By20→ 1Bx17→ 1Bx20→ 1Bx13→ 1Bx6→ 1Bx7→ 1Ax2*→ 1Ax1→ 1Dx5→ 1Dx4→ 1Dx3→ 1Dx2,标准出峰时间依次为9.39、9.69、10.30、11.70、11.89、12.09、12.22、12.36、12.62、12.83、13.08、13.18、13.50、13.73、14.04、14.24、14.46和14.73 min, RSD<0.2%。以1Bx17为分界线, 9.39 min到12.36 min为y型亚基区域, 12.36 min到14.76 min为x型亚基区域。结合亚基迁移顺序、标准出峰时间及图谱特点可对小麦相关HMW-GS进行HPCE快速鉴定。本研究获得的分离体系及鉴定图谱可用于小麦HMW-GS定性分析和种质资源筛选。     

Allelic variation of the HMW glutenin subunits in Aegilops tauschii accessions detected by sodium dodecyl sulphate (SDS-PAGE), acid polyacrylamide gel (A-PAGE) and capillary electrophoresis

Variability of high molecular weight glutenin subunits (HMW-GS) was studied in198 accessions of Ae. Tauschii (2n=2x=14, DD) by sodium dodecyl sulphate(SDS-PAGE) and acid polyacrylamide gel electrophoresis (A-PAGE) and capillary electrophoresis (CE). A high allelic variation of HMW-GS, including some novel x- and y-type subunits and variable subunit combinations were observed. One accession(TD159) showed a x-type null form. The results by A-PAGE analysis revealed that the subunits Dx5 ^t and Dy10 ^t encoded by Glu-D ^t 1 locus in Ae. tauschii were different in relative mobilities in comparison with the subunits Dx5 and Dy10 found in bread wheats, whereas they had the same mobilities, respectively, when separated by SDS-PAGE. The higher resolution of Ae. tauschii HMW-GS separated by CE method showed two clear peaks in accordance with x- and y-type subunits, respectively,except the accession TD151 which possessed only subunit Dy12.1^*t. The electro elution time of the x-type and y-type subunits were about 13–14 and 7–8minutes, respectively. Characterization of wheat HMW-GS was facilitated by using CE which provides high resolution and increases the speed of analysis in conjunction with the traditional gel electrophoretic methods. A total of 42HMW-GS alleles were identified, among which were several alleles not presently detected in bread wheats. Hence Ae. tauschii is potentially a valuable genetic resource for quality improvement of bread wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

High molecular weight glutenin subunits of current bread wheats grown in Iran

Variation of high molecular weight gluteninsubunits (HMW-GS) at the Glu-1 lociwas studied using SDS-PAGE method in 43advanced lines or cultivars commonly grownin Iran. Fourteen alleles and 21 alleliccompositions were detected. Among the 43bread wheats analysed only five showed aunique HMW-GS composition. The mostfrequent HMW-GS patterns were 2^*, 7+8,2+12 and 2^*, 17+18, 2+12 which wereobserved in 13 and six cultivars,respectively. In the remainings, each twoto three lines or cultivars showed a commonHMW-GS pattern. Therefore, allelicvariation at Glu-1 loci is of limitedvalue for cultivar identification comparedwith loci controlling gliadins. Sevencultivars were observed to consist of twoto three biotypes with different alleles.In cultivar Mahdavi a biotype showedinactivity at the Glu-B1 locus. Analready reported rare subunit pair'2^***+12' at Glu-D1locus was found in cultivar Kavir. Theresults of scoring cultivars for theirquality based on the HMW-GS compositionswith an average score of eight, wasgenerally good. Cultivars Inia, Tajan, andadvanced lines N-75-11 and N-75-15 showedquality score equal to 10, whereas Alamootand C-75-5 showed quality scores equal tofive. The quality of former and latterlines and cultivars were considered highestand lowest, respectively. The resultsobtained in this study are useful inbreeding programs to improve bread makingquality, developing uniformity andimproving heterogeneous cultivars by meansof selection of the best genotypes.     

小麦高分子量谷蛋白亚基功能的体外鉴定

通过SDS-PAGE方法回收纯化小麦高分子量谷蛋白亚基1Ax1、1Dx5、1Dy10、1Bx7、1By8和1By9,然后利用微量配粉方法将单个亚基分别添加到对照“京411”面粉中,经过揉混仪分析单个亚基对揉面特性的影响,进而确定各个亚基的功能特性。根据揉面时间、稳定时间等参数的变化,6个小麦高分子量谷蛋白亚基对面粉品质的影响表现为1Dy10 > 1Ax1 > 1Dx5 > 1By8 > 1Bx7 > 1By9。研究结果表明,通过揉混仪进行体外配粉检测是快速鉴定高分子量谷蛋白亚基功能的一种有效方法。     

小麦Glu-B1位点1Bx14+1By18新亚基对材料的创制及其对加工质量的影响分析

以小偃54为轮回亲本,以引进品种Prinqual为1Bx17+1By18的供体,培育高分子量麦谷蛋白亚基（HMW-GS）近等基因系,以期客观评价1Bx14、1By15、1Bx17和1By18等亚基（对）对小麦加工品质的贡献。近等基因系回交至BC₄代时发现一个1Bx17亚基不表达的重组体,该重组体经自交分离纯合后获得了携带新亚基对1Bx14+1By18且可稳定遗传的品系012912。采用SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）、酸性聚丙烯酰胺凝胶电泳（A-PAGE）、小麦SDS微量沉降值和面粉揉面仪等方法。对小偃54和012912的谷蛋白及醇溶蛋白组成、总蛋白质含量、沉降值和揉面特性等指标进行了比较,结果表明,012912（1Ax1, 1Bx14+1By18, 1Dx2+1Dy12）与其轮回亲本小偃54（1Ax1, 1Bx14+1By15, 1Dx2+1Dy12）在谷蛋白成分上的唯一差别是以1By18替换了小偃54的1By15亚基,且1By18基因的表达量比小偃54的1By15表达量提高29%;012912品系的沉降值比小偃54高2.5 mL,揉面特性比小偃54好,说明1By18基因比1By15对面团加工品质的正向效应大。     

Allelic variation and genetic diversity of HMW glutenin subunits in Chinese wheat (Triticum aestivum L.) landraces and commercial cultivars

Wheat landraces have abundant genetic variation at the Glu-1 loci, which is desirable germplasms for genetic enhancement of modern wheat varieties, especially for quality improvement. In the current study, we analyzed the allelic variations of the Glu-1 loci of 597 landraces and 926 commercial wheat varieties from the four major wheat-growing regions in China using SDS-PAGE. As results, alleles Null, 7+8, and 2+12 were the dominant HMW-GSs in wheat landraces. Compared to landraces, the commercial varieties contain higher frequencies of high-quality alleles, including 1, 7+9, 14+15 and 5+10. The genetic diversity of the four commercial wheat populations (alleles per locus (A) = 7.33, percent polymorphic loci (P) = 1.00, effective number of alleles per locus (Ae) = 2.347 and expected heterozygosity (He) = 0.563) was significantly higher than that of the landraces population, with the highest genetic diversity found in the Southwestern Winter Wheat Region population. The genetic diversity of HMW-GS is mainly present within the landraces and commercial wheat populations instead of between populations. The landraces were rich in rare subunits or alleles may provide germplasm resources for improving the quality of modern wheat.     

以EMS诱变创制软质小麦宁麦9号高分子量谷蛋白亚基突变体

本研究旨在创制遗传背景一致的不同高分子量谷蛋白亚基(HMW-GS)缺失系, 为小麦品质研究和育种提供材料。将软质小麦品种宁麦9号, 用0.4% EMS溶液处理10 000粒种子, 获得3781个M1代单株, 采用“半粒法”对每个株系进行SDS-PAGE鉴定, 从中筛选出299个(7.91%) HMW-GS突变株, 包括HMW-GS缺失和分子量突变两种类型。其中, HMW-GS缺失突变176株, 突变频率为4.65%, 缺失亚基涉及Ax1、Bx7、By8、Dx2和Dy12, 突变频率为0.24%~3.28%; 分子量突变130株, 突变频率为3.44%。将突变体的具胚端种子于温室中繁殖获得M2代, 再次鉴定各株系的HMW-GS, 并经M3代验证, 最终获得Ax1、Dx2、Bx7、By8和Dy12缺失突变体, 及Ax1+By8双缺失突变体。用高效液相色谱(HPLC)分析这些突变体的谷蛋白大聚体(GMP)和谷蛋白/醇溶蛋白(GLU/GLI)比值, 发现不同缺失突变体的GMP含量都有不同程度的降低, 尤以Bx7亚基缺失突变体中GMP含量降幅最大, 高达42%。另外, 不同缺失突变体的谷蛋白总量和GLU/GLI比值也低于对照, Ax1+By8双缺失突变体的GLU/GLI比值最小。     

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     

新疆小麦品种高分子量谷蛋白亚基及相关品质基因的分子标记检测

高分子量麦谷蛋白亚基、1B·1R易位系、多酚氧化酶(PPO)活性及黄色素含量等基因对小麦加工品质有重要影响,准确、快速鉴定这些基因对品质改良有重要意义.本研究对新疆当地及国内外引进的321份小麦品种进行SDS-PAGE分析,利用特异性分子标记对高分子量谷蛋白亚基中的Dx5、Bx7、By8、By9亚基、1B·1R易位系、PPO和黄色素含量基因进行鉴定.进一步验证分子标记检测的可靠性和准确性,为优质小麦分子标记辅助育种提供材料和方法.SDS-PAGE分析表明,供试材料有21种亚基类型,其中Glu-A1位点有3种类型,以null为主;Glu-B1位点有10种类型,Bx7+By8和Bx7+By9亚基占主导地位;Glu-DJ位点有8种类型,Dx2+Dy12和Dx5+Dy10占主导地位.分子标记检测表明,Dx5亚基的频率为38.3%,Bx7亚基的频率为85.7%,By8亚基的频率为38.9%,Bx9亚基的频率为42.7%;特异性PCR标记扩增与SDS-PAGE鉴定结果的吻合率分别为97.2%、98.4%、93.4%和97.2%.在新疆当地品种、其他国内品种和国外品种中,1B·1R易位系材料的频率分别为22.0%、31.5%和25.0%,Psy-A1b的频率分别为9.0%、10.8%和5.4%,Ppo-A1b的频率分别为38.0%、43.8%和45.7%,Ppo-Dla的频率分别为48.0%、66.9%和40.2%.同时含Ppo-A1b和Ppo-D1a的材料有74份,占23.0%.本试验中采用的基因特异性标记重复性好、准确率高,可有效用于小麦品质分子标记辅助选择.     

小麦编码高分子量谷蛋白亚基基因的转化

以小麦的幼穗和幼胚作为转化受体,首次用抗除草剂草甘麟的EPSPs基因作为选择标记,通过基因枪法共转化,将小麦编码高分子量谷蛋白亚基的基因1Dx5和1Dy10转移到普通小麦京花1号中,获得33株再生植株,经PCR初步检测有12株同时扩增到了3个处在不同质粒上的外源基因EPSPs, 1Dx5和1Dy10的目标片段。将部分PCR检测为阳性的转化体     

Relationship Between High Molecular Weight Glutenin Subunit Composition and Gluten Quality in Ethiopian-grown Bread and Durum Wheat Cultivars and Lines

The high molecular weight glutenin subunit (HMW-GS) composition of 42 Ethiopian-grown bread wheat and 31 durum wheat cultivars and lines were examined using SDS-PAGE. Low variability in HMW-GS composition was present in both classes of wheat. A total of 10 variants with 14 different HMW patterns and seven variants with six different patterns were identified in bread and durum wheat, respectively, reflecting the limited ability of HMW-GS for cultivar identification. The most predominant alleles were 2*, 7 + 9 and 5 + 10 in bread wheat and nul and 7 + 8 in durum wheat. The Glu-1 quality scores for bread wheat ranged from 6 to 10, with an average value of 8.7. The variation in HMW-GS significantly correlated with and accounted for 44 % of the total variation in gluten quality, measured by the sodium dodecyl sulphate sedimentation test. In durum wheat, HMW-GS variation at Glu-B1 explained about 25 % of the variation in gluten quality. The high frequency of the 7 + 8 alleles among the landraces and the significant contribution of Glu-B1 alleles to the total variation in gluten quality indicate the potential benefit of Ethiopian tetraploid landraces in the development of lines suitable for both bread and pasta production.     

Characterization of HMW-GS and evaluation of their diversity in morphologically elite synthetic hexaploid wheats

High molecular weight glutenin subunit composition and variation in 95 Elite-1 synthetic hexaploid (SH) wheats (Triticum turgidum/Aegilops tauschii; 2n = 6× = 42; AABBDD) were determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis method (SDS-PAGE). Twenty two different alleles at Glu-1 loci in SHs were observed. Forty four different patterns of HMW-GS in synthetics were found. This higher HMW glutenin composition was due to higher proportion of D-genome encoded subunits in these SHs. 8% urea/SDS-PAGE better discriminated subunit 2* than 12% gels. However 12% urea/SDS-PAGE allowed differentiated mobility of Glu-D^t1 subunits. Genetic variability at Glu-D^t1 locus was greater than Glu-A1 and Glu-B1 loci. The relative high frequency of superior alleles, Glu-B1b and Glu-D^t1d indicated the superior bread making quality attributes embedded in these synthetic hexaploid wheats. Of the 95 Elite-1 SHs 27.1% possessed superior alleles at Glu-A1 and 51% had superior alleles at Glu-B1 locus. At Glu-D^t1 frequency of inferior allele 1Dx2 + 1Dy12 was very low (5.26%) and nine different rare alleles along with the higher frequency (22.1%) of D-genome encoded subunit, 1Dx5 + 1Dy10, were observed. These superior alleles shall form the priority selective sieve for their usage in wheat improvement efforts.     

高分子量谷蛋白5+10亚基和1B/1R易位分子标记辅助选择在小麦品质育种中的应用

谷蛋白亚基组成对小麦加工品质具有重要作用。以豫麦34、藁城8901和中优9507为优质亲本,以轮选987、石4185和周麦16为农艺回交亲本,采用5+10亚基和1B/1R易位分子标记结合田间农艺性状选择,育成4个BC₂F₄群体共125个高代品系。2008—2009年度,将这些高代品系分别种植于北京和河南安阳,分析了5+10亚基和1B/1R易位对蛋白质含量、和面时间和峰值曲线面积等品质参数的影响。4个群体中蛋白质含量与和面时间、峰值曲线面积等参数变幅较大,后代品系间品质差异明显,5+10亚基可显著增加和面时间和峰值曲线面积,1B/1R易位对和面时间和峰值曲线面积的作用则受遗传背景的影响。和面时间和峰值曲线面积等主要品质参数还受亚基表达量的影响,和面时间和峰值曲线面积与低分子量谷蛋白亚基含量显著正相关(r = 0.38~0.74,P < 0.05),导入5+10亚基可显著增加高分子量和低分子量谷蛋白亚基含量;Glu-B3位点等位基因的变化对高分子量谷蛋白亚基含量的影响不显著,对低分子量谷蛋白亚基含量的影响则因组合而异。通过有限回交,育种早代在室内采用5+10优质亚基和1B/1R易位分子标记辅助选择,结合田间农艺性状选择,可以加速培育优质新品种。     

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.     

小麦高分子量麦谷蛋白亚基多重PCR扩增体系的建立

小麦的加工品质与小麦高低分子量麦谷蛋白的组成和含量密切相关,特别是Glu-1位点编码的高分子量麦谷蛋白亚基(HMW-GS)的组成和含量.其中与好的加工品质相关的优质亚基如1Dx5、1Ax2*和1Bx7OB(over-expression)特异的分子标记已经被建立并应用于品质改良的分子育种.本研究的目的在于通过摸索PCR反应组分和循环参数对多重PCR反应结果的影响,建立一次反应能够同时鉴定1Ax2*、1Bx7OE和1Dx5亚基的多重PCR反应体系.结果表明:反应体系中的三种亚基的引物浓度比例和反应的Tm值是多重PCR成败的关键因素,当引物浓度比例为1Dx5:1Bx7CEL:1Ax2*=0.1:0.2:0.3或1Dx5:1Bx7OE:1Ax2*=0.1:0.2:0.4,Tm=60℃时1Ax2*、1Bx7OE和1Dx5亚基的多重PCR结果最好.利用HMW-GS的多重PCR的方法可以快速高效的鉴定多个亚基,可以进行小麦品质育种的复合分子标记辅助选择.     

Diversity of five glutenin loci within durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husn.) germplasm grown in Algeria

The HMW and B‐LMW glutenin subunits composition of 120 durum wheat germplasm grown in Algeria was examined using SDS‐PAGE. All together, 39 glutenin patterns were detected, including eight for HMW and 21 for B‐LMW glutenin subunits. Twenty‐six different alleles were identified for the five glutenin loci studied, that is, Glu‐A1 (3), Glu‐B1 (7), Glu‐A3 (5), Glu‐B3 (9) and Glu‐B2 (2). Two new alleles were found at Glu‐B3 locus: Glu‐B3new₁ encodes for five subunits (7 + 8 + 14 + 16 + 18) and Glu‐B3new₂ codes for five subunits (4 + 6* + 12 + 15 + 15*), of which subunit 15* with mobility between bands 15–16 was not described previously. At the Glu‐1 loci, the Glu‐A1c/Glu‐B1e allelic composition was predominant. For the B‐LMW glutenins, the most common allelic composition was Glu‐A3a/Glu‐B3a/Glu‐B2a. The collection analysed shows glutenin alleles and allele combinations related to high gluten strength. This information could be useful to select varieties with improved quality and also as a source of genes to develop new lines when breeding for quality.