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黄河中游粗山羊草三种y-型高分子量谷蛋白亚基的鉴定、克隆及系统进化分析 总被引:2,自引:0,他引:2
粗山羊草(Aegilops tauschii, 2n = 2x = 14, DD)是六倍体普通小麦的祖先之一,其高分子量谷蛋白亚基(HMW-GS)变异类型丰富,是小麦品质改良的重要基因资源。利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了黄河中游地区161份粗山羊草的HMW-GS,发现3种编码序列未知的y-型亚基,即Dy10.5t、Dy10.4t和Dy10.5*t亚基。通过AS-PCR扩增、克隆、测序和氨基酸序列推导,发现3种未知序列均具有典型HMW-GS的序列结构特征且较为相似,仅Dy10.4t与Dy10.5t亚基存在一个氨基酸重复单元的缺失,Dy10.5t与Dy10.5*t亚基在信号肽部位有一个氨基酸的替换(L-F)。通过对这3种HMW-GS与32个已知氨基酸序列的HMW-GS多序列比对和系统进化关系分析,证实Dy10.5t、Dy10.4t和Dy10.5*t 3个亚基是D基因组编码的高分子量谷蛋白y-型亚基家族的新成员。 相似文献
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粗山羊草(Aegilops tauschii, 2n = 2x = 14, DD)是六倍体普通小麦的祖先之一,其高分子量谷蛋白亚基(HMW-GS)变异类型丰富,是小麦品质改良的重要基因资源。利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了黄河中游地区161份粗山羊草的HMW-GS,发现3种编码序列未知的y-型亚基,即Dy10.5t、Dy10.4t和Dy10.5*t亚基。通过AS-PCR扩增、克隆、测序和氨基酸序列推导,发现3种未知序列均具有典型HMW-GS的序列结构特征且较为相似,仅Dy10.4t与Dy10.5t亚基存在一个氨基酸重复单元的缺失,Dy10.5t与Dy10.5*t亚基在信号肽部位有一个氨基酸的替换(L-F)。通过对这3种HMW-GS与32个已知氨基酸序列的HMW-GS多序列比对和系统进化关系分析,证实Dy10.5t、Dy10.4t和Dy10.5*t 3个亚基是D基因组编码的高分子量谷蛋白y-型亚基家族的新成员。 相似文献
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为了探明小麦收获前测定高分子量谷蛋白亚基的最佳时间,实现高分子量谷蛋白亚基与综合育种目标性状的同时选择.从小麦开花期始,对21个小麦品种(以中国春为对照)每5天用SDS.PAGE法测定其HMW-GS以寻找最佳测定时间,并对21个杂交组合后代进行连年选择.结果表明,乳熟中期HMW-GS谱带始达清晰,是当代测定当代选择的最佳测定时期.通过连年高分子量谷蛋白亚基的选择,部分高分子量谷蛋白优质亚基出现的频率得到显著的提高,选择效果良好.选育出高产优质小麦新苗头品系宝麦35,证明当代测定高分子量谷蛋白亚基的同时结合综合育种目标性状进行选择,对于选育优质,高产、抗逆新品种效果是良好的. 相似文献
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小麦收获前HMW-GS最佳测定时间的探讨及在育种上的应用 总被引:1,自引:1,他引:0
摘 要:为了寻找小麦收获前测定高分子量谷蛋白亚基的最佳时间,实现高分子量谷蛋白亚基与综合育种目标性状的同时选择。从小麦开花期始,对21个小麦品种(以中国春为对照)每5天用SDS-PAGE法测定其HMW-GS以寻找最佳测定时间,并对21个杂交组合后代进行连年选择。试验结果表明,乳熟末期HMW-GS谱带始达清晰,是当代测定当代选择的最佳测定时期。通过连年高分子量谷蛋白亚基的选择,部分高分子量谷蛋白优质亚基出现的频率得到显著的提高,选择效果良好。本研究选育出高产优质小麦新苗头品系宝麦35,证明当代测定高分子量谷蛋白亚基的同时结合综合育种目标性状进行选择,对于选育优质,高产、抗逆新品种效果是良好的。 相似文献
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《分子植物育种》2017,(10)
高分子量谷蛋白(HMW-GS)是小麦的重要储藏蛋白之一,决定小麦烘烤品质,其中HMW-GS Glu-D1位点与面包烘烤品质的关系最为密切。人工合成小麦(synthetic hexaploid wheat,SHW)继承了节节麦D基因组丰富的遗传变异,是高效利用野生祖先种优良基因的重要桥梁资源;近年来随着人工合成小麦在小麦遗传育种中的不断应用,来自节节麦类群的大量高分子量谷蛋白亚基类型越来越多的涌向普通小麦。而传统SDS-PAGE无法区分节节麦HMW-GS Dtx5和普通小麦Dx5亚基,鉴于此本研究利用节节麦HMW-GS Dtx5亚基和普通小麦HMW-GS Dx5亚基DNA序列之间存在的几个SNPs差异开发出等位特异PCR(allele specific polymerase chain reaction,AS-PCR)引物,通过PCR方法来区分两种不同来源的5亚基。结果显示,所设计出的两对引物都能够扩增出清晰稳定的目标条带,并且两对引物的扩增结果一致,含HMW-GS Dx5亚基的表现为阳性带,含节节麦来源的Dtx5亚基表现为阴性;同源性分析表明Dtx5亚基与Dx2亚基氨基酸序列的同源性要高于Dtx5与Dx5之间的同源性。 相似文献
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陇东地区部分冬小麦品种(系)HMW-GS的组成与遗传变异分析 总被引:1,自引:1,他引:0
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. 相似文献
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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. 相似文献
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小麦地方品种高分子量谷蛋白亚基多样性分析 总被引:3,自引:0,他引:3
采用SDS-PAGE方法,对我国9个麦区的76份代表性地方品种的高分子量谷蛋白亚基(HMW-GS)组成比较分析,并探讨其与环境因素(平均海拔、年平均降雨量和年平均温度)的相关性。结果表明,25.0%的品种具有异质性,分别包含2~4种不同HMW-GS组合;在Glu-1位点共检测到14个等位变异,其中Glu-A1、Glu-B1和Glu-D1等位变异数分别为2、7和5;发现了3个新等位变异,包括Glu-B1位点2个和Glu-D1位点1个。所有等位变异构成16种不同的亚基组合类型,以(N, 7+8, 2+12)为主,频率为69.7%。在Glu-1位点上,不同麦区遗传多样性分布存在一定的不均衡性,年平均降雨量和年平均温度与麦区多样性指数呈负相关。推测环境压力可能是地方品种多样性地区分化的重要因素。 相似文献
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明确 2004s-47品系 1Dx基因的序列,为寻找新的优质 HMW-GS类型、实现小麦品质的改良及育种提供理论依据。应用 SDS-PAGE对 2004s-47品系中 HMW-GS的亚基组成进行分析,用 PCR技术克隆1Dx亚基基因并用 DNAMAN软件进行序列分析。结果从 2004s-47品系中扩增出了大小为 2514 bp的基因。该基因具有典型的小麦 HMW-GS x-型基因序列结构特征。又因为 1D比 1A和 1B基因组中的 x-型和 y-型基因易表达,结合 SDS-PAGE的结果,所以 2004s-47品系中 HMW-GS的亚基组成为(Null, 7+8, ?+10)。序列比对表明, 2004s-47品系的 1Dx?基因与节节麦(Aegilops tauschii) 1Dx2.1t (AF480486)同源性最高。将该序列提交到 NCBI,获得序列登陆号为: KP702118。2004s-47品系中 1Dx?序列的确定,为原核表达确定1Dx?是否为新的基因提供了基础,也为品质改良及育种提供了依据。 相似文献
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中国特有小麦与斯卑尔脱小麦和密穗小麦高分子量谷蛋白亚基多态性比较分析 总被引:3,自引:0,他引:3
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法对中国特有小麦(新疆稻麦、西藏半野生小麦和云南铁壳麦)高分子量谷蛋白亚基组成进行了研究,并综合前人的研究结果,比较分析了中国特有小麦、斯卑尔脱小麦和密穗小麦高分子量谷蛋白亚基的多态性,结果显示,斯卑尔脱小麦在高分子量谷蛋白亚基组成上具有明显特点,在Glu-A1位点上以1亚基为优势亚基,在Glu-B1位点上以13+16亚基居多,而这两种亚基在其他六倍体小麦中出现的频率相对较低。新疆稻麦在Glu-1D位点上具有特殊的HMW-GS类型。结合核基因组和叶绿体基因组的研究结果对中国特有小麦的起源演化进行了探讨。 相似文献
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X. L. An X. H. Li X. J. Xiong Y. M. Yan Y. Z. Zhang L. Y. Gao A. L.Wang K. Wang F. J. Zeller S. L. K. Hsam 《Plant Breeding》2009,128(1):41-45
A new x-type HMW glutenin subunit, designated as 1Dx1.6t from Aegilops tauschii was identified and characterized by SDS-PAGE and MALDI-TOF-MS. This subunit is located between 1Dx2 and 1Dx1.5t and possesses a molecular mass ( M r ) of 88565.8 Da. Its complete coding sequence was amplified via allele-specific PCR (AS-PCR), and the amplified product was cloned and sequenced. The authenticity of the cloned 1Dx1.6 t gene was confirmed by successful expression of its open reading frame in Escherichia coli. The molecular characterization of 1Dx1.6 t gene showed that its coding region consisted of 2541 bp encoding a polypeptide of 845 amino acid residues. Sequence comparison to previously characterized 1Dx1.5t subunit which is related to good dough quality of bread wheat indicated that the 1Dx1.6t subunit displayed high homology, but possesses 14 residue substitutions and a nonapeptide insertion. A total of 12 single-nucleotide polymorphisms (1 per 212 bp) was identified in the 1Dx1.6 t allele (11 in repetitive domain and 1 in the C-terminal domain), which could facilitate the design of AS-PCR markers. 相似文献
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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. 相似文献
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High-molecular-weight (HMW) glutenin subunits in wheat Triticumaestivum L., allelic variation for which affects bread-making quality, areencoded by Glu-1 homoeoloci located on the homoeologous group1 chromosomes. Many alleles at Glu-B1 and Glu-D1 producetwo subunits, an x-type of low electrophoretic mobility in polyacrylamidegels, and a y-type of high mobility. In the current study, a combination ofnear isogenic lines of cultivar `Sicco' has been used to characterise theeffect upon quality of the absence of individual subunits 7 (Glu-B1x-type), 12 (Glu-D1 y-type) and, assuming an additive model ofsubunit action, 2 (Glu-D1 x-type). Absence of subunit 7 gave amoderate reduction in SDS-sedimentation volume, indicating its associationwith lower gluten strength (confirmed by Farinogram and Extensogramstudies), yet, from a full mixing input bake, a moderate increase in loafvolume and a considerable improvement in loaf score (an overall evaluationof loaf quality). Absence of subunit 12 gave a slightly larger reduction inSDS-volume, yet no change in loaf volume or score. Absence of bothsubunits 2+12 gave a larger reduction again in SDS-volume, a moderatereduction in loaf volume and a large reduction in loaf score. Absence ofsubunit 2 alone is therefore predicted to reduce SDS-volume, loaf volumeand score such that loss of this x-type subunit would lead to larger changesin quality parameters than loss of y-type subunit 12. A general conclusionof the study is that, while deficiency for HMW glutenin subunits generallyleads to reduced gluten strength and viscoelasticity, the resultantintermediate gluten strength may on occasions lead to improvements in loafperformance in situations where the base gluten strength is high. Theremay, then, be contexts in breeding programmes where selection fordeficiency would be a possible strategy for improving bread-making quality,adding to the flexibility available to the breeder. Somewhat unexpectedly,additional analysis found that, in the genetic background of cultivar `Sicco'used in this study, subunits 7+8 at Glu-B1 were indistinguishablefrom their allelic counterparts subunits 7+9 for virtually all characters, andthat subunits 2+12 at Glu-D1, while inferior in performance formixing properties compared to subunits 5+10, were associated with goodloaf characteristics. 相似文献
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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。 相似文献