小麦骨干亲本碧蚂4号系谱种HMW-GS组成分析

利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,对小麦骨干亲本碧蚂4号80个衍生品种、系谱当中涉及的18个中间亲本、碧蚂4号6个姊妹系品种及其亲本共106个品种(系)的HMW-GS组成进行分析.结果表明,碧蚂4号HMW-GS组成为Null、7+8、2+12.在Glu-B1位点,碧蚂4号的7+8亚基与一代衍生品种的3个亲本均不同,其在衍生一代中的遗传受到了显著的选择,遗传频率达到84.6%;6个(54.5%)二代衍生品种的亲本含有与碧蚂4号相同亚基,该代7+8亚基的遗传频率仍最高,达到64.1%;三代和四代衍生品种的5个亲本中只有北京6号和矮秆早含有7+8亚基,这两个世代7+8亚基遗传频率明显降低,7+9成为主要类型,频率分别达到了69.2%和64.3%.在Glu-A1和Glu-D1位点,各有11个(64.7%)中间亲本具有与碧蚂4号相同的亚基,它们分布于不同子代涉及的中间亲本中,Null和2+12亚基在其衍生的4个世代品种的遗传频率均不小于76.9%.系谱分析发现,在三代和四代,中间亲本HMW-GS组成遗传频率显著增加,其中以洛夫林10的Null、7+9、2+12亚基为主.由于HMW-GS组成不是碧蚂4号系谱品种育成时期的选择目标,对骨干亲本碧蚂4号和洛走林10号被选择的Glu-B1位点进一步研究有助于解析骨干亲本的形成原因.     

小麦骨干亲本碧蚂4号系谱品种HMW-GS组成分析

利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,对小麦骨干亲本碧蚂4号80个衍生品种、系谱当中涉及的18个中间亲本、碧蚂4号6个姊妹系品种及其亲本共106个品种(系)的HMW-GS组成进行分析。结果表明,碧蚂4号HMW-GS组成为Null、7+8、2+12。在Glu-B1位点,碧蚂4号的7+8亚基与一代衍生品种的3个亲本均不同,其在衍生一代中的遗传受到了显著的选择,遗传频率达到84.6%;6个(54.5%)二代衍生品种的亲本含有与碧蚂4号相同亚基,该代7+8亚基的遗传频率仍最高,达到64.1%;三代和四代衍生品种的5个亲本中只有北京6号和矮秆早含有7+8亚基,这两个世代7+8亚基遗传频率明显降低,7+9成为主要类型,频率分别达到了69.2%和64.3%。在Glu-A1和Glu-D1位点,各有11个(64.7%)中间亲本具有与碧蚂4号相同的亚基,它们分布于不同子代涉及的中间亲本中,Null和2+12亚基在其衍生的4个世代品种的遗传频率均不小于76.9%。系谱分析发现,在三代和四代,中间亲本HMW-GS组成遗传频率显著增加,其中以洛夫林10的Null、7+9、2+12亚基为主。由于HMW-GS组成不是碧蚂4号系谱品种育成时期的选择目标,对骨干亲本碧蚂4号和洛夫林10号被选择的Glu-B1位点进一步研究有助于解析骨干亲本的形成原因。     

小麦HMW-GS基因及其AS-PCR分子标记研究进展

小麦高分子量谷蛋白亚基(HMW-GS)与小麦品质性状,尤其是沉降值性状显著相关。利用其做分子标记选育聚合优质亚基的品种,具有快速、简单、实用、有效的特点。目前,普通小麦基因组中已有15个已命名Glu-1基因被克隆和测序,这使设计引物序列、进行等位基因的特异性扩增成为可能。对普通小麦高分子量谷蛋白亚基基因组成特点及分子标记现状进行了分析,并针对国内利用高分子量谷蛋白亚基进行分子标记辅助育种做了展望。     

小麦—中间偃麦草衍生材料农艺性状、HMW-GS及GISH鉴定

本研究分析了143个小麦—中间偃麦草种质材料的农艺性状、高分子量麦谷蛋白亚基及部分代表性材料的染色体构成,旨在为小麦育种中广泛有效地利用这些种质提供有用信息。结果表明,小麦—中间偃麦草种质主要农艺性状变异丰富,其在穗长、小穗数和分蘖数性状上明显优于主栽品种,分别有142(99.3%)、125(87.4%)和62(43.4%)个小麦—中间偃麦草材料的穗长、分蘖数和小穗数大于主栽品种的平均值。供试材料在Glu-1的3个基因位点上共检测到12个等位变异,形成15种亚基组合类型,以(2*,7+8,5+10)为主,占所有材料的25.7%;Glu-A1(1和2*)、Glu-B1(7+8)和Glu-D1(5+10)位点的优质亚基比例分别达到了68.4%、68.4%和52.0%,有102(71.3%)个材料在Glu-1的2或3个位点同时具有优质亚基;有17个材料的优质亚基组合为(2*,7+8,5+10)或(1,7+8,5+10),且在穗长、小穗数和分蘖数性状上均优于主栽品种。进一步对30个代表性材料GISH分析发现,8个为八倍体小偃麦,其他为非整倍体。研究结果表明这些材料可以作为改良普通小麦的有益基因资源。     

西南冬麦区地方品种HMW-GS组成遗传多样性研究

采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对西南冬麦区(云南、贵州、四川)3个省份共计560份小麦地方品种的高分子量谷蛋白亚基(HMW-GS)组成进行了研究。结果表明:Glu-1位点共有22种等位基因,其中Glu-A1位点4种、Glu-B1位点11种、Glu-D1位点7种;亚基null、7 8和2 12在各自位点的频率最高,分别为89.64%、68.21%和96.43%。亚基组成类型共有46种,以null/7 8/2 12和null/7 9/2 12为主,频率分别为50.89%和11.79%。在这些材料中筛选出一些含有1、2*、17 18、14 15、5 10等优质亚基的材料,其中有52份材料含有优质亚基组合。     

小麦骨干亲本阿夫及其衍生品种（系）的高分子量麦谷蛋白亚基演变分析

阿夫是我国小麦育种的骨干亲本之一,由其衍生的品种(系)有171个.为了了解阿夫对衍生品种在品质育种上的贡献,本文利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳技术(SDS-PAGE)对阿夫及其衍生品种(系)的高分子量麦谷蛋白亚基(HMW-GS)的组成与演变进行了分析.结果表明,阿夫HMW-GS组成为1/14+15/2+12,品质得分8.5分;而在衍生品种(系)中,携带与阿夫HMW-GS组成相同的品种占7.01%,平均品质得分7.68分,低于亲本阿夫.其中,由阿夫衍生的子一代到子四代品种中,携带与阿夫相同的14+15优质亚基的品种分别占28.30%、19.74%、9.09%和7.69%,平均品质得分分别是7.74分、7.84分、7.56分和7.23分,均低于亲本阿夫.结果说明,骨干亲本阿夫对其衍生后代在品质改良上的贡献很小,而主要贡献可能是当时育种目标需求的抗条锈病和丰产性等性状.     

谷蛋白聚合体大小分布与面粉揉面特性的初步研究

用单向一步SDS-PAGE方法分析表明小麦品种Suneca和Cook在麦谷蛋白5个亚基位点（Glu-B1,Glu-D1,Glu-A3,Glu-B3和Glu-D3)均含不同等位基因。选用Suneca×Cook的F4代群体中麦谷蛋白亚基位点均为纯合基因的60个系,研究麦谷蛋白基因型不同的株系间谷蛋白聚合体粒度大小分布（用SE-HPLC测定）和面粉揉面特性的变异。结果表明,不同的谷蛋白基因型,其谷蛋白聚合体粒度大小相对分布（用不溶谷蛋白聚合体占总谷蛋白聚合体含量的百分数表示,即UPP%）和面团形成时间（即揉面仪曲线图峰值的和面时间,简写PTM）均有显差异;面粉的揉面曲线形状与其UPP%值密切相关;UPP%与PTM呈极显正相关,与揉面仪曲线图峰高（PHM）呈显负相关;与面粉蛋白质含量（FP%）相比,UPP%对PTM和PHM的影响更大些,可作为育种早代品质性状选择一个指标。     

抗病基因Pi-ta在骨干亲本及其衍生品种的传递

利用Pi-ta的显性分子标记对寒地稻区水稻骨干亲本合江20号及其衍生品种进行Pi-ta抗性基因传递分析。结果表明,抗病基因Pi-ta在亲本合江20号衍生子一代出现的频率为63%,子二代出现的频率为33%、子三代出现的频率为9%,抗病基因Pi-ta的传递与合江20号衍生系谱一致。抗谱分析表明,Pi-ta抗病基因在子代中出现与衍生品种的抗谱正相关,这可能与后代选择过种中抗病基因的丢失有关,这也可能是决定不同水稻品种的稻瘟病发生程度的主要原因之一。     

部分小麦高分子量谷蛋白亚基组成分析

利用十二烷基硫酸钠聚丙烯胺凝胶电泳（SDS－PAGE）分析了85个小麦材料的高分子量谷蛋白亚基的构成,其结果表明：（1）目前生产中应用的优质小麦品种,大部分具有1A上的优质亚基1,1B上的14＋15／17＋18或1D上的5＋10,个别品种还同时聚合有1A,1B,1D上的优质亚基;（2）在所分析的28个八倍体小偃麦中,多数材料含有1,2^*和5＋10等优质亚基;（3）在本实验室创造的材料中,来源于中间偃麦草和普通小麦杂交的后代材料中大部分具有14＋15亚基。此外,个别种质材料还含有Payne亚基命名系统中未命名的一些稀有的高分子量谷蛋白亚基。     

西北春麦区小麦地方品种高分子量麦谷蛋白亚基组成分析

为了给品质改良提供基础材料,并了解西北春麦区小麦地方品种的遗传多样性,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493份小麦地方品种的高分子量麦谷蛋白亚基(HMW-GS)的组成.结果表明:在供试材料中,Glu-1位点共有26个等位基因,其中Glu-A1位点3个,Glu-B1位点9个,Glu-D1位点14个,亚基null、7+8、2+12在各自的位点上出现频率最高,分别达到了94.53%、92.92%、86.24%;亚基组成类型共有30种,主要为null/7+8/2+12,频率达79.76%;同时筛选出一些含有1、2*、13+16、14+15、5+10、1.5+10等优质亚基或亚基对的材料,可作为优质基因源;西北春麦区小麦地方品种间Glu-1位点的遗传多样性,以Glu-D1位点最高,其次是Glu-B1位点,Glu-A1位点最低.     

西藏半野生小麦高分子量麦谷蛋白亚基组成分析

应用SDS-PAGE分析了50份西藏半野生小麦(Triticum aestivum ssp.tibetanum Shao)的高分子量麦谷蛋白亚基等位基因组成。结果表明,43份材料的HMW-GS组成是同质的,7份材料为异质。供试材料共有7种HMW GS组合,以Null、7 8、2 12为主要类型,占所分析材料的68．4％。在Glu-1位点共检测到10种等位基因,Glu- A1位点2种,Glu～B1位点4种,Glu～D1位点4种。Null(96％)、7 8(80．4％)和2 12(94．9％)分别是Glu-A1、 Glu-B1和Glu～D1位点上主要的等位基因。在Glu-B1位点还新发现2个亚基,暂时分别命名为8*和7**。说明西藏半野生小麦中存在着较广泛的HMW-GS等位基因变异,是小麦品质育种潜在的可利用的遗传资源。     

High Molecular Weight Glutenin Subunit Composition of Indian Wheats and Their Relationships with Dough Strength

One hundred and seventy two wheat varieties including twenty-five durum wheat cultivars were evaluated for high molecular weight glutenin subunit (HMW-GS) composition using SDS-PAGE. The relationship between HMW-GS and sedimentation tests for dough strength was studied. Three alleles were present at the Glu-A1 locus, eight at Glu-B1 and two at Glu-D1 in bread wheat. The data indicated the prevalence of the Glu-A1b allele (63.5%) at the Glu-A1 and Glu-D1a (71.4%) at Glu-D1 loci. Three alleles, namely Glu-B1b (30.61%), Glu-B1c (25.85%) and Glu-B1i (34.00%) represented about 90% of the alleles at Glu-B1 locus. The combination of Glu-A1b, Glu-B1i and Glu-D1d alleles exhibited highest dough strength as measured by sedimentation value in comparison to other combinations (p<0.001). However, this combination was present only in 7% of the samples evaluated. In durum wheat, the null allele (Glu-A1c) was observed more frequently (76%) than the Glu-A1b allele (24%). Glu-B1f and Glu-B1e alleles represented equally (32% each). Protein subunits 13+16 and 6+8 were found correlated positively (p<0.05) with improved dough strength as compared to subunit 20 in durum wheat. This information can be a valuable reference for designing breeding programme for the improvement of bread and pasta making quality of bread and durum wheats, respectively in India.     

New DNA markers for high molecular weight glutenin subunits in wheat

End-use quality is one of the priorities of modern wheat (Triticum aestivum L.) breeding. Even though quality is a complex trait, high molecular weight (HMW) glutenins play a major role in determining the bread making quality of wheat. DNA markers developed from the sequences of HMW glutenin genes were reported in several previous studies to facilitate marker-assisted selection (MAS). However, most of the previously available markers are dominant and amplify large DNA fragments, and thus are not ideal for high throughput genotyping using modern equipment. The objective of this study was to develop and validate co-dominant markers suitable for high throughput MAS for HMW glutenin subunits encoded at the Glu-A1 and Glu-D1 loci. Indels were identified by sequence alignment of allelic HMW glutenin genes, and were targeted to develop locus-specific co-dominant markers. Marker UMN19 was developed by targeting an 18-bp deletion in the coding sequence of subunit Ax2* of Glu-A1. A single DNA fragment was amplified by marker UMN19, and was placed onto chromosome 1AL. Sixteen wheat cultivars with known HMW glutenin subunits were used to validate marker UMN19. The cultivars with subunit Ax2* amplified the 362-bp fragment as expected, and a 344-bp fragment was observed for cultivars with subunit Ax1 or the Ax-null allele. Two co-dominant markers, UMN25 and UMN26, were developed for Glu-D1 by targeting the fragment size polymorphic sites between subunits Dx2 and Dx5, and between Dy10 and Dy12, respectively. The 16 wheat cultivars with known HMW glutenin subunit composition were genotyped with markers UMN25 and UMN26, and the genotypes perfectly matched their subunit types. Using an Applied Biosystems 3130xl Genetic Analyzer, four F₂ populations segregating for the Glu-A1 or Glu-D1 locus were successfully genotyped with primers UMN19, UMN25 and UMN26 labeled with fluorescent dyes.     

Allelic variation of high molecular weight glutenin subunits of bread wheat in Hebei province of China

In common wheat (Triticum aestivum L.), allelic variations of Glu-1 loci have important influences on grain end-use quality. The allelic variations in high molecular weight glutenin subunits (HMW-GSs) were identified in 151 hexaploid wheat varieties representing a historical trend in the cultivars introduced or released in Hebei province of China from the years 1970s to 2010s. Thirteen distinct alleles were detected for Glu-1. At Glu-A1, Glu-B1 and Glu-D1, we found that the most frequent alleles were the 1 (43.0%), 7+8 (64.9%), 2+12 (74.8%) alleles, respectively, in wheat varieties. Twenty two different HMW-GS compositions were observed in wheat. Twenty-five (16.6%) genotypes possessed the combination of subunits 1, 7+8, 2+12, 25 (16.6%) genotypes had subunit composition of 2*, 7+8, 2+12; 20 (13.2%) genotypes had subunit composition of null, 7+8, 2+12. The frequency of other subunit composition was less than 10%. The Glu-1 quality score greater than or equal to 9 accounted for 20.6% of the wheat varieties. The percentage of superior subunits (1 or 2* subunit at Glu-A1 locus; 7+8, 14+15 or 17+18 at Glu-B1 locus; 5+10 or 5+12 at Glu-D1 locus) was an upward trend over the last 40 years. The more different superior alleles correlated with good bread-making quality should be introduced for their usage in wheat improvement efforts.     

Development of primers specific for LMW-GS genes located on chromosome 1D and molecular characterization of a gene from Glu-D3 complex locus in bread wheat

Glutenins are multimeric aggregates of high molecular weight (HMW) and low molecular weight (LMW) subunits, which determine the quality in wheat. Development of locus-specific primers is an important step toward cloning specific LMW glutenin subunits (LMW-GS) by PCR method. Based on the publicly available, a pair of primer, namely primer 3 (5' TTGTAGAAACTGCCATCCTT 3') and primer 4 (5' GTCACCGCTGCAT CGACATA 3') was designed and verified to specific for LMW-GS genes located on chromosome 1D in this study. The LMW-GS gene located at the Glu-D3 locus in bread wheat cultivar Xiaoyan 6 was cloned using this pair of primer. The clone designated as XYGluD3-LMWGS1 (AY263369), contains the endosperm-specific-expression promoter and the entire coding region. Nucleotide sequence comparison of the XYGluD3-LMWGS1 with other reported LMW-GS genes located at different Glu-3 loci showed the degree of identity among them ranged from 59.57% to 99.78%. The LMW-GS genes at the same locus showed more similar to each other than to the gene at different locus. Comparison of the deduced amino acid sequence of the XYGluD3-LMWGS1 with the sequences of 12 group LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequence was nearly the same to LMW-GS group 10 (identity 99.67%). The deduced LMW-GS contains nine cystine residues, which contained one more cystine residue in the C-terminal conserved domain than previous reported. This was the first LMW-GS gene encoding for a LMW-GS with 9 cystine residues that has been discovered so far.     

湖北推广小麦品种(系)高分子量麦谷蛋白亚基组成分析

应用SDS-PAGE技术分析了45份湖北推广小麦品种(系)籽粒的高分子量麦谷蛋白亚基组成。40份材料的高分子量麦谷蛋白亚基组成为同质,5份为异质。在Glu-1位点共检测到9种等位基因变异类型,其中Glu-A1位点有“1、2^ 、Null”3种变异类型,Glu-B1位点有“7、7 8、7 9、14 15”4种,Glu-D1位点有“2 12、5 10”2种。“Null、7 8、2 12”是主要亚基,它们的频率分别是62．5％、60％和72．5％。亚基组合类型有12种,其中(Null,7 8,2 12)亚基组合占30．0％,(1,7 8,2 12)、(1,14 15,2 12)、(Null,7 9,2 12)、(Null,7 8,5 10)4种组合的频率都在10％以上,这5种亚基组合占总组合的72．5％。供试小麦材料品质评分在5～10之间,平均评分为7．0。含5 10亚基的品种(系)所占比例低,是湖北小麦烘烤品质较差的部分原因。     

Identification of LMW glutenin-like genes from Secale sylvestre host

Three low-molecular-weight (LMW) glutenin-like genes (designated as Ssy1, Ssy2 and Ssy3) from Secale sylvestre Host were isolated and characterized. The three genes consist of a predicted highly conservative signal peptide with 20 amino acids, a short N-terminal region with 13 amino acids, a highly variable repetitive domain and a less variable C-terminal domain. The deduced amino acid sequences of the three genes were the LMW-m type due to a methionine residue at the N-terminus. The phylogenic analysis indicated that the prolamin genes could be perfectly clustered into five groups, including HMW-GS, LMW-GS, alpha/beta-, gamma- and omega-prolamin. The LMW glutenin-like genes of S. sylvestre were more orthologous with the LMW-GS genes of wheat and B hordein genes of barley, which also had been confirmed by the homology analysis with the LMW-GS of wheat at Glu-A3, Glu-B3 and Glu-D3 loci. These results indicated that a chromosome locus (designated as Glu-R3) might be located on the R genome of S. sylvestre with the functions similar to the Glu-3 locus in wheat and its related species.     

小麦高分子量麦谷蛋白亚基鉴定及其品质效应研究进展

高分子量谷蛋白亚基(HMW-GS,high molecular weight glutenin subunits)是小麦子粒贮藏蛋白的重要组成成分,其组成、搭配、表达水平及含量决定面团弹性和面包加工品质。本文主要介绍了小麦HMW-GS编码基因的克隆、分子特征、分子标记开发及其在小麦育种中的应用,并综述了不同HMW-GS与面粉加工品质之间的关系,以及HMW-GS基因遗传转化、微量配粉和突变体培育等方面的研究进展,分析了目前研究中存在的主要问题,认为通过分子标记辅助选择和转基因技术聚合优质亚基,培育优质面包小麦品种和明确各个HMW-GS基因的品质效应是今后的研究重点。     

Development of isohomoeoallelic lines within the wheat cv. Courtot for high molecular weight glutenin subunits: transfer of the <Emphasis Type="Italic">Glu-D1</Emphasis> locus to chromosome 1A

Wheat quality depends on protein composition and grain protein content. High molecular weight glutenin subunits (HMW-GS) play an important role in determining the viscoelastic properties of gluten. In an attempt to improve the bread-making quality of hexaploid wheat by elaborating novel HMW-GS combinations, a fragment of wheat chromosome 1D containing the Glu-D1 locus encoding the Dx2+Dy12 subunits was translocated to the long arm of chromosome 1A using the ph1b mutation. The partially isohomoeoallelic line selected was characterized using cytogenetical and molecular approaches to assess the amount of chromatin introgressed in the translocated 1A chromosome. Triple-target genomic in situ hybridization indicated that the translocated 1A chromosome had a terminal 1D segment representing 25% of the length of the recombinant long arm. The translocation was also identified on the long arm using molecular markers, and its length was estimated with a minimum of 91 cM. Proteome analysis was performed on total endosperm proteins. Out of the 152 major spots detected, 9 spots were up-regulated and 4 spots were down-regulated. Most of these proteins were identified as α-, β-, γ-gliadins assigned to the chromosomes of homoeologous groups 1 and 6. Quantitative variations in the HMW-GS were only observed in subunit Dy12 in response to duplication of the Glu-D1 locus.