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1.
粗山羊草高分子量谷蛋白亚基组成分析   总被引：1，自引：0，他引：1       下载免费PDF全文
为了发现能够用于小麦品质改良的优异高分子量谷蛋白亚基(HMW-GS),应用SDS-PAGE技术分析了47份粗山羊草Glu-Dt1位点的HMW-GS组成,分别检测到5种x-型亚基(1.1t、1t、1.5t、2t、3t)和4种y-型亚基(10.1t、11t、12t、12.4t).其中,1Dx1.1t是1种新的x-型亚基,它的迁移率比普通小麦的1Ax1亚基稍慢,这是目前在粗山羊草中发现的分子量最大的HMW-GS.供试粗山羊草中有8种HMW-GS组合类型:1.1t 11t、1t 11t、1.5t 12t、2t 10.1t、2t 11t、2t 12t、2t 12.4t和3t 11t.其中,1.1t 11t和2t 12.4t未见报道.  相似文献   

2.
为了解不同遗传背景的黑麦染色体对普通小麦主要高分子量谷蛋白亚基(HMW-GS)和醇溶蛋白组成的影响,采用SDS-PAGE和A-PAGE分析了两套小麦-黑麦(Holdfast-King和中国春-Impire) 双二倍体和不同附加系的HMW-GS和醇溶蛋白组成.结果表明:(1)两份双二倍体的HMW-GS都表达出小麦亲本带型和一条迁移率与双亲不一致的条带,醇溶蛋白呈共显性表达.(2)Holdfast-King的1R附加系HMW-GS带型与其双二倍体带型一致,醇溶蛋白呈共显性表达;3R附加系的HMW-GS缺失受体亲本条带并表达出一条迁移率与双亲有差异的条带,其醇溶蛋白缺失受体亲本条带;其余附加系(2R,4R～7R)的HMW-GS和醇溶蛋白带型与其受体亲本带型一致.(3)中国春-Impire的1R附加系HMW-GS带型与其双二倍体一致,醇溶蛋白只表现受体小麦亲本带型,2R到7R附加系的HMW-GS和醇溶蛋白带型都与小麦亲本带型一致.这表明小麦-黑麦异源双二倍体中小麦和黑麦的基因组会相互影响,引起籽粒贮藏蛋白组成表达上的差异;同一种黑麦的不同染色体对籽粒贮藏蛋白组成表达的影响也有差异.  相似文献   

3.
21份印度小麦高分子谷蛋白亚基、醇溶蛋白及品质分析   总被引：1，自引：0，他引：1  
为挖掘优良的小麦种质资源,以引进的21份印度小麦种质为材料,利用SDS-PAGE和A-PAGE技术,分析了其高分子量谷蛋白亚基和醇溶蛋白组成。结果表明,在21份印度小麦材料中出现了12种HWM-GS亚基类型和14种亚基组合,其中有3种亚基类型(Null、1、2*)在Glu-A1位点,4种亚基类型(7+8,17+18,7,7+9,13+16)在Glu-B1位点,4种亚基类型(5+10,2+12,4+12,5+12)在Glu-D1位点。1、2+12优质亚基的比例相对较高,均为47.6%。小麦HWM-GS亚基组合1/7+9/2+12在所有亚基组合类型中出现频率高达19.0%。大多数材料的品质得分为7、8分,平均7.52分。共分离出32种迁移率不同的醇溶蛋白谱带,2和3号带出现频率最高,分别为95.24%和90.48%。DA 7200近红外分析仪初步分析结果表明,这21份印度小麦种质资源品质指标相对偏低。  相似文献   

4.
人工合成六倍体小麦的高分子量谷蛋白亚基组成分析   总被引：5，自引：2，他引：5  
利用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳技术(SDS-PAGE)对103份人工合成六倍体小麦(2n=6x=42,AABBDD)的高分子量谷蛋白亚基组成进行了分析。结果发现,所分析材料具有丰富的等位变异。其中,在Glu-A1住点上共有5种变异类型,在Glu-B1和Glu-D1位点上均出现了12种等住变异类型;有些材料在Glu-D1位．占、上只表达1Dy类型亚基,而在1Dx亚基位置上出现空位形式;同时还发现了一些新的亚基组合形式和一未知新亚基。这些人工合成六倍体小麦材料中丰富的高分子量谷蛋白亚基变异,可能对小麦品质改良具有重要的应用价值。  相似文献   

5.
抗赤霉病小麦地方品种的贮藏蛋白分析   总被引：3，自引：0，他引：3  
采用A-PAGE和SDS-PAGE方法,对来自不同地方的23个抗赤霉病小麦地方品种的醇溶蛋白和谷蛋白亚基进行了分析。结果表明,在A-PAGE电泳分析中,23个供试品种具有23种不同的醇溶蛋白带型,共分离出39条相对迁移率不同的谱带,其中31条具有多态性,占86.8％,每份材料可电泳出14～23条带,存在着广泛的等位基因变异。在SDS-PAGE电泳分析中,出现了9种不同的高分子量谷蛋白亚基及6种亚基组合类型,优质亚基及亚基组合所占的比例较少,品质评分较低,其变幅为5～9分,平均为6.8分。  相似文献   

6.
小麦高分子量谷蛋白亚基1Dx5的AS-PCR分子鉴定   总被引：4，自引：1，他引：4       下载免费PDF全文
小麦高分子量谷蛋白亚基Glu—D1位点上1Dx5—1Dy10和1Dx2—1Dy12分别紧密连锁，与小麦面包加工品质的优劣密切相关。为了加速小麦品质育种进程，利用1Dx5亚基特异PCR标记鉴定了67份小麦品种（系）的谷蛋白Glu—D1位点，有22个品种扩增出478bp特异片段，表明这些品种具有1Dx5亚基；45份品种未扩增出478bp特异片段，表明其不具有1Dx5亚基，1Dx5亚基出现频率为32．8％。PCR标记的鉴定结果与SDs-PAGE电泳结果一致，通过比较，初步建立了鉴定1Dx5亚基的稳定扩增体系。  相似文献   

7.
为了探明杂交小麦F1与F2代品质的组配规律及高分子量谷蛋白亚基组成规律,选取不同品质类型的14个亲本材料,以8种设计,按照强筋×强筋、强筋×中筋、强筋×弱筋、中筋×强筋、中筋×中筋、弱筋×强筋、弱筋×中筋、弱筋×弱筋等方式组配不同杂交组合,分别测定其中8种F2代的品质以及亲本、F1代、F2代的高分子量谷蛋白亚基组成,结果表明,以优质和含有优质亚基的品种作母本对杂交小麦F2品质改良有重要作用,同时在亚基中出现了5 10亚基,其对品质有重要的影响。F1代籽粒中出现了高分子量谷蛋白亚基共显现象和不完全表达或缺失现象。经观察发现,缺失的带基本都是父本的亚基带。  相似文献   

8.
为了发掘新的小麦高分子量谷蛋白亚基用于品质育种,测定了小麦/长穗偃麦草体细胞杂种F6代共322个株系的高分子量谷蛋白亚基（HMWGS）组成,并进行了品质评分。结果表明,体细胞杂种株系在GluA1、GluB1和GluD1位点上的遗传变异率分别为0.52、0.58和0.46,一共出现27种不同的亚基组合形式,其中Null、7+9、2+12（同亲本小麦济南177）出现的频率最高,而2*、7+8+9、2+5+12出现的频率最低;5+12出现的频率约为33.5%;在与优质可能相关的亚基组合当中,13+16出现的频率约为31.1%。由于5+12尚未有品质评分,因此部分株系无法给出分数。本研究表明,通过体细胞杂交可以产生大量的高分子量谷蛋白亚基/组合变异,这有助于今后小麦品质育种工作。  相似文献   

9.
小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达   总被引：1，自引：0，他引：1  
为了给小麦优质亚基研究奠定基础,将高分子量谷蛋白亚基1Dx5基因的核苷酸序列与载体pRSET A重组,将构建好的质粒转化大肠杆菌BL21(DE3)pLysS菌株,通过IPTG使其得到了成功表达.大肠杆菌中表达的1Dx5亚基在SDS-PAGE上与小麦品种钱尼中的1Dx5亚基具有相似的迁移率.用Western blot技术成功检测到了目的基因产物.通过改变IPTG浓度、诱导时间、培养基成分及初始菌液浓度研究了最适表达条件.结果表明,最适表达条件为:LB培养基,菌液初始浓度(OD600)0.4～0.6,IPTG浓度0.4mmol/L,诱导时间为3～5 h.  相似文献   

10.
为了给小麦育种提供新的种质资源,对4份圆锥小麦-乌拉尔图小麦双二倍体(AABBA^uA^u)的农艺性状、染色体组成及高分子量谷蛋白亚基组成进行了鉴定。农艺性状调查结果表明,4份双二倍体的株高介于108.45~129.43 cm,分蘖数介于7.3~17.5个,穗长介于10.23~12.17 cm,小穗数介于16.26~22.06,自交结实率介于37.77%~70.46%;4份双二倍体对目前的流行条锈菌混合生理小种(条中31、条中32、条中33、条中34和水源11-4)均表现为高抗。利用寡核苷酸序列探针Oligo-pSc119.2-1、Oligo-pTa535-1、Oligo-pTa71-2、pTa-713和简单重复序列探针(AAC)5进行FISH分析,可区分出圆锥小麦-乌拉尔图小麦双二倍体的42条染色体。花粉母细胞染色体配对观察表明,在这些双二倍体的减数分裂中期I,染色体大多配对成二价体,仅有少量的单价体、三价体和四价体。SDS-PAGE分析表明,来自于乌拉尔图小麦TA#831的1Ay亚基在双二倍体Syn-TAU-2中得到了表达,但其迁移率发生了变化;来自于PI428270和PI428274的1Ax和1Ay亚基分别在双二倍体Syn-TAU-3和Syn-TAU-4中得到了表达。这些双二倍体可作为新的资源材料用于普通小麦的遗传改良。  相似文献   

11.
Electrophoretic (urea SDS–PAGE) and chromatographic (RP–HPLC) analysis was performed on 8 allelic variants of HMW glutenin subunits derived fromTriticum tauschiiand from the D genome of a hexaploid wheat species (Triticum macha) and hexaploid landraces. These subunits had been previously identified using SDS–PAGE. The characterisation revealed that subunits Dy10tand Dy12tfromT. tauschiicould be differentiated from their bread wheat counterparts using both urea SDS–PAGE and RP–HPLC. In the latter case, theT. tauschiiy-type subunits were clearly more hydrophobic than the Dy type subunits of bread wheat. The characterisation also suggested that subunit Dx5t, derived from two separateT. tauschiiaccessions, did not contain the extra cysteine residue characteristic of Dx5 from bread wheat. RFLP analysis of the genes encoding the HMW glutenin subunits of interest suggested that the absence of Dx-type HMW glutenins in two hexaploid landraces was due to lack of expression of their encoding genes. The relationship betweenHindIII DNA fragment size and protein subunit size, as measured by electrophoretic mobility, is examined and discussed. Finally, the solubility properties of a HMW protein designated T1 (derived fromT. tauschiiaccession AUS 18913) suggested that it was not a HMW glutenin subunit as was previously thought. Further studies are needed to clarify the identity of this subunit.  相似文献   

12.
The x- and y-type high molecular weight (HMW) glutenin subunits are conserved seed storage proteins in wheat and related species. Here we describe investigations on the HMW glutenin subunits from several Pseudoroegneria accessions. The electrophoretic mobilities of the HMW glutenin subunits from Pd. stipifolia, Pd. tauri and Pd. strigosa were much faster than those of orthologous wheat subunits, indicating that their protein size may be smaller than that of wheat subunits. The coding sequence of the Glu-1St1 subunit (encoded by the Pseudoroegneria stipifolia accession PI325181) was isolated, and found to represent the native open reading frame (ORF) by in vitro expression. The deduced amino acid sequence of Glu-1St1 matched with that determined from the native subunit by mass spectrometric analysis. The domain organization in Glu-1St1 showed high similarity with that of typical HMW glutenin subunits. However, Glu-1St1 exhibited several distinct characteristics. First, the length of its repetitive domain was substantially smaller than that of conventional subunits, which explains its much faster electrophoretic mobility in SDS-PAGE. Second, although the N-terminal domain of Glu-1St1 resembled that of y-type subunit, its C-terminal domain was more similar to that of x-type subunit. Third, the N- and C-terminal domains of Glu-1St1 shared conserved features with those of barley D-hordein, but the repeat motifs and the organization of its repetitive domain were more similar to those of HMW glutenin subunits than to D-hordein. We conclude that Glu-1St1 is a novel variant of HMW glutenin subunits. The analysis of Glu-1St1 may provide new insight into the evolution of HMW glutenin subunits in Triticeae species.  相似文献   

13.
The high-molecular weight (HMW) glutenin subunits of bread wheat are major determinants of end-use quality. The objective of this study was to determine the 1Dx and 1Dy subunits present in 43 synthetic hexaploid wheat (SHW) lines derived by crossing durum ‘Langdon’ to 43 Aegilops tauschii accessions. Protein samples were initially electrophoresed multiple times on SDS-PAGE gels to arrange subunits into similar groups and then were electrophoresed on urea/SDS-PAGE gels. Initial results with SDS-PAGE gels indicated that there were six 1Dx and six 1Dy subunits in these SHW lines. However, results of the urea/SDS-PAGE indicated that some of the subunit groups could be further differentiated into additional subunits. A total of eleven 1Dx and eight 1Dy subunits including the newly designated subunits 1Dx2t-1, 1Dx2t-2, 1Dx2t-3, 1Dx1.5t-1, 1Dx2.1t-1, 1Dy10t-1, and 1Dy12t-1 were identified, and they composed 17 1Dx and 1Dy combinations in the SHW lines. Eight of the combinations included at least one novel subunit and hence they were novel Glu-D1 alleles. Our results indicated that urea/SDS-PAGE can be very useful in identifying new HMW glutenin subunits. Quality testing of the SHW lines will determine if any of the alleles are useful in improving wheat-baking quality.  相似文献   

14.
The high molecular weight (HMW) glutenin subunits Dtx1.5+Dty10 from Aegilops tauschii are a novel pair of subunits not detected previously in common wheat (Triticum aestivum). The partial DNA sequences of the x-type HMW glutenin alleles from A. tauschii and synthetic hexaploid wheat samples with different HMW glutenin subunits were charcterised. Five samples were found to contain the HMW glutenin subunit Dtx1.5 that may affect bread-making quality. Polymorphisms of the DNA sequences were detected among alleles encoding different HMW glutenin subunits and within an allele encoding the same HMW glutenin subunit, such as the Dtx1.5 subunit. Three single nucleotide polymorphisms (SNPs) that can distinguish the Dxt1.5 from Dtx2, Dtx5, Dx2 and Dx5 alleles were identified. Allelic specific (AS)-PCR primers were developed based the SNPs located at the non-repetitive N-terminal of the HMW glutenin subunits. The AS-PCR primers can accurately identify accessions containing the Dtx1.5 subunit from those containing other studied subunits by PCR analysis, suggesting the usefulness of AS-PCR for identifying different HMW glutenin subunits of A. tauschii and synthetic hexaploid wheat. The AS-PCR primers developed based on SNPs in this study are valuable in wheat breeding for effective selection of special HMW glutenin subunits.  相似文献   

15.
野生二粒小麦高分子量谷蛋白亚基的多态性分析   总被引：7，自引：2，他引：7  
采用十二烷基磺酸钠 -聚丙烯酰胺凝胶电泳 (SDS- PAGE)方法 ,对 172份来自以色列的野生二粒小麦的高分子量谷蛋白亚基 (HWM- GS)组成进行了鉴定和分析。结果发现 ,这 172份野生二粒小麦 1B染色体上的HMW- GS有 19种变异类型 ,比普通小麦 1B染色体上 HMW- GS的变异类型丰富得多 ,其中有 8种 (6 * 8* ,6 8* ,2 2 ,6 * 8,7* 9,7* 9* ,7* 8* ,7* 8)为普通小麦中不常见类型 ,各个亚基的分布频率不同 ,17 18亚基分布频率最高 (2 9.6 5 % ) ,并且存在 3种普通小麦中不存在的特殊亚基 (图 1中加 ?的类型 ) ;1A染色体上的 HMW- GS有4种变异类型 ,其中一个亚基 (自定义为 1* )以前未见报道。发现野生二粒小麦 HMW- GS组成中 Glu- B1位点的 17 18亚基和 Glu- A1位点的 1亚基出现频率比普通小麦中出现的频率高。这些试验数据可为野生二粒小麦上特异HMW- GS的进一步研究与利用提供理论依据。  相似文献   

16.
Cloning and functional analysis of high molecular weight wheat glutenin subunit (HMW-GS) 1By8 from Italy durum cultivar Simeto was carried out in this study. All HMW-GS from Simeto were separated and characterized by appropriate electrophoresis methods, reversed-phased high performance liquid chromatography (RP-HPLC) and mass spectrometry (MS). The complete gene encoding 1By8 subunit was amplified by allele-specific PCR primers, including an upstream sequence of 857 bp and an open reading frame (ORF) of 2166 bp encoding a mature protein of 720 amino acid residues. The promoter sequence, containing -300 element (cereal glutenin gene control element) and enhancer was highly conserved among HMW-GS genes. Comparison with the sequence of subunit 1By9 from bread wheat demonstrated 99% identity with the main difference being that the 1By8 subunit possesses an additional insertion of 15 amino acid residues (QYPASQQQPA QGQQG) at position 342 and two residue substitutions at position 78 (leucine/proline) and 442 (arginine/glutamine). The molecular weight differences between MALDI-TOF-MS and deduced amino acid sequence of the coding gene revealed the possibility of some kinds of post-translational modifications present in 1By8 subunit. The protein subunit expressed in Escherichia coli showed a very similar mobility to the endogenous 1By8 of Simeto on SDS-PAGE. The function of the isolated protein on wheat processing quality was determined by 10 g Mixgraph analysis. Results demonstrated that addition of y-type HMW glutenin subunits into the base flour had significant positive effects on main mixing parameters and significant difference in effects were observed among different y-type subunits.  相似文献   

17.
Cloning and functional analysis of high molecular weight wheat glutenin subunit (HMW-GS) 1By8 from Italy durum cultivar Simeto was carried out in this study. All HMW-GS from Simeto were separated and characterized by appropriate electrophoresis methods, reversed-phased high performance liquid chromatography (RP-HPLC) and mass spectrometry (MS). The complete gene encoding 1By8 subunit was amplified by allele-specific PCR primers, including an upstream sequence of 857 bp and an open reading frame (ORF) of 2166 bp encoding a mature protein of 720 amino acid residues. The promoter sequence, containing -300 element (cereal glutenin gene control element) and enhancer was highly conserved among HMW-GS genes. Comparison with the sequence of subunit 1By9 from bread wheat demonstrated 99% identity with the main difference being that the 1By8 subunit possesses an additional insertion of 15 amino acid residues (QYPASQQQPA QGQQG) at position 342 and two residue substitutions at position 78 (leucine/proline) and 442 (arginine/glutamine). The molecular weight differences between MALDI-TOF-MS and deduced amino acid sequence of the coding gene revealed the possibility of some kinds of post-translational modifications present in 1By8 subunit. The protein subunit expressed in Escherichia coli showed a very similar mobility to the endogenous 1By8 of Simeto on SDS-PAGE. The function of the isolated protein on wheat processing quality was determined by 10 g Mixgraph analysis. Results demonstrated that addition of y-type HMW glutenin subunits into the base flour had significant positive effects on main mixing parameters and significant difference in effects were observed among different y-type subunits.  相似文献   

18.
In this work we report the effects of the HMW-GS 1Ax1, 1Dx5 and 1Dy10 on the breadmaking quality of the bread wheat cultivar Anza that contains the HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8, and is null for the Glu-A1 locus. This allows the characterization of individual subunits 1Dx5 and 1Dy10 in the absence of subunit 1Dx5, and the interactions between these subunits and subunits 1Dx2 and 1Dy12 to be determined. Three transgenic lines termed T580, T581 and T590, containing, respectively, the HMW-GS 1Ax1, 1Dx5 and 1Dy10 were characterized over 3 years using a range of widely-used grain and dough testing methods. The transgenic subunits 1Ax1, 1Dx5 and 1Dy10 accounted for 25.2%, 20.3% and 17.9%, respectively, of the total HMW-GS in the three transgenic lines. Although lines T581 and T590 expressed similar levels of subunits 1Dx5 and 1Dy10 they had different effects on other aspects of protein composition, including changes in the ratios of glutenin/gliadin, of HMW/LMW-GS, the 1Dx2/1Dy12, the x-type/y-type HMW-GS and the proportions of high molecular mass glutenin polymers. In contrast, lines transformed to express subunits 1Ax1 and 1Dx5 showed similar changes in protein composition, with higher protein contents and decreased ratios of glutenin/gliadin and 1Dx2/1Dy12. In addition, both transgenic lines showed similar increases in the ratio of x-type/y-type subunits compared to the control line. The transgenic lines were analysed using Farinograph, Mixograph and Alveograph. This confirmed that the expression of all three subunits resulted in increased dough strength (and hence breadmaking quality) of the cultivar Anza. A beneficial effect of subunit 1Dx5 has not been reported previously, transgenic wheat lines expressing this subunit giving overstrong dough unsuitable for breadmaking. However, the expression of subunit 1Dy10 had a greater effect on breadmaking quality than subunits 1Ax1 and 1Dx5. The Farinograph parameters such as dough stability and peak time were increased by 9.2-fold and 2.4-fold, respectively, in line T590 (expressing 1Dy10) with respect to the control line. Similarly, the Mixograph mixing time was increased by four-fold and the resistance breakdown decreased by two-fold in line T590 compared with the control line. The Alveograph W value was also increased by 2.7-fold in line T590 compared to the control line. These transgenic lines are of value for studying the contribution of specific HMW-GS to wheat flour functional properties.  相似文献   

19.
为了克隆节节麦中的HMW-GS基因,利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了节节麦的HMW-GS,发现1种编码序列未知的y-型亚基,即Dy8.1t亚基。通过对目的片段的AS-PCR扩增、克隆、测序和氨基酸序列推导,发现这种未知序列具有典型HMW-GS的序列结构特征。通过与已知亚基序列比较发现,Dy8.1t与Dy10t、Dy10.4t、Dy10.5t和Dy12t在N-端序列有一个氨基酸差别(R-G)。与已知氨基酸序列的HMW-GS多序列比对和系统进化关系分析,证实Dy8.1t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   

20.
Many studies have suggested that the true variation of high-molecular-weight (HMW) glutenin subunits in wheat could be under-estimated due to the anomalous mobility of these proteins in one-dimensional electrophoresis (SDS-PAGE). An alternative technique used is the specific PCR amplification of these genes, however the scarce variation in size among the different alleles could also under-estimate the true variability. In this study, the variability of HMW glutenin subunits found in one Spanish collection of rivet wheat (Triticum turgidum L. ssp. turgidum) was evaluated by a combination of two techniques (PCR amplification and digestion with endonucleases). The data allowed detection of allelic variations that were not clearly detected by SDS-PAGE or PCR analysis alone. The approach used in the current study could allow identification of alleles contributing to achieve desirable quality in modern wheat.  相似文献   

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