Genetic diversity of high-molecular-weight glutenin subunit compositions in landraces of hexaploid wheat from Japan

The high-molecular-weight (HMW) glutenin subunit composition of seed storage protein of 174 Japanese hexaploid wheat (Triticum aestivum) landraces have been examined by using sodium dodecyl sulfate polyacrylamide gel electrophoresis system. Twenty four different, major glutenin HMW subunits were identified, and each of the landraces contained three to five subunits and 17 different glutenin subunit patterns were observed for 13 alleles in the landraces. On the basis of HMW glutenin subunits composition, Japanese landraces showed a specific allelic variation, close to Japanese commercial wheats in HMW glutenin subunits, different from those in alien hexaploid wheats. Further, it could be concluded that all common glutenin alleles can be found in the 174 landraces originated from Japan. The variation detected in the glutenin subunits is useful for variety identification, has a bearing on our understanding of hexaploid wheat genetic resource evolution in Japan, and raises questions concerning the nature of this genetic variation. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

High molecular weight glutenin subunit composition of Chinese bread wheats

Summary The endosperm storage proteins of 205 Chinese bread wheat cultivars and advanced lines were fractionated by SDS-PAGE to determine their high molecular weight (HMW) glutenin subunit composition. Seventeen alleles were identified: three at Glu-A1, eight at Glu-B1, and six at Glu-D1. The most common alleles were Null, 1, 7+8, 7+9, and 2+12. The results indicate that wheats from different regions differ in their frequencies of HMW glutenin subunits, however, none of the subunits could be related to specific environments. The glutenin quality scores of Chinese wheats ranged from 3 to 10, with an average of 6.7. Increasing quality scores have implications in improving steam-bread making quality for Chinese consumers. On the basis of HMW glutenin subunit composition, Chinese wheats are close to European wheats, especially Italian wheats because several Italian introductions are widely distributed in the pedigrees of Chinese wheat.     

Variation in high molecular weight glutenin subunits in landraces of hexaploid wheat from Afghanistan

Summary Variation for high molecular weight (HMW) glutenin subunits is reported in Afghan hexaploid wheat landraces from different locations in the country ranging in altitude from 395 to 3170 metres. The variation appeared to be independent of the altitude and geographical location of the landraces. Studies of a number of samples from each of five sites revealed that at some sites there was allelic variation at theGlu-A1 andGlu-B1 loci coding from HMW glutenin subunits, but there was no variation at theGlu-D1 locus within and between sites.     

Transfer of the Glu-D1 Gene from Chromosome 1D to Chromosome 1A in Hexaploid Triticale

To complement previously developed recombinant chromosomes 1R.1D, two series of translocations involving the Glu-D1 gene from chromosome ID to chromosome 1A were produced in hexaploid triticale. These series involve seven independent transfers of allele d encoding for high molecular weight glutenin subunits 5+10 and ten independent transfers involving allele a encoding for HMW glutenin subunits 2 + 12. The frequency of homoeologous recombination between chromosomes 1A and 1D was within the range observed for pairs of homologues in wheat, supporting earlier observations that homoeologous recombination in triticale is frequent. Recombined chromosomes 1A.1D can be used to introduce the Glu-D1 gene to durum wheats, and to manipulate the dosage of Glu-D1 in hexaploid triticale and bread wheat.     

Molecular structure of a novel y-type HMW glutenin subunit gene present in Triticum tauschii

An unusually small y-type high molecular weight (HMW) glutenin subunit gene from Triticum tauschii was sequenced. This gene, encoded at the Glu-D^t1 locus was designated 12.4^t and is the smallest HMW glutenin subunit gene described so far in Triticum species. Oligonucleotide primers based on published sequences of HMW glutenin genes were designed to amplify the encoding region and the central repetitive domain of the gene, which produced fragments of 1.4 and 0.85 kb, respectively. PCR products were cloned and sequenced. The derived amino acid sequence was compared with the amino acid sequences of the HMW glutenin subunits Dy12^t, from T. tauschii, and subunits Dy10 and Dy12 of T. aestivum. The amino acid sequence deduced from the nucleotide sequence demonstrated that deletions of hexapeptides and nonapeptides were responsible for the reduction in the size of this HMW glutenin subunit. The estimated molecular weight of the Dy12.4^t subunit, calculated on the basis of the deduced amino acid sequence, was 45,228 Daltons. There were also single amino acid differences in the N-, C-terminal and central repetitive domains of this gene in comparison to the three other y-type subunits encoded at the Glu-D1 locus. The Dy12.4^t subunit showed the highest similarity to the Dy12 subunit present in the hexaploid wheat Chinese Spring.     

Variation of high-molecular-weight glutenin subunits amongst cultivars of Triticum turgidum L. from Portugal

Summary Variation in high-molecular-weight (HMW) glutenin subunit composition amongst 63 varieties of Triticum turgidum L. from Portugal was investigated using SDS-PAGE. A total of thirteen Glu-A1 and Glu-B1 alleles were identified, and three of them were found to be different from those previously described in the literature. A number of the tetraploid wheats examined contained subunits known to have a beneficial effect of the bread-making properties of T.aestivum. Camara, a tetraploid wheat cultivar carrying a 1D/1B chromosomal substitution, is proposed as a bridge for the transfer of Glu-D1 alleles and of other protein fractions controlled by the 1D chromosomes (Gli-D1 locus) from hexaploid to tetraploid wheat.     

Detection of HMW glutenin subunit variations among 205 cultivated emmer accessions (Triticum turgidum ssp. dicoccum)

The high molecular weight glutenin subunits (HMW‐GS) encoded by Glu‐1 loci among 205 accessions of cultivated emmer wheat (Triticum turgidum ssp. dicoccum Schrank) collected from different regions of Europe and China were separated and characterized by SDS‐PAGE in combination with two‐dimensional gel electrophoresis (A‐PAGE × SDS‐PAGE) and acidic capillary electrophoresis. High genetic polymorphisms in HMW‐GS compositions were found. A total of 40 alleles (6 for Glu‐A1 and 34 for Glu‐B1) and 62 subunit combinations (genotypes) were detected, some of which were not previously described. At Glu‐A1 locus, two novel alleles, designated Glu‐A1x coding for the subunit 1A × 1.1 and Glu‐A1y coding for the subunit 1A × 2.1′ were found while seven new subunits (1B × 17*, 1B × 6′, 1B × 13′, 1B × 20*, 1By9*, 1By14.1 and 1By8.1) and 20 novel alleles at Glu‐B1 locus were detected. In particular, some additional protein components were detected, which probably were 1Ay subunits encoded by Glu‐A1 locus. The introduction of both Ax and Ay subunits from tetraploid wheats into hexaploid wheats may increase the genetic variability of gluten genes and consequently improve flour technological properties.     

Introduction of high molecular weight glutenin subunits 5 + 10 for the improvement of the bread-making quality of hexaploid triticale

In an earlier study, chromosome 1D of the hexaploid breadwheat cultivar ‘Chinese Spring’ was introduced into hexaploid triticale to improve its bread‐making quality. That specific chromosome, 1D, carried the a allele at the Glu‐D1 locus coding for high molecular weight (HMW) glutenin subunits 2 + 12, and since subunits 2 + 12 are associated with poor bread‐making quality in wheat, in the present study hexaploid 1D substitution triticale was crossed with octoploid triticale with the d allele at the Glu‐D1 locus encoding HMW glutenin subunits 5 + 10. Following backcrosses to different triticale varieties, 1D substitution lines were established that had Glu‐D1 allele a or d in an otherwise genetically similar background, and the influence of these two different alleles on bread‐making quality of hexaploid triticale was compared. The agronomic performance of 76 selected lines was evaluated in a field trial. The Zeleny sedimentation value was determined as a parameter for bread‐making quality, and related to the presence of chromosome 1D, the different glutenin alleles and the nature of the substitution. The presence of chromosome 1D had a significant and positive effect on the Zeleny sedimentation value, but the difference between the two glutenin alleles 2 + 12 and 5 + 10 was not as obvious as in wheat. Owing to its high cytological stability and minimal effect on agronomic performance, substitution 1D(1A) appears to be the most desirable one to use in triticale breeding.     

Introduction to bread wheat (Triticum aestivum L.) and assessment for bread-making quality of alleles from T. boeoticum Boiss. ssp. thaoudar at Glu-A1 encoding two high-molecular-weight subunits of glutenin

Two alleles, Glu-A1r encoding high-molecular-weight (HMW) glutenin subunits 39+40 and Glu-A1s encoding HMW glutenin subunits 41+42, were introgressed to bread wheat (Triticum aestivum L.) cv. Sicco from two accessions of T. boeoticum Boiss. ssp. thaoudar (A genome species, 2n=2x=14). Alleles at Glu-A1 in current commercial bread wheats encode zero or one subunit, and alleles at the homoeoloci Glu-B1 and Glu-D1 encode a maximum of two subunits; hence the maximum number of subunits found in commercial wheats is five, whereas the lines incorporating Glu-A1r and Glu-A1s carry six. Using near-isogenic lines, the current results demonstrated that the introduction of Glu-A1r resulted in diminished dough stickiness and improved stability during mixing compared with Glu-A1a encoding subunit 1, and a small improvement in gluten strength as shown by the SDS- sedimentation test. The introduction of Glu-A1a also resulted in a small improvement in gluten strength predicted by the SDS-sedimentation test. Thus the alleles are of potential value in breeding programmes designed to improve bread-making quality.     

Relationship between common wheat (Triticum aestivum L.) gluten proteins and dough rheological properties

This paper reports the correlation between the rheological properties of bread wheat dough and the types and quantities of endosperm proteins in 28 common wheat cultivars. Different methods were used to analyse the allelic composition of these cultivars and the relative quantities of the different proteins contributing to the gluten structure. Neither dough strength (W) nor tenacity/extensibility (P/L) correlated with allelic composition. Different wheats with the same allelic composition (i.e., with respect to glutenins) showed different rheological properties. The glutenins were the most influential components of W and P/L, especially the high molecular weight (HMW) glutenin subunits and in particular the type x form. These proteins seem to increase W and are the main constituents of the gluten network. The gliadins and low molecular weight (LMW) glutenin subunits appear to act as a “solvent”, and thus modify the rheological properties of the dough by either interfering with the polymerisation of the HMW glutenin subunits, or by altering the relative amounts of the different types of glutenin available. Thus, the protein subunits coded for by the alleles Glu-B1x7 and Glu-D1x5 stabilised the gluten network, whereas those coded for by Glu-B1x17 and Glu-D1x2 had the opposite effect. Dough properties therefore appear to depend on the glutenin/gliadins balance, and on the ratio of the type x and type y HMW proteins. The influence of external factors seems to depend on the allelic composition of each cultivar.     

对小麦不同HMW麦谷蛋白亚基质量评分系统的评价

小麦HMW麦谷蛋白亚基组成不同是造成各品种间烘烤品质差异的主要原因。不同研究者分别依据亚基间沉淀值（Payne等，1987）、评价值（傅宾孝等．1989）、抗延伸力（赵友梅等，1990）以及包括沉淀值在内的多个性状（赵和等，1994）的差异。先后建立了各自的评分系统。比较研究结果表明，前3种亚基评分系统对亚基的质量评分具有一定的共性，个别亚基的评分则存在差异；赵和等（1994）的评分系统则侧重考虑1D控制的亚基对烘烤品质的决定性作用，1A和1B控制的亚基作用甚小，因而与其余3种评分方法存在较大分歧。相关分析结果表明，就评价中国小麦品种的品质而言，国内制订的评分系统优于国外的。除容重、灰分等性状外，HMW麦谷蛋白亚基组成与大多数品质性状呈显著或极显著相关关系．     

小麦品质的麦谷蛋白亚基评定标准研究

测定了233份小麦面粉样品的高分子量谷蛋白亚基(HMW-GS)含量、Zeleny-沉降值和谷蛋白大聚体(GMP)含量,并根据SDS-PAGE结果计算了其Payne亚基品质评分.结果表明,不同HMW-GS含量差异显著,不同HMW-GS所对应品种的HMW谷蛋白总量、沉降值、GMP含量、Payne品质评分平均值也存在显著差异,说明不同亚基对品质的影响存在显著差异.HMW谷     

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.     

The high-molecular-weight glutenin subunit composition of Soviet wheat varieties

Summary One hundred and twenty-eight wheat varieties bred in the Soviet Union were screened for the composition of high-molecular-weight (HMW) subunits of glutenin. In general, variability was low compared to that seen in varieties from other countries. However, varieties from different regions showed distinctive patterns, in some cases clearly due to the use of particular parents in certain breeding programmes, but in others possibly due to the adaptive value of particular alleles to the environmental conditions under which the varieties were bred. For example, among spring varieties, the Glu-D1 allele encoding subunits 2+12 was more common in varieties from areas with limited rainfall than was the allele encoding subunits 5+10.The pattern of HMW glutenin subunits amongst varieties with superior bread-making quality showed few differences from that amongst bread-making varieties of lower quality.Abbreviations BMQ bread-making quality - HMW high-molecular-weight - LMW low-molecular-weight - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Relationship between electrophoretic pattern of storage proteins and gluten strength in durum wheat landraces from Turkey

A collection of 180 Triticum durum lines from Turkey and 22 Italian cultivars were investigated by electrophoretic techniques and evaluated for grain parameters such as protein content and sodium dodecyl sulphate (SDS)-sedimentation test. A total of 14 different patterns were observed for high-molecular-weight (HMW) glutenin subunits resulting from the combination of three alleles from the A genome and seven from the B genome. Three new allelic variants, one at Glu-A1 and two at Glu-B1, were found. Five different alleles were detected at the Glu-B1 locus. Genotypes with the same allelic variant at Gli-B1 showed inconsistency of composition in B group of low-molecular-weight (LMW) glutenin subunits. A total of 11 different B-subunit patterns were identified in the material analysed. A substantial proportion (78%) of the variation in gluten properties could be explained in terms of protein composition, with the LMW glutenin subunits making the largest contribution (54%). Results indicate that, in addition to Glu-B3 encoded proteins, variation at other loci coding for some intermediate and fastest B subunits might also influence rheological properties.     

Generation and characterization of a high molecular weight glutenin 1Bx14-deficient mutant in common wheat

Significant progress has been made in understanding the structure of high molecular weight (HMW) glutenin subunits and their role in determining the end use quality of wheat grains. However, few reports have dealt with the development and characterization of knock out mutants for HMW glutenin subunit genes. Here, the molecular analysis of MB14, a mutant derived from an elite Chinese wheat variety Xiaoyan 54 through chemical mutagenesis is described. SDS‐PAGE and Western blot experiments revealed that, in the seeds of homozygous MB14 plants, the expression of the 1Bx14 subunit was specifically blocked whereas the remaining four subunits (1Ax1, 1By15, 1Dx2, 1Dy12) accumulated to levels comparable to those in the wild type plants. The 5′‐flanking region and the open reading frame (ORF) of the mutant 1Bx14 allele were amplified and compared to the corresponding regions of wild type 1Bx14. The nucleotide sequences of the 5′‐flanking regions from the mutant and wild type 1Bx14 alleles were identical. However, the ORF of the mutant allele differed from that of the wild type 1Bx14 by three point substitutions, one of which resulted in a premature stop codon in the mutant ORF. Interestingly, the mutant 1Bx14 allele was still transcribed in the developing seeds, but no truncated translation product could be detected by Western blot analysis. Potential application of the 1Bx14 knock out mutant in studying the biological function of 1Bx14 and its contribution to the end use quality control in hexaploid wheat is discussed.     

Allelic variation at the <Emphasis Type="Italic">Glu-1</Emphasis> and <Emphasis Type="Italic">Glu-3</Emphasis> loci,presence of the 1B.1R translocation,and their effects on mixographic properties in Chinese bread wheats

Allelic variations at the Glu-1 and Glu-3 loci play an important role in determining dough properties and bread-making quality. Two hundred and fifty-one cultivars and advanced lines from four major Chinese wheat-producing zones in the autumn-sown wheat regions were used to investigate the high-molecular-weight glutenin subunits (HMW GS) and low-molecular-weight glutenin subunit (LMW GS) composition controlled by the Glu-1 and Glu-3 loci, respectively, as well as the presence of the 1B.1R translocation, and to determine the association of storage protein composition with protein content, SDS sedimentation value, and dough-mixing properties measured by mixograph. Three, nine, and four allelic variations were present at Glu-A1, Glu-B1, and Glu-D1, respectively. Subunits 1, N, 7+8, 7+9, and 2+12 are the dominant HMW GS, with frequencies of 51.3, 39.4, 38.2, 45.0, and 59.8%, respectively. Five and eight allelic variations were present at the Glu-A3 and Glu-B3 loci (data of Glu-D3 were not available), Glu-A3a, Glu-A3d, Glu-B3j (presence of the 1B.1R translocation), and Glu-B3d are the dominant LMW GS, with frequencies of 37.1, 31.7, 44.6, and 20.3%, respectively. The frequencies of allelic variation at Glu-1 and Glu-3 differ greatly in different regions. The effects of HMW GS and LMW GS on SDS sedimentation value, mixing time, and mixing tolerance were significant at P = 0.01, with Glu-D1 and Glu-B3 showing the largest contributions to mixing time and mixing tolerance. Averaged data from two locations showed that the quality effects of glutenin loci could be ranked as Glu-B3 > Glu-B1 > Glu-A1 > Glu-D1 > Glu-A3 for SDS sedimentation value, Glu-D1 > Glu-B3 > Glu-A1 = Glu-B1 = Glu-A3 for mixing time, and Glu-D1 > Glu-B3 = Glu-B1 > Glu-A3 > Glu-A1 for mixing tolerance, respectively. The significant and negative effect of the 1B.1R translocation on dough properties was confirmed. It was concluded that the high frequency of undesirable HMW GS and LMW GS and the presence of the 1B.1R translocation are responsible for the weak gluten property of Chinese germplasm; hence, reducing the frequency of the 1B.1R translocation and integration of desirable subunits at Glu-1 and Glu-3 such as 1, 7+8, 14+15, 5+10, Glu-A3d, and Glu-B3d, could lead to the improvement of gluten quality in Chinese wheats.     

Variation at Glu-1 Loci in Club Wheats

Hexaploid club wheats (Triticum aestivum L.) possess unique end-use quality characteristics and are grouped as a U.S. market subclass of soft white common wheat. Although there have been many reports on associations among high-molecular-weight glutenin storage protein (HMW-Glu) sub-units with end-use quality in hard wheats; there has been very limited work done on surveying the club wheats for these subunits. The HMW-Glu subunits, spike types and grain color were determined for 41 U.S. club wheat cultivars and 79 club accessions obtained from the National Small Grains Collection (NSGC), USDA-ARS. Accession ‘Harlan JR 35’ (PI 420948), which appears morphologically to be a hexaploid club wheat, was determined to be tetraploid. Egyptian line ‘Maya II-Tel's’ (PI 422288) was shown to have previously undescribed HMW-Glu subunits. In the U.S. club wheats the most common HMW-Glu subunits were: null, 49 % (Glu-A1); 6, 37 % (Glu-B1); and 2 + 12, 94 % (Glu-D1). In the NSGC group the most common HMW-Glu sub-units were: 2*, 52 % (Glu-A1); 7 + 8, 31 % (Glu-B1); and 2 + 12, 92 % (Glu-D1). The high frequency of subunits 2 + 12 in the club wheat groups has not been observed previously in numerous surveys of diverse wheat cultivars. The Glu-B1 subunits 6 (without subunit 8) and 20 which have not been reported in US red wheats, were shown to be common in both club wheat groups. A comparison of diversity indices, including previous studies on HMW-Glu subunit frequencies on all market classes of wheat, showed that the club groups are as diverse for the Glu-A1 and Glu-B1 loci and less diverse for the Glu-D1 locus.     

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.