Characterization of high-molecular-weight glutenin subunit genes from Elytrigia elongata

Three high‐molecular‐weight glutenin subunit (HMW‐GS) genes from Elytrigia elongata (Host) Nevski were characterized by determining the coding sequences of two x‐type subunit genes Ee2.1 and Ee1.9, and one y‐type subunit gene Ee1.8 with 2082, 1938 and 1788 bp, respectively. The numbers of amino acids in the central repeat domains of Ee2.1, Ee1.9 and Ee1.8 were considerably fewer than the other known HMW‐GSs with substitutions, insertions and/or deletions involving a single or more amino acid residues. Moreover, an extra cysteine (Cys) was found in the repeated domain of x‐type subunit Ee2.1. The difference in the number and position of Cys residues might be associated with the good dough quality. The extra Cys in the good‐quality subunit Dx5 was at the beginning of the repetitive domain, while the extra Cys in Ee2.1 was at the central repetitive domain. Evolutionary relationships between the HMW‐GSs from E. elongata and the known HMW‐GSs were estimated. It transpired that Ee1.9, Ee2.1 and Bx7 were clustered into one subgroup with high bootstrap values, while Ee1.8 was closely related to Ay.     

Identification of low-molecular weight glutenin subunits of wheat associated with bread-making quality

Although it is known that the compositions of low‐molecular weight glutenin subunits (LMW‐GSs) are important factors for bread‐making quality of wheat, it is still not clear which LMW‐GSs confer improved bread‐making quality and how those LMW‐GSs interact with high‐molecular weight (HMW) GSs. Using a hard red winter wheat line with good bread‐making quality and a Japanese wheat cultivar with poor quality as well as their progeny we identified LMW‐GSs associated with the bread‐making quality. One such LMW‐GS, KS2, which had a molecular weight of 42 kDa and was allelic to HS1, was associated with bread‐making quality. Furthermore, by using four recombinant inbred lines with different HMW‐GS and LMW‐GS combinations, KS2 and HMW‐GS 5+10 showed interaction effects on the bread‐making quality. Two‐dimensional polyacrylamide gel electrophoresis (Page) analysis showed that KS2 consists of two protein components and that HS1 is composed of three components. The N‐terminal amino acid sequences of the five components were identical to the most frequently analysed sequence of LMW glutenin components.     

The HMW glutenin suhunit composition of bread-wheat varieties bred in Pakistan

A total of 50 bread-wheat varieties bred in Pakistan were characterized for the composition of the high-molecular-weight glutenin subunit (HMW-GS). The glutenin subunits, as revealed by sodium dodecyl-sulfate-polyacrylamide-gel electrophoresis (SDS-PAGE) were numbered according to Payne's numbering system and, varieties were accordingly assigned theoretical quality scores. All varieties were found to possess either subunit 1 or 2* coded by the A genome. Subunits 17+18 and 7 + 9 of the B genome and 5+10 of the D genome were predominantly found in this set of varieties. The frequency of the appearance of Glu-l alleles in the varieties was different from that seen in other countries, especially in terms of the absence of the ‘null’ form of the A genome and the presence of novel subunits at the Glu-Bl and Glu-Dl loci. The compositions of HMW-GS were generally of good quality, with more than 50% of the varieties achieving quality scores of 9–10 with a high (8.6) average score. The results in this study indicate that wheat varieties bred in Pakistan have a narrow genetic base in terms of HMW glutenin subunits. It appears that the breeders have been selecting wheat genotypes of glutenin subunits associated with good quality without knowing the actual composition of these subunits. The database established on the basis of these results is useful for wheat-improvement programmes aimed at varietal identification and breeding for good quality parameters.     

Effect of the leaf rust resistance gene Lr28 on grain yield and bread-making quality of wheat

Near‐isogenic lines carrying the Lr28 gene developed in five genetic backgrounds were tested for 2 years with and without fungicide treatment. The Lr28 gene increased grain yield, 1000‐grain weight and number of effective tillers per plant under heavy leaf rust infection with no negative effects on yield and bread‐making quality in rust‐free plots. Although a reduction in dough development time was found to be associated with Lr28, it can still be used extensively in wheat breeding programmes.     

Rapid screening of selected European winter wheat varieties and segregating populations for the Glu-D1d allele using PCR

Forty-two winter wheat varieties and 193 F₂ and BC₁F₂ seeds were screened for Glu-D1d allele encoding the HMW glutenin subunits 5 + 10 using polymerase chain reaction (PCR). The segregating populations originated from crosses involving wheat parents with good and poor bread-making quality. A clear PCR product of 450 bp, representing 1Dx5 of the Glu-D1d allele was identified in 24 varieties and 111 hybrid seeds. Four different Glu-D1 alleles: a (2 + 12), b (3 + 12), c (4 + 12) and d (5 + 10) were detected using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Only genotypes possessing Glu-D1d gave a positive PCR signal, hexaploid triticale and 4 × wheat lacking Glu-D1 locus gave a negative signal. The efficiency of PCR selection for bread-making quality in early generations using half seed is discussed.     

Genetic analysis of bread-making quality in wheat and spelt

Bread‐making quality in wheat and spelt reflects the combination of several, mostly quantitatively inherited parameters. The aim was to find molecular markers linked to quantitative trait loci (QTL) for quality parameters. Zeleny sedimentation values (Zel), protein (Prot), kernel hardness (KH) and 1000‐kernel weight (TKW) of 226 F₅ recombinant inbred lines (RILs) from a cross between wheat and spelt were assessed in different environments. The dough properties of 204 RILs were assessed with an alveograph. Based on a genetic map of 187 loci, nine QTL were found for Zel and Prot, explaining 47% and 51% of the phenotypic variance, respectively. Fifty‐four per cent of the variance was explained by 10 QTL for KH and eight for TKW. For the alveograph parameters 10 QTL were found for baking strength, nine for tenacity, seven for configuration ratio, and four for elasticity index and extensibility. The phenotypic variance explained ranged from 25% to 48%. The population mean of the dough parameters was shifted towards the spelt parent. It is concluded that non‐additive effects are crucial in the expression of high bread‐making quality of wheat. The consequences for wheat and spelt breeding programmes are discussed.     

2··, a new high molecular weight glutenin subunit coded by Glu-A1: its predicted structure and its impact on bread-making quality

The suitability of wheat varieties for bread‐making depends on their glutenin subunits. The amino acid composition of these gluten building‐blocks have a strong influence on the rheology of the dough and, thus, on the suitability of the variety for bread‐making. This study reports a new x‐type high molecular weight glutenin subunit coded by the locus Glu‐A1 and named 2··. To investigate the impact of this allele on 10 quality parameters, a doubled haploid (DH) population of Triticum aestivum, segregating for Glu‐A1, was created. The statistical analysis demonstrates that, at Glu‐A1, the subunit 2·· is as favourable for quality as the subunit 2*. This is in accordance with results showing that the 2·· open reading frame still has the same number of cysteines as 2*. The small differences in the length of the central domain had no detectable effect on the elasticity, tenacity and baking quality, of the dough.     

Effect of variation at Glu-D1 on club wheat end-use quality

Club wheats (Triticum aestivum L.), having the allele at the C locus conferring short spike rachis internodes and giving compact appearance of spikes, which have unique and highly desirable soft white wheat end-use quality characteristics are a vital submarket class of soft white wheat in the US Pacific Northwest. Two important varieties, ‘Tyee’ and ‘Albit’, are heterogeneous for high molecular weight glutenin subunits 2 + 12 and 5 + 10 encoded by the Glu-D1 locus. Replicated near-isogenic lines (NILs) of club wheats ‘Tyee’ and ‘Albit’ were grown in four field environments and used to determine the effect of Glu-D1 coded high molecular weight glutenin subunits (HMWGS) 5 + 10 and 2 + 12 on various end-use quality traits. The greatest effect of variation at this locus was observed for mixing time to peak, where there was significant variation (P < 0.01) between each 5 + 10 and 2 + 12 NIL group in each environment. Mixing time values for the 2 + 12 NILs for both ‘Albit’ and ‘Tyee’ ranged from 0.60 to 1.23 min lower than the 5 + 10 NILs. Mean values for traits mixing time to peak, cake volume, and viscosity were more favourable for the 2 + 12 NIL groups for all genotypes in all environments. No effects of these HMWGS were detected for test weight, kernel hardness, whole wheat protein, flour yield, ash, flour protein or cookie diameter. Selection for HMWGS 2 + 12 in club wheat breeding programmes should have positive effects on end-use quality.     

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.     

Additive and epistatic effects of allelic variation at the high molecular weight glutenin subunit loci in determining the bread-making quality of breeding lines of wheat

Summary The relation has been studied between the high molecular weight glutenin (HMWg) subunit alleles and the bread-making quality of 226 lines of winter wheat (T. aestivum L.), grown in The Netherlands. The lines represented a wide range of genetic backgrounds, and had not been selected for quality, in contrast to the established varieties used by other authors.The variation in HMWg subunit genotypes accounted for about 20% of the total variation in loaf volume among the lines. Most important was the allelic variation at the Glu-D1 locus. The Glu-D1 allele encoding the subunits 5+10 was superior to its allelic counterpart, encoding 2+12. The difference in average of loaf volume between groups of lines containing 5+10 or 2+12 was negatively related with protein content of the flours. When protein content was below 9.2%, no effect of allelic variation at the Glu-D1 locus was present. Epistatic effects between the Glu-I loci also contributed to the variation in loaf volume of the lines: i.e. the effect of allelic variation at Glu-A1 and Glu-B1 depended on the allele present at the Glu-D1. The contribution of the epistatic effects was about half the contribution of the additive effects, and should therefore be included in predictive models for bread-making quality.     

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.     

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     

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.     

Genetic control of some polymeric glutenin fractions in hexaploid wheat (Triticum aestivum)

Reciprocal crosses were made between seven different hexaploid wheat genotypes. Hybrid kernels and their parents were used to determine the amount of polymeric glutenin fractions by size-exclusion high-performance liquid chromotography (SE-HPLC) analysis. Quantitative aspects of the genetic control for various glutenin fractions were investigated through diallel cross analysis. The association between the potential effect on hexaploid wheat quality of various allelic types and the quantitative expression of the polymeric glutenin fractions was confirmed. Significant average heterosis effects were demonstrated for insoluble glutenin (pFi), total soluble and insoluble glutenin (pF1 + pF2 + pFi) and the ratio of soluble to insoluble glutenin, (pF1 + pF2)/pFi. Some genotypes showed significant positive or negative combining abilities and general reciprocal effects for the glutenin fractions studied. For gluten quality, ‘Qualital’ was the best combiner for determining an optimal glutenin composition (high values for pFi and pF1 + pF2 + pFi and low values for pF1/pF2 and (pF1 + pF2)/pFi, respectively). These results should be of great interest in breeding programmes aimed at improving hexaploid wheat quality.     

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 high-molecular-weight glutenin subunit Loci, Glu-A1, Glu-B1 and Glu-D1, in Japanese and Chinese hexaploid wheats

Variation in the electrophoretic banding patterns of high-molecular-weight (HMW) glutenin subunits of 274hexaploid wheat (Triticum aestivum) varieties from China was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 27 different major HMW glutenin subunits were identified. Each variety contained three to five subunits and 29different glutenin subunit patterns were observed in274 Chinese hexaploid wheats. Seventeen alleles were identified based on the comparison of subunits mobility with that previously identified in a set of standard hexaploid wheats. The Chinese hexaploid wheats exhibited allelic variation in HMW glutenin subunit composition and the variation differed from that of Japanese and hexaploid wheats of other countries. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

SDS-PAGE of the high-molecular-weight subunits of wheat glutenin and the characterization of 1R (1B) substitution and 1BL/1RS translocation lines

Summary The high-molecular-weight subunits of glutenin from wheat 1R(1B) substitution and 1BL/1RS translocation lines were fractionated by SDS-PAGE. Two new subunits denoted R₁ and R₂ were characterized in 1R(1B) substitution, but not in 1BL/1RS translocation lines. R₁ and R₂ were proved to be rye proteins by 2d electrophoresis (NEPHGE x SDS-PAGE).In contrast to literature citations it was demonstrated that the cultivar Winnetou is a 1R(1B) substitution line and the cultivars Clement and Mildress both are 1BL/1RS translocation lines.     

Influence of high molecular weight glutenin subunit substitution on rheological behaviour and bread-baking quality of near-isogenic lines developed from Chinese wheats

Three near‐isogenic lines (NILs) of wheat involving Glu‐B1 and Glu‐D1 alleles were used to study the genetic contribution of high molecular weight glutenin subunits (HMW‐GS) to gluten strength. The HMW‐GS composition of each NILs was determined by SDS‐PAGE. No significant differences were found in grain protein contents among the NILs. Gluten strength and dough‐mixing properties were measured by the Farinograph, the Extensograph, and SDS‐sedimentation (SDS‐SE). Results indicated that line 2, containing the Glu‐1B 14 + 15 and Glu‐1D 5 + 10 combination of subunits, had higher values for flour quality, dough rheological parameters, and bread‐baking quality when compared with lines 8 and 13. Line 8, containing Glu‐1B 7 + 9 and Glu‐1D 5 + 10, was better than line 13 with the Glu‐1B 14 + 15 and Glu‐1D 10 combination. Some major parameters appeared significantly different. The presence of Glu‐1B 14 + 15 was associated with higher dough strength based on SDS‐SE volume and several rheological parameters when compared with Glu‐1B 7 + 9. Lines with subunit 10 at Glu‐D1 performed significantly worse than those with 5 + 10 in gluten index, SDS‐SE volume, Farinograph stability time, Extensograph area and bread‐baking quality.     

Dough properties and bread-making quality-related characteristics of Yumechikara near-isogenic wheat lines carrying different Glu-B3 alleles

We investigated the relationships of three allelic variations in Glu-B3 (ab, g, and h) with dough properties and bread-making quality-related characteristics using near-isogenic lines (NILs) of ‘Yumechikara’ that commonly carry Glu-A1a, Glu-B1b, Glu-D1d, Glu-A3f, Glu-B3ab and Glu-D3a. Measurement of peak time (PT) in a 2-g mixograph indicated that Glu-B3g was the most effective for a strong dough property, followed by Glu-B3ab, with Glu-B3h being the least effective. The results of measurement of mixing time during bread-making were similar to those for PTs, i.e., the lines carrying Glu-B3g showed the longest mixing time, followed by those of Glu-B3ab, and those of Glu-B3h showed the shortest mixing time. Since two parameters of bread-making quality, loaf volume (LV) and specific loaf volume (SLV), were affected by flour protein contents in all groups of the Glu-B3 genotype, we compared the effects of the three Glu-B3 alleles on those parameters using analysis of covariance (ANCOVA) to remove the effect of protein content. The results indicated that the Glu-B3h group showed the largest SLV, followed by the Glu-B3ab group, and the Glu-B3g group showed the smallest SLV. These results suggest that the introduction of Glu-B3h into ‘Yumechikara’ makes it possible to breed varieties with good bread-making quality-related characteristics.