Genetic Variance for Gluten Strength Contributed by High Molecular Weight Glutenin Proteins

A total of 162 doubled haploid (DH) lines were produced from a cross between Triticum aestivum L. ‘AC Karma’ and line 87E03‐S2B1 to study the genetic contribution of high molecular weight (HMW) glutenin subunits to gluten strength. HMW glutenin subunit composition of each DH line was determined by SDS‐PAGE. The population was grown in the field at one location in 1999 and at three locations in 2000. Gluten strength and dough mixing properties were measured by mixograph test and SDS‐sedimentation test. Variance components were estimated for each measurement to determine the variability contributed by HMW glutenin subunits. Results indicated significant environmental impact on tested mixograph parameters, SDS‐sedimentation volumes and grain and flour protein concentration. Significant main effects of Glu‐1D loci encoded subunits were obtained for mixograph development time, energy to peak, slope after peak, and first minute slope. Lines containing 5+10 combination of subunits had higher values for mixograph development time and energy to peak, while slope after peak and first minute slope were lower as compared with 2+12 containing lines. Low intergenomic interactions were observed for bandwidth energy (BWE), total energy (TEG), and SDS‐sedimentation test, involving B and D genomes only. A portion of the genetic variability for gluten strength was accounted for overexpression of Bx7 subunit originating from the cultivar Glenlea derived line 87E03‐S2B1. There was no significant effect of Glu‐A1 encoded subunits on any of the tested parameters. Estimated genetic variability for gluten strength contributed by Glu‐B1 and Glu‐D1 encoded HMW glutenins was 55% for mixing development time and 51% for energy to peak.     

Relationship Between the Qualitative and Quantitative Compositions of Gluten Protein Types and Technological Properties of Synthetic Hexaploid Wheat Derived from Triticum durum and Aegilops tauschii

The contribution of the diploid wheat species Aegilops tauschii (Coss.) Schmall to the technological properties of bread wheat (Triticum aestivum L.) was previously studied by the investigation of synthetic hexaploids derived from tetraploid durum wheat (T. turgidum L.) and three diploid Ae. tauschii lines. The results indicated that bread volume, gluten index, SDS‐sedimentation volume, and maximum resistance of gluten were significantly influenced by the Ae. tauschii lines. To determine the relationship between technological properties and qualitative and quantitative compositions of gluten proteins, the flours of parental and synthetic lines were extracted using a modified Osborne fractionation. Gliadin and glutenin fractions were then characterized by reversed‐phase (RP) HPLC on C₈ silica gel. The HPLC patterns revealed typical differences between synthetic and parental lines. The gliadin patterns of three synthetic lines and the glutenin patterns of two synthetic lines were more similar to that of the diploid Ae. tauschii parents involved in the hybrids. In the glutenin pattern of one synthetic line, characteristics from both Ae. tauschii and the durum wheat parents were observed. The amount of total gliadin and gliadin types of the synthetic lines was mostly intermediate between those of the durum and Ae. tauschii parents. The amounts of total glutenin and glutenin types (HMW and LMW subunits) of the synthetic lines were generally higher than those of the parental lines, and the ratio of gliadins to glutenins was significantly decreased. High positive correlations were found between the amount of total glutenins, HMW, and LMW subunits and bread volume, maximum resistance and extension area of gluten, and SDS‐sedimentation volume. The ratio of gliadins to glutenin subunits had a strong negative influence on these properties. The protein content of the flours and the amount of total gluten proteins were not correlated with any of the technological properties. Results on the relationship between biochemical characteristics and the breadmaking properties indicated that wheat prebreeding would benefit from studies on protein types and quantification in the choice of parents. In addition, the potential of the diploid Ae. tauschii for improvement of breadmaking quality should be further exploited.     

Breadmaking Quality of Selected Durum Wheat Genotypes and Its Relationship with High Molecular Weight Glutenin Subunits Allelic Variation and Gluten Protein Polymeric Composition

Twenty‐seven durum wheat genotypes originating from different geographical areas, all expressing LMW‐2 at Glu‐B3, and five bread wheats were evaluated for flour mixing properties, dough physical characteristics, and baking performance. Gluten polymeric composition was studied using size‐exclusion HPLC of unreduced flour protein extracts. As a group, durum wheats had poorer baking quality than bread wheats in spite of higher protein and total polymer concentrations. Durum wheats exhibited weaker gluten characteristics, which could generally be attributed to a reduced proportion of SDS‐unextractable polymer, and produced less extensible doughs than did bread wheats. However, substantial variation in breadmaking quality attributes was observed among durum genotypes. Better baking performance was generally associated with greater dough extensibility and protein content, but not with gluten strength related parameters. Extensibility did not correlate with gluten strength or SEHPLC parameters. Genotypes expressing high molecular weight glutenin subunits (HMW‐GS) 6+8 exhibited better overall breadmaking quality compared with those expressing HMW‐GS 7+8 or 20. Whereas differences between genotypes expressing HMW‐GS 6+8 and those carrying HMW‐GS 7+8 could only be attributed to variations in extensibility, the generally inferior baking performance of the HMW‐GS 20 group relative to the HMW‐GS 6+8 group could be attributed to both weaker and less extensible gluten characteristics.     

Interactions of Genotype and Glutenin Subunit Composition on Breadmaking Quality of Durum 1AS•1AL‐1DL Translocation Lines

Dual‐purpose durum (Triticum turgidum L. subsp. durum) wheat, having both good pasta and breadmaking quality, would be an advantage in the market. In this study, we evaluated the effects of genotype and varying HMW and LMW glutenin subunit composition on durum breadmaking quality. Genotypes included five near‐isogenic backgrounds that also differed by variability at the Glu‐D1d (HMW subunits 1Dx5+1Dy10), Glu‐B1 (presence or absence of subunit 1By8), and Glu‐B3 (LMWI or LMWII pattern) loci. Quality tests were conducted on genotypes grown at five North Dakota locations. Genotype had a stronger influence on free asparagine content than glutenin subunit composition. Genotypes carrying Glu‐D1d had higher glutenin content than lines that did not carry Glu‐D1d. Among Rugby translocation genotypes, lines carrying LMWI had higher gliadin content and better loaf volume than genotypes carrying LMWII. Absence of 1By8 produced major reductions in loaf volume in nontranslocation lines regardless of whether LMWI or LMWII was present. In contrast, the presence of Glu‐D1d compensated well for the absence of 1By8 regardless of which LMW pattern was present. The durum genotypes did not have loaf volumes equal to bread wheat cultivars, and results suggest that improved extensibility is needed to improve durum breadmaking quality.     

Effect of the Molecular Weight Distribution of Glutenin Protein from an Extra‐Strong Wheat Flour on Rheological and Breadmaking Properties Through Reconstitution Studies

Ten glutenin fractions were separated by sequential extraction of wheat gluten protein with dilute hydrochloric acid from defatted glutenin‐rich wheat gluten of the Canadian hard red spring wheat (HRSW) cultivar Glenlea. The molecular weight distribution (MWD) of 10 different soluble glutenin fractions was examined by multistacking SDS‐PAGE under nonreduced conditions. Also, the subunit composition of the different glutenin fractions was determined by SDS‐PAGE under reduced conditions. The MWD of the fractions (especially HMW glutenins) varied from fraction to fraction. From early to later fractions, the MWD shifted from low to high. The early extracted fractions contained more LMW glutenin subunits (LMW‐GS) and less HMW glutenin subunits (HMW‐GS). The later extracted fractions and the residue fraction contained much more HMW‐GS (2*, 5, and 7 subunits) than the early extracted fractions. The trend in the amounts of 2*, 5, and 7 subunits in each fraction from low to high matched the extraction solvent sequence containing from lower to higher levels of HCl. The influence of glutenin protein fractions from the extra‐strong mixing cultivar, Glenlea, on the breadmaking quality of the weak HRSW, McVey, was assessed by enriching (by 1%) the McVey base flour with isolated glutenin protein fractions from Glenlea. The mixograph peak development times and loaf volumes of enriched flour were measured in an optimized baking test. The results indicated that the higher content in Glenlea glutenin of HMW‐GS with higher molecular weight, such as 2*, 5, and 7, seem to be the critical factor responsible for the strong mixing properties of Glenlea. Our results confirmed that subunit 7 occurred in the highest quantity of all the HMW‐GS. Therefore, it seems that the greater the content of larger molecular weight glutenin subunits, the larger the glutenin polymers and the stronger the flour.     

Synergistic and Additive Effects of Three High Molecular Weight Glutenin Subunit Loci. II. Effects on Wheat Dough Functionality and End‐Use Quality

Understanding the relationship between basic and applied rheological parameters and the contribution of wheat flour protein content and composition in defining these parameters requires information on the roles of individual flour protein components. The high molecular weight glutenin subunit (HMW‐GS) proteins are major contributors to dough strength and stability. This study focused on eight homozygous wheat lines derived from the bread wheat cvs. Olympic and Gabo with systematic deletions at each of three HMW‐GS encoding gene loci, Glu‐A1, Glu‐B1, and Glu‐D1. Flour protein levels were adjusted to a constant 9% by adding starch. Functionality of the flours was characterized by small‐scale methods (2‐g mixograph, microextension tester). End‐use quality was evaluated by 2‐g microbaking and 10‐g noodle‐making procedures. In this sample set, the Glu‐D1 HMW‐GS (5+10) made a significantly larger contribution to dough properties than HMW‐GS coded by Glu‐B1 (17+18), while subunit 1 coded by Glu‐A1 made the smallest contribution to functionality. These differences remained after removing variations in glutenin‐to‐gliadin ratio. Correlations showed that both basic rheological characteristics and protein size distributions of these flours were good predictors of several applied rheological and end‐use quality tests.     

Influence of High Molecular Weight Glutenins on Viscoelastic Properties of Intact Wheat Kernel and Relation to Functional Properties of Wheat Dough

High and low molecular weight glutenin subunits (HMW‐GS and LMW‐GS, respectively) are the main factors determining the viscoelastic properties of wheat dough. The mechanical and viscoelastic properties of 29 samples of wheat kernels differing in HMW‐GS were evaluated with load‐compression tests. Samples were grouped by genotypes differing in HMW‐GS composition (allelic variants: Glu‐A1: null, 1, 2*; Glu‐B1: 7, 7+8, 7+9, 13+16, and 17+18; Glu‐D1: 5+10, 2+12). Groups representing Glu‐A1 1 and 2*; Glu‐B1 7, 7+9 and 17+18; and Glu‐D1 5+10 generally possessed hard grain and showed the largest kernel elasticity values, while those representing subunits Glu‐A1 null; Glu‐B1 7+8; and Glu‐D1 2+12 had soft kernels and showed lower elastic work values. Genotypes possessing HMW‐GS 1, 17+18 and 5+10 gave large SDS‐sedimentation values and better dough viscoelastic properties than those with allelels: null, 7+8, and 2+12. Kernel hardness showed significant correlation with the dough‐strength‐related parameters: SDS‐sedimentation; dough mixing time; and the alveographic parameters, W and P. There was a negative correlation between kernel plastic work and dough mixing time and the dough tenacity/extensibility parameters, P/L. The significant relationship between sedimentation tests and kernel elastic work seems to indicate that elastic work is related to genotype (protein composition). The general tendency was that higher values in kernel elastic work and size corresponded to better dough rheological quality. Mechanical properties of the kernel were significantly related to the elastic behavior measured in a single wheat kernel. The use of the compression test on individual kernels is easy, rapid and nondestructive and therefore seems to show potential use as a rapid tool in breeding to improve wheat quality.     

Influence of Low‐Molecular‐Weight Glutenin Subunit Haplotypes on Dough Rheology in Elite Common Wheat Varieties

The low‐molecular‐weight glutenin subunits (LMW‐GSs) are a class of wheat seed storage proteins encoded by a multigene family located at the Glu‐3 loci that influences wheat end‐use quality. Owing to ambiguities in the LMW‐GS allele nomenclature and to the complexity of the Glu‐3 loci organization, a clear relationship between LMW‐GS alleles and wheat end‐use quality has not been adequately determined. In the present study, four sets of elite common wheat varieties were analyzed for their LMW‐GS genic profile, along with their dough rheology and end‐product baking properties. Among these varieties, variation at the Glu‐A3 locus had a major impact on the analyzed dough rheology parameters, followed by the Glu‐B3 and Glu‐D3 loci. Also, the genes located at the linkage groups Glu‐A3‐3, Glu‐B3‐3, and Glu‐D3‐5 were more highly associated with dough strength, mixing, and extensibility properties. Results obtained in this study clearly indicate that there are specific LMW‐GS haplotypes that are more highly associated than others to variation in dough rheology.     

Effects of Nitrogen Fertilizer on Protein Quantity and Gluten Strength Parameters in Durum Wheat (Triticum turgidum L. var. durum) Cultivars of Variable Gluten Strength

Field studies were conducted over three years at two locations in Saskatchewan, Canada, to determine the effect of nitrogen fertilizer on protein quantity and protein strength in 10 cultivars of durum wheat (Triticum turgidum L. var. durum) representing a range of gluten strength. Increasing nitrogen fertilizer resulted in increased protein content in all cultivars across environments. Cultivars were clearly differentiated on the basis of gluten strength using a gluten index (GI), SDS sedimentation (SDSS), alveograph indices of overpressure (P) and deformation energy (W), mixograph energy to peak (ETP), and mixograph bandwidth energy (BWE) at all fertilizer levels. Variable cultivar response to nitrogen fertilizer was observed only for protein content, GI, and alveograph W. The nature of the cultivar‐by‐fertilizer interaction for GI suggested that the conventional strength cultivars would benefit more from nitrogen fertilizer than the extra‐strong types, which showed no change or slight decreases in GI with nitrogen fertilizer despite an increase in total gluten. SDSS increased with nitrogen fertilizer, following similar trends as protein. Gluten strength rankings of the cultivars by SDSS were maintained with increased fertilizer. Fertilizer had little effect on alveograph P, mixograph ETP, and mixograph BWE. Overall, GI values were more stable across increasing levels of nitrogen fertilizer and resultant increased protein content compared with SDSS, mixograph development time, and alveograph W and L, suggesting it is a good test for estimating intrinsic gluten strength for cultivars with a wide range of protein content.     

Effects of Allelic Variations in Wx‐1, Glu‐D1, Glu‐B3, and Pinb‐D1 Loci on Flour Characteristics and White Salted Noodle‐Making Quality of Wheat Flour

Doubled haploid wheat lines developed from a cross between a hard white winter wheat variety of normal starch endosperm and a waxy wheat variety were used to determine the effects of allelic variation in Wx‐1, Glu‐D1, Glu‐B3, and Pinb‐D1 loci on physiochemical properties of flour, noodle dough properties, and textural quality of cooked noodles. Milling yield, damaged starch content, protein content, and SDS sedimentation volume of flour were influenced the most by allelic composition of Pinb‐D1 loci, less by Wx‐1 loci, and least by Glu‐B3. Wheat lines carrying Pinb‐D1b or Glu‐B3h alleles exhibited higher milling yield and damaged starch content of flour than those with Pinb‐D1a and Glu‐B3d alleles. Wheat lines carrying the Pinb‐D1b allele were higher in protein content and SDS sedimentation volume than those carrying Pinb‐D1a. Mixograph water absorption was largely influenced by allelic composition of Wx‐1 loci, whereas mixograph mixing time and mixing tolerance were predominantly determined by allelic composition of Glu‐D1 loci. Amylose content and pasting properties of starch were mainly determined by allelic composition of Wx‐1 loci with little influence by allelic compositions of Glu‐D1, Glu‐B3, and Pinb‐D1 loci. Allelic composition of Wx‐1 loci contributed 53.4% of the variation in optimum water absorption of noodle dough and 26.7% of the variation in thickness of the noodle dough sheet. The variation of 7.8% in optimum water absorption of noodle dough was contributed by the allelic composition of Pinb‐D1 loci. Allelic composition of Wx‐1 loci was responsible for 73.2, 74.4, and 59.6% in the variation of hardness, springiness, and cohesiveness of cooked noodles, respectively. Cohesiveness of cooked noodles was also influenced by the allelic compositions of Glu‐B3 and Pinb‐D1 loci to a smaller extent.     

Synergistic and Additive Effects of Three High Molecular Weight Glutenin Subunit Loci. I. Effects on Wheat Dough Rheology

The high molecular weight glutenin subunits (HMW‐GS) play an important role in governing the functional properties of wheat dough. To understand the role of HMW‐GS in defining the basic and applied rheological parameters and end‐use quality of wheat dough, it is essential to conduct a systematic study where the effect of different HMW‐GS are determined. This study focuses on the effect of HMW‐GS on basic rheological properties. Eight wheat lines derived from cvs. Olympic and Gabo were used in this study. One line contained HMW‐GS coded by all three loci, three lines were each null at one of the loci, three lines were null at two of the loci and one line null at all three loci. The flour protein level of all samples was adjusted to a constant 9% by adding starch. In another set of experiments, in addition to the flour protein content being held at 9%, the glutenin‐to‐gliadin ratio was maintained at 0.62 by adding gliadin. Rheological properties such as elongational, dynamic, and shear viscometric properties were determined. The presence of Glu‐D1 subunits (5+10) made a significantly larger contribution to dough properties than those encoded by Glu‐B1 (17+18), while subunit 1, encoded by Glu‐A1, made the least contribution to functionality. Results also confirmed that HMW‐GS contributed to strength and stability of dough.     

Allelic Variation at Glu-D1 Locus for High Molecular Weight (HMW) Glutenin Subunits: Quantification by Multistacking SDS-PAGE of Wheat Grown Under Nitrogen Fertilization

Two biotypes of an Australian wheat cultivar, Warigal, differing only in the Glu-D1 high molecular weight (HMW) glutenin subunits 5+10 and 2+12 were used in this study. The objective was to examine the effects of nitrogen fertilization and allelic variation at the Glu-D1 locus on the characteristics of glutenin polymers. Unreduced proteins containing the SDS-soluble glutenins and the other protein classes were analyzed by multistacking SDS-PAGE which separates the glutenin into six distinctly different-sized aggregates. The results showed that nitrogen fertilization significantly increased protein quantity, ratio of polymers to monomeric proteins, and sizes of SDS-soluble glutenins. Nitrogen fertilization affected the proportions of HMW subunits in both SDS-soluble and SDS-insoluble glutenin polymers and the ratio of x to y subunits in SDS-insoluble glutenin polymers. Nitrogen fertilization, however, did not cause a significant change in ratio of SDS-soluble to SDS-insoluble glutenins. SDS-insoluble glutenins had a greater ratio of HMW to LMW and x to y subunits, especially with a higher increase of 1Dx subunits, than SDS-soluble glutenins. The HMW/LMW subunit ratio and the x/y subunit ratio may be used to predict sizes of glutenin polymers. The biotype with 5+10 subunits had a greater x/y subunit ratio in the SDS-insoluble glutenins than the 2+12 type. A greater proportion of subunit 5 was formed than subunit 2 in the SDS-insoluble glutenin polymers. Both nitrogen fertilization and allelic variation at Glu-D1 loci could affect the characteristics of glutenin polymers.     

Composition of HMW and LMW Glutenin Subunits and Their Effects on Dough Properties,Pan Bread,and Noodle Quality of Chinese Bread Wheats

Knowledge of composition of high molecular weight glutenin subunits (HMW‐GS) and low molecular weight glutenin subunits (LMW‐GS) and their associations with pan bread and noodle quality will contribute to genetically improving processing quality of Chinese bread wheats. Two trials including a total of 158 winter and facultative cultivars and advanced lines were conducted to detect the allelic variation at Glu‐1 and Glu‐3 loci by SDS‐PAGE electrophoresis and to understand their effects on dough properties, pan bread, and dry white Chinese noodle (DWCN) quality. Results indicate that subunits/alleles 1 and null at Glu‐A1, 7+8 and 7+9 at Glu‐B1, 2+12 and 5+10 at Glu‐D1, alleles a and d at Glu‐A3, and alleles j and d at Glu‐B3 predominate in Chinese germplasm, and that 34.9% of the tested genotypes carry the 1B/1R translocation (allelic variation at Glu‐D3 was not determined because no significant effects were reported previously). Both variations at HMW‐GS and LMW‐GS/alleles and loci interactions contribute to dough properties and processing quality. For dough strength related traits such as farinograph stability and extensigraph maximum resistance and loaf volume, subunits/alleles 1, 7+8, 5+10, and Glu‐A3d are significantly better than those of their counterpart allelic variation, however, no significant difference was observed for the effects of d, b, and f at Glu‐B3 on these traits. For extensigraph extensibility, only subunits 1 and 7+8 are significantly better than their counterpart alleles, and alleles d and b at Glu‐B3 are slightly better than others. For DWCN quality, no significant difference is observed for HMW‐GS at Glu‐1, and Glu‐A3d and Glu‐B3d are slightly better than other alleles. Glu‐B3j, associated the 1B/1R translocation, has a strong negative effect on all quality traits except protein content. It is recommended that selection for subunits/alleles 1, 7+8, 5+10, and Glu‐A3d could contribute to improving gluten quality and pan bread quality. Reducing the frequency of the 1B/1R translocation will be crucial to wheat quality improvement in China.     

Specific Glu‐D1ƒ Allele Frequency of Japanese Common Wheat Compared with Distribution of Glu‐1 Alleles in Chinese Wheat

The quality of wheat (Triticum aestivum L.) grain favored in breadmaking is strongly affected by components of seed storage protein, particularly high molecular weight glutenin subunits (HMW‐GS). The HMW‐GS 2.2 controlled by the Glu‐D1ƒ allele is frequently found in Japanese cultivars and landraces. In the investigation into the factors affecting the distribution of the allele, the available data on HMW‐GS of common wheats from Japan were analyzed and compared with the data for intensity of winter habit and wheat flour hardness. We show that the main factors affecting the Glu‐D1ƒ allele frequency in Japanese wheat were the intensity of natural selection for winter habit and artificial selection for flour hardness. According to a study of the worldwide distribution of Glu‐1 alleles, the Glu‐D1ƒ allele is rare. However, Glu‐D1ƒ allele was the most common Japanese wheat seed storage protein allele. It is well known that Chinese wheat contributed to Japanese landraces, and Japanese landraces contributed to modern cultivars from Japan. However, common Japanese and Chinese wheats differ in the frequencies of Glu‐D1ƒ allele. These results may be explained either by the founder effect or by a selective bottleneck in Japanese common wheat genetic resources.     

Comparing Small‐Scale Testing Methods for Predicting Wheat Gluten Strength Across Environments

Gluten strength is the main factor determining the rheological and processing properties of wheat. Rapid, small‐scale tests that can indirectly predict gluten strength are extremely important for wheat‐breeding selection, particularly when using pedigree methodology. The efficiency and reliability of three small‐scale tests (SDS sedimentation volume [SDSS], swelling index of glutenin [SIG], and lactic acid retention capacity [LARC]) across three environments (E1, no stress; E2, drought stress; and E3, heat stress) were evaluated by using 15 common wheat and nine durum wheat cultivars. In the case of common wheat, SIG highlighted its advantage for predicting gluten strength, even under stress environments, compared with LARC and SDSS, whereas SDSS showed the best relationship with bread loaf volume. For durum wheat, SIG showed the best predicting value in E1 and E3; however, under drought stress, SDSS, SIG, and LARC all lost their good ability for predicting gluten strength in durum wheat, which needs further investigation. Also, the comparison between two mixograph parameters (mixograph peak time and mixograph peak integral) for predicting gluten strength and the suitability of testing SIG and LARC with whole meal (or semolina) instead of refined flour were also investigated.     

Isolation and Characterization of High‐Molecular‐Weight (HMW) Gliadins from Wheat Flour

The aim of this study was to isolate high‐molecular‐weight (HMW) gliadins from wheat flour and to characterize the protein components that contribute to HMW gliadins. Wheat flour Akteur was extracted with a modified Osborne procedure, and the fraction soluble in 60% ethanol (total gliadins) was separated by gel‐permeation HPLC, yielding three fractions, GP1–GP3. GP1 (21.5%) consisted of oligomeric HMW gliadins, GP2 (15.2%) of ω5‐gliadins, and GP3 (63.3%) of ω1,2‐, α‐, and γ‐gliadins. Two‐dimensional SDS‐PAGE of HMW gliadins showed that interchain disulfide bonds were present in HMW gliadins. The molecular mass distribution of HMW gliadins determined by gel‐permeation HPLC was in a range from 66,000 to 680,000 with an average degree of polymerization of 13. Reduced HMW gliadins were further separated by preparative reversed‐phase HPLC into four subfractions (RP1, RP2, RP3, and RP4), which were characterized by SDS‐PAGE and semiquantitative N‐terminal sequencing. HMW gliadins of the wheat flour Akteur contained all types of gluten proteins: 48% low‐molecular‐weight glutenin subunits, 18% γ‐gliadins, 13% α‐gliadins, 9% ω1,2‐gliadins, 8% HMW glutenin subunits, and 4% ω5‐gliadins. We postulate that the existence of HMW gliadins can be explained by the presence of terminators, which interrupt the polymerization of glutenin subunits during biosynthesis and lead to polymers of limited size (oligomers) that are still soluble in aqueous ethanol.     

Expression,Purification, and Functional Analysis of Three Low‐Molecular‐Weight Glutenin Subunits from Wheat Cultivar Cheyenne

Glutenins, which form the network of gluten protein, are of great importance for the quality of flour products. Glutenins can be divided into HMW and LMW subunits according to molecular weight. Three genes for LMW glutenin subunits (LMW‐GS), named lmw‐cnd1, lmw‐cnd2, and lmw‐cnd3 with open reading frames of 1,053, 903, and 969 bp, respectively, were cloned from wheat cultivar Cheyenne. Heterologous expression vectors of the three LMW‐GS were constructed, and the recombinant proteins LMW‐CND1, LMW‐CND2, and LMW‐CND3 were overexpressed in Escherichia coli. After cell disruption with ultrasound, target proteins of high purity were obtained by using Ni²⁺ affinity chromatography. Farinograph and TAPlus measurements were used to investigate the effects of the three LMW‐GS on the characteristics of flour and dough. The results showed that the addition of each LMW‐GS can lead to an increase in the elasticity of the dough. Moreover, LMW‐CND2 and LMW‐CND3 promoted the strength of the dough. All three LMW‐GS caused a decrease of hardness and increase of springiness and cohesiveness of dough according to texture profiling results. Consequently, all three LMW‐GS have positive effects on the processing characteristics of dough and can improve bread quality to different extents.     

Influence of Low Molecular Weight Glutenins on Viscoelastic Properties of Intact Wheat Kernels and Their Relation to Functional Properties of Wheat Dough

The mechanical and viscoelastic properties of intact wheat kernels of 36 wheat cultivars differing in low molecular weight glutenin subunit (LMW‐GS) composition (loci Glu‐A3, Glu‐B3, and Glu‐D3) were evaluated using load‐compression tests. Comparison among genotypic groups representing Glu‐3 allelic variants showed that groups representing the alleles Glu‐A3 b, c, and d; Glu‐B3 d, g, and h; and Glu‐D3 a, b, and d, had harder kernel texture, higher kernel elastic work and larger gluten strength‐related parameters than those possessing alleles Glu‐A3 e; Glu‐B3 f, i and j (translocation 1B/1R); and Glu‐D3 d. Modulus of elasticity (stress to strain ratio) showed low values (111.9–168.8 MPa) for allelic groups possessing poor elastic properties (Glu‐A3 e; Glu‐B3 f, i, and j; and Glu‐D3 d), and high values (179.8–222.6 MPa) for allelic groups possessing high kernel elastic properties (Glu‐A3 b c, and d; Glu‐B3 d, g, and h; and Glu‐D3 a, b and c). The highest values for gluten strength‐related parameters (SDS‐sedimentation, dough mixing time, and dough strength [W]) corresponded to allelic groups Glu‐A3 d; Glu‐B3 d and g; and Glu‐D3 d, while the lowest corresponded to Glu‐A3 e and Glu‐B3 j. No significant differences were observed among groups with regard to gluten extensibility parameters; however, the highest P/L value (least extensibility) corresponded to Glu‐B3 j, which indicates presence of 1B/1R translocation. Except for the Glu‐B3 j (translocation 1B/1R) allele, which presented more variation within samples, a general relationship between kernel viscoelastic properties and dough viscoelastic properties was observed; samples showing higher elastic work to plastic work ratio (E/P) tended to possess better gluten strength than cultivars with low E/P ratio.     

Protein and Quality Characterization of Complete and Partial Near‐Isogenic Lines of Waxy Wheat

The objective of this study was to evaluate protein composition and its effects on flour quality and physical dough test parameters using waxy wheat near‐isogenic lines. Partial waxy (single and double nulls) and waxy (null at all three waxy loci, Wx‐A1, Wx‐B1, and Wx‐D1) lines of N11 set (bread wheat) and Svevo (durum) were investigated. For protein composition, waxy wheats in this study had relatively lower albumins‐globulins than the hard winter wheat control. In the bread wheats (N11), dough strength as measured by mixograph peak dough development time (MDDT) (r = 0.75) and maximum resistance (R_max) (r = 0.70) was significantly correlated with unextractable polymeric protein (UPP), whereas in durum wheats, moderate correlation was observed (r = 0.73 and 0.59, respectively). This may be due to the presence of high molecular weight glutenin subunits (HMW‐GS) Dx2+Dy12 at the Glu‐D1 locus instead of Dx5+Dy10, which are associated with dough strength. Significant correlation of initial loaf volume (ILV) to flour polymeric protein (FPP) (r = 0.75) and flour protein (FP) (r = 0.63) was found in bread wheats, whereas in durum wheats, a weak correlation of ILV was observed with FP (r = 0.09) and FPP (r =0.51). Significant correlation of ILV with FPP in bread wheats and with % polymeric protein (PPP) (r = 0.75) in durum lines indicates that this aspect of end‐use functionality is influenced by FPP and PPP, respectively, in these waxy wheat lines. High ILV was observed with 100% waxy wheat flour alone and was not affected by 50% blending with bread wheat flour. However, dark color and poor crumb structure was observed with 100% waxy flour, which was unacceptable to consumers. As the amylopectin content of the starch increases, loaf expansion increases but the crumb structure becomes increasingly unstable and collapses.     

Effects of High and Low Molecular Weight Glutenin Subunits on Rheological Dough Properties and Breadmaking Quality of Wheat

High molecular weight (HMW) or low molecular weight (LMW) subunits of different chemical state (reduced, reoxidized with KBrO₃, or KIO₃) or gliadins were added in 1% amounts to a base flour of the wheat cultivar Rektor and mixed with water. The corresponding doughs were then characterized by microscale extension tests and by microbaking tests and were compared to doughs from the base flour without additives. The maximum resistance of dough was strongly increased by HMW subunits in a reduced state and by HMW subunits reoxidized with KBrO₃. A moderate increase of resistance was caused by HMW subunits reoxidized with KIO₃ and by LMW subunits reoxidized with KBrO₃ or KIO₃. This resistance was strongly lowered by LMW subunits in a reduced state and by gliadins. The extensibility of dough was significantly increased only by gliadins and reduced HMW subunits; HMW subunits reoxidized with KBrO₃ had no effect, and all other fractions had a decreasing effect. In particular, glutenin subunits reoxidized with KIO₃ induced marked decrease of extensibility, resulting in bell‐shaped curve extensigrams, which are typical for plastic properties. The effect of reoxidized mixtures (2:1) of HMW and LMW subunits on maximum resistance depended on the oxidizing agent and on the conditions (reoxidation separated or together); extensibility was generally decreased. Bread volume was increased by addition of HMW subunits (reduced or reoxidized with KBrO₃) and decreased by LMW subunits (reoxidized with KBrO₃ or KIO₃) and by a HMW‐LMW subunit mixture (reoxidized with KBrO₃). The volume was strongly decreased by addition of reduced LMW subunits. A high bread volume was related to higher values for both resistance and extensibility.