High Molecular Weight Glutenin Subunits Alleles Research Articles

Molecular Characterization and Marker Development of the HMW-GS Gene from Thinopyrum elongatum for Improving Wheat Quality

The quality of wheat primarily depends on its storage protein quality, especially in regards to gluten content and high-molecular-weight glutenin subunits (HMW-GS). The number of HMW-GS alleles is limited in bread wheat (Triticum aestivum L.), whereas it is abundant in wheat relatives. Therefore, HMW-GS alleles from wheat relatives could provide a potential for improving quality in wheat breeding. Thinopyrum elongatum (EE) is one of the relatives of wheat. The E genome is closely related to the ABD genome in wheat; therefore, Th. elongatum is often used as an excellent exogenous gene donor for wheat genetic improvement. In this study, the high-molecular glutenin subunit gene was cloned and sequenced from Th. elongatum. A specific molecular marker for identifying the Glu-1Ey subunit gene was developed and applied to detected wheat-Th. elongatum alien introgression lines. Quality analysis indicated that the substitution and addition lines containing Th. elongatum alleles significantly (p < 0.05) increased grain protein content by 3.76% to 5.11%, wet-gluten content by 6.55% to 8.73%, flour 8-MW by 0.25% to 6.35%, and bread volume value by 33.77 mL to 246.50 mL, in comparing it with Chinese Spring. The GMP content and lactic acid SRC showed significant positive correlations with flour processing quality and might be used as indicators for wheat quality. The results were expected to provide a novel route for improving processing quality in wheat quality breeding.

A Rapid, Reliable RP-UPLC Method for Large-Scale Analysis of Wheat HMW-GS Alleles.

High-molecular-weight glutenin subunits (HMW-GS) account for only 10% of total wheat storage proteins, but play an important role in the processing quality of wheat flour. Therefore, identifying HMW-GS alleles associated with good end-use quality provides important information for wheat breeders. To rapidly, accurately and reproducibly identify HMW-GS, we established an optimized reversed-phase ultra-performance liquid chromatography (RP-UPLC) method. Separation parameters were optimized using an ACQUITY UPLC Protein BEH C4 column and stepwise ACN gradient, and the separation patterns and retention times (RTs) of 22 subunits were comparatively analyzed in 16 standard wheat cultivars. All HMW-GS proteins were well separated within about 5.5 min, and all analyses were complete within 12 min. We distinguished the 16 subunits based on RT, although three subunits in 1Bx (1Bx7/1Bx7OE and 1Bx17) and three subunits in 1By (1By8*, 1By9 and 1By15) had overlapping RTs; these were differentiated by SDS-PAGE. To distinguish 1Bx7 and 1Bx7OE, which differ in protein abundance, RP-UPLC was combined with PCR analysis of DNA junction markers. The optimized method was successfully applied to determine HMW-GS alleles in a large collection of bread wheat germplasm (1787 lines). This protocol is an appropriate option for selecting lines harboring favorable HMW-GS alleles in wheat breeding.

Interaction of B-type starch with gluten skeleton improves wheat dough mixing properties by stabilizing gluten micro-structure

Some recent studies have revealed individual and the combined interactions of gluten and starch affecting dough mixing properties. However, the combined influence of high-molecular-weight glutenin subunits (HMW-GS) and starch on dough mixing and rheological properties requires elucidation. Thus four recombinant inbred lines, SS 1, SS 2, ZZ 1 and ZZ 2, were selected based on their HMW-GSs compositions. Compared to ZZ 1 and ZZ 2, both SS 1 and SS 2 carried superior HMW-GS alleles, and exhibited extended dough development and stability time, indicating their significant dough mixing characteristics. The gluten skeleton of the wheat lines SS 2 and ZZ 2 with higher B-type starch proportions exhibited fewer breakages along with the rise of dough temperature during mixing. Higher content of B-type starch strengthens interaction between starch and gluten skeleton at the dough heating stage, suggesting a specific range of B-type starch proportion can improve dough mixing characteristics.

High-throughput analysis of high-molecular weight glutenin subunits in 665 wheat genotypes using an optimized MALDI-TOF-MS method.

The online version contains supplementary material available at 10.1007/s13205-020-02637-z.

Characterization of high- and low-molecular-weight glutenin subunits from Chinese Xinjiang wheat landraces and historical varieties.

Landraces and historical varieties are necessary germplasms for genetic improvement of modern cereals. Allelic variations at the Glu-1 and Glu-3 loci in 300 common wheat landraces and 43 historical varieties from Xinjiang, China, were evaluated by Sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and allele-specific molecular markers. Among the materials investigated, three, nine, and seven alleles were identified from the Glu-A1, Glu-B1, and Glu-D1 loci, respectively, and a total of 26 high-molecular-weight glutenin subunit (HMW-GS) combinations were found, of which 18 combinations were identified in landraces and historical varieties. Allelic frequency of HMW-GS combinations null, 7 + 8, 2 + 12 was found to be the highest in both the landraces (63.3%) and historical varieties (39.5%). Besides, some distinctive HMW-GS alleles, such as the novel Glu-B1 allele 6.1* + 8.1* and Glu-D1 alleles 2.6 + 12, 2.1 + 10.1, and 5** + 10 were observed in Xinjiang wheat landraces. Among the Glu-A3 and Glu-B3 loci of landraces and historical varieties, a total of eight and nine alleles were found, respectively. At each locus, two novel alleles were identified. A total of 33 low-molecular-weight glutenin subunit (LMW-GS) combinations of Glu-A3 and Glu-B3 were identified, with 31 and 14 combinations occurring in landraces and historical varieties, respectively, but only 10 combinations shared by both of them. As Glu-D1, Glu-A3, and Glu-B3 have highest contribution to the end-use quality and processing properties as compared to Glu-A1, Glu-B1, and Glu-D3 locus, the novel or distinctive HMW-GS and LMW-GS alleles in these loci could potentially be utilized for the improvement in the quality of modern wheat.

SNP markers for early identification of high molecular weight glutenin subunits (HMW-GSs) in bread wheat.

A set of eight SNP markers was developed to facilitate the early selection of HMW-GS alleles in breeding programmes. In bread wheat (Triticum aestivum), the high molecular weight glutenin subunits (HMW-GSs) are the most important determinants of technological quality. Known to be very diverse, HMW-GSs are encoded by the tightly linked genes Glu-1-1 and Glu-1-2. Alleles that improve the quality of dough have been identified. Up to now, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of grain proteins is the most widely used for their identification. To facilitate the early selection of HMW-GS alleles in breeding programmes, we developed DNA-based molecular markers. For each accession of a core collection (n = 364 lines) representative of worldwide bread wheat diversity, HMW-GSs were characterized by both genotyping and SDS-PAGE. Based on electrophoresis, we observed at least 8, 22 and 9 different alleles at the Glu-A1, Glu-B1 and Glu-D1 loci, respectively, including new variants. We designed a set of 17 single-nucleotide polymorphism (SNP) markers that were representative of the most frequent SDS-PAGE alleles at each locus. At Glu-A1 and Glu-D1, two and three marker-based haplotypes, respectively, captured the diversity of the SDS-PAGE alleles rather well. Discrepancies were found mainly for the Glu-B1 locus. However, statistical tests revealed that two markers at each Glu-B1 gene and their corresponding haplotypes were more significantly associated with the rheological properties of the dough than were the relevant SDS-PAGE alleles. To conclude, this study demonstrates that the SNP markers developed provide additional information on HMW-GS diversity. Two markers at Glu-A1, four at Glu-B1 and two at Glu-D1 constitute a useful toolbox for breeding wheat to improve end-use value.

A wheat-Aegilops umbellulata addition line improves wheat agronomic traits and processing quality.

Wheat processing quality is mainly correlated with high-molecular-weight glutenin subunits (HMW-GS) of grain endosperm. In bread wheat, the number of HMW-GS alleles are limited. However, wheat relative species possess numerous HMW-GS genes. In our previous study, a pair of novel HMW-GS 1Ux3.5+1Uy1.9 was characterized in Aegilops umbellulata. In this work, a novel wheat-Ae. umbellulata addition line, GN05, carrying a pair of 1U chromosome was developed and identified via cytogenetic analysis. Protein composition analysis indicated that GN05 carried HMW-GS of Ae. umbellulata. Accumulation of glutenin macropolymer (GMP) showed that GN05 had a much higher GMP content than the recurrent parent Chinese Spring. Rheological characteristics were analyzed by mixing test and the dough quality of GN05 was significantly improved compared to Chinese Spring. The results presented here may provide a valuable resource for the improvement of bread wheat quality.

Rapid Development and Characterization of Chromosome Specific Translocation Line of Thinopyrum elongatum with Improved Dough Strength.

The protein content and its type are principal factors affecting wheat (Triticum aestivum) end product quality. Among the wheat proteins, glutenin proteins, especially, high molecular weight glutenin subunits (HMW-GS) are major determinants of processing quality. Wheat and its primary gene pool have limited variation in terms of HMW-GS alleles. Wild relatives of wheat are an important source of genetic variation. For improvement of wheat processing quality its wild relative Thinopyrum elongatum with significant potential was utilized. An attempt was made to replace Th. elongatum chromosome long arm (1EL) carrying HMW-GS genes related to high dough strength with chromosome 1AL of wheat with least or negative effect on dough strength while retaining the chromosomes 1DL and 1BL with a positive effect on bread making quality. To create chromosome specific translocation line [1EL(1AS)], double monosomic of chromosomes 1E and 1A were created and further crossed with different cultivars and homoeologous pairing suppressor mutant line PhI. The primary selection was based upon glutenin and gliadin protein profiles, followed by sequential genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH). These steps significantly reduced time, efforts, and economic cost in the generation of translocation line. In order to assess the effect of translocation on wheat quality, background recovery was carried out by backcrossing with recurrent parent for several generations and then selfing while selecting in each generation. Good recovery of parent background indicated the development of almost near isogenic line (NIL). Morphologically also translocation line was similar to recipient cultivar N61 that was further confirmed by seed storage protein profiles, RP-HPLC and scanning electron microscopy. The processing quality characteristics of translocation line (BC4F6) indicated significant improvement in the gluten performance index (GPI), dough mixing properties, dough strength, and extensibility. Our work aims to address the challenge of limited genetic diversity especially at chromosome 1A HMW-GS locus. We report successful development of chromosome 1A specific translocation line of Th. elongatum in wheat with improved dough strength.

Evaluation of HMW-GS 20 and 2.2 from near isogenic lines of wheat variety HD2329 for bread quality improvement.

Hexaploid wheat (Triticum aestivum L.) dominates the list of the most important human food sources ever. Its complex genetic background is the reason behind the wide diversity that exists in nutritional as well as food end-product quality. High-molecular-weight glutenin sub-units (HMW-GS) are the main grain storage proteins in the endosperm of wheat and related species. It is well established that the composition and quantity of allelic variation in HMW-GS genes substantially affect the taste and appearance of dough products and therefore work in this area is highly desired. A significant positive effect on wheat dough quality traits was observed among near isogenic lines of HMW-GS sub-units 20 and 2.2 in wheat variety HD2329 during quality evaluation of data generated over 2 years. A remarkably significant (P < 0.01) effect was observed on dough quality parameters like ratio of wet gluten/dry gluten, SDS sedimentation, farinogram parameters, and bread/chapatti traits whereas flour protein and dry gluten content showed an insignificant effect. HMW-GS 20 was found to be superior to HMW-GS 2.2 in terms of dough quality and both the near isogenic lines developed by us were found to be highly superior to the recurrent parent HD2329. As we know that the improvement of flour quality based on superior HMW-GS alleles is necessary to meet changing consumer demand, the study can be of immense use to future researchers who can target these HMW sub-units 20 and 2.2 in breeding programmes for the improvement of wheat end-product quality. © 2017 Society of Chemical Industry.

Characterization of a Triticum aestivum–Dasypyrum villosum T1VS·6BL translocation line and its effect on wheat quality

Bread wheat quality is mainly correlated with protein quality, particularly the glutenin content and high molecular weight glutenin subunits (HMW-GS) of grain endosperm. The number of HMW-GS alleles and loci are limited in bread wheat cultivars, though ideally, a large amount of HMW-GS alleles in wheat-related grasses should be exploited. In this study, a novel wheat-Dasypyrum villosum GP005 translocation line, NAU425, carrying a pair of T1VS·6BL translocation chromosomes was developed and assessed via molecular cytogenetic analysis. Grain quality analysis indicated that NAU425 has a positive effect on protein content, Zeleny sedimentation value, wet gluten content, and the rheological characteristics of wheat flour dough compared to the same qualities of recurrent parent ‘Chinese Spring’ attributed to the additional HMW-GS donated by 1VS of D. villosum. The protein content of T1VS·6BL was significantly improved compared to ‘Chinese Spring’ likely owing to the dramatic increase in glutenin content. Considering the importance of glutenin content for bread wheat end-use quality, and T1VS·6BL line with good plant vigor, full fertility and cytogenetic stability, NAU425 may be valuable in bread wheat quality improvement. Our results presented here may provide an approach to improve bread wheat quality through additional alien HMW-GS introgression.

Combination of null alleles with 7 + 9 allelic pair at Glu-B1 locus on the long arm of group 1 chromosome improves wheat dough functionality for tortillas

Deletion of one or more high molecular weight glutenin subunit (HMW-GS) alleles alters gluten functionality in ways that can be exploited to design wheat for specialty products. In this work, we investigated whether combining 7 + 9 allelic pair at Glu-B1 locus with deletion of other HMW-GS alleles at Glu-A1or Glu-D1 locus would produce ideal gluten functionality for tortillas. Flours from 17 wheat lines were evaluated for insoluble polymeric protein (IPP) content and gluten properties. Tortillas were produced by hot-press method and evaluated during 16 days storage. Compared to non-deletion lines, deletion line generally had lower IPP (362 vs 414 mg/g); produced more extensible dough (44.9 vs 33.4 mm), which required lower equilibrium force to compress (4.12 vs 7.55 N); and produced larger diameter tortillas (180 vs 165 mm). In general, deletion lines produced tortillas with poor flexibility (score <3.0) after 16 days. However, combination of 7 + 9 allelic pair at Glu-B1 locus with deletions at Glu-A1 or Glu-D1 consistently produced large diameter tortillas that retained good flexibility during 16 days of storage. The 7 + 9 allelic pair at Glu-B1 plays a crucial role in wheat gluten functionality essential for tortillas, and may be a good target for genetic improvement of wheat for flatbreads.

Identification and evaluation of disease resistance and HMW-GS composition of Aegilops geniculata Roth

Aegilops geniculata Roth is an important germplasm resource for the transfer of beneficial genes from alien species of wheat into common wheat (Triticum aestivum L.). In this study, we identified and evaluated agronomic characters, powdery mildew resistance, stripe rust resistance, and high molecular weight glutenin subunit (HMW-GS) compositions of 44 Ae.geniculata accessions. The average growth period (254 days) was longer than that of common wheat (240 days). Coefficients of variation (CVs) of all examined agronomic characters except growth period were >10 %. The largest CV, 23.80 %, was that of 1,000-grain weight. An assay for disease resistance revealed that 37 Ae.geniculata accessions (85.09 %) were resistant to powdery mildew, of which 33 were immune, 1 was highly resistant, and 3 were moderately resistant. This assay also indicated that 33 of the accessions (75 %) were resistant to stripe rust, with 25 showing immunity, 4 highly resistant, and 4 moderately resistant. Additionally, 19 accessions (43.18 %) were immune to both powdery mildew and stripe rust. HMW-GSs in the 44 accessions were analyzed by SDS polyacrylamide-gel electrophoresis, which indicated rich germplasm diversity. A total of 21 different HMW-GS alleles were identified at Glu-1 loci. The most frequent subunits were 1Dx2, 1Dx1.1, 1Dx2.2, 1Dx1.5, 1Bx20, 1Bx22, 1Dy10, and 1Dy12, found in 36.36, 54.55, 40.91, 34.09, 38.64, 25.00, 47.73, and 20.45 % of accessions, respectively. The data uncovered for the 44 accessions should facilitate their use as beneficial germplasm donors in wheat breeding.

Polymorphism of high M r glutenin subunits in wild emmer Triticum turgidum subsp. dicoccoides: chromatographic, electrophoretic separations and PCR analysis of their encoding genes

Triticum turgidum subsp. dicoccoides (Korn. ex Asch. et Graebn.) Thell. (AABB), the immediate progenitor of tetraploid and hexaploid wheats, is a species characterised by a wide range of protein polymorphism and by high protein content. Surveys on polymorphism and genetic control of the high molecular weight glutenin subunits (HMW-GS) present in this species, in two forms x- and y-type at the Glu-A1 and Glu-B1 loci, are still considered useful, both to improve technological properties of breeding varieties and to study the genome evolutionary process in wheats. Comparative Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoretic and Reversed Phase High Performance Liquid Chromatographic analyses (SDS-PAGE, RP-HPLC) of the HMW-GS present in several accessions of T. turgidum subsp. dicoccoides allowed the detection of new alleles of Glu-A1 and Glu-B1 loci, with x- and y-type glutenin subunits, apparently similar to those present in cultivated wheats in molecular weight, but different in surface hydrophobicity. In addition, changes in the number of x- and y-type subunits at the glutenin loci were also ascertained. The y-type subunits at the Glu-A1 locus, which are never expressed in cultivated bread and durum wheats, and single y-type expressed glutenin subunits at the Glu-B1 locus were also identified in several accessions. DNA extracted from samples, differing in number or type of HMW-GS and corresponding to x- and y-type genes at Glu-1 loci, were amplified using specific primers, two of which were constructed within the transposon-like sequence of Chinese Spring DNA and analysed by polymerase chain reaction. The results showed this insertion in some accessions of T. turgidum subsp. dicoccoides and also the presence of silent Ax, Bx and By type genes. The usefulness for breeding of these comparative analyses carried out on different HMW-GS alleles detected in Triticum turgidum subsp. dicoccoides, is discussed.

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.

Diversity of seed storage proteins in common wheat (Triticum aestivum L.)

The objective of our study was to determine the composition of high-molecular weight-glutenin subunits (HMW-GS) in 120 cultivars of common wheat (Triticum aestivum L.). Fourteen alleles and 34 allelic compositions were detected using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The most frequent HMW-GS alleles at the Glu-A1, Glu-B1 and Glu-D1 loci were null (57.1%), 7+9 (43.3%) and 5+10 (61.9%), respectively. However, low-frequency HMW-GS alleles were also observed, such as 13+16, 20, 21, 7 and 18, encoded by the Glu-B1 locus, and 4+12, encoded by the Glu-D1 locus. The wheat–rye 1BL.1RS translocation was identified in 25 cultivars, using acid polyacrylamide gel electrophoresis. The Glu-score varied greatly, and some lines reached the maximum value of 10.

Generation of high frequency of novel alleles of the high molecular weight glutenin in somatic hybridization between bread wheat and tall wheatgrass

Somatic hybridization between bread wheat and tall wheatgrass (Agropyron elongatum) has generated fertile introgression progenies with novel combinations of high molecular weight glutenin subunits (HMW-GS). Most of these novel HMW-GS alleles were stably inherited. Sixteen HMW-GS sequences were PCR amplified from three introgression progeny lines and sequenced. The alignment of these sequences indicated that five, probably derived from point mutations of the parental genes, whereas eight likely represent the product of replication slippage. Three Glu-1Ay sequences appear to have lost the transposon presented in the parental gene. Two subunits carry an additional cysteine residue, which may be favorable to the quality of end-use product. We demonstrate that novel HMW-GS alleles can be rapidly generated via asymmetric somatic hybridization.

Evaluation of genetic variation in high molecular weight glutenin subunits of seed storage protein using landraces of common wheat from Pakistan

SDS-PAGE was used to determine the variation in high molecular weight glutenin subunits (HMW-GS) alleles present in 170 landraces of common wheat from Sindh, Balochistan and NWFP in Pakistan. Three alleles for Glu-A1, six alleles for Glu-B1 and four alleles for Glu-D1 were detected. These included three new alleles, namely Glu-B1e, Glu-B1h and Glu-D1n, which had not previously been found in landraces of Pakistani wheat. Furthermore, Glu-D1m and Glu-D1n, which are considered to be endemic alleles in Pakistan and Afghanistan, were both detected from Balochistan. Twenty genotypes were identified based on combinations of alleles at the three Glu-1 loci. The present study showed that Pakistani landraces of common wheat have maintained a broad diversity of HMW-GS alleles, and that they may serve as genetic resources to improve wheat varieties adapted to Pakistani climatic and edaphic conditions.

Characterizing HMW-GS alleles of decaploid Agropyron elongatum in relation to evolution and wheat breeding

Bread wheat quality is mainly correlated with high molecular weight glutenin subunits (HMW-GS) of endosperm. The number of HMW-GS alleles with good processing quality is limited in bread wheat cultivars, while there are plenty of HMW-GS alleles in wheat-related grasses to exploit. We report here on the cloning and characterization of HMW-GS alleles from the decaploid Agropyron elongatum. Eleven novel HMW-GS alleles were cloned from the grass. Of them, five are x-type and six y-type glutenin subunit genes. Three alleles Aex4, Aey7, and Aey9 showed high similarity with another three alleles from the diploid Lophopyrum elongatum, which provided direct evidence for the Ee genome origination of A. elongatum. It was noted that C-terminal regions of three alleles of the y-type genes Aey8, Aey9, and Aey10 showed more similarity with x-type genes than with other y-type genes. This demonstrates that there is a kind of intermediate state that appeared in the divergence between x- and y-type genes in the HMW-GS evolution. One x-type subunit, Aex4, with an additional cysteine residue, was speculated to be correlated with the good processing quality of wheat introgression lines. Aey4 was deduced to be a chimeric gene from the recombination between another two genes. How the HMW-GS genes of A. elongatum may contribute to the improvement of wheat processing quality are discussed.

Genetic diversity of HMW glutenin subunit in Chinese common wheat (Triticum aestivum L.) landraces from Hubei province

The high molecular weight (HMW) glutenin subunit composition of 111 common landraces of bread wheat collected from Hubei province, China has been determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Ninety six of the accessions were homogeneous for HMW glutenin subunit composition and 15 were heterogeneous. For the Glu-1 loci, 16 alleles were detected, 3 at the Glu-A1locus, 9 at the Glu-B1and 4 at the Glu-D1. Three novel alleles were identified, two at the Glu-B1 and one at the Glu-D1locus. Combination of these 16 alleles resulted in 14 different HMW subunit patterns. The distribution of HMW glutenin subunit alleles in a subset of 105 of the 111 accessions representing six populations was assessed both at the individual population and whole population levels. The results demonstrated that the distribution of allelic patterns varied among populations. Taken together, 62.5% of the alleles detected were considered to be rare alleles while the Glu-A1c (null), Glu-B1b (1Bx7 + 1By8) and Glu-D1a (1Dx2 + 1Dy12) alleles were found most frequently in the six populations. The subset exhibited relatively high genetic diversity (A = 5.33, P = 1.00, Ae = 1.352 and He = 0.238) with 81.5% of the diversity being within populations and 18.5% between populations.

Protein Heterogeneity in European Wheat Landraces and Obsolete Cultivars: Additional Information II

The endosperm storage protein of 46 European wheat (Triticum aestivum L.) landraces and obsolete cultivars have been fractionated by SDS-PAGE to determine the composition of high molecular weight glutenin subunits (HMW-GS) composition. It has been discovered that about 46% of the wheats were heterogeneous, comprising 2–11 different glutenin profiles. Eighteen of them were observed to be homogeneous. A total of 13 HMW-GS alleles, including 3 at the Glu-A1, 8 at the Glu-B1, and 3 at the Glu-D1 loci were revealed. HMW-GS null controlled by locus Glu-A1, subunits 7 + 8 by Glu-B1, and 2 + 12 by Glu-D1 predominated. However low frequented alleles such as 17 + 18, 20, 6, and 7 were observed. Furthermore, other new alleles encoding HMW-GS at the locus Glu-B1 have been found in one of France cultivar (Saumur d’Automne). The glutenin-based quality score ranged from 4 to 10.