Influence of variation in high-molecular-weight glutenin subunits composition at Glu-B1 and Glu-D1 loci on gluten microstructure and northern Chinese steamed bread quality

doi:10.1016/j.jia.2025.12.020

Advanced Online Publication | Current Issue | Archive | Adv Search

Ke Ding^{1, 2*}, Weihong Huang^1*, Kehui Sun^1*, Huali Tang¹, Xi Li¹, Surong Wang¹, Hao Peng¹, Zhiyang Han, Xiaohui Li^2#,Yueming Yan^2#, Ke Wang^1#

¹State Key Laboratory of Crop Gene Resources and Breeding/National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China

²College of Life Science, Capital Normal University, Beijing 100048, China

Highlights

1. Near-isogenic and chromosome substitution lines with uniform genetic backgrounds were employed to systematically reveal the effects of gluten proteins on gluten structure, flour properties, and northern Chinese steamed bread (NCSB) quality.

2. A comprehensive evaluation of wheat processing quality was achieved by integrating multiple analytical techniques, highlighting the advantages of confocal laser scanning microscopy (CLSM) and laser confocal microscopy Raman (LCM-Raman).

3. Thermogravimetric analysis (TGA) revealed weight loss patterns at 150°C and 600°C that closely correlated with steamed bread quality and overall bread quality, respectively.

4. Superior steamed bread quality is not necessarily dependent on higher gluten strength.

Abstract
References

Download: PDF in ScienceDirect
Export: BibTeX | EndNote (RIS)

摘要

高分子量谷蛋白亚基（HMW-GS）对小麦加工品质具有关键影响，但其对中国北方馒头（NCSB）品质的贡献机制尚未系统阐明。本研究利用Glu-B1和Glu-D1位点不同HMW-GS组成的近等基因系和染色体代换系，通过多维度分析手段全面探究谷蛋白组成对其结构特性、面粉品质及馒头品质的影响。采用不可提取性聚合蛋白含量（% UPP）、二硫键含量、热重分析（TGA）、快速黏度分析（RVA）、面筋聚集特性以及馒头蒸制实验评估面筋结构与功能特性。激光共聚焦扫描显微镜（CLSM）与激光共聚焦拉曼光谱（LCM-Raman）分析表明，CB037B和SL/B显著增加了面筋蛋白网络交联点数量、蛋白面积及β-折叠含量，同时降低了孔隙率和α-螺旋含量。馒头制作实验结合面团流变学特性证实，虽然CB037B属于强筋小麦，但其制备的馒头品质却低于CB037A，印证了优质馒头并非必然依赖于更强的面筋强度的观点。热重分析进一步揭示，150°C时的失重规律（CB037A>CB037B>SL/B>CS）与馒头品质实验结果一致，而600°C的失重情况与面包品质相关联。本研究系统揭示了HMW-GS在小麦中的结构-功能关系，为预测馒头与面包品质提供了新的评价指标，为我国北方小麦加工品质改良的育种实践提供了理论依据。

Abstract

High-molecular-weight glutenin subunits (HMW-GS) play a key role in determining wheat processing quality, but their contribution to NCSB quality has not been systematically elucidated. In this study, near-isogenic lines and chromosome substitution lines differing in HMW-GS compositions at the Glu-B1 and Glu-D1 loci were utilized to comprehensively assess the impact of gluten proteins on gluten microstructure, flour functionality, and NCSB quality through multiple analytical methods. Unextractable polymeric protein content (%UPP), disulfide bond content, TGA, rapid viscosity analyzer (RVA) profiles, gluten aggregation behavior, and steamed bread-making trials were used to evaluate gluten structure and functional performance. CLSM and LCM-Raman analyses demonstrated that CB037B and SL/B significantly increased junction density, protein network area, and β-sheet content, while reducing lacunarity and α-helix content. Steamed bread-making trials combined with dough rheological assessments revealed that although CB037B exhibited strong-gluten characteristics, the resulting steamed bread quality was lower than that of CB037A, supporting the concept that superior NCSB quality does not necessarily depend on higher gluten strength. TGA further indicated that the weight loss pattern at 150°C (CB037A>CB037B>SL/B>CS) aligned with steamed bread quality rankings, whereas weight loss at 600°C corresponded more closely with conventional bread quality. Overall, this work provides valuable insights into the structure–function relationships of HMW-GS in wheat and introduces novel indicators for predicting both steamed bread and bread quality, offering guidance for breeding programs aimed at improving wheat processing performance in northern China.

Keywords: HMW-GS gluten molecular structure thermal stability Confocal laser scanning microscopy LCM-Raman steamed bread quality

Online: 08 December 2025

Fund:

This work was supported by the National Natural Science Foundation of China (32472182) and the Science and Technology Department of Ningxia, China (2022BBF02039).

About author: #Correspondence Ke Wang, Tel/Fax: 86-10-82105173, E-mail: wangke03@caas.cn; Yueming Yan, Tel/Fax: 86-10-68902777, E-mail: yanym@cnu.edu.cn; Xiaohui Li, Tel/Fax: 86-10-68901842, E-mail: lixiaohui@cnu.edu.cn. *These authors contributed equally to this article

	Service
	E-mail this article
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

Ke Ding, Weihong Huang, Kehui Sun, Huali Tang, Xi Li, Surong Wang, Hao Peng, Zhiyang Han, Xiaohui Li#, Yueming Yan, Ke Wang. 2025. Influence of variation in high-molecular-weight glutenin subunits composition at Glu-B1 and Glu-D1 loci on gluten microstructure and northern Chinese steamed bread quality. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2025.12.020

Barak S, Mudgil D, Khatkar B S. 2015. Biochemical and functional properties of wheat gliadins: A review. Critical Reviews in Food Science and Nutrition, 55, 357-368.

Bekes F, Cavanagh C R, Martinov S, Bushuk S, Wrigley C W F. 2008. Part II. Composition table for the HMW subunits of glutenin. In: The Gluten Composition of Wheat Varieties and Genotypes. American Association of Cereal Chemists International, St. Paul, MN, USA.

Brönneke V, Zimmermann G, Killermann B. 2000. Effect of high molecular weight glutenins and D-zone gliadins on bread-making quality in German wheat varieties. Cereal Research Communications, 28, 187–194.

Dong L, Li N, Lu X, Prodanović S, Xu Y, Zhang W, Yan Y. 2016. Quality properties and expression profiling of protein disulfide isomerase genes during grain development of three spring wheat near isogenic lines. Genetika, 48, 249–269.

Gadaleta A, Blechl A E, Nguyen S, Cardone M F, Ventura M, Quick J S, Blanco A. 2008. Stably expressed D-genome-derived HMW glutenin subunit genes transformed into different durum wheat genotypes change dough mixing properties. Molecular Breeding, 22, 267–279.

Gao X, Liu T, Ding M, Wang J, Li C, Wang Z, Li X. 2018. Effects of HMW-GS Ax1 or Dx2 absence on the glutenin polymerization and gluten micro structure of wheat (Triticum aestivum L.). Food Chemistry, 240, 626-633.

Gao X, Tong J, Guo L, Yu L, Li S, Yang B, Cao X. 2020. Influence of gluten and starch granules interactions on dough mixing properties in wheat (Triticum aestivum L.). Food Hydrocolloids, 106, 105885.

Goel S, Yadav M, Singh K, Jaat R S, Singh N K. 2018. Exploring diverse wheat germplasm for novel alleles in HMW-GS for bread quality improvement. Journal of Food Science and Technology, 55, 3257-3262.

Hong T, Tan Z, Xu D, Jin Y, Wu F, Huang G, Zhong X, Zhang J, Xu X. 2025. Influence of zein on viscoelastic properties and gluten network development during dough formation. Food Chemistry: X, 27, 102437.

Hu J, Zhang J, Wang W, Liu J, Tang H, Wan Y, Wang K. 2025. Effects of a dramatic composition change of high-molecular-weight glutenin subunits by gene editing on flour-processing quality in wheat. Journal of Integrative Agriculture, (DOI: 10.1016/j.jia.2025.05.017).

Humphris A D, McMaster T J, Miles M J, Gilbert S M, Shewry P R, Tatham A S. 2000. Atomic force microscopy (AFM) study of interactions of HMW-GS subunits of wheat glutenin. Cereal Chemistry, 77, 107-110.

Jiang P, Xue J, Duan L, Gu Y, Mu J, Han S, Li X. 2019. Effects of high-molecular-weight glutenin subunit combination in common wheat on the quality of crumb structure. Journal of the Science of Food and Agriculture, 99, 1501-1508.

Jin H, Zhang Y, Li G, Mu P, Fan Z, He Z. 2013. Effects of allelic variation of HMW-GS and LMW-GS on mixograph properties and Chinese noodle and steamed bread qualities in a set of Aroona near-isogenic wheat lines. Journal of Cereal Science, 57, 146-152.

Kolster P, Krechting C F, Van Gelder W M J. 1988. Variation in high molecular weight glutenin subunits of Triticum aestivum and T. turgidum ssp. dicoccoides. Euphytica, 39, 141-145.

Kłosok K, Welc R, Fornal E, Nawrocka A. 2021. Effects of physical and chemical factors on the structure of gluten, gliadins and glutenins as studied with spectroscopic methods. Molecules, 26, 508.

Li M, Deng X, Xu X, Liu N, Wang Z, Yan Y. 2019. Effects of water deficit and different nitrogen fertilizer treatments on the quality of wheat for Chinese fresh white noodles and steamed bread and the composition of storage proteins. Journal of the Science of Food and Agriculture, 99, 6431-6443.

Li X, Liu T, Song L, Zhang H, Li L, Gao X. 2016. Influence of high-molecular-weight glutenin subunit composition at Glu-A1 and Glu-D1 loci on secondary and micro structures of gluten in wheat (Triticum aestivum L.). Food Chemistry, 213, 728-734.

Liu J, Zhang X, Li M, Wen L, Jiang W, Zhang Y, Gao D. 2021. Quality analysis of Yangmai series wheat varieties for Biscuit-Making. Journal of Triticeae Crops, 41, 50-60. (in Chinese)

Miles M J, Carr H J, McMaster T C, I'Anson K J, Belton P S, Morris V J, Tatham A S. 1991. Scanning tunneling microscopy of a wheat seed storage protein reveals details of an unusual supersecondary structure. Proceedings of the National Academy of Sciences of the United States of America, 88, 68-71.

Ma, F., Baik, B. K. (2017). Qualitative effect of added gluten on dough properties and quality of Chinese steamed bread. Cereal Chemistry, 94, 827-833.

Naeem H A, Paulon D, Irmak S, MacRitchie F. 2012. Developmental and environmental effects on the assembly of glutenin polymers and the impact on grain quality of wheat. Journal of Cereal Science, 56, 51-57.

Nawrocka A, Krekora M, Niewiadomski Z, Szymańska-Chargot M, Krawęcka A, Sobota A, Miś A. 2020. Effect of moisturizing pre-treatment of dietary fibre preparations on formation of gluten network during model dough mixing–a study with application of FT-IR and FT-Raman spectroscopy. Food Science and Technology, 121, 108959.

Nawrocka A, Szymańska-Chargot M, Miś A, Wilczewska A Z, Markiewicz K H. 2017. Aggregation of gluten proteins in model dough after fibre polysaccharide addition. Food Chemistry, 231, 51-60.

Payne P I, Holt L M, Krattiger A F, Carrillo J M. 1988. Relationships between seed quality characteristics and HMW glutenin subunit composition determined using wheats grown in Spain. Journal of Cereal Science, 7, 229-235.

Popineau Y, Cornec M, Lefebvre J, Marchylo B. 1994. Influence of high Mr glutenin subunits on glutenin polymers and rheological properties of glutens and gluten subfractions of near-isogenic lines of wheat Sicco. Journal of Cereal Science, 19, 231-241.

Roman-Gutierrez A D, Guilbert S, Cuq B. 2002. Description of microstructural changes in wheat flour and flour components during hydration by using environmental scanning electron microscopy. Food Science and Technology, 35, 730-740.

Shewry P, Gilbert S, Savage A, Tatham A, Wan Y F, Belton P, Halford N. 2003. Sequence and properties of HMW subunit 1Bx20 from pasta wheat (Triticum durum) which is associated with poor end use properties. Theoretical and Applied Genetics, 106, 744-750.

Sivam A S, Sun-Waterhouse D, Perera C O, Waterhouse G I N. 2012. Exploring the interactions between blackcurrant polyphenols, pectin and wheat biopolymers in model breads: A FTIR and HPLC investigation. Food Chemistry, 131, 802-810.

Wang H, Wu J, Kan J. 2016. Comparison of structure characteristics of gliadin and glutenin in highland barley and wheat. Food Science, 3, 43-48.

Wang P, Xu L, Nikoo M, Ocen D, Wu F, Yang N, Xu X. 2014. Effect of frozen storage on the conformational, thermal and microscopic properties of gluten: Comparative studies on gluten-, glutenin- and gliadin-rich fractions. Food Hydrocolloids, 35, 238-246.

Wang P, Xu X. 2015. Physicochemical alterations of wheat gluten proteins upon dough formation and frozen storage – a review from gluten, glutenin and gliadin perspectives. Trends in Food Science and Technology, 46, 189-198.

Wang S, Yu Z, Cao M, Shen X, Li N, Li X, Yan Y. 2013. Molecular mechanisms of HMW glutenin subunits from 1S^l genome of Aegilops longissima positively affecting wheat breadmaking quality. PLoS ONE, 8, e58947.

Wieser H. 2007. Chemistry of gluten proteins. Food Microbiology, 24, 115-119.

Wu J, Lu X, Yu Z, Han C, Li X, Prodanović S, Yan Y. 2017. Effects of Glu-1 and Glu-3 allelic variations on wheat glutenin macropolymer (GMP) content as revealed by size-exclusion high performance liquid chromatography (SE-HPLC). Genetika, 49, 677-691.

Yang T, Ge J, Wang P, Zhong Y, Zhou Q, Wang X, Cao W. 2023. Effect of high-molecular weight glutenin subunits (HMW-GS) on gluten polymerization during biscuit making: Insights from experimental and molecular dynamics simulation study. Journal of Agricultural and Food Chemistry, 71, 8150-8163.

Yang T, Wang P, Zhou Q, Jiang D, Jiang H. 2025. Effect of high-molecular-weight glutenin subunit deletion on gluten functionality and Chinese southern-type steamed bread quality. Food Chemistry, 464, 141664.

Zhang J, Luo F, Sun H, Wang J, Duan W, Yan Y. 2023. Wheat Glu-A1a encoded 1Ax1 subunit enhances gluten physicochemical properties and molecular structures that confer superior breadmaking quality. International Journal of Biological Macromolecules, 225, 701-714.

Zhang J, Ma D Y, Zhang Y P, Ai Z L. 2015. The effect of HMW-GS of wheat on the quality of flour and steamed bun. Food and Fermentation Industries, 41, 74-79. (in Chinese)

Zhang P, He Z, Chen D, Zhang Y, Larroque O R, Xia X. 2007. Contribution of common wheat protein fractions to dough properties and quality of northern-style Chinese steamed bread. Journal of Cereal Science, 46, 1-10.

Zhang P, Jondiko T O, Tilley M, Awika J M. 2014. Effect of high molecular weight glutenin subunit composition in common wheat on dough properties and steamed bread quality. Journal of the Science of Food and Agriculture, 94, 2801-2806.

Zhang X, Li J, Zhao J, Mu M, Jia F, Wang Q, Wang J. 2021. Aggregative and structural properties of wheat gluten induced by pectin. Journal of Cereal Science, 100, 103247.

Zhang X, Zhang B Q, Wu H Y, Lu C B, Lü G F, Liu D T, Gao D R. 2018. Effect of high-molecular-weight glutenin subunit deletion on soft wheat quality properties and sugar-snap cookie quality estimated through near-isogenic lines. Journal of Integrative Agriculture, 17, 1066-1073.

Zhang Y, Hong T, Yu W, Yang N, Jin Z, Xu X. 2020. Structural, thermal and rheological properties of gluten dough: Comparative changes by dextran, weak acidification and their combination. Food Chemistry, 330, 127154.

Zhang Z, Jiang X, Wang Z, Zhu Q, Liu M, Ru Z. 2019. Genetic diversity of HMW-GS and its relationship with quality of wheat germplasms from different regions. Acta Agriculturae Boreali-Sinica, 34, 75-81. (in Chinese).

Zhao J, Wang T, Zhao J, Qiao L, Wu B, Hao Y, Zheng X. 2024. Effects of high-molecular-weight glutenin subunit on hard-steamed bread quality. Frontiers in Genetics, 15, 1473518.

Zhao J, Zheng X, Qiao L, Ge C, Wu B, Zhang S, Zheng J. 2020. Effects of HMW-GS on quality related traits in wheat (Triticum aestivum L.) under different water regimes. PLoS ONE, 15, e0237711.

[1]	WANG Yan, GUO Zhen-ru, CHEN Qing, LI Yang, ZHAO Kan, WAN Yong-fang, Malcolm J. HAWKESFORD, JIANG Yun-feng, KONG Li, PU Zhi-en, DENG Mei, JIANG Qian-tao, LAN Xiu-jin, WANG Ji-rui, CHEN Guo-yue, MA Jian, ZHENG You-liang, WEI Yu-ming, QI Peng-fei. Effect of high-molecular-weight glutenin subunit Dy10 on wheat dough properties and end-use quality[J]. >Journal of Integrative Agriculture, 2023, 22(6): 1609-1617.
[2]	JIANG Yun, WANG De-li, HAO Ming, ZHANG Jie, LIU Deng-cai. *Development and characterization of wheat–Aegilops* kotschyi 1U^k(1A) substitution line with positive dough quality parameters** [J]. >Journal of Integrative Agriculture, 2023, 22(4): 999-1008.
[3]	LIU Da-tong, ZHANG Xiao, JIANG Wei, LI Man, WU Xu-jiang, GAO De-rong, BIE Tong-de, LU Cheng-bin. *Influence of high-molecular-weight glutenin subunit deletions at the Glu-A1* and Glu-D1 loci on protein body development, protein components and dough properties of wheat (Triticum aestivum L.)**[J]. >Journal of Integrative Agriculture, 2022, 21(7): 1867-1876.
[4]	ZHANG Xiao, ZHANG Bo-qiao, WU Hong-ya, LU Cheng-bin, Lü Guo-feng, LIU Da-tong, LI Man,. Effect of high-molecular-weight glutenin subunit deletion on soft wheat quality properties and sugar-snap cookie quality estimated through near-isogenic lines[J]. >Journal of Integrative Agriculture, 2018, 17(05): 1066-1073.
[5]	ZHAO Pei, WANG Ke, LIN Zhi-shan, LIU Hui-yun, LI Xin, DU Li-pu, YAN Yue-ming, YE Xing-guo. A genetic evidence of chromosomal fragment from bridge parent existing in substitution lines between two common wheat varieties[J]. >Journal of Integrative Agriculture, 2016, 15(1): 10-17.

No Suggested Reading articles found!

Viewed

Full text

Abstract

Cited

Shared

Discussed

Cite this article:

About JIA

Editorial board

For authors