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The aim of this study was to clarify the effects of the high‐molecular‐weight glutenin subunits (HMW‐GSs) 1Dx3+1Dy12 (3+12) and 1Dx4+1Dy12 (4+12) at the Glu‐D1 locus on gluten and Chinese steamed bread (CSB) quality. The grain protein content and composition, gluten content and gluten index, farinograph properties, and CSB quality were investigated using four wheat near‐isogenic lines (NILs) carrying HMW‐GSs 1Dx2+1Dy12 (2+12), 3+12, 4+12 and 1Dx5+1Dy10 (5+10), respectively. The unextractable polymeric protein (UPP) and glutenin macropolymer (GMP) content, gluten index, dough development time, stability time, and farinograph quality number of four NILs all ranked as 5+10 > 3+12 > 2+12/4+12, such as the gluten index ranked as 5+10(44.88%) > 3+12(40.07%) > 2+12(37.46%)/4+12(35.85%); however, their contributions to the quality of CSB were ranked as 3+12 > 5+10 > 2+12/4+12, such as the specific volume ranked as 3+12(2.64 mL/g) > 5+10(2.49 mL/g) > 2+12(2.36 mL/g)/4+12(2.35 mL/g), which indicated that a suitable gluten strength (3+12) was crucial to making high‐quality CSB. In addition, subunits 4+12 had a similar quality performance to low‐quality subunits 2+12. All these findings suggested that, except for the acknowledged high‐quality subunits 5+10, the introduction of 3+12 at the Glu‐D1 locus is an efficient way for quality improvement of gluten as well as CSB. [ABSTRACT FROM AUTHOR]

Low-molecular-weight glutenin subunits (LMW-GS) account for 40% of the total wheat grain gluten protein fraction, which plays a significant role in the formation of noodle processing quality. The goal of this study was to clarify the effects of the major LMW-GS encoded by Glu-A3 on gluten and Chinese fresh noodle (CFN) quality. Four near-isogenic lines (NILs) were used as materials in this study, respectively carrying alleles Glu-A3a, Glu-A3b, Glu-A3c, and Glu-A3e, against the background of wheat variety Xiaoyan 22. The grain protein and its component contents and the gluten content, gluten index, farinograph properties, cooking quality, and textural quality of CFN were investigated. The results show that the ratios of glutenin to gliadin (Glu/Gli) in the NILs ranked them as Glu-A3b > Glu-A3c/Glu-A3a > Glu-A3e, and the unextractable polymeric protein content (UPP%), gluten index (GI), and farinograph quality in the NILs ranked them as Glu-A3b > Glu-A3c > Glu-A3a/Glu-A3e. Compared to Glu-A3b and Glu-A3a, the NILs carrying alleles Glu-A3c and Glu-A3e had better cooking and texture properties in CFN. All these findings suggest that the introduction of alleles Glu-A3c or Glu-A3e is an efficient method for quality improvement in CFN, which provides an excellent subunit selection for improving CFN quality. [ABSTRACT FROM AUTHOR]

Background: The composition of glutenin protein significantly affects protein–starch interactions and starch digestion characteristics in wheat dough matrices. To elucidate the effects of high molecular weight glutenin subunits at the Glu‐B1 locus on dough processing quality, the detailed structural changes of protein, starch, and their complexes were compared in Mixolab dough samples of two near isogenic lines 7 + 8 and 7 + 9. Results: The results showed that the degree of protein aggregation increased continuously during dough processing, as did the destruction and rearrangement of the gluten network. Compared to 7 + 8, the stronger and more stable protein network formed in 7 + 9 dough induced intensive interactions between protein and starch, primarily through hydrogen bonds and isomeric glycosidic bonds. In 7 + 9 dough, the more compact and extensive protein–starch network significantly inhibited starch gelatinization during dough pasting, while during the dough cooling stage [from C4 (82.8 °C) to C5 (52.8 °C)], more protein–starch complexes composed of monomeric proteins and short‐chain starch were generated, which remarkably inhibited starch retrogradation. All protein–starch interactions in the 7 + 9 dough improved the starch digestion resistance, as reflected by the high content of resistant starch. Conclusion: The more extensive and intensive protein–starch interactions in the 7 + 9 dough inhibited the gelatinization and enzymatic hydrolysis of starch, thereby producing more slowly digestible starch and resistant starch. These findings demonstrate the feasibility of optimizing the texture and digestibility of wheat‐based food products by regulating the behavior and interactions of proteins and starch during dough processing. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

AP2/ERF (APETALA2/ethylene responsive factor) is a family of plant-specific transcription factors whose members are widely involved in many biological processes, such as growth, development, and biotic and abiotic stress responses. Here, 20 AP2/ERF genes were identified based on wheat RNA-seq data before and after drought stress, and classified as AP2, ERF, DREB, and RAV. The analysis of gene structure revealed that about 85% of AP2/ERF family members had lost introns, which are presumed to have been lost during the formation and evolution of the wheat genome. The expression of 20 AP2/ERF family genes could be verified by qRT-PCR, which further supported the validity of the RNA-seq data. Subsequently, subcellular localization and transcriptional activity experiments showed that the ERF proteins were mainly located in the nucleus and were self-activating, which further supports their functions as transcription factors. Furthermore, we isolated a novel ERF gene induced by drought, salt, and cold stresses and named it TaERF-6-3A. TaERF-6-3A overexpression increased sensitivity to drought and salt stresses in Arabidopsis, which was supported by physiological and biochemical indices. Moreover, the expression of stress- and antioxidant-related genes was downregulated in TaERF-6-3A-overexpressing plants. Overall, these results contribute to the further understanding of the TaERF-6-3A gene function in wheat. [ABSTRACT FROM AUTHOR]

Drought is the main abiotic stress factor limiting the growth and yield of wheat (Triticum aestivum L.). Therefore, improving wheat tolerance to drought stress is essential for maintaining yield. Previous studies have reported on the important role of TaNRX1 in conferring drought stress tolerance. Therefore, to elucidate the regulation mechanism by which TaNRX1 confers drought resistance in wheat, we generated TaNRX1 overexpression (OE) and RNA interference (RNAi) wheat lines. The results showed that the tolerance of the OE lines to drought stress were significantly enhanced. The survival rate, leaf chlorophyll, proline, soluble sugar content, and activities of the antioxidant enzymes (catalase, superoxide dismutase, and peroxidase) of the OE lines were higher than those of the wild type (WT); however, the relative electrical conductivity and malondialdehyde, hydrogen peroxide, and superoxide anion levels of the OE lines were lower than those of the WT; the RNAi lines showed the opposite results. RNA-seq results showed that the common differentially expressed genes of TaNRX1 OE and RNAi lines, before and after drought stress, were mainly distributed in the plant–pathogen interaction, plant hormone signal transduction, phenylpropane biosynthesis, starch and sucrose metabolism, and carbon metabolism pathways and were related to the transcription factors, including WRKY, MYB, and bHLH families. This study suggests that TaNRX1 positively regulates drought stress tolerance in wheat. [ABSTRACT FROM AUTHOR]

Photoperiod (Ppd) genes play an important role in the adaptation of wheat to the ecological environment. However, the transcriptional regulation mechanism of photoperiodic genes has remained elusive. This study isolated a full-length promoter of Ppd-D1b (2518 bp) from the common wheat genome. Several essential core cis-acting elements and numerous light-responsive cis-acting regulatory elements were identified in Ppd-D1b promoter by the in-silico analysis. Ten 5'-deleted length fragments of the Ppd-D1b promoter fused with GUS were constructed and named D0 ~ D9, then transferred them into Arabidopsis thaliana. GUS gene driven by full-length (D0) in transgenic Arabidopsis thaliana showed the same rhythm with Ppd-D1b in wheat under short-day conditions (SDs, 8-h light/16-h dark). The expression of GUS gene in D0 reached its peak at 3 h after dawn, then decreased to the lowest and remained stable. Analysis of the series of 5'-deleted fragments showed that at 3 h after dawn, GUS gene expression activity decreased significantly in D7a due to removal of CHEBS (CCA1 HIKING EXPEDITION binding site). Moreover, yeast one-hybrid (Y1H) and dual-luciferase (dual-LUC) assays revealed that TaTCP20 could bind to the Ppd-D1b promoter to increase its transcriptional activity. This study revealed a transcription factor, TaTCP20, which activated Ppd-D1b by binding to CHEBS, provided a foundation for the theoretical research on wheat's photoperiodic response mechanism. [ABSTRACT FROM AUTHOR]

Flowering time is one of the most important agronomic traits in wheat production. A proper flowering time might contribute to the reduction or avoidance of biotic and abiotic stresses, adjust plant architecture, and affect the yield and quality of grain. In this study, TaTOE1-B1 in wheat produced three transcripts (TaTOE1-B1-1, TaTOE1-B1-2, and TaTOE1-B1-3) by alternative splicing. Compared to the longest transcript, TaTOE1-B1-1, TaTOE1-B1-3 has a deletion in the sixth exon (1219–1264 bp). Under long-day conditions, the heterologous overexpression of the TaTOE1-B1-3 gene delayed flowering, prolonged the vegetative growth time, and enlarged the vegetative body of Arabidopsis, but that of TaTOE1-B1-1 did not. As typical AP2 family members, TaTOE1-B1-1 and TaTOE1-B1-3 are mainly located in the nucleus and have transcriptional activation activities; the transcriptional activation region of TaTOE1-B1-3 is located in the C-terminal. In TaTOE1-B1-3 overexpression lines, the expression of flowering-related AtFT and AtSOC1 genes is significantly downregulated. In addition, this study confirms the protein–protein interaction between TaTOE1-B1-3 and TaPIFI, which may play an important role in flowering inhibition. These results provide a theoretical basis for the precise regulation of wheat flowering time. [ABSTRACT FROM AUTHOR]

The WRKY transcription factor family has been associated with a variety of plant biological processes, such as biotic and abiotic stress responses. In this study, 13 wheat TaWRKY DEGs in transcriptome data before and after drought stress, namely TaWRKY1 to TaWRKY8 , including various copies, were identified and classified as Group I, II, or III. TaWRKY1-2D overexpression enhanced drought tolerance in transgenic Arabidopsis. Moreover, the AtRD29A , AtP5CS1 , AtPOD1 , AtCAT1 , and AtSOD (Cu/Zn) genes, which are related to the stress response and antioxidant system, were significantly upregulated in TaWRKY1-2D transgenic Arabidopsis under drought stress. TaWRKY1-2 silencing in wheat increases the MDA content, reduces the contents of proline and chlorophyll and the activities of antioxidant enzymes, and inhibits the expression levels of antioxidant (TaPOD , TaCAT , and TaSOD (Fe))- and stress-related genes (TaP5CS) under drought stress. Yeast two-hybrid screening revealed TaDHN3 as an interaction partner of TaWRKY1-2D; their interaction was further confirmed using yeast two-hybrid and bimolecular fluorescence complementation. Furthermore, TaWRKY1-2D may play essential roles in wheat drought tolerance through posttranslational regulation of TaDHN3. Overall, these findings contribute to our knowledge of the WRKY family in wheat and identify TaWRKY1-2D as a promising candidate gene for improving wheat breeding to generate drought-tolerant wheat. [ABSTRACT FROM AUTHOR]

[Display omitted] • Gluten network goes through disassembly and repolymerization during processing. • β-sheets are positively associated with gluten polymerization and dough viscosity. • The stronger dough gluten inhibits starch pasting more significantly. • Covalent protein-starch interactions are found in strong-gluten dough. • Additional protein-starch complexes in dough matrix retard starch retrogradation. To clarify the detailed behaviors of protein, starch and interactions during complex dough processing, structural changes in dough protein and starch during continuous Mixolab processing were investigated using wheat near-isogenic lines carrying high-molecular-weight glutenin subunits 1Dx5 + 1Dy10 (5 + 10) or 1Dx2 + 1Dy12 (2 + 12) at the Glu-D1 locus. A more stable gluten network including disulfide bonds and hydrophobic interactions, was formed in the 5 + 10 dough before dough weakening at 53.5 °C, than in the 2 + 12 dough. Thereafter, thermo-mechanical treatment induced the depolymerization of gluten until starch gelatinization peak at 74.6 °C; however, from the peak to trough viscosity at 82.8 °C, additional monomeric proteins were incorporated into the repolymerized proteins characterized by increased disulfide bonds, hydrogen bonds, and β-sheets. Generally, the protein aggregates of 5 + 10 showed a higher degree of polymerization and better stability than those of 2 + 12 during dough processing, which significantly slowed starch gelatinization and recyclization. Moreover, stronger interactions between monomeric proteins and amylose/short-branch starch via glycosidic and hydrogen bonds were found in 5 + 10 dough during starch pasting and retrogradation. The findings demonstrate the feasibility of optimizing the texture and digestibility of wheat-based food products by regulating the behaviors and interactions of proteins and starch during dough processing. [ABSTRACT FROM AUTHOR]

• A total of 26 VDAC genes were identified and characterized in wheat (Triticum aestivum L). • TaVDAC1-B OE lines appeared tolerant to salt stress and sensitive to drought stress. • The interaction between TaVDAC1-B and TaNRX-D1 protein was identified. Voltage-dependent anion channels (VDACs) are major transport proteins localized in the outer membrane of mitochondria and play critical roles in regulating plant growth and responding to stress. In this study, a total of 26 VDAC genes in common wheat (Triticum aestivum L.) were identified. TaVDACs that contained β -barrel structures were classified into three groups with phylogenetic and sequence alignment. Additionally, the gene structure and protein conserved motif composition varied among diverse subfamilies but were relatively conserved within the same subfamily. The basic elements that were stress- and hormone-related, including TATA-box, CAAT-box, MBS, LTR, TC-rich repeats, ABRE, P-box and TATC-box, were predicted within the promoter region of TaVDAC genes. TaVDAC expression patterns differed among tissues, organs and abiotic stress conditions. Overexpression (OE) of TaVDAC1-B conferred high tolerance to salinity and less resistance to drought stress in Arabidopsis thaliana. TaVDAC1-B interacted with Nucleoredoxin-D1 (TaNRX-D1) protein. Furthermore, compared with WT lines, salinity stress further upregulated the level of AtNRX1 (homologous gene of TaNRX-D1 in Arabidopsis) expression and the activity of superoxide dismutase in TaVDAC1-B OE lines, which led to a decrease in superoxide radical accumulation; drought stress further downregulated AtNRX1 expression and superoxide dismutase activity in TaVDAC1-B OE lines, resulting in the accumulation of superoxide radicals. Our study not only presents comprehensive information for understanding the VDAC gene family in wheat but also proposes a potential mechanism in response to drought and salinity stress. [ABSTRACT FROM AUTHOR]