Abstract Background Wheat flour maturation affects the aggregation and structural stability of proteins. The number of high-molecular-weight glutenin subunits (HMW-GSs) differs in various wheat varieties. The effects of Dx2 absence on the protein aggregation characteristics and thermal stability of flour were investigated during 120 d of maturation using near-isogenic lines (NILs). Results The absence of Dx2 delayed and decreased the protein aggregation of flours during maturation, i.e. the maturation-induced increases were later and smaller for glutenin, glutenin macropolymer (GMP), glutenin/gliadin ratio, β-sheets, and β-sheet/α-helix ratio in HMW-D1a without Dx2 than in HMW-D1p with Dx2; these differences were ascribed to the weaker interactions between the sulfhydryl (-SH) groups, disulfide bonds (-S-S-), and hydrophobicity in the flours without Dx2. Flour maturation caused the dough microstructures to be more compact and denser, thereby increasing the flour thermal stability as observed by a higher denaturation peak temperature (Tp), enthalpy of thermal transition (ΔH), and degradation temperature (Td), These changes led to better dough properties such as dough development time, dough stability time, and protein weakening, but the optimal stage in HMW-D1a without Dx2 was reached later. Conclusion These findings deepen the understanding of how HMW-GS Dx2 modifies protein structures during flour maturation.
SUMMARY Plants have developed various protective mechanisms to survive drought stress. Previously, it was shown that a wheat bZIP transcription factor gene TaFD‐Like2‐1A ( TaFDL2‐1A ) can confer drought tolerance in Arabidopsis. However, the biological functions related to drought stress tolerance of TaFDL2‐1A in wheat ( Triticum aestivum L.) remain unclear. In the present study, overexpression of TaFDL2‐1A in the wheat cultivar Fielder improved drought resistance and conferred abscisic acid (ABA) hypersensitivity. Further analysis showed that overexpression of TaFDL2‐1A increased the hypersensitivity of stomata to drought stress and endogenous ABA content under drought conditions. Genetic analysis and transcriptional regulation analysis indicated that TaFDL2‐1A binds directly to the promoter fragments of TaRAB21s and TaNCED2s via ACGT core cis ‐elements, thereby activating their expression, leading to enhanced ABA responses and endogenous ABA accumulation. In addition, our results demonstrate that overexpression of TaFDL2‐1A results in higher SOD and GPX activities in wheat under drought conditions by promoting the expression of TaSOD1 and TaGPx1‐D , indicating enhanced reactive oxygen species (ROS) scavenging. These results imply that TaFDL2‐1A positively regulates ABA biosynthesis, ABA responses, and ROS scavenging to improve drought stress tolerance in transgenic wheat. Our findings improve our understanding of the mechanisms that allow the wheat bZIP transcription factor to improve drought resistance and provide a useful reference gene for breeding programs to enhance drought resistance.
In this study, we investigated the effect of high-molecular-weight glutenin subunits (HMW-GSs) on gluten quality and glutenin synthesis based on the cytological, physicochemical, and transcriptional levels using Xinong1718 and its three near-isogenic lines (NILs). Cytological observations showed that the endosperm of Glu-1Bh with Bx14+By15 accumulated more abundant and larger protein bodies at 10 and 16 days after anthesis than the other NILs. Glu-1Bh exhibited higher nitrogen metabolism enzyme gene expression and activity levels. The transcriptional levels of genes encoding HMW-GSs, protein folding, and transcription factors differed significantly among the NILs, and they were highest in Glu-1Bh. Our results demonstrate that variations in the expression patterns of nitrogen metabolism and glutenin synthesis-related genes may account for the differences in the accumulation of glutenin, glutenin macropolymers, and protein bodies, thereby affecting the structural and thermal stability of gluten. These findings provide novel insights into how different HMW-GSs might improve the quality of wheat.
Absence of high-molecular-weight glutenin subunit (HMW-GS) Dx2 weakens the gluten quality, but it is unclear how the absence of Dx2 has these effects. Thus, we investigated the gluten quality in terms of cytological, physicochemical, and transcriptional characteristics using two near-isogenic lines with Dx2 absent or present at Glu-D1 locus. Cytological observations showed that absence of Dx2 delayed and decreased the accumulation of protein bodies (PBs), where fewer and smaller PBs formed in the endosperm. The activity and gene expression levels of nitrogen assimilation and proteolysis enzymes were lower in HMW-D1a without Dx2 than HMW-D1p with Dx2, and thus less amino acid was transported for protein synthesis in the grains. The expression pattern of genes encoding Glu-1Dx2+1Dy12 was similar to those of three transcription factors, where these genes were significantly down-regulated in HMW-D1a than HMW-D1p. Three genes involving with glutenin polymerization were also down-regulated in HMW-D1a. These results may explain the changes in the glutenin and glutenin macropolymer (GMP) levels during grain development. Therefore, we suggest that the lower nitrogen metabolism capacity and expression levels of glutenin synthesis-related genes in HMW-D1a accounted for the lower accumulation of glutenin, GMP, and PBs, thereby weakening the structural‒thermal properties of gluten.
Abstract Genetic strategies can be effective in improving wheat (Triticum aestivum L.) drought stress tolerance, but accumulating evidence suggests that overexpressing drought-resistance genes, especially genes related to the abscisic acid (ABA) signaling pathway, can retard plant growth. We previously characterized the positive roles of the wheat bZIP transcription factor TaFD-Like2-1A (TaFDL2-1A) in drought stress tolerance and ABA biosynthesis and response, whereas a dwarfing shoot exhibited under normal conditions. This study determined the underlying mechanisms that allow TaFDL2-1A to affect shoot growth. Overexpressing TaFDL2-1A decreased cell length, cell width, leaf size, shoot length, and biomass in wheat. The results of RNA-seq showed that multiple differently expressed transcripts are enriched in the auxin signaling pathway. Further analysis indicated higher expression levels of Gretchen Hagen3 (GH3) genes and lower indole-3-acetic acid (IAA) concentrations in the TaFDL2-1A overexpression lines. Exogenous IAA treatment restored the phenotypes of the TaFDL2-1A overexpression lines to wild-type levels. Transcriptional regulation analysis suggested that TaFDL2-1A enhances the expression of auxin metabolism genes, such as TaGH3.2-3A, TaGH3.2-3B, TaGH3.8-2A, and TaGH3.8-2D, by directly binding to ACGT core cis-elements. Furthermore, tafdl2 knock-out plants had lower expression levels of these GH3 genes and higher IAA levels than Fielder wheat. These GH3 gene expression and IAA levels were induced and reduced in Fielder wheat and tafdl2 knock-out plants treated with exogenous ABA. Our findings elucidate mechanisms underlying the functional redundancy of TaFDL2-1A in the crosstalk between ABA and IAA to affect shoot growth and provide insights into the balance between drought resistance and yield in wheat.
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