Contributions of individual and combined Glu-B1x and Glu-B1y high-molecular-weight glutenin subunits to semolina functionality and pasta quality

Durum wheat is an important food crop used primarily for pasta production. High-molecular-weight glutenin subunits (HMW-GS) encoded by the closely linked genes Glu-B1x and Glu-B1y are known for their combined effects on pasta quality, but their individual contributions and interactions remain poorly understood. In this study, we show that individual loss-of-function mutants of Glu-B1x (ΔBx6) and Glu-B1y (ΔBy8) were associated with significant reductions in gluten strength compared to the wildtype, with stronger effects in the ΔBxy double mutant. Reductions in gluten strength were reflected in reduced mixograph and alveograph parameters, gluten index, faster extrusion flow rates and increased cooking loss. Interestingly, the Glu-B1x mutation was also associated with significant increases in grain and semolina protein content, increased pasta firmness, reduced starch viscosity and increased amylose in ΔBx6 and ΔBxy. In general, the ΔBx6 mutation had stronger effects than the ΔBy8 mutation, and significant interactions between the two genes were frequent. In addition to the basic knowledge gained on the individual effects of the Bx6 and By8 subunits and their interactions, the genetic stocks developed in this study provide useful tools to study the effects of natural or synthetic HMW-GS on pasta quality parameters in a background lacking endogenous HMW-GS.     

Distribution of bioactive compounds in maize fractions obtained in two different types of large scale milling processes

Maize kernels contain different bioactive compounds that are important for human health. The aim of this study was to analyze the distribution of the bioactive compounds in maize fractions derived from two industrial dry-milling processes, characterized by a dry-degermination (DD) system and a tempering-degermination (TD) system.The bioactive compounds in maize resulted unevenly distributed in the milling fractions of the kernel. By-products such as the germ and the animal feed flour, had higher total antioxidant capacity (TAC), total polyphenol content (TPC) and total dietary fibre content (TDF) than the whole grains, while xanthophyll and resistant starch resulted to be higher in the fractions derived from the vitreous endosperm. The germ fraction showed also the highest folate content. Results also showed that the type of degermination process influences the bioactive compound contents in the milling fraction, in accordance to the effectiveness of the germ and bran removal from the endosperm fractions. In particular, the animal feed flour obtained by means of TD system resulted in a higher TAC, TPC and TDF than the same fraction obtained by means of the DD system. Conversely, the extraction rate do not affect the recovery of bioactive components in particular fractions.     

The agronomic performance and nutritional content of oat and barley varieties grown in a northern maritime environment depends on variety and growing conditions

Warmer temperatures and increasing interest in high provenance food and drink products are creating new opportunities for cereal growing in northern Europe. Nevertheless, cultivation of oats and barley in these areas for malting and milling remains a challenge, primarily because of the weather, and there are few reports of their nutritional content from this region. In this study, trials in Orkney compared agronomic characteristics and nutritional content of recommended UK oat and barley varieties with Scandinavian varieties over three years. For a subset of varieties, nutritional content was compared with samples cultivated in more southerly sites. For Orkney, barley was considered a more suitable crop than oats because varieties matured earlier. In both crops, Scandinavian varieties matured earlier than UK varieties and some produced comparable yields. The range of values for macronutrients and minerals in oats and barley in Orkney were similar to those reported previously for other locations, but there were some significant differences attributable to variety and year. Compared with grain samples from more southerly locations, oats in Orkney had a significantly lower β-glucan and higher sodium content. The lower β-glucan may have resulted from higher rainfall and lower temperatures during the months of grain filling and maturation.     

Effect of physicochemical and empirical rheological wheat flour properties on quality parameters of bread made from pre-fermented frozen dough

The objective of this study was to examine the influence of flour quality on the properties of bread made from pre-fermented frozen dough. The physicochemical parameters of 8 different wheat flours were determined, especially the protein quality was analysed in detail by a RP-HPLC procedure. A standardized baking experiment was performed with frozen storage periods from 1 to 168 days. Baked bread was characterised for specific loaf volume, crumb firmness and crumb elasticity. The results were compared to none frozen control breads. Duration of frozen storage significantly affected specific loaf volume and crumb firmness. The reduction of specific loaf volume was different among the used flours and its behaviour and intensity was highly influenced by flour properties. For control breads wet gluten, flourgraph E7 maximum resistance and RVA peak viscosity were positively correlated with specific loaf volume. However, after 1–28 days of frozen storage, wet gluten content was not significantly influencing specific loaf volume, while other parameters were still significantly correlated with the final bread properties. After 168 days of frozen storage all breads showed low volume and high crumb firmness, thus no significant correlations between flour properties and bread quality were found. Findings suggest that flours with strong gluten networks, which show high resistance to extension, are most suitable for frozen dough production. Furthermore, starch pasting characteristics were also affecting bread quality in pre-fermented frozen dough.     

Breeding for increased grain protein and micronutrient content in wheat: Ten years of the GPC-B1 gene

To provide food and nutrition security for a growing world population, continued improvements in the yield and nutritional quality of agricultural crops will be required. Wheat is an important source of calories, protein and micronutrients and is thus a priority to breed for improvements in these traits. The GRAIN PROTEIN CONTENT-B1 (GPC-B1) gene is a positive regulator of nutrient translocation which increases protein, iron and zinc concentration in the wheat grain. In the ten years since it was cloned, the impacts of GPC-B1 allelic variation on quality and yield traits have been extensively analyzed in diverse genetic backgrounds in field studies spanning forty environments and seven countries. In this review, we compile data from twenty-five studies to summarize the impact of GPC-B1 allelic variation on fifty different traits. Taken together, the results demonstrate that the functional copy of the GPC-B1 gene is associated with consistent positive effects on grain protein, Fe and Zn content with only marginally negative impacts on yield. We conclude that the GPC-B1 gene has the potential to increase nutritional and end use quality in a wide range of modern cultivars and environments and discuss the possibilities for its application in wheat breeding.     

Responses of glutamine synthetase activity and gene expression to nitrogen levels in winter wheat cultivars with different grain protein content

Glutamine synthetase (GS) plays a central role in plant nitrogen (N) metabolism, which improves crops grain protein content. A pot experiment in field condition was carried out to evaluate GS expression and activity, and grain protein content in high (Wanmai16) and low grain protein (Loumai24) wheat cultivars under two N levels (0.05 and 0.15 g N kg⁻¹ soil). High nitrogen (HN) resulted in significant increases in GS1 and GS2 expression at 10 days after anthesis (DAA), and higher GS activity during the entire grain filling stage. HN also significantly increased yield, grain protein content and protein fraction (except for glutenin of Luomai24) in two wheat cultivars, which indicated that it increased grain yield and protein content by improving nitrogen metabolism. Wanmai16 showed higher grain protein content, gliadin and glutenin content, and had higher expression level of GS2 both in flag leaves and grains at early grain filling stage. However, Luomai24 had greater yield and higher expression level of GS1. The difference expression of GS2 and GS1 genes indicates they had various contributions to the accumulation of protein and starch in wheat grains, respectively. The results suggest that GS2 would be serving as a potential breeding target for improving wheat quality.     

Combined mutations in five wheat STARCH BRANCHING ENZYME II genes improve resistant starch but affect grain yield and bread-making quality

Increases in the proportion of amylose in the starch of wheat grains result in higher levels of resistant starch, a fermentable dietary fiber associated with human health benefits. The objective of this study was to assess the effect of combined mutations in five STARCH BRANCHING ENZYME II (SBEII) genes on starch composition, grain yield and bread-making quality in two hexaploid wheat varieties. Significantly higher amylose (∼60%) and resistant starch content (10-fold) was detected in the SBEII mutants than in the wild-type controls. Mutant lines showed a significant decrease in total starch (6%), kernel weight (3%) and total grain yield (6%). Effects of the mutations in bread-making quality included increases in grain hardness, starch damage, water absorption and flour protein content; and reductions in flour extraction, farinograph development and stability times, starch viscosity, and loaf volume. Several traits showed significant interactions between genotypes, varieties, and environments, suggesting that some of the negative impacts of the combined SBEII mutations can be ameliorated by adequate selection of genetic background and growing location. The deployment of wheat varieties with increased resistant starch will likely require economic incentives to compensate growers and millers for the significant reductions detected in grain and flour yields.     

Characterization and bioactivities of young rice protein hydrolysates

Immature rice was reported to contain higher quantities of bioactive compounds than mature rice. Young rice protein is easy to digest and has hypoallergenic potential, with protein content of 7.2–11.5% compared to rice bran at 9.8%. Few studies have reported on bioactivities and characterization of young rice proteins and their hydrolysates. Bioactivities of native protein and protein hydrolysates of two rice varieties (white rice and colored rice) were characterized and investigated for four development stages (flowery, milky, dough, and mature). Degree of hydrolysis of young rice protein was considerably higher than at the mature stage. Highest DPPH and iron chelating activity were found in alcalase® protein hydrolysate during the flowery-to-milky stage. Iron chelating activity was constant in all development stages because of the low polar amino acid content in rice. The ACE activity of alcalase® protein hydrolysate was higher than native protein at the same development stage, as observed in the milky and dough stages. Inhibitory activity of young rice hydrolysate HepG2 cells was concentration-dependent and not correlated with protein molecular size.     

Use of fermented milling by-products as functional ingredient to develop a low-glycaemic index bread

Selected Lactobacillus plantarum DSM 32248 and Lactobacillus rossiae DSM 32249, isolated and identified from wheat germ, were used to ferment a milling by-products mixture. Lactic acid bacteria metabolisms improved the functional properties of wheat bran and germ, which are considered important sources of functional compounds. Wheat breads were manufactured using 15% (w/w) of fermented (and unfermented) milling by-products, and compared to baker’s yeast wheat bread manufactured without the addition of milling by-products. The use of the fermented ingredient improved the biochemical, functional, nutritional, textural, and sensory features of wheat bread, showing better performances compared to the solely use of wheat flour. Protein digestibility, nutritional indexes, and the rate of starch hydrolysis markedly improved using fermented milling by-products as ingredient. Enriched bread was also characterized by high content of dietary fibre and low glycaemic index determined in vivo.This study exploited the potential of fermented milling by products as functional ingredient. According to the Regulations the bread made under this study conditions can be defined as “high fibre content” and “low glycaemic index”. A number of advantages encouraged the manufacture of novel and healthy and functional leavened baked goods.     

Gluten proteome comparison among durum wheat genotypes with different release date

In order to deepen insight into durum wheat prolamin composition in relation to both end use quality and gluten related disorders, a phenotyping of four genotypes of different release date and technological performance was carried out. A proteomic approach integrated with the evaluation of protein aggregation level, amino acid composition and reactivity to G12 monoclonal antibody, was adopted. The degree of polymerization, estimated as unextractable polymeric proteins (UPP), was positively influenced by Cys-rich proteins (Glu-B3 LMW-GS), with Saragolla showing up to 40% higher values than other genotypes. The proteomic assessment allowed to identify proteins involved in gluten related disorders. In particular, ω5-gliadin involved in wheat allergy (WA), resulted markedly over-expressed in the old landrace Dauno III, up to four-fold than in modern Saragolla. A marked influence of genotype on the expression level of gliadins containing toxic epitopes (TECP) was observed with the more recent genotypes showing lower values (−53%). Also, reactivity to G12 moAb resulted higher in the two old genotypes consistently to their higher celiac disease (CD) toxicity evaluated by the proteomic approach. In conclusion a better gluten composition for processing seems to be associated to a lower expression level of CD toxic peptides and Tri a 19.     

Dough rheological properties and noodle-making performance of non-waxy and waxy whole-wheat flour blends

Dough rheological properties and noodle-making performance of non-waxy whole-wheat flour (WWF) with partial- or full-waxy (PW- or FW-) WWF substitution were studied. The substitution levels were 0, 250, 500, 750, and 1000 g/kg, respectively. FW-WWF reduced the peak viscosity and pasting temperature of WWF blends as its substitution level was increased due to its higher proportions of B-type starch granules and short amylopectin chains, while PW-WWF increased peak viscosity with the increasing substitution level because of its higher amylopectin content. As demonstrated by farinograph and rheometer measurements, FW-WWF interfered with gluten development because of the increased competition for water by arabinoxylans and amylopectin; however, PW-WWF enhanced dough strength due primarily to its increased protein content. Consequently, FW-WWF showed a detrimental effect on cooked noodle texture as the cooked noodle hardness was reduced by 50% at the 1000 g/kg substitution level. In contrast, PW-WWF enhanced noodle integrity and elasticity by increasing cooked noodle cohesiveness and resilience by 10.1% and 14.8%, respectively, at the 1000 g/kg substitution level. The results suggest that with waxy WWF substitution, the changes in starch composition, arabinoxylans, and protein content could modify the interactions among flour components and influence the quality characteristics of noodle products.     

Genetic responses in milling,flour quality,and wheat sensitivity traits to grain yield improvement in U.S. hard winter wheat

A rising global population necessitates continued genetic improvement of wheat (Triticum spp.), but not without monitoring of unintended consequences to processors and consumers. Our objectives were to re-establish trends of genetic progress in agronomic and milling traits using a generational meter stick as the timeline rather than cultivar release date, and to measure correlated responses in flour quality and human wheat-sensitivity indicators. Grain yield and kernel size showed stepwise increases over cycles, whereas wheat protein content decreased by 1.1 g/100 g. Reduced protein content, however, did not result in lower dough strength pertinent to bread baking. A novel method of directly testing gluten elasticity via the compression-recovery test indicated a general increase in gluten strength, whereas the ratio of total polymeric to total monomeric proteins remained stable. Also showing no change with genetic progress in yield were flour levels of gluten epitopes within the key immunotoxic 33-mer peptide. The oligosaccharide fructan, present in milled and wholemeal flours, increased with increasing grain yield potential. While yield improvement in U.S. bread wheat was not accompanied by a decline in gluten strength or systematic shift in a key wheat sensitivity parameter, the unanticipated rise in total fructans does implicate potentially new dietary concerns.     

Impact of ethanol,succinic acid,and the combination thereof at levels produced during sponge fermentation on hard wheat,soft wheat,and durum wheat farinograph rheology

The sponge and dough mixing process is one of the most common in the world, yet the mechanistic understanding of this process has yet to be sufficiently explored. In this study, aqueous solutions of ethanol, succinic acid, and their combination were prepared at concentrations intended to replicate fermentation times of 3, 4 and 6 h. These solutions were added to a farinograph mixer to make dough using hard wheat, soft wheat, and durum wheat flour. The results indicate that these yeast metabolites (ethanol, succinic acid) impact the mixing resistance, peak mixing value, and dough mixing stability in each of the flour types, likely primarily affected by the ratios of gliadin to glutenin and LMW glutenin in each flour type. Results suggest a stabilizing non-covalent interaction imparted by gliadin at peak mixing time, a stabilizing effect of HMW glutenin during break down, and synergistic effects of ethanol and succinic acid that leads to a faster rate of breakdown in later stages of mixing. It also suggests an increase in mixing resistance when acidulants are added to durum wheat dough. Taken together, this study adds new insights on the sponge and dough mixing process in a way that has not previously been conducted.     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Effects of wheat tempering and stone rotational speed on particle size,dough rheology and bread characteristics for a stone-milled weak flour

The poor technological performance of weak wheat flours means that they are usually considered difficult to be transformed into satisfactory bread. During milling, there are several settings that can affect flour characteristics. In this study, we tested two operative parameters that have the potential to affect flour quality – stone rotational speed and wheat tempering. Tempering moistures were set at 11%, 13%, 15%, and 17%, while stone rotational speeds were set at 173, 260, and 346 rpm. Both factors were found to affect operative milling parameters, notably flour yield, process productivity and specific energy consumption. Grain moisture had a significant effect on both dough rheology and bread characteristics (dough stability, tenacity, and extensibility). Dough stability was maximum at 13% moisture. Dough tenacity decreased as moisture increased, while extensibility increased as moisture increased. Bread specific volume and crumb specific volume were improved at 13% and 15% moisture. In conclusion, wheat tempering can be used to improve the potential of a weak flour and bread characteristics, while stone rotational speed affects operative parameters and white flour yield.     

Characterization of the FODMAP-profile in cereal-product ingredients

Cereal-based products, such as bread, are staple foods in the western diet. Due to the nature of their basic ingredients and the diversity of recipes, the amount of fermentable short-chain carbohydrates (FODMAPs) in those products may be high. This study characterized the FODMAP-profiles of a broad range of cereal-product ingredients, serving as a basis for low FODMAP product development. Different cereals, pseudo-cereals, gluten-free flours, pulses, pulse protein ingredients, commercial sprouts, and other cereal-product ingredients were analyzed, using anion-exchange chromatography with electrochemical detection. Wheat and related cereals were high in fructans. Pulses, such as peas contained high galactooligosaccharides (GOS) amounts. Whereas GOS levels in pulse protein ingredients varied, depending on their production. Gluten-free flours, for instance, rice-flour, showed low FODMAP-profiles. Amongst those, buckwheat, which does not contain any of the FODMAPs investigated, contained high amounts of other soluble non-digestible carbohydrates, namely fagopyritols; these may have a similar effect on a sensitive gut as GOS. Finally, ingredients contained mainly high levels of fructans and GOS. Yet, the analysis of commonly consumed commercial cereal products, including bread, pasta, crackers and biscuits, highlighted the relevance of lactose, fructose in excess of glucose and polyols. These products serve as benchmarks for further product development.     

Transgenic sorghum with suppressed synthesis of kafirin subclasses: Effects on flour and dough rheological characteristics

Arising from work showing that conventionally bred high protein digestibility sorghum types have improved flour and dough functionality, the flour and dough properties of transgenic biofortified sorghum lines with increased protein digestibility and high lysine content (TG-HD) resulting from suppressed synthesis of several kafirin subclasses, especially the cysteine-rich γ-kafirin, were studied. TG-HD sorghums had higher flour water solubility at 30 °C (p < 0.05) and much higher paste viscosity (41% higher) than their null controls (NC). TG-HD doughs were twice as strong as their NC and dynamic rheological analysis indicated that the TG doughs were somewhat more elastic up to 90 °C. CLSM of doughs and pastes indicated that TG-HD had a less compact endosperm protein matrix surround the starch compared to their NC. The improved flour and dough functional properties of the TG-HD sorghums seem to be caused by reduced endosperm compactness resulting from suppression of synthesis of several kafirin subclasses which modifies protein body and protein matrix structure, and to improved protein-starch interaction through hydrogen bonding specifically caused by reduction in the level of the hydrophobic γ-kafirin. The improved flour functionality of these transgenic biofortified sorghums can increase their commercial utility by complementing their improved nutritional quality.     

Prediction of bread wheat baking quality using an optimized GlutoPeak®-Test method

The GlutoPeak^®-Test (GPT) as a rapid small-scale technique was optimized to evaluate the gluten aggregation properties and to predict the loaf volume, on the basis of a multiyear and multilocation analysis of wheat samples, using different solvents. 5 % lactic acid and 1 % sodium chloride displayed significant GPT responses. Relationships between protein content, sedimentation value, GPT parameters and loaf volume were investigated. With 1 % sodium chloride, the torque 15 s before maximum torque (AM) presented the highest correlation with loaf volume of samples from 2013 to 2014 (r = 0.77, r = 0.63, p < 0.001, respectively). A multiple regression analysis indicated that the best prediction of loaf volume was a linear function of protein content and AM, explaining the variation in loaf volume by 63 % and providing an uncertainty of ±39 ml. The accuracy of the validation of the linear function leads to 64 % correct and to 36 % incorrect predictions of the loaf volume. This emphasizes that the application of the linear function of protein content and AM cannot replace the actual measurement of loaf volume, but it could be a useful rapid screening test in breeding for improved baking quality in bread wheat.     

The impact of Septoria tritici blotch in bread making quality among argentinean wheat cultivars

Septoria tritici blotch (STB), caused by Zymoseptoria tritici is a relevant foliar wheat disease worldwide. Several reports show the importance of STB on grain yield, their components and grain protein while little is known about its effect on the rheological properties of the wheat flour. The scarce literature found, only mentions the effect of the complex of foliar diseases on wheat quality, without individualizing the effect of the different pathogens separately. This study analyze the influence of increasing doses of inoculum of Zymoseptoria tritici, on the bread making quality of ten Argentinean wheat cultivars and its possible variation according to their quality group. The increase of inoculum concentration augmented the area under disease progress curve, decreased green flag leaf area duration and green leaf area duration. Cultivars K. Flecha and B.75 Aniversario had the lowest green flag leaf area duration causing higher reduction in grain filling period and higher reductions in P, indicating a lower gliadin/glutenin ratio. STB decreased P/L and E while L, W, D, SV and bread volume increased. Cultivars differed in rheological parameters according to their quality group. Gluten/protein relationship was significant in quality group 1 and non-significant in cultivars belonging to quality group 2 and 3.