Vitreosity,its stability and relationship to protein content in durum wheat

High quality requirements are set on durum wheat (Triticum durum) from semolina mills and pasta producers. For the production of semolina and pasta with good cooking quality, high grain protein content and vitreosity is required. The dependency of vitreosity on protein content as well as its stability under the influence of humidity was not well investigated up to now. We (1) compared two methods to determine vitreosity, (2) investigated the relationship between vitreosity and protein content, (3) developed a method to analyze vitreosity under humidity, and (4) examined the relationship between protein content and agronomical as well as quality traits in durum wheat. The results showed that the formation of vitreous kernels greatly depends on the protein content. To evaluate the stability of vitreosity under the influence of humidity a new method was elaborated and employed to assess the durum germplasm under study. This revealed that vitreosity of a durum wheat variety depends on the potential to form vitreous kernels but also to maintain this vitreosity under the influence of humidity. Our results further show that protein content is a central trait in durum wheat that strongly influences important traits like grain yield, vitreosity, and b-value.     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

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.     

Variation in gluten quality parameters of spring wheat varieties of different origin grown in contrasting environments

The aim of this study was to investigate variation in protein content and gluten viscoelastic properties in wheat genotypes grown in two mega-environments of contrasting climates: the southeast of Norway and Minnesota, USA. Twelve spring wheat varieties, nine from Norway and three HRS from Minnesota, were grown in field experiments at different locations in Norway and Minnesota during 2009–2011. The results showed higher protein content but lower TW and TKW when plants were grown in Minnesota, while the gluten quality measured as Rmax showed large variation between locations in both mega-environments. On average, Rmax of the samples grown in Minnesota was higher than those grown in Norway, but some locations in Norway had similar Rmax values to locations in Minnesota. The data showed inconsistent relationship between the temperature during grain filling and Rmax. Our results suggest that the weakening effect of low temperatures on gluten reported in this study are caused by other environmental factors that relate to low temperatures. The variety Berserk showed higher stability in Rmax as it obtained higher values in the environments in Norway that gave very weak gluten for other varieties.     

Alkylresorcinol composition allows the differentiation of Triticum spp. having different degrees of ploidy

Total alkylresorcinol (AR) content and homologue composition were assessed in whole grain flours of 15 varieties each of bread wheat, durum, spelt, emmer, and einkorn grown in four different environments. Bread wheat (761 ± 92 μg/g DM) and spelt (743 ± 57 μg/g) belonging to the hexaploid species showed higher AR concentrations than the tetraploid durum (654 ± 48 μg/g, p < 0.05), while the concentrations found in the diploid einkorn (737 ± 91 μg/g) and the tetraploid emmer (697 ± 94 μg/g) did not significantly differ from the other species. The AR content showed a remarkable heritability and, thus, seemed to be mainly determined by genetic factors. If ARs were assumed to be deposited within a specific AR-rich layer of the kernel, AR levels of all varieties would easily surpass their minimal inhibitory concentrations against fungal pathogens within this barrier layer. Although the AR carrying a C21:0 side chain was the main homologue in all species, the levels of all AR homologues and their relative composition significantly differed between hexaploid (bread wheat and spelt), tetraploid (durum and emmer) and diploid (einkorn) species. Consequently, a clear-cut differentiation of Triticum species and derived whole grain flours according to their degrees of ploidy was established based on concentrations of saturated C₁₇-, C₁₉-, C₂₁-, C₂₃-, and C₂₅-substituted ARs.     

Study of the mechanism of improvement due to waxy wheat flour addition on the quality of frozen dough bread

Waxy wheat flour (WWF) was substituted for 10% regular wheat flour (RWF) in frozen doughs and the physicochemical properties of starch and protein isolated from the frozen doughs stored for different time intervals (0, 1, 2, 4 and 8 weeks) were determined to establish the underlying reasons leading to the effects observed in WWF addition on frozen dough quality. Using Nuclear Magnetic Resonance (NMR), Differential Scanning Calorimeter (DSC) and X-ray Diffraction (XRD) among others, the gluten content, water molecular state, glutenin macropolymer content, damaged starch content, starch swelling power, gelatinization properties, starch crystallinity and bread specific volume were measured. Compared to RWF dough at the same frozen storage condition, 10% WWF addition decreased dry gluten and glutenin macropolymer contents and T₂₃ proton density of frozen dough, but increased the wet gluten content, T₂₁ and T₂₂ proton density. 10% WWF addition also decreased damaged starch content, but increased starch swelling power, gelatinization temperature and enthalpy, crystallinity of starch and bread specific volume of frozen dough. Results in the present study showed that the improvement observed due to WWF addition in frozen dough bread quality might be attributed to its inhibition of redistribution of water molecules bound to proteins, increase in damaged starch content and decrease in starch swelling power.     

Molecular characterization of storage proteins for selected durum wheat varieties grown in different environments

The variations of the amounts of individual high molecular weight glutenin subunits (HMW-GS), of the ratios HMW-GSy to HMW-GSx and HMW-GS to low molecular weight glutenin subunits (LMW-GS) and of protein content were evaluated for eight durum wheat cultivars in two regions using four fertilizer combinations during two successive years. All measured parameters showed significant variation with genotypes (G), environments (E) and fertilizers (F). The interaction E × G × F was highly significant for glutenin amount variation. Amongst cultivars possessing HMW-GS 20, landraces seem to better value the N-fertilizer use for the accumulation of HMW-GSy than high yielding cultivars. Both HMW-GSy to HMW-GSx and HMW-GS to LMW-GS ratios were found to be positively correlated (p < 0.05) with total protein content.     

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.     

Effect of allelic variation at the Glu-3/Gli-1 loci on breadmaking quality parameters in hexaploid wheat (Triticum aestivum L.)

Low molecular weight glutenin subunits (LMW-GS) encoded by the Glu-3 loci are known to contribute to wheat breadmaking quality. However, the specific effect of individual Glu-3 alleles is not well understood due to their complex protein banding patterns in SDS-PAGE and tight linkage with gliadins at the Gli-1 locus. Using DNA markers and a backcross program, we developed a set of nine near isogenic lines (NILs) including different Glu-A3/Gli-A1 or Glu-B3/Gli-B1 alleles in the genetic background of the Argentine variety ProINTA Imperial. The nine NILs and the control were evaluated in three different field trials in Argentina. Significant genotype-by-environment interactions were detected for most quality parameters indicating that the effects of the Glu-3/Gli-1 alleles are modulated by environmental differences. None of the NILs showed differences in total flour protein content, but relative changes in the abundance of particular classes of proteins cannot be ruled out. On average, the Glu-A3f, Glu-B3b, Glu-B3g and Glu-B3i_Man alleles were associated with the highest values in gluten strength-related parameters, while Glu-A3e, Glu-B3a and Glu-B3i_Chu were consistently associated with weak gluten and low quality values. The value of different Glu-3/Gli-1 allele combinations to improve breadmaking quality is discussed.     

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.     

Influence of allelic prolamin variation and localities on durum wheat quality

Thirty-seven varieties of a Mediterranean durum wheat collection grown in Tunisia and Spain were analysed for their allelic composition in prolamins, as well as their protein concentration, sodium dodecyl sulphate sedimentation (SDSS) test and mixograph parameters. Genotype was a greater source of variation in all measurements than locality. Uncommon high and low molecular glutenin subunits (HMW-GS and LMW-GS) were found (V and 2•• subunits at Glu-A1, 13 + 16 at Glu-B1, 5* subunit and ax allele at Glu-A3). The rare combinations 2 + 4+14 + 18 and 8 + 9+13 + 16+18 subunits at the Glu-B3 locus were found. Glu-A3ax had a positive influence on SDSS and mixograph parameters. Of all the prolamins, those that have the B-LMW-GS composition aaa (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d gave the best semolina quality. By contrast, semolina quality is poor when this same composition is associated with the Glu-A1c and Glu-B1e and even poorer when associated with the Glu-A1c and Glu-B1f. In addition, the cultivars with B-LMW-GS allelic composition aab (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d, gave high quality, whereas when associated with the Glu-A1c and Glu-B1e or with Glu-A1o and Glu-B1f, the quality was very poor.     

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.     

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.     

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.     

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.     

Influence of the cultivar,environment and management on the grain yield and bread-making quality in winter wheat

The genotype, environment and their interaction play an important role in the grain yielding and grain quality attributes. The main aim of this study was to determine the contributions of the genotype, environment and their interaction to the variation in bread-making traits. The data that were used for the analyses performed in this study were obtained from 3 locations in Poland from post-registration multi-environment trials with winter wheat in 2009 and 2010. The experimental factors were the cultivar (7 cultivars) and the crop management level (low input and high input). In the multi-environment trials, 17 traits were investigated that characterize grain, flour and dough quality. Most of the traits were affected much more strongly by environmental factors (i.e., year and location) than by genotype. The variance components revealed an especially strong effect of the year on the baking score, loaf volume and water absorption, as well a strong effect of the location on dough development and protein content. The obtained results demonstrate that the grain quality as measured by the parameters based on the protein content and quality may be substantially improved by crop management practices, especially by N fertilization level.     

Effect of high molecular weight glutenins and rye translocations on soft wheat flour cookie quality

The influence of high molecular weight glutenin subunits (HMW-GS) on wheat breadmaking quality has been extensively studied but the effect of different Glu-1 alleles on cookie quality is still poorly understood. This study was conducted to analyze the effect of HMW-GS composition and wheat-rye translocations on physicochemical flour properties and cookie quality of soft wheat flours. Alleles encoded at Glu-A1, Glu-B1 and Glu-D1 locus had a significant effect over physicochemical flour properties and solvent retention capacity (SRC) profile. The null allele for Glu-A1 locus presented the highest cookie factor observed (CF = 7.10), whereas 1BL/1RS and 1AL/1RS rye translocations had a negative influence on CF. The three cultivars that showed the highest CF (19, 44 and 47) had the following combination: Glu-A1 = null, Glu-B1 = 7 + 8, Glu-D1 = 2 + 12 and no secalins. Two prediction equations were developed to estimate soft wheat CF: one using the HMW-GS composition and the other using physicochemical flour parameters, where SRCsuc, SRC carb, water-soluble pentosans, damaged starch and protein turned out to be better CF predictors. This data suggests that grain protein allelic composition and physicochemical flour properties can be useful tools in breeding programs to select soft wheat of good cookie making quality.     

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.     

Real-time quantitative PCR based method for the quantification of fungal biomass to discriminate quantitative resistance in barley and wheat genotypes to fusarium head blight

Fusarium head blight (FHB) caused by Fusarium graminearum is one of the devastating diseases of small grain crops, including barley and wheat. Breeding for resistance is one of the best and ecofriendly strategies to manage the FHB. However, the existing methods used for screening genotypes, both under field and greenhouse conditions, often resulted in high experimental error, leading to inconsistent ranking of genotypes over years. In the postgenomic era, precise assessment of resistance is crucial to identify candidate genes. Here, we report a pathogen inoculation procedure and a real-time quantitative polymerase chain reaction (qPCR) based protocol for the quantification and discrimination of quantitative resistance among barley and wheat genotypes to FHB. Using Fusarium specific primer pair Tri6_10, for the trichothecene biosynthetic cluster (Tri6) gene, we successfully quantified the relative fungal biomass in both spikelets and rachis. A qPCR of spikelets and rachis collected on 6 dpi, from inoculated three alternate spikelet regions, discriminated resistance with less experimental error than those based on the proportion of spikelets diseased (PSD) at 9 dpi. This method can be applied for medium to high-throughput barley and wheat breeding programmes to discriminate quantitative resistance among genotypes against FHB.