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.     

Genome-wide association mapping of starch granule size distribution in common wheat

Starch is a crucial component in wheat endosperm and plays an important role in processing quality. Endosperm of matured wheat grains contains two distinct starch granules (SG), referred to as larger A- and smaller B-granules. In the present study, 166 Chinese bread wheat cultivars planted in four environments were characterized for variation in SG size. A genome-wide association study (GWAS) using the 90 K SNP assay identified 23 loci for percentage volumes of A- and B-granules, and 25 loci for the ratio of A-/B-granules volumes, distributing on 15 chromosomes. Fifteen MTAs were associated with both the percentage volumes of A-, B-granules and the ratio of A-/B-granules volumes. MTAs IWB34623 and IWA3693 on chromosome 7A and IWB22624 and IWA4574 on chromosome 7B associated with the percentage volumes of A- and B-granules consistently identified in multiple environments were considered to be stable. Linear regression analysis showed a significantly negative correlation of the number of favorable alleles with the percentage volumes of A-granules and a significantly positive correlation between the number of favorable alleles and the percentage volumes of B-granules, respectively. The loci identified in this study and associated markers could provide basis for manipulating SG size to obtain superior noodle quality in wheat.     

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.     

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.     

Immunomodulatory activities of non-prolamin proteins in wheat germ and gluten

Albumin (Alb), globulin (Glo), glutelin (Gll) and glutenin (Gln) were separately extracted from wheat germ and wheat gluten. Amino acisd composition, molecular weight distribution, solubility, in vitro digestibility, and immunomodulatory activities were all analyzed. Gll and Gln have similar molecular weight distributions, which differed from those of Alb and Glo. Alb showed the highest solubility at various pH values (except pH 4.0), whereas Glo showed the highest in vitro digestibility. Glo and Gll have the highest proportion of essential to total amino acids, while Alb and Gll have the highest protein digestibility-corrected amino acid scores. Gll had the strongest immunomodulatory effects in terms of stimulation of RAW 264.7 cells to produce IL-6, TNF-α, and IL-10, and good stimulatory effects on splenocyte proliferation, production of IL-2, phagocytosis, and secretion of nitric oxide in RAW 264.7 cells. Gll can be considered a good protein source for use in health foods.     

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.     

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.     

Puroindoline genes introduced into durum wheat reduce milling energy and change milling behavior similar to soft common wheats

Grain physical characteristics and milling behavior of a durum wheat line in which both wild-type puroindoline genes were translocated and stabilized after backcrossing (Svevo-Pin) were compared with the parent line (Svevo). The only observed differences between grain characteristics were the mechanical resistance and starchy endosperm porosity revealed through vitreosity measurement. A significant increase of flour and a decrease of semolina yield and break milling energy were observed from Svevo-Pin in comparison with the non-recombinant parent line in accordance to the lower grain mechanical resistance and higher porosity measurements. Moreover, the particle size distribution shown for Svevo-Pin flour appeared consistent with a lower adhesion between starch granules and the protein matrix attributed to the presence of wild-type puroindolines. Coarse bran yield was conversely increased. This appeared to be due to a lower starchy endosperm recovery as a higher proportion of grain starch was found in this bran fraction. Flour from the durum parent line was inversely enriched in phytic acid, a cellular marker of the aleurone layer. Starch damage was also lower in Svevo-Pin flours in comparison with Svevo. All of the observed differences between translocation and parent lines were confirmed independent of the culture growth conditions (n = 12).     

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.     

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.     

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.     

Modification of protein structure and dough rheological properties of wheat flour through superheated steam treatment

Native (NF, 13.5% w.b) and moistened (MF, 27% w.b) wheat flours were treated with superheated steam (SS) at 170 °C for 1, 2 and 4 min, and their protein structure as well as dough rheological properties were analyzed. Confocal laser scanning microscopy (CLSM) and SDS-PAGE patterns indicated the formation of protein aggregates with reduced SDS extractability after treatment. Farinograph and dynamic rheometry measurements showed that the strength as well as elastic and viscous moduli of the dough made from SS-treated flours progressively increased with SS treatment time. And both the improvements were more pronounced for superheated steam-treated moistened flours (SS-MF) than for superheated steam-treated native flours (SS-NF). Size-exclusion high performance liquid chromatography (SE-HPLC) analysis demonstrated that dough rheological parameters have positive correlations with SDS unextractable polymeric proteins (UPP) contents. SS treatment on flours led to a transition of protein secondary structures to more ordered form (α-helix and β-sheet). Additionally, free sulfhydryl (SH) contents decreased after treatment, which implied that disulfide bonds accounted for protein extractability loss and dough rheological properties improvement. Elevated moisture level promoted the modification of both protein structure and dough behaviors of flours during SS treatment.     

Fluorimetric studies of the interactions of wheat puroindolines with polar lipids on the surface starch granules

The interactions of puroindolines with polar lipids were investigated using polarization of fluorescence probes preincorporated into a liposomal bilayer containing PC, PI, PS, MGDG, DGDG, and sulfolipids. The intrinsic fluorescence of Trp residue method was also used. Regardless of the kind of lipid used for liposome preparation, proteins interacted with the liposomes. Conformational changes of the proteins were observed simultaneously with the change in the molecule packing in the lipid bilayer of the liposomes. Puroindoline interactions with the surface of the liposomes have explicit importance for the net charge of this surface. The strong interaction between the proteins and lipids takes place in the presence of a ligand with a negative charge. The obtained results confirm that lipids take part in puroindoline–starch granule surface interactions.     

Interplay between starch and proteins in waxy wheat

Most of the unique properties of waxy wheat have been associated with the lack of amylose, that in turn may affect the mutual interactions between starch and proteins. To address this particular aspect, we carried out molecular, rheological, and calorimetric studies on flours from two waxy wheat lines that were compared with a non-waxy one. Dough thermal properties and water binding capacity were investigated by Differential Scanning Calorimetry (DSC) and by thermogravimetric analysis, respectively. Protein solvation, aggregation, and thiol accessibility were also investigated, together with dough mixing properties and stickiness. Proteins in waxy wheat samples needed more water to complete solvation, likely because of the water-retaining capacity of waxy wheat starch. In waxy wheat dough, water was tightly bound to starch, and DSC studies indicated an increase in gelatinization temperature. Moreover, the low water mobility in waxy wheat resulted in low and retarded gluten hydration and in high stickiness. In samples with the highest stickiness, protein aggregates were stabilized mainly by hydrophobic interactions. Differences between waxy wheat lines may be attributed to a different structural organization of components within each class of biopolymers.     

The molecular structure features-immune stimulatory activity of arabinoxylans derived from the pentosan faction of wheat flour

Recently, the immune stimulation properties of cereal arabinoxylans (AX) have been reported. The aims of this study were to identify the molecular features and potential immune stimulation activities of AX and enzymatic modified arabinoxylan (AXE) from the pentosan fraction of wheat flour.The results of molecular characterization of AX and AXE show that AXE contains a larger portion (85.7%) of low Mw arabinoxylans (≤25 kDa) and has a higher degree of branch substitution (0.81 ± 0.01) compared to AX (49.5% and 0.62 ± 0.02). In in vitro testing, the ability of AX and AXE to stimulate immune cells, as measured by NO₂⁻ production by U937 cells and IL-8 secretion by Caco-2 cells were found to be dose–dependent in the range tested (100–5000 μg/mL p ≤ 0.05). AXE showed a greater activity at each concentration (100–5000 μg/mML) than AX (p ≤ 0.05).In conclusion, the greater immune stimulating activity of AXE may be associated with its low Mw (≤25 kDa) and a higher degree of branch substitution (0.81 ± 0.01).     

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.     

LC-MS analysis reveals the presence of graminan- and neo-type fructans in wheat grains

This is the first study describing the fine structure of the main, individual fructan oligosaccharides present in wheat grains. Wheat grain fructan structure was investigated in developing wheat grains and in different tissues of mature grains with liquid chromatography-mass spectrometry. Fructan oligosaccharides with a low degree of polymerization (<5) were mainly of the graminan- and inulin-type in developing wheat grains during the first week after anthesis. Starting from 14 days after anthesis, neo-type fructans, fructans with an internal glucose, were observed for the first time. Several neo-type fructan structures were identified and their portion in the total fructan pool gradually increased during grain development. In the mature kernel, almost no differences were noted between the fructan distributions of wheat flour and two wheat bran fractions enriched in either pericarp or aleurone tissue. Results are related to wheat fructan metabolizing enzymes and the nutritional implications are discussed.     

Whole wheat bread: Effect of bran fractions on dough and end-product quality

Consumption of whole-wheat based products is encouraged due to their important nutritional elements that benefit human health. However, the use of whole-wheat flour is limited because of the poor processing and end-product quality. Bran was postulated as the major problem in whole wheat breadmaking. In this study, four major bran components including lipids, extractable phenolics (EP), hydrolysable phenolics (HP), and fiber were evaluated for their specific functionality in flour, dough and bread baking. The experiment was done by reconstitution approach using the 2⁴ factorial experimental layout. Fiber was identified as a main component to have highly significant (P < 0.05) and negative influence on most breadmaking characteristics. Although HP had positive effect on farinograph stability, it was identified as another main factor that negatively impacted the oven spring and bread loaf volume. Bran oil and EP seemed to be detrimental to most breadmaking characteristics. Overall, statistical analysis indicates that influence of the four bran components are highly complex. The bran components demonstrate multi-way interactions in regards to their influence on dough and bread-making characteristics. Particularly, Fiber appeared to have a high degree of interaction with other bran components and notably influenced the functionality of those components in whole wheat bread-making.