Effects of drought on the morphological and physicochemical characteristics of starch granules in different elite wheat varieties

In this study, scanning electron microscopy (SEM) revealed the formation of pits and pores on the surfaces of starch granules in response to drought stress, with substantially more pronounced effects in the ordinary yield potential wheat cv. Xindong 23 than the excellent yield potential wheat cv. Xindong 20. Drought induced a significant reduction in starch granule sizes in both wheat varieties, though the reduction observed in Xindong 23 was six times more pronounced than that observed for Xindong 20. Amyloglucosidase and α-amylase treatment of starch from wheat grown in drought conditions released significantly more reducing sugars compared with samples from irrigated controls. SEM and confocal laser scanning microscopy (CLSM) revealed that starch granules from the two wheat varieties grown under drought conditions had substantially increased fluorescence after treatment with proteolytic enzymes and staining with methanolic merbromin and 3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde dyes. Analysis of pasting properties showed significant increases of peak viscosity, trough viscosity, break down, and setback following drought stresses. Furthermore, drought induced a significant reduction in the water binding capacity and increased damage to starch only in Xindong 23. These results provide insight into the potential mechanisms through which drought influences the ultrastructures and physicochemical properties of starch in wheat.     

The influence of abiotic stress conditions on dough mixing characteristics of two hard red spring wheat cultivars

Two commercial hard red spring wheat cultivars were exposed to high and low temperatures, as well as drought stress when the main tiller kernels were at the soft dough stage. The trial was done in the greenhouse for two consecutive seasons to determine the effects of these stress conditions on protein content, SDS sedimentation and selected Mixsmart characteristics. Heat stress had the largest effect on mixing characteristics. Heat and drought stress caused a significant increase in flour protein content of both cultivars and had similar effects on mixing characteristics. The Mixsmart characteristics associated with dough strength were increased by heat and drought stress. Cold stress caused a slight increase in protein content of the cultivars, but in general caused a reduction in dough strength as measured with Mixsmart characteristics. The reaction of Mixsmart characteristics to heat and drought stress was much larger in Duzi than in Kariega, confirming that there is a large genotype effect in rheological characteristics.     

Barley genotype expressing “stay-green”-like characteristics maintains starch quality of the grain during water stress condition

The identification of “stay-green” in sorghum and its positive correlation with yield increases has encouraged attempts to incorporate “stay-green”-like traits into the genomes of other commercially important cereal crops. However, knowledge on the effects of “stay-green” expression on grain quality under extreme physiological stress is limited. This study examines impacts of “stay-green”-like expression on starch biosynthesis in barley (Hordeum vulgare L.) grain under mild, severe, and acute water stress conditions induced at anthesis. The proportions of long amylopectin branches and amylose branches in the grain of Flagship (a cultivar without “stay-green”-like characteristics) were higher at low water stress, suggesting that water stress affects starch biosynthesis in grain, probably due to early termination of grain fill. The changes in long branches can affect starch properties, such as the rates of enzymatic degradation, and hence its nutritional value. By contrast, grain from the “stay-green”-like cultivar (ND24260) did not show variation in starch molecular structure under the different water stress levels. The results indicate that the cultivar with “stay-green”-like traits has a greater potential to maintain starch biosynthesis and quality in grain during drought conditions, making the “stay-green”-like traits potentially useful in ensuring food security.     

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.     

Metabolite profiling of the response to high-nitrogen fertilizer during grain development of bread wheat (Triticum aestivum L.)

Wheat yield and quality are dependent largely on nitrogen (N) availability. In this study, we performed the first metabolomic analysis of the response to high-N fertilizer during wheat grain development using non-targeted gas chromatography-mass spectrometry (GC–MS). Quality parameter analyses demonstrated that high-N fertilizer application led to a significant increase in grain protein content and improvement in starch and bread-making quality. Comparative metabolomic profiling of six grain developmental stages resulted in identification of 74 metabolites, including amino acids, carbohydrates, organic acids and lipids/alcohol, which are primarily involved in carbon and N metabolism. Under high-N fertilizer treatment, numerous metabolites accumulated significantly during grain development. Principal component analysis revealed two principal components as being responsible for the variances resulting from N-fertilizer treatments. Metabolite–metabolite correlation analysis demonstrated that the high-N treatment group had a greater number of positive correlations among metabolites, suggesting that high-N fertilizer treatment induced a concerted metabolic change that resulted in improved grain development. Particularly, the high-N treatment-mediated significant accumulation of metabolites involved in the TCA cycle, starch and storage protein synthesis could be responsible for the improvement of grain yield and quality. Our results provide new insight into the molecular mechanisms of wheat grain development and yield and quality.     

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).     

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.     

Suitability of tef varieties in mixed wheat flour bread matrices: A physico-chemical and nutritional approach

Wheat flour replacement from 0 to 40% by single tef flours from three Ethiopian varieties DZ-01-99 (brown grain tef), DZ-Cr-37 (white grain tef) and DZ-Cr-387 (Quncho, white grain tef) yielded a technologically viable ciabatta type composite bread with acceptable sensory properties and enhanced nutritional value, as compared to 100% refined wheat flour. Incorporation of tef flour from 30% to 40% imparted discreet negative effects in terms of decreased loaf volume and crumb resilience, and increase of crumb hardness in brown tef blended breads. Increment of crumb hardness on aging was in general much lower in tef blended breads compared to wheat bread counterparts, revealing slower firming kinetics, especially for brown tef blended breads. Blended breads with 40% white tef exhibited similar extent and variable rate of retrogradation kinetics along storage, while brown tef-blended breads retrograded slower but in higher extent than control wheat flour breads. Breads that contains 40% tef grain flour were found to contain five folds (DZ-01-99, DZ-Cr-387) to 10 folds (DZ-Cr-37) Fe, three folds Mn, twice Cu, Zn and Mg, and 1.5 times Ca, K, and P contents as compared to the contents found in 100% refined wheat grain flour breads. In addition, suitable dietary trends for lower rapidly digestible starch and starch digestion rate index were met from tef grain flour fortified breads.     

A rapid,automated method for measuring α-amylase in pre-harvest sprouted (sprout damaged) wheat

The quality of wheat for baking is critically dependent on the level of α-amylase (1,4-α-D-glucan glucanohydrolase, EC 3.2.1.1), which can be present as “late maturity α-amylase” (LMA), or due to pre-harvest sprouting due to high rainfall and humidity at the time of harvesting. The most commonly used method to measure α-amylase in wheat grain is the Hagberg Falling Number method, but values are also influenced by rheological properties of starch in the grain. In this study we describe a simple, rapid, automated method (Amylase SD) for measurement of α-amylase in pre-harvest sprouted (sprout damaged) wheat grain. The method (Amylase SD) measures the release of p-nitrophenol from 4,6-O-ethylidene-α-4-nitrophenyl-maltoheptaoside by α-amylase in the presence of α-glucosidase. The absorbance of p-nitrophenolate measured at 405 nm in a ChemWell^®-T auto-analyser is directly related to the level of α-amylase activity present in the milled wheat grain extract. The Amylase SD method generated <6%CV and correlation to the Falling Number method was represented by an inflection point at ∼160 s. The precision, sensitivity and speed of this method provides an ideal alternative to the Falling Number method for measurement of α-amylase (sprout damage) in wheat grain in wheat breeding programmes or at grain receival points.     

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.     

Non-cellulosic cell wall polysaccharides are subject to genotype × environment effects in sorghum (Sorghum bicolor) grain

Sorghum is a staple food for half a billion people and, through growth on marginal land with minimal inputs, is an important source of feed, forage and increasingly, biofuel feedstock. Here we present information about non-cellulosic cell wall polysaccharides in a diverse set of cultivated and wild Sorghum bicolor grains. Sorghum grain contains predominantly starch (64–76%) but is relatively deficient in other polysaccharides present in wheat, oats and barley. Despite overall low quantities, sorghum germplasm exhibited a remarkable range in polysaccharide amount and structure. Total (1,3;1,4)-β-glucan ranged from 0.06 to 0.43% (w/w) whilst internal cellotriose:cellotetraose ratios ranged from 1.8 to 2.9:1. Arabinoxylan amounts fell between 1.5 and 3.6% (w/w) and the arabinose:xylose ratio, denoting arabinoxylan structure, ranged from 0.95 to 1.35. The distribution of these and other cell wall polysaccharides varied across grain tissues as assessed by electron microscopy. When ten genotypes were tested across five environmental sites, genotype (G) was the dominant source of variation for both (1,3;1,4)-β-glucan and arabinoxylan content (69–74%), with environment (E) responsible for 5–14%. There was a small G × E effect for both polysaccharides. This study defines the amount and spatial distribution of polysaccharides and reveals a significant genetic influence on cell wall composition in sorghum grain.     

Antioxidant properties of wheat and rye bran extracts obtained by pressurized liquid extraction with different solvents

Rye and wheat brans are valuable sources of bioactive compounds, which could be used for the development and commercialization of high added value functional ingredients such as dietary antioxidants. The aim of this study was to evaluate antioxidant potential of rye and wheat bran using different polarity solvents. Cereal brans were ground to four different particle size fractions and extracted at 10.3 MPa pressure and 80 °C temperature by consecutive application of hexane, acetone and methanol:water (80:20%). The highest extract yield was obtained from rye bran using methanol-water; particle size in most cases had a significant effect. Antioxidant potential of extracts was assessed by ABTS^+•, DPPH^• scavenging, ORAC and total phenols content (TPC) assays. Extraction solvent had a major influence on TPC and antioxidant activity of the extracts. The most active extracts were obtained using methanol:water; rye bran extracts, in general, were stronger antioxidants than wheat bran extracts. For the majority of assays, reduction of particle size resulted in higher antioxidant activity values. However, ABTS^+• scavenging was found to decrease by decreasing particle size of rye bran used for extraction.     

Effects of post-anthesis fertilizer on the protein composition of the gluten polymer in a US bread wheat

Both genetic and environmental factors influence the types and amounts of wheat proteins that link together to form polymers essential for flour quality. To understand how plant growth conditions might influence gluten polymer formation, protein fractions containing small and large polymers were separated from flour from the US wheat Butte 86 grown in the absence or presence of post-anthesis fertilizer. Proteins in the polymer fractions were analyzed by quantitative two-dimensional gel electrophoresis (2-DE). The ratio of high molecular weight glutenin subunits (HMW-GS) to low molecular weight glutenin subunits (LMW-GS) increased in both fractions in response to fertilizer, due in part to small increases in the proportions of individual HMW-GS. There were also changes within the LMW-GS. In particular, omega and alpha chain terminators increased in proportion in both polymer fractions, but changes were more pronounced in the large polymer fractions. Serpins also increased in both polymer fractions. Additionally, the study revealed differences in the proportions of traditional LMW-GS in small and large polymer fractions. LMW-s type proteins were more abundant in the large polymers while LMW-i type proteins were more prevalent in the small polymers, suggesting that these proteins may play different roles in the gluten polymer.     

Pre-maturity α-amylase in wheat: The role of abscisic acid and gibberellins

The occurrence of pre-maturity α-amylase (PMA) is a major cause of poor bread-making quality (low Hagberg Falling Number) in wheat grain. In susceptible genotypes, it involves the excessive accumulation of high isoelectric point (pI) α-amylase in mature grain prior to germination and in the absence of pre-harvest sprouting. Several factors regulate PMA formation in developing grain, including genotype, agronomy, and environmental conditions. In particular, a cold period during mid-grain development has been found to be a major stimulus for PMA induction. Although the factors affecting the PMA occurrence are well known, little is known about the molecular mechanism governing its induction. The plant hormones abscisic acid (ABA) and gibberellins (GAs) influence various aspects of grain development, and it has been suggested that PMA involves changes in the amount of these hormones or the sensitivity of the grain to these hormones. This review summarizes recent studies investigating the role of ABA and GAs in PMA induction and PMA occurrence.     

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.     

Effect of enzymatic and technological treatments on solubilisation of arabinoxylans from brewer's spent grain

Brewer's spent grain (BSG), the most abundant brewing by-product, has hidden and underexploited nutritional potential. In order to valorize BSG, the effects of three commercial xylanases and a peptidase on water unextractable arabinoxylans (WUAX) were studied. Comparing all treatments, higher addition of xylanase resulted in an increase in water extractable arabinoxylans (WEAX). In the most efficient treatment, xylanase alone was able to solubilise 23.7% of WUAX, while the peptidase showed no effect. However, when added together with xylanase, peptidase increased the solubilisation of WUAX up to 1.6 folds. A positive correlation between particle size reduction and solubilisation of WUAX was also proved through milling BSG. These results suggest that access to xylan backbone increases with proteolytic activities, proving a synergistic effect of these specific enzymes. Therefore, if properly treated before being added as ingredient, BSG could add health functionalities to foodstuff while reducing the environmental impact of brewing industries.     

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.     

A single amino acid substitution in puroindoline b impacts its self-assembly and the formation of heteromeric assemblies with puroindoline a

Puroindolines (PINs) A and B were purified from soft (Paledor) and hard (Recital, Courtot) wheat cultivars. Their purity and heterogeneity due to post-translational processing were characterized by SDS- and acid-PAGE, reversed-phase HPLC and mass spectrometry. By using dynamic light scattering (DLS), asymmetrical flow field-flow fractionation (AF4) and size exclusion chromatography (SEC), we showed that the size distributions of PINA are similar for the three varieties and that, in solution, they self-assembled into small aggregates, mainly dimers. Conversely, PINB isolated from hard varieties (PINB-D1b and PINB-D1d) are assembled into large aggregates while PINB-D1a formed small aggregates, mainly monomers. Mixed solutions of PINA and PINB formed heteromeric aggregates. The large PINB-D1b aggregates were retained even at a high (4:1) PINA/PINB weight ratio. Reversible dissociation of large aggregates into small aggregates suggested that weak interactions control the self-assembly of PINs. The aggregative properties of PINs have now to be taken into account when studying their interactions with other components to decipher the causal relationships between these proteins and grain hardness.