Characterisation of starch during germination and seedling development of a rice mutant with a high content of resistant starch

To acquire a better understanding of whether RS influences the dynamics of in vivo starch digestion and seed vigour, the high-RS rice mutant RS4 (RS ca. 10%) and the wild type R7954 were used to investigate total amylase activity, seedling vigour, starch content and starch granule structure during germination. RS4 exhibited similar seed vigour to R7954. Amylose and amylopectin in R7954 showed synchronous degradation throughout the whole process, while amylopectin was hydrolysed significantly faster than amylose in RS4 during the earlier germination stages. The starch residues of RS4 after germination (GD) lost endotherm peaks and showed a special X-ray diffraction pattern with only two peaks at around 16.90° and 21.62°, probably due to remnants of amylopectin and its tight crosslinking with the cell wall. The remaining starch after 10 GD, primarily amylopectin may make a critical contribution to total resistant starch content. These results indicated that RS had no negative impact on seed vigour in rice lines, although RS cannot be hydrolysed by α-amylase from human and animal in vitro. By appropriately increasing the special amylopectin fraction, a new breeding programme of high RS crops and improvement in the eating quality of high RS rice varieties might be achieved.     

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.     

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.     

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.     

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.     

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.     

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.     

In vitro starch digestibility and predicted glycaemic indexes of buckwheat,oat, quinoa,sorghum, teff and commercial gluten-free bread

The in vitro starch digestibility of five gluten-free breads (from buckwheat, oat, quinoa, sorghum or teff flour) was analysed using a multi-enzyme dialysis system. Hydrolysis indexes (HI) and predicted glycaemic indexes (pGI) were calculated from the area under the curve (AUC; g RSR/100g TAC*min) of reducing sugars released (RSR), and related to that of white wheat bread. Total available carbohydrates (TAC; mg/4 g bread “as eaten”) were highest in sorghum (1634 mg) and oat bread (1384 mg). The AUC was highest for quinoa (3260 g RSR), followed by buckwheat (2377 g RSR) and teff bread (2026 g RSR). Quinoa bread showed highest predicted GI (95). GIs of buckwheat (GI 80), teff (74), sorghum (72) and oat (71) breads were significantly lower. Significantly higher gelatinization temperatures in teff (71 °C) and sorghum flour (69 °C) as determined by differential scanning calorimetry (DSC) correlated with lower pGIs (74 and 72). Larger granule diameters in oat (3–10 μm) and sorghum (6–18 μm) in comparison to quinoa (1.3 μm) and buckwheat flour (3–7 μm) as assessed with scanning electron microscopy resulted in lower specific surface area of starch granules. The data is in agreement with predictions that smaller starch granules result in a higher GI.     

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.     

Modification of physicochemical properties and in vitro digestibility of wheat flour through superheated steam processing

Native and moistened wheat flours (moisture contents were 13.5 and 27.0%, respectively) were treated with superheated steam (SS) at different temperatures (140 and 170 °C) and times (1, 2 and 4 min). Their physicochemical and digestive properties were analyzed. For native flour, SS treatment altered the starch molecular structure and behavior slightly. While for moistened flour, crystalline degree, gelatinization enthalpy, amylose leaching (AML) and falling number significantly decreased, but thermal transition temperatures increased with the rise of treating severity. Clumping of starch granules, aggregation of proteins and formation of amylose-lipid complexes occurred in both native and moistened flours. Broader pasting temperature ranges and higher viscosities were found on SS-modified flours. Additionally, SS treatment on moistened flours increased resistant and slowly digestible starch contents. In general, SS treatment induced changes in starch molecular structure and reactions among flour components leading to more stable structures, thus affecting their pasting behavior, thermal properties and in vitro digestion.     

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.     

Investigating the impact of α-amylase, α-glucosidase and glucoamylase action on yeast-mediated bread dough fermentation and bread sugar levels

Wheat flour is generally supplemented with α-amylases to increase maltose levels in bread dough and increase loaf volume. While the preference of yeast for glucose and fructose over maltose as substrate for fermentation is well documented, the impact of maltose versus glucose producing enzymes on bread dough fermentation kinetics and bread sugar levels is ill documented. Hence the impact of α-amylase, α-glucosidase and glucoamylase action on both aspects was investigated. Glucoamylase and α-amylase increase the total fermentable sugar content of dough, while α-glucosidase only affects the glucose/maltose ratio. Due to their effect on total fermentable sugar levels, addition of α-amylase or glucoamylase prolongs the total productive fermentation time, while this is not the case for α-glucosidase. In contrast to α-amylase, both glucoamylase and α-glucosidase supplementation leads to higher CO₂ production rates during the initial stages of fermentation. In the final bread product, different sugar levels are observed depending on the dosage and type of starch-degrading enzyme. The results of this study imply that long and short fermentation processes benefit from α-amylase and α-glucosidase addition, respectively, while glucoamylase supplementation is suitable for both long and short fermentation times.     

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.     

Effects of selected extrusion parameters on physicochemical properties and in vitro starch digestibility and β-glucan extractability of whole grain oats

Whole grain oat flour was extruded under different moisture contents (15%, 18%, 21%), barrel temperatures (100 °C, 130 °C), and screw speeds (160 rpm, 300 rpm, 450 rpm), and selected physicochemical properties, in vitro starch digestibility, and β-glucan extractability of the extrudates were analyzed. An increase in screw speed resulted in an increase in radial expansion index, water absorption index, and water solubility index. Screw speed significantly affected slowly and rapidly digestible starch. Moderate screw speed (300 rpm) led to higher slowly digestible starch with an accompanying decrease in rapidly digestible starch. Low moisture conditions (15%) resulted in the highest resistant starch and water-extractable β-glucan. Under the conditions used in this study, extrusion did not result in changes in water-extractable β-glucan molecular weight. Thus, extrusion might be beneficial in improving functionality and consumer acceptability by affecting physicochemical properties, in vitro starch digestibility, and β-glucan extractability of oat extrudates.     

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.     

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.     

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.     

High pressure processing manipulated buckwheat antioxidant activity,anti-adipogenic properties and starch digestibility

Effect of high pressure processing (HPP) on buckwheat nutritional properties is investigated in this study. The results indicated that the digestion of buckwheat starches of untreated and HPP treated at room temperature (RT) showed an “internal corrosion” pattern. However, an “exocorrosion” digestion behavior was observed for the starches of HPP treated sample at 45 °C, which might be due to the formation of amylose–lipid complexes during the process as revealed by X-ray diffraction (XRD). HPP treatment at 45 °C increased the antioxidant activity of the buckwheat as measured using 1,1′-diphenyl-2-picrylhydrazylvalue (DPPH) and iron chelating capacity (ICC) compared to untreated sample (P < 0.05), which might be associated with the release of bound phenolic compounds. More importantly, this is the first study to investigate the ability of buckwheat extract for inhibiting the formation of fat droplets using a C3H10T1/2 mesenchymal stem cell line, and found that the extract of buckwheat after HPP treatment at 45 °C demonstrated significant anti-adipogenic effects. This study suggested the HPP treatment at an increased temperature (45 °C) achieved a better nutritional value of the buckwheat than both untreated and treated at RT.     

Determination of volatile compounds in heat-treated straight-grade flours from normal and waxy wheats

Volatile compounds formed during heat-treatment of wheat flour influence the application of treated flour. In this study, normal and waxy hard wheat flours before and after dry-heat treatment were subjected to headspace analysis by solid-phase microextraction of volatiles followed gas chromatography–mass spectrometry (GC/MS). The untreated waxy wheat flour contained higher levels of odor-active compounds than normal wheat flour including aldehydes, alcohols, furans, and ketones. Lipid oxidation appears to play major role in producing such odor compounds. Heat treatments, depending on the severity, alter the profile of volatile compounds. Low temperature (100–110 °C) treatments effectively eliminated cereal odor (aldehyde) and did not introduce additional odors, providing a possible way to produce low-odor flours. Heat treatments at 120 °C and higher temperatures elevated the content of pyrazines, furans, and sulfur-containing compounds which together gave a roasty aroma to the flours. Considering organoleptic properties, treatments of flours at 140 °C was superior to 160 °C. The waxy wheat flour was more prone to produce odor-active compounds than normal wheat flour during the same heat treatment.