High frequency of abnormal high molecular weight glutenin alleles in Chinese wheat landraces of the Yangtze-River region

A total of 485 common landraces of bread wheat were collected from the Yangtze-River region of China. Their high molecular weight glutenin subunit (HMW-GS) composition was analyzed by Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry (MALDI-TOF-MS). Among all landraces tested, 453 were homogeneous for HMW-GS, 32 were heterogeneous, and 37 contained abnormal subunits. A total of 22 alleles were detected, including 3 at Glu-A1, 13 at Glu-B1 and 6 at Glu-D1, respectively. Higher variations occurred at the Glu-B1 locus compared with Glu-A1 and Glu-D1. Glu-A1c (74.0%), Glu-B1b (40.4%), Glu-D1a (84.9%) appeared to be the most frequent alleles at Glu-A1, Glu-B1 and Glu-D1, respectively. Two alleles ("null" and 1) at the Glu-A1 locus, three allele compositions (7 + 8, 7OE + 8, 7 + 9) at the Glu-B1 locus, and two (2 + 12 and 5 + 10) at the Glu-D1 locus appeared to be the common types in the 485 landraces. Sixteen new alleles represented by abnormal subunits were identified at the Glu-B1 and the Glu-D1 locus.     

Properties of bread made from frozen dough containing waxy wheat flour

The quality of bread made from frozen dough is diminished, and staling rate is increased by changes that occur during freezing and storage. New cultivars of waxy wheat flour (WWF), containing higher levels of amylopectin, may help improve the quality of baked products. Bread quality and staling were investigated for bread containing 0–45% WWF and 55–65% water after freezing and 90-day frozen storage. The specific volume was highest with 15% WWF substitution and 60% water in bread made from both unfrozen and frozen dough. With higher levels of WWF and lower water content, bread staling rates decreased. Bread with higher levels of WWF were darker and had greater color variation. ¹H NMR studies showed that bread with greater WWF and water had higher transverse relaxation (T₂) times (9–11 ms), but less change in T₂ during storage. This research demonstrated that specific combinations of WWF and water produced a better quality of bread after dough freezing.     

Quality characteristics of northern-style Chinese steamed bread prepared from soft red winter wheat flours with waxy wheat flour substitution

Quality characteristics of northern-style Chinese steamed bread (CSB) prepared from two soft red winter (SRW) wheat flours blended with 0–30% waxy wheat flour (WWF) were analyzed to estimate the influence of starch amylose content. The increased proportion of WWF in blends raised mixograph absorption with insignificant changes in protein content and dough strength-related parameters. WWF incorporation generally increased specific volume and crumb softness of CSB. The analysis of covariance revealed that CSB quality attributes were little affected by protein content and dough strength-related parameters, indicating that starch amylose content was largely responsible for the changes in CSB quality. Flour blends with 5–10% WWF, of which starch amylose content was 22.4–24.7%, produced CSB with superior crumb structure compared to other blends, but insignificant changes in surface smoothness, stress relaxation and total score compared to the respective control wheat flours. Flour blends with 15% WWF to produce a starch amylose content of 21.4–22.7% exhibited reduced staling of CSB with total scores comparable to the respective control wheat flours. CSB prepared from blends with more than 10% WWF exhibited a higher soluble starch content, indicative of reduced starch retrogradation, than that prepared from wheat flours without WWF during storage for 3 days.     

Functionality of native tetraploid wheat starches: Effects of waxy loci alleles and amylose concentration in blends

Partial waxy (reduced amylose) and fully waxy (amylose-free) tetraploid durum wheats (Triticum turgidum L. var. durum) were used to investigate the relationships between both intra- and inter-granular variation in amylose concentration and starch functionality. Starches isolated from each genotype (intra-granular amylose variants) were compared to those of commercially available hexaploid wild-type and waxy starches, and functionalities compared to blends (inter-granular variants) of durum waxy and wild-type starches of 0, 6, 12, 18, 24, & 30% amylose content. Starch particle size distributions were not significantly different amongst partial waxy and wild-type genotypes; waxy samples had significantly smaller mean starch granule size. Few significant differences for crystallite melting and related intrinsic heat as determined by differential scanning calorimetry (DSC) were observed. The detected differences in starch gel color or gel strength generally were observed for the waxy samples relative to wild-type. Numerous significant differences were observed via Rapid-Visco Analysis (RVA). Pasting peak viscosity and breakdown were inversely proportional to % amylose. Wx-B1 null final viscosity differed significantly from that of all other blends and genotypes, demonstrating that genotypic differences exist amongst the partial waxy types, independent of amylose concentration per se.     

Comparative analyses of LMW glutenin alleles in bread wheat using allele-specific PCR and SDS-PAGE

One hundred and eighty-two bread wheat cultivars developed in India were characterized for low molecular weight (LMW) glutenins using SDS-PAGE and allele-specific polymerase chain reaction (PCR) to assess allelic diversity encoded by Glu-3 loci, as well as their utility for correctly identifying different alleles. SDS-PAGE indicated Glu-A3c is present in 64.6% of the cultivars, Glu-A3b in 13.8%, Glu-A3d in 12.7% and Glu-A3e/f in 8.8%. Seven types of alleles were present at the Glu-B3 locus: Glu-B3b (29.3%), Glu-B3g (27.0%), Glu-B3h (13.8%), Glu-B3i (16.1%), Glu-B3j (12.1%), Glu-B3c (0.6%) and Glu-B3d (1.1%). SDS-PAGE found three types of Glu-D3 alleles: Glu-D3a (30.2%), Glu-D3b (67.1%) and Glu-D3c (2.7%). However, PCR found two different alleles in cultivars classified as carrying Glu-D3a and three alleles in those identified as carrying Glu-D3b cultivars, indicating a more complex nature of the Glu-D3 locus. In conclusion, the data found greater consistency between the SDS-PAGE and PCR amplification patterns of alleles such as Glu-A3c, Glu-A3d, Glu-B3g, Glu-B3h and Glu-B3i, and less consistency between those same patterns in the Glu-A3b, Glu-A3e/f and Glu-B3b alleles. More studies are needed in order to achieve unambiguous identification of the Glu-3 alleles and thereby allow their greater utility in germplasm evaluation and breeding.     

Influencing factors of sodium dodecyl sulfate sedimentation in bread wheat

Bread wheat elite lines and F4 populations were evaluated to determine the influence of genotype and environment on variation in sodium dodecyl sulphate (SDS) sedimentation, and the relation between SDS sedimentation and other quality characteristics. Nine intermediate hard red wheat elite lines and two checks were evaluated for three years over eight locations, and six F4 populations and two hard red wheat checks were evaluated at three locations. In both sets of material, the genotype and location main effect, and genotype × location interaction were highly significant. The genotype component contributed 85.96% of the total variation in SDS sedimentation in the F4 material, and the genotype × location component only 12.87%. In the elite material the contribution of genotype was high enough to make effective selection for SDS sedimentation possible. The genotype × year effect was large, indicating that testing genotypes across years may be more important than across locations. SDS sedimentation was significantly positively correlated with protein content and mixograph development time, and negatively with yield. Selection of higher SDS sedimentation may lead to overly strong dough and lower yields. Therefore a careful approach should be taken in the selection process, balancing the different objectives in a breeding program.     

Volume, texture, and molecular mechanism behind the collapse of bread made with different levels of hard waxy wheat flours

Physico-chemical properties of bread baked by partially replacing normal wheat (Triticum aestivum L.) flour (15, 30, and 45%) with two hard waxy wheat flours were investigated. Substitution with waxy wheat flour resulted in higher loaf volume and softer loaves. However, substitution at >30% resulted in excessive post-bake shrinkage and a ‘key-hole’ shape with an open crumb structure. Bread crumb microstructure indicated a loss of starch granule rigidity and fusing of starch granules. The cells in the interior of the bread did not become gas-continuous and as a result, shrunk as the loaf cooled. Soluble starch content was significantly higher in bread crumb containing waxy wheat flour than in control bread. Debranching studies indicated that the soluble starch in bread made with 30-45% hard waxy wheat flour was mostly amylopectin. Incorporation of waxy wheat flour resulted in softer bread immediately after baking but did not retard staling upon storage.     

Phenolics in the bran of waxy wheat and triticale lines

The present study was designed to determine total phenolic acid contents (TPC) and compositions of bran from newly developed near-isogenic waxy wheat and triticale translocation lines. Two waxy wheat sets, Svevo (durum) and N11 (bread wheat), consisting of partial and waxy null lines and four sets of triticales (GDS7, Trim, Rhino and Rigel) having translocations at 1A.1D and 1R.1D with high molecular weight glutenin subunits (HMW-GS) 5 + 10 and 2 + 12 were investigated. Similar to non-waxy wheat, ferulic acid was the predominant phenolic acid found in waxy wheats analyzed. Two other major phenolics include p-coumaric and vanillic acids followed by lesser quantities of syringic acid. Waxy lines had higher TPC than the parent line in the N11 set, whereas the Svevo set showed the opposite trend. TPC of waxy bread wheats were correlated with amylose fractions in which the order was complete waxy < double waxy nulls < single waxy nulls. Lines with HMW-GS 2 + 12 have lower TPC than other lines in each group of triticales, except the Trim set. TPC was negatively correlated (r = −0.41; p > 0.1) with bran yields in triticale lines studied, indicating that variation in phenolics was not only due to bran yields but also to genotypic differences.     

Influence of puroindoline alleles on milling performance and qualities of Chinese noodles,steamed bread and pan bread in spring wheats

Two trials with a total of 75 spring bread wheat cultivars and advanced lines, were used to evaluate single kernel characterization system hardness, puroindoline alleles, milling yield, flour ash content, flour colour, and end-use qualities for Chinese noodles, steamed bread and pan bread. The results indicated that all International Maize and Wheat Improvement Center hard wheat lines surveyed were Pina-D1b genotype, whereas Pinb-D1b was the most common allele in the remaining cultivars. Genotypes with Pinb-D1b possess significantly lower flour ash content and higher milling yield than those of genotypes with Pina-D1b. For steamed bread, mean scores for loaf volume, crumb colour, width, structure and total score of Pinb-D1b genotypes were significantly higher than those of genotypes with Pina-D1b and wild type. For Chinese fresh white noodles, means for noodle a*, colour score, viscoelasticity and total score with Pinb-D1b were significantly higher than those of Pina-D1b and wild type. Means of loaf volume, texture and total score for Pinb-D1b genotypes were significantly higher than those of Pina-D1b genotypes. This study further illustrated the superiority of the Pinb-D1b allele on milling and processing qualities for various end products and provides useful information for wheat quality improvement.     

Genetic control of processing quality in a bread wheat mapping population grown in water-limited environments

End-use quality of bread wheat (Triticum aestivum L.) is affected by both genetic and environmental variation. Current understanding of the genetic control of wheat quality traits is mainly based on genetic experiments conducted using grain produced in favourable conditions. The objective of this research was to extend the genetic analysis of these traits by using grain produced in water-limited environments. Grain samples harvested from a mapping population grown in field experiments at two locations in Australia were used to assess characteristics of the grain, flour, dough and bread. Quantitative trait loci (QTLs) were mapped. The parents of the population, RAC875 and Kukri, differ at several loci that are known to affect grain quality or plant phenology. Of these, a high-molecular-weight glutenin locus (Glu-B1) affected dough properties, the puroindoline-encoding Ha locus affected grain hardness, flour and loaf properties and a photoperiod response locus (Ppd-D1) affected flour extraction and protein content. Similarly, several previously reported quantitative trait loci (not associated with specific genes) also had effects in the stress environments used here. In addition, novel loci were detected for bread wheat quality traits; their effects may be specific to materials grown in water-limited environments.     

Biochemical and sensory evaluation of wheat bran supplemented sorghum bread

Addition of wheat bran to sorghum flour (Dabar cultivar) at two extraction rates 72% and 80% resulted in lowering reducing sugars. The percent decreasewas 75.6% compared with the control at the end of fermentation period. There was a highly significant (p 0.05) increase in crude fiber content as a result of addition of wheat bran. The increase was from 0.8 to 5.2 and from 0.5 to 5.3% for the 80% S/WB and 72% S/WB blends, respectively. Sorghum bread containing wheat bran was lower in reducing sugars and showed a significant increase (p 0.05) in starch content. Sorghum bread containing wheat bran resulted in a lower in vitro protein and starch digestibilities.     

Effect of the three waxy null alleles on enzymes associated to wheat starch granules using proteomic approach

In bread wheat (Triticum aestivum L.), waxy genes are present at three loci (Wx-A1, Wx-B1 and Wx-D1) and are responsible for amylose synthesis in the grain. Near isogenic lines (NILs) of the agronomic cultivar Tremie with either one, two or three waxy null alleles were used to study the genome response to these null alleles through proteomic analysis of starch granule associated proteins of mature grain. Among the 352 spots that were revealed, 86 varied significantly between NILs. The main differences were observed for the granule bound starch synthase (GBSS), product of the waxy genes. A strong relation (R² = 0.94) between the amount of GBSS per gram of starch granule and the amylose quantity in the grain was revealed through quantification of the GBSS in each NIL. The analysis of GBSS quantity for each NIL invalidated the additive hypothesis for the expression of this enzyme in (Wx-A1a Wx-D1a Wx-B1b) (Wx-D1a Wx-B1a Wx-A1b) and the normal form of Tremie. In addition, four proteins identified as inactive GBSS were increased (+144.9%) in the triple null isogenic form compared to the normal form of Tremie. The quantitative variations of the proteins present in starch granules support the idea of regulatory mechanisms between genomes.     

Effect of heating temperature on the particle size distribution in waxy wheat flour

The particle size of waxy (amylose-reduced) wheat (Triticum aestivum L.) starch was determined at isothermal temperatures by laser diffraction analysis. Flour samples were suspended in deionized water at temperatures ranging from 30 to 90 °C for 20–60 min. At 30 °C, all of the flour particles exhibited trimodal size distributions, i.e., the particles in the first, second, and third modes were <10 μm, 10–50 μm, and 51–300 μm, respectively. Control experiments with isolated starch indicated that the first and second modes were associated mainly with starch granules, whereas the third mode may have been related to gluten and gluten adhesion. The particle size distributions of waxy segregant wheat flours were temperature dependent. At 60 °C, there were significant changes in the particle size and distribution of waxy flours, which indicated the swelling of starch granules in response to elevated temperature. As the temperature increased, the peak particle size of waxy segregant wheat flours increased in different ways. The results suggest that variations in the swelling properties of selected waxy genotype flours may be due to the strength of starch–protein interaction and the capacity for starch granule gelatinization.     

Association analysis reveals effects of wheat glutenin alleles and rye translocations on dough-mixing properties

The glutenin loci of wheat (Triticum aestivum L.) are important determinants of bread-making quality, although the effects of alleles at those loci are incompletely understood. We applied an association analysis method to assess the effects of glutenin alleles and 1RS wheat–rye (Secale cereale L.) translocations on dough-mixing properties in 96 wheat cultivars and advanced lines grown at three Colorado locations while accounting for population structure and relatedness of individuals in the population. The results indicated that (1) in the majority of cases, controlling relatedness of individuals reduced the significance of associations between glutenin loci and Mixograph traits; (2) the Glu-D1 and Glu-B3 loci and 1RS translocations had greater impacts on dough-mixing properties compared to other glutenin loci; (3) Glu-B1w, Glu-D1d, and Glu-B3b were consistently associated with greater (more favorable) Mixograph peak time (MPT) than other alleles at the respective loci, whereas Glu-B1e, Glu-D1a, and Glu-B3c were associated with reduced MPT; (4) the 1BL.1RS translocation was associated with a decrease in Mixograph properties. Our results indicate that taking multiple-level relatedness of individuals into account can improve the results of association analysis for wheat-quality traits.     

Effect of wx genes on amylose content,physicochemical properties of wheat starch,and the suitability of waxy genotype for producing Chinese crisp sticks

The effect of wx genes on amylose content, physicochemical properties of wheat starches, and the quality of Chinese crisp stick were investigated using near-isogenic lines (NILs) with null wx alleles in Yangmai 17 and Yangmai 01-2 backgrounds. wx genes showed significant effects on amylose content and other traits. The triple-null genotype had the lowest amylose content among eight genotypes, followed by double-null, single-null, and wild-type genotypes. The triple-null also showed lower flour yield, higher percentage of type B-granules on a volume basis and higher crystallinity than non-waxy genotypes, and showed significant differences in all pasting and thermal transition parameters compared to non-waxy genotypes, except for degree of retrogradation at day 14. For the quality of Chinese crisp stick, the hardness, crispness, fracturability, and specific volume of waxy genotype were 3.91 kg, 11.0, 1.85 mm and 104.4 ml, whereas the corresponding ranges for non-waxy genotypes were 5.39–5.70 kg, 0.5–0.9, 0.69–0.86 mm and 49.5–57.6 ml, respectively, in Yangmai 17 background. This indicates that waxy genotypes showed significantly better crisp stick quality than non-waxy genotypes. A similar trend was also observed in Yangmai 01-2 background. This indicated the potential utilization of waxy wheat for producing traditional products.     

Effects of two novel Wx-A1 alleles of common wheat (Triticum aestivum L.) on amylose and starch properties

Common wheat (Triticum aestivum L.) has three Wx proteins (Wx-A1, Wx-B1, and Wx-D1), which are granule-bound starch synthases I and are responsible for the amylose synthesis of flour starch. The effects of two novel Wx-A1 proteins identified by gel electrophoresis on amylose content and starch properties were analyzed. The variant Wx-A1 protein coded by the Wx-A1i allele was present in smaller amounts and produced less amylose (7.3%) compared to standard lines with the Wx-A1a allele (21.5%). Wx-A1i generated altered starch properties; greater swelling power (SP), glucoamylase digestibility, starch paste clarity, and gelatinization enthalpy in differential scanning calorimetry. The starch from Wx-A1i also showed an altered pasting profile on a Rapid Visco-Analyzer, greater peak viscosity, smaller final viscosity, and lower pasting temperature. The Wx-A1i allele is a novel genetic resource for reducing amylose content in wheat. The other Wx-A1 protein coded by the Wx-A1j allele showed a more basic isoelectric point compared to Wx-A1a on an electrophoretic gel. The amylose content of Wx-A1j did not differ from standard Wx-A1a. Starch SP, paste clarity, and glucoamylase digestibility also suggested that Wx-A1j produced amylose as much as Wx-A1a.     

Chemical changes in wheat pan bread during storage and how it affects the sensory perception of aroma,flavour, and taste

Wheat bread and whole wheat bread were stored for up to 3 weeks and specific chemical reactions were related to perceptual flavour changes as analysed by sensory profiling. Volatile and non-volatile compounds were quantified using GC–MS and HPLC. Results were examined by multivariate data analysis.     

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.     

Effects of ozone treatment on the molecular properties of wheat grain proteins

Ozone is a powerful and highly reactive oxidizing agent, which has found increasing applications in the field of grain processing. However, in some cases, O₃ can potentially promote oxidation and/or degradation of the chemical constituents of grains. Experiments were carried out to evaluate the specific effects of gaseous ozone on the molecular properties of wheat grain proteins and their consequences on the bread-making quality of the resulting flours.Ozonation causes a significant reduction in the SDS solubility of the wheat prolamins, which can reasonably be attributed to conjugate effects of an increase in molecular dimensions and an increase in the compactness of the protein polymers initially present. In fact, our results demonstrate that this general reinforcement of the aggregative status of prolamins due to ozonation of wheat grains results from (i) the formation of new intermolecular S-S bonds, (ii) to a lesser extent, the formation of other types of intermolecular covalent cross-links (dityrosine cross-links) and finally, (iii) significant changes in secondary structure. By significantly affecting the molecular properties of wheat grain prolamins, ozone leads to profound changes in the rheological properties (i.e. increase in the tenacity and a great limitation of the extensibility) of the flours and/or doughs obtained.     

Quantitative proteomic analysis of wheat grain proteins reveals differential effects of silencing of omega-5 gliadin genes in transgenic lines

Novel wheat lines with altered flour compositions can be used to decipher the roles of specific gluten proteins in flour quality. Grain proteins from transgenic wheat lines in which genes encoding the omega-5 gliadins were silenced by RNA interference (RNAi) were analyzed by quantitative 2-dimensional gel electrophoresis (2-DE). The precise effects of the genetic modifications on the proteome were assessed in four homozygous lines generated with the same RNAi construct. Two of the lines showed >80% decreases in omega-5 gliadins with only small changes in the levels of other gluten proteins. In the other two lines, omega-5 gliadins were not detectable by 2-DE. However, there were notable reductions in all omega-1,2 gliadins. Small decreases in several other gluten proteins were also detected in one of the lines. The other line showed notable decreases in three HMW-GS and an s-type LMW-GS, increases in two m-type LMW-GS and several alpha gliadins, as well as increases in both serpins and triticin. The study demonstrates that the same RNAi construct can have differential effects on the wheat grain proteome and highlights the importance of detailed proteomic analyses of transgenic grain prior to selecting lines for further assessment of flour quality and allergenic potential.