Effect of genetic variation on phenolic acid and policosanol contents of Pegaso wheat lines

Total phenolic acid and policosanol contents and compositions of bran from an Italian bread wheat variety Pegaso and its 11 near-isogenic lines were measured. The near-isogenic wheat lines differed at one or more loci coding for storage proteins. The genetic variation included deletions, additions and/or combinations of variations. Almost 95% or more of phenolic acids were in the bound form. Ferulic acid was the predominant bound phenolic acid present in wheat bran samples. Other phenolic acids were p-coumaric, vanillic and syringic acids. Tetracosanol, docosanol, hexacosanol, octacosanol, tricosanol and heneicosanol were found as major policosanol compounds in their decreasing order. Highly significant genotypic differences were observed in total phenolic acid and policosanol concentrations. None of the genetic lines had higher phenolic acid contents than the parent line Pegaso, whereas some of the lines had more policosanol levels. In general, both total phenolic acid composition and contents were higher with genetic lines that varied at Glu-1 loci with 2+Dy high molecular weight glutenin subunit (HMW-GS) (Pegaso 184), variation at Gli-D2 loci (Pegaso 219) and single null A1 (variation at Gli-1/Glu-3 loci; Pegaso 30). Highest total policosanol content was observed with the double null at Glu-A1/Glu-D1 loci (Pegaso 236). These findings may lead to new opportunities for wheat breeders and eventually commercial wheat growers to promote the production of wheat with enhanced levels of health beneficial compounds.     

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.     

Identification and quantification of phenolic acids and anthocyanins as antioxidants in bran,embryo and endosperm of white,red and black rice kernels (Oryza sativa L.)

Whole rice grain comprising endosperm, embryo (or germ), and bran has potential beneficial health effects in addition to provision of nutrients. The distribution of phenolic acids and anthocyanins in endosperm, embryo, and bran of white, red, and black rice grains was investigated in this study. The total phenolic content (TPC) was highest in the bran, averaging 7.35 mg GAE/g and contributing 60%, 86% and 84% of phenolics in white, red and black rice. The average TPC of the embryo and endosperm were 2.79 and 0.11 mg GAE/g accounting for 17% and 23%, 4% and 10% and 7% and 9% in white, red and black rice, respectively. The antioxidant capacity determined using DPPH and ORAC displayed a trend similar to TPC. Free/conjugated phenolic acids in white, red and black rice bran accounted for 41%, 65% and 85% of total acids. Bound phenolic acids in rice bran accounted for 90% of total acids in whole grain. Cis-p-coumaric was detected in bound form in bran while cis-sinapic acid was detected in the free/conjugated form in embryo and bran. Cyanidin-3-O-glucoside and peonidin-3-O-glucoside were identified mainly in black rice bran as the total anthocyanins. Cyanindin-3-O-rutinoside was also detected in black rice bran.     

PCR-based markers for identification of HMW-GS at Glu-B1x loci in common wheat

The bread wheat elasticity, which is very important for bread-making quality, is largely determined by the composition of high-molecular-weight glutenin subunits (HMW-GS). The HMW-GS encoded by Glu-B1 loci are highly polymorphic and the combinations 17+18 and 14+15 are good for bread making. Thus it is very important to identify the alleles at Glu-B1 loci for wheat quality improvement. In this study, the five common HMW-GS types encoded by Glu-B1x locus carried by 18 Chinese bread wheat cultivars (or lines) were analyzed by SDS-PAGE. Two pairs of PCR primers which could distinguish the Glu-Blx alleles of the five common HMW-GS types were designed based on the Glu-B1x gene sequences (Reddy and Appels, 1993; Genbank accession: X13927; Genbank accession:AY367771). 22 recombinant inbred lines (RILs) derived from Jing711 (contains 17 subunit on Glu-B1x) and Pm97034 (contains 14 subunit on Glu-B1x) were used to validate the accuracy of the primers, which showed that the two specific markers could be used together to distinguish alleles at Glu-B1x locus and accelerate wheat quality breeding by marker assisted selection.     

Phenolic acids composition,total polyphenols content and antioxidant activity of Triticum monococcum, Triticum turgidum and Triticum aestivum: A two-years evaluation

Wheat is a good source of polyphenols, plant metabolytes with beneficial effects on human health. However, little information is available on phenolic acid composition and concentration in different Triticum species, as well as on possible environmental effects. To shed some light on this issue, thirty-nine wheat accessions cropped for two years and belonging to different Triticum species (Triticum monococcum, Triticum turgidum ssp. dicoccum, turanicum and durum, Triticum aestivum ssp. spelta and aestivum) were assessed for phenolic acids (ferulic, p-coumaric, vanillic, syringic, p-hydroxybenzoic, caffeic acids and syringaldehyde), total polyphenols and antioxidant activity in soluble conjugated and insoluble bound extracts. Ferulic acid was the most abundant compound in both extracts. Insoluble bound phenolic acids represented >90% of total phenolic acids. Einkorn showed the maximum concentration of conjugated phenolic acids (50.5 mg/kg DM), while durum and bread wheats presented the highest content of bound phenolic acids (651.8 and 629.2 mg/kg DM, respectively). Cropping year influenced the concentration of conjugated but not of bound phenolic acids. A survey of phenolic acid distribution in the kernel showed that they are rare in endosperm, but abundant in germ and bran. Total polyphenols and antioxidant activity were highly correlated to phenolic acid content.     

Effects of region and cultivar on alkylresorcinols content and composition in wheat bran and their antioxidant activity

This study evaluated the effects of cultivar and region on the composition of alkylresorcinols (ARs) of 24 wheat bran samples from 6 cultivars grown in four locations (Bath, Nairn, Palmerstone and Ridgetown) in Ontario (ON), Canada, using gas chromatography/mass spectrometry (GC/MS). Total phenolic content (TPC) of wheat bran extracts was determined by the Folin–Ciocalteau method and the antioxidant activity of wheat bran extracts was measured by 2, 2-Dipheny-1-picryhydrazyl radical (DPPH) scavenging activity and oxygen radical absorbance capacity (ORAC). The highest ARs content (μg/g) was found in cultivars Emmit (1522), Harvard (1305), Warthog (1170), and Superior (853), grown in Ridgetown. The relative saturated and unsaturated ARs (%) were 89 and 11, respectively. Total ARs content, their composition, TPC and antioxidant activity of wheat bran extracts were significantly (P < 0.05) affected by location and cultivar and their interaction. TPC, %DPPH_dis, and ORAC values for different wheat bran sample extracts ranged from 3 to 58 (mg FAE/g), 5 to 68 (%), and 6 to 94 (μmol TE/g), respectively. Our work provides a detailed examination of region and cultivar effects on potential of ARs in wheat bran and the results can be used for screening and breeding purposes.     

Differential feeding by grasshoppers and levels of foliar diseases in various cultivars of spring cereals

Sixty-four cultivars and strains of cereals were evaluated under field conditions for differential feeding by grasshoppers, (Orthoptera:: Acrididae) and for their resistance to foliar disease. Grasshoppers actively fed on all triticale and durum wheat lines, on 11 of 12 bread wheat lines, and on 11 of 24 barley lines. Both the incidence and severity of foliar disease were high in all cereals. There was no correlation between grasshopper damage and resistance to foliar disease (r+ −0·059) in the barley but a positive trend appeared in wheat lines (bread r= +0·487; durum r= +0·295). The data suggest that present trends in breeding disease-resistant cereal cultivars will not introduce increased susceptibility to grasshopper damage.     

Wx-1基因变异和优质HMW-GS组合对中筋小麦品质的影响

高分子量谷蛋白亚基(HMW-GS)和Wx蛋白是两个影响小麦理化品质和制成品品质的重要因素。为研究HMW-GS和Wx基因变异对小麦理化品质、面条感官评分和质构特性的影响,以镇麦9号和扬糯麦1号为亲本的2个高代系为试验材料,研究了优质HMW-GS 5+10及Wx基因缺失对小麦理化品质及面条加工品质的效应。结果表明,品系1与品系2的湿面筋含量和SDS沉淀值与镇麦9号相近,但显著好于扬糯麦1号。相较于镇麦9号,品系1与品系2的直链淀粉含量分别降低了1.5%和2.5%,其峰值黏度和稀懈值均显著高于镇麦9号,膨胀势也高于镇麦9号。5+10亚基显著提高了面条质构参数中的硬度和咀嚼性,而Wx基因缺失显著提高了面条的软硬度评分和光滑性评分。综合来看,具有5+10亚基和 Wx-D1缺失型的品系2具有较高的SDS沉淀值、面团形成时间和稳定时间、较低的直链淀粉含量、较高的峰值黏度、稀懈值和膨胀势,蛋白质和淀粉综合品质表现较好。品系2面条总评分最高,显著高于镇麦9号和扬糯麦1号,其主要体现在适中的面条软硬度和较好的光滑性。推测Wx基因缺失和优质HMW-GS聚合可显著提高小麦淀粉品质和蛋白质品质,有效改善面条的蒸煮品质和感官评分。     

Variation in mineral micronutrient concentrations in grain of wheat lines of diverse origin

150 lines of bread wheat representing diverse origin and 25 lines of durum, spelt, einkorn and emmer wheat species were analysed for variation in micronutrient concentrations in grain. A subset of 26 bread wheat lines was grown at six sites or seasons to identify genetically determined differences in micronutrient concentrations. Substantial variation among the 175 lines existed in grain Fe, Zn and Se concentrations. Spelt, einkorn and emmer wheats appeared to contain higher Se concentration in grain than bread and durum wheats. Significant differences between bread wheat genotypes were found for grain Fe and Zn, but not Se concentration; the latter was influenced more by the soil supply. Grain Zn, but not Fe, concentration correlated negatively with grain yield, and there was a significant decreasing trend in grain Zn concentration with the date of variety release, suggesting that genetic improvement in yield has resulted in a dilution of Zn concentration in grain. Both grain Zn and Fe concentrations also correlated positively and significantly with grain protein content and P concentration, but the correlations with kernel size, kernel weight or bran yield were weak. The results from this study are useful for developing micronutrient biofortification strategies.     

Stacking HMW-GS transgenes in bread wheat: Combining subunit 1Dy10 gives improved mixing properties and dough functionality

We have determined the technological properties of four lines containing combinations of three HMW-GS transgenes, encoding HMW-GS 1Ax1, 1Dx5 and 1Dy10. These lines were produced by conventional crossing of three single transgenic lines of the bread wheat cultivar Anza that contains the endogenous HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8 and is null for the Glu-A1 locus. Consequently, the total number of HMW-GS ranged from 4 in the control line Anza to 7 in line T618 which contains all three HMW-GS transgenes. The lines were studied over two years using a range of widely used grain and dough testing methods. All lines with transgenic subunits showed higher levels of glutenin proteins than the Anza control, and these differences were highly significant for lines T616, T617 and T618, containing, respectively, the transgenes encoding HMW-GS 1Ax1 and 1Dy10, 1Dx5 and 1Dy10 and 1Ax1, 1Dx5 and 1Dy10. These increases in glutenin levels are compensated by lower levels of gliadins present in transgenic lines. These changes affected the ratio of polymeric to monomeric gluten proteins (poly:mono), the ratio of HMW-GS to LMW-GS (HMW:LMW) and the contents of individual 1Ax, 1Bx, 1By, 1Dx and 1Dy subunits. Transgenic lines expressing subunit 1Dy10 together with x-type subunits (T616, T617 and T618) were superior to line T606, which had only increases in x-type subunits. In particular, the combination of transgenic subunits 1Dx5 and 1Dy10 (line T617) gave better dough rheological properties than the other combinations of transgenic subunits. For example, dough development time and stability were increased by 3.5-fold and 8.5-fold, respectively, while the mixing tolerance index (MTI) was decreased by 3.3-fold in line T617 with respect to the control line. Alveograph analyses showed that all four transgenic combinations had increased P values compared to the Anza control but subunit 1Dx5 greatly reduced the extensibility (L). These results show that stacking HMW-GS transgenes by conventional crossing is a valid strategy for the improvement of wheat quality, with different effects being related to the different HMW-GS combinations.     

Effects of microfluidization on antioxidant properties of wheat bran

Phenolic compounds present in native wheat bran are majorly bound to polysaccharides and entrapped in the fibre matrix. Recently, it was demonstrated that the microfluidization process could significantly improve physicochemical properties of wheat bran due to particle size reduction and microstructure modification. The current study provides further evidence that the process also significantly increased the contents of surface-reactive, alkaline and acid hydrolysable phenolics by 280%, 60% and 20%, respectively, after a total of 8 passes through the IC₂₀₀ and IC₈₇ chambers. Accordingly, the associated antioxidant capacity increased with increase in the extent of the treatment. However, there was a decrease in solvent extractable phenolic contents due to their dispersion in water and loss during the treatment. It is also worth noting that the residues after alkaline and acid hydrolysis still contained a high content of surface-reactive phenolics, which might indicate a significant underestimation of the total phenolic content and antioxidant capacity of wheat bran when using the conventional method based on solvent extraction and alkaline and/or acid hydrolysis.     

Low molecular phytochemicals of Indian dwarf (Triticum sphaerococcum Percival) and Persian wheat (T. carthlicum Nevski) grain

Ancient wheat grain is considered by consumers to be more natural, pro-healthy and better tolerated, so these genotypes are being steadily reintroduced to cultivation. This study presents the content and composition of phenolic acids, alkylresorcinols, sterols, tocols and carotenoids in the grain of Indian dwarf and Persian wheats, extended by characteristics of their kernels (weight, dimensions and colour). To compare these features, four other wheat species (bread, spelt, durum and einkorn wheats) were used. It was found that the grain of Indian dwarf and Persian wheat is similar in weight and dimensions to grain of einkorn wheat, while in colour to bread wheat. Among the tested samples, grain of both new genotypes was the richest source of total low molecular phytochemicals, especially phenolic acids and alkylresorcinols, while being the weakest source of carotenoids. For these wheats, an enhanced share of ferulic acid (93–95%) was found, accompanied by higher quantitative and qualitative variability of homologues within sterols, tocols and carotenoids. In turn, the alkylresorcinol composition was related to wheat ploidy level.     

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.     

Effects of allelic variation of HMW-GS and LMW-GS on mixograph properties and Chinese noodle and steamed bread qualities in a set of Aroona near-isogenic wheat lines

High-molecular-weight glutenin (HMW-GS) and low-molecular-weight glutenin (LMW-GS) subunits play an important role in determining wheat quality. To clarify the contribution of each subunit/allele to processing quality, 25 near-isogenic lines with different HMW-GS and LMW-GS compositions grown at two locations during the 2010 cropping season were used to investigate the effects of allelic variation on milling parameters, mixograph properties, raw white Chinese noodle (RWCN) and northern style Chinese steamed bread (NSCSB) qualities. The results showed that Glu-B1 and Glu-B3 made a large contribution to determining mixograph properties and processing quality, respectively. Subunit pairs 17 + 18 and 5 + 10, and alleles Glu-A3b, Glu-A3d, Glu-B3g and Glu-D3f made significant contributions to mixograph properties and no significant difference was detected on most parameters of RWCN and NSCSB for the allelic variation of HMW-GS and LMW-GS. The allelic interactions among glutenin loci had significant effects on wheat quality. The line with 1, 17 + 18, 2 + 12, Glu-A3c, Glu-B3b, Glu-D3c associated with superior mixograph properties, the line with 1, 7 + 9, 2 + 12, Glu-A3c, Glu-B3d, Glu-D3c had superior viscoelasticity of RWCN, and the line with 1, 7 + 9, 2 + 12, Glu-A3e, Glu-B3b, Glu-D3c had the highest total score of NSCSB. These results provide useful information for genetic improvement of the qualities of traditional Chinese wheat products.     

Antioxidant activity of small grain cereals caused by phenolics and lipid soluble antioxidants

In this study, the content of soluble, free forms of phenolic compounds (total phenolics, flavonoids, PVPP (polyvinylpolypyrrolidone) bound phenolics, proanthocyanidins and phenolic acids), as well as the content of carotenoids and tocopherols, were determined in whole grains of bread and durum wheat, rye, hull-less barley and hull-less oat, each represented with four genotypes. Antioxidant activity was evaluated as radical scavenging activity with DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent, as well as by hydrogen transfer reaction (reduction power) based on the reduction of Fe³⁺. Generally, a considerable variation in antioxidant activities and phytochemical contents was observed between the cereals. Remarkably higher DPPH radical scavenging ability and reducing power were detected in hull-less barley, followed by rye and hull-less oat and durum and bread wheat, indicating that small grain species have different major antioxidants with different properties. Hull-less barley had the highest content of total free phenols, flavonoids, PVPP bound phenolics and contained flavan-3-ols, not found in other species. Hull-less oat had the highest content of tocopherols, very high content of yellow pigments and PVPP bound phenolics. Ferulic acid was the major free phenolic acid in small grain cereals tested. The relationship between the content of soluble phenols, as well as reducing power and DPPH^• scavenging activity are also considered.     

Predicting wheat quality – consequences of the ascorbic acid improver effect

Genotype and environment sets of wheat cultivars and breeding lines were tested for bread making, dough mixing, dough rheology, protein composition and thiol/disulphide composition to find methods that could identify wheat with high baking quality (high-BQ) and moderate work input (moderate-WI) requirement using the mechanical dough development system. Wheat with these properties generally had a high baking response to ascorbic acid (AA) and a GluD1a (HMW-GS 2 + 12) allelic composition. Strong wheat with high WI and high-BQ generally had low baking response to AA and a GluD1d (HMW-GS 5 + 10) allelic composition. Using protein composition data to identify wheat of high-BQ (with AA) and moderate-WI, it was best to select wheat with as high as possible percent of SDS-unextractable polymeric protein (%UPP) in flour and as low as possible %UPP in total polymeric protein. Using a dough extension test for identifying wheat of high-BQ (with AA) and moderate-WI, it was best to select wheat with intermediate values for maximum resistance to extension (Rmax) and for values of extension at Rmax as high as possible within the intermediate Rmax range. Cysteine content of protein fractions and glutathione content of flour gave mostly poor to weak correlations with all baking and mixing properties.     

Agronomic and quality characteristics of triticale (X Triticosecale Wittmack) with HMW glutenin subunits 5+10

Sets of triticale (X Triticosecale Wittmack) lines derived from the cv. Presto with HMW glutenin allele Glu-D1d (subunits 5+10) translocated from bread wheat (Triticum aestivum L.) chromosome 1D to chromosome 1R were evaluated for agronomic and grain quality characteristics in 2002–2005. Two different translocation types were used: (a) single translocation 1R.1D₅₊₁₀-2 where the long arm of 1R carries the wheat segment from 1DL with the Glu-D1d replacing a secalin locus Sec-3, (b) double translocation Valdy where the long arm of 1R has the translocation 1R.1D₅₊₁₀-2 and the short arm has a segment from 1DS carrying wheat loci Gli-D1 and Glu-D3. The presence of Glu-D1d was determined by polyacrylamide gel electrophoresis (PAGE-ISTA) and DNA markers. The tested lines of triticale were compared with the check triticale cv. Presto and with wheat cultivars of different bread making quality (E-C quality classes). Single translocation 1R.1D₅₊₁₀-2 reduced grain yield by 16% and Valdy translocation by 24% as compared with cv. Presto. The Valdy translocation had substantially shortened spike length and reduced specific weight in comparison with check cv. Presto. Wet gluten content (according to the Perten method) was 12% in both translocation types, 8% in check Presto and on average 24% in wheat. Translocations increased the Zeleny sedimentation value (Valdy — 27 ml, 1R.1D₅₊₁₀-2 – 25 ml, cv. Presto — 23 ml). Triticale had a very low Hagberg falling number (FN) of 62–70 s without significant differences, while wheat had on average 301 s. The translocations did not significantly increase loaf volume; however, they improved loaf shape (height/width ratio): Valdy — 0.61, 1R.1D₅₊₁₀-2 – 0.56, cv. Presto 0.44, wheat on average 0.70. The dough was non-sticky in Valdy, slightly sticky in 1R.1D₅₊₁₀-2 and sticky in cv. Presto. Problems with a low FN for improving bread making quality of triticale are discussed. Higher bread making quality can be influenced by appropriate combination with donors of low α-amylase activity.     

The effect of infrared stabilized rice bran substitution on physicochemical and sensory properties of pan breads: Part I

Infrared stabilized rice bran (SRB) substitution to white wheat, wheat bran and whole grain wheat breads at the levels of 2.5, 5.0 and 10.0% was evaluated in terms of proximate composition, crumb color, dietary fiber, texture and sensory attributes. An increasing tendency was observed in crude fat and ash content of the breads. Redness (+a*), yellowness (+b*) chroma and redness (a*/b*) values were increased gradually with the addition of SRB. Crumb color was found to be darker when 10% of SRB was added to the breads. SRB inclusion did not affect the content of soluble dietary fiber while it significantly increased the insoluble and total dietary fiber contents of the breads (p < 0.05). In general, whole grain wheat bread differed from the other bread types in terms of textural behavior. Based on the overall acceptability scores, white wheat and wheat bran breads were sensory accepted up to 10% of flour replacement with rice bran, while substitution levels higher than 2.5% negatively affected the sensory scores of whole grain wheat bread.     

Polymerization of glutenin during grain development in near-isogenic wheat lines differing at Glu-D1 and Glu-B1 in greenhouse and field

The polymerization of glutenin polymers was monitored by measuring the Unextractable Polymeric Protein (UPP) at 3-day intervals after anthesis for four pairs of near-isogenic wheat lines. Two pairs, the variety Lance, differing at the Glu-D1 locus (HMW-GS 5+10 or 2+12) and the variety Halberd, differing at the Glu-B1 locus (HMW-GS 7+9 or 20x+20y) were grown in the field (2000) and twice in the greenhouse (2000 and 2001). Two other pairs, the varieties Warigal and Avocet, differing at the Glu-D1 locus (HMW-GS 5+10 or 2+12) were grown in the greenhouse in 2001. The behavior of all lines was consistent from greenhouse and field plantings in that the lines possessing strength-associated HMW-GS (5+10 at Glu-D1 and 7+9 at Glu-B1) showed an increase in accumulation of larger glutenin polymers (measured by UPP) at an earlier stage during grain filling than the lines with allelic counterparts (HMW-GS 2+12 at Glu-D1 and 20x+20y at Glu-B1). In all cases, the increases were maintained until maturity, paralleling the greater dough strength of flour from these lines, measured by mixograph dough development time.     

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.