小麦籽粒发育时期Puroindolines蛋白与硬度的关系

为探讨Puroindolines蛋白的表达特点与籽粒硬度的关系,采用改进的SDS-PAGE凝胶分析了不同硬度小麦品种的籽粒在各个发育时期Puroindolines蛋白的表达.结果表明,不同硬度的小麦籽粒中总Puroindolines(PinA和PinB)蛋白的表达量差异不大,但与胚乳淀粉颗粒结合的Puroindolines蛋白量差异非常明显:在籽粒发育的不同时期,软质小麦籽粒淀粉粒表面结合的Puroindolines蛋白量显著高于硬质小麦;基因型同为野生型但硬度有差异的品种,籽粒较软的材料其淀粉粒表面结合的Puroindolines蛋白量也明显高于较硬的材料,说明该蛋白的结合特性是决定籽粒硬度的直接原因.结果还表明,胚乳中水溶性戊聚糖与籽粒硬度关系密切.     

Arabinoxylans and Endoxylanases in Wheat Flour Bread-making

For the past 50 years the function of arabinoxylans in bread-making has been the subject of much debate and controversy. In the last decade, these molecules have been put in the spotlight again inter alia because of the increasing use of endoxylanases in European wheat and rye flour bread-making processes. This renewed interest has led to considerable advances in the understanding of both arabinoxylan and endoxylanase functionalities in bread-making. We here present a survey of the relevant work. Knowledge of both the substrate (arabinoxylan) and the enzyme (endoxylanase), as well as of recently discovered endoxylanase inhibitors, is summarised. Facts on arabinoxylans and endoxylanases in wheat flour bread-making are presented and integrated in an up-to-date view on their functionality in bread-making.     

Water-deficit and high temperature affected water use efficiency and arabinoxylan concentration in spring wheat

This study was conducted in a controlled environment to evaluate the combined effects of water-deficit (imposed at the stem elongation stage) and high temperature (imposed at the booting stage) on the water use efficiency (WUE) and arabinoxylan concentration of two spring wheat varieties (‘Superb’ and ‘AC Crystal’) commonly grown in Canada. The temperature treatments were 22/12 (day/night, T1) and 32/22 °C (T2). Overall, time to maturity under high temperature was 10 days shorter for ‘Superb’ than for ‘AC Crystal’, indicating that ‘Superb’ was more sensitive to high temperature stress. Leaf relative water content (RWC) and specific leaf area (SLA) were more sensitive to drought than to high temperature for both varieties. Drought and high temperature decreased (P < 0.05) biomass, water use and grain yield but increased WUE of ’Superb’ and ‘AC Crystal’. Without temperature stress, significant drought and variety effects were found on CID (carbon isotope discrimination) which was negatively correlated with WUE. All gas exchange parameters declined under drought and high temperature. High temperature increased the grain arabinoxylan concentration (especially the water-extractable arabinoxylans). The different arabinoxylan fractions were positively correlated with WUE suggesting that arabinoxylans can be increased by selecting for increased WUE.     

Fractionation and characterisation of arabinoxylans from brewers' spent grain and wheat bran

Two agro-industrial co-products, brewers' spent grain and wheat bran, were fractionated by sequentially extracting with alkali of increasing strength. Over 60% of the brewers' grain biomass was solubilised by these treatments, compared with only 25% for wheat bran. The carbohydrate and phenolic composition of the solubilised fractions were determined, highlighting two compositionally different sets of fractions. In both co-products arabinoxylan was the main polysaccharide released. The degree of arabinose substitution of the extracted arabinoxylan diminished as the alkali strength increased. Insoluble residues contained both cellulose and non-cellulosic polysaccharides. In spent grain, the composition of the arabinoxylan in the residue was similar to that of the starting material. In wheat bran, the residual xylan was very highly substituted with arabinose. Both ferulic acid and three forms of diferulic acid (5,5′, 8-O-4′ and 8,5′) were present in the solubilised material, even after treatment with 4 M KOH. Esterified acetate was also present on polymers solubilised with KOH at concentrations up to 1 M. The more soluble fractions of spent grain represented a heterogeneous aggregation of feruloylated arabinoxylans with a broad molecular mass range, but the fractions extracted with stronger base were separated into low molecular weight moieties, presumably due to cleavage of covalent cross-links. Potential food and non-food uses for the isolated fractions from the cereal co-products are discussed.     

Dietary fiber in triticale grain: Variation in content, composition, and molecular weight distribution of extractable components

Triticale, a man made cereal is mainly used in animal feed but also in food and as a renewable crop for energy. In this study, eight triticale cultivars and reference rye and wheat were grown at two locations in Sweden (Svalöv and Kölbäck). The harvested grains were characterized for dietary fiber (DF) content, molecular weight distribution of extractable DF components, and some general quality parameters (bulk density, thousand kernel weight, and content of crude protein, crude fat, starch, maltose, sucrose, neutral detergent fiber, and acid detergent fiber). Total DF content (13-16%) was affected by both location and cultivar. Arabinoxylan (average 6.8%), fructan (average 2.3%), cellulose (average 2.1%), Klason lignin (average 1.6%), and β-glucan (average 0.7%) were main components of DF. Cultivars grown at Kölbäck contained extractable arabinoxylan with significantly lower weight-average and number-average molecular weights, probably due to rainy weather and sprouting in the ears. The Calcofluor average molecular weight of extractable β-glucan was highly reduced at this site. About 80% of the fructan had a low degree of polymerization (3-9). Our results show that modern triticale grain has a DF profile and molecular weight distribution of extractable DF components that are much more similar to wheat than rye.     

Relationships Between Mixograph Parameters and Indices of Wheat Grain Quality

A 10-gram Mixograph was used to analyse 39 bread wheat cultivars having various agricultural characteristics and origins. Eleven parameters obtained during the first 8 minutes of the mixing procedure were selected using a repeatability analysis in order to investigate variations between genotypes. Several flour protein fractions characterised by their extractability properties and size distributions were quantified. In addition grain hardness, water-extractable arabinoxylan contents and relative arabinoxylan viscosity were also determined and correlated with dough formation during mixing. Dough consistency, as measured by the height of the Mixogram, was correlated with grain hardness, as expressed by the starch damage content and the proportion (%) of flour particles larger than 37·8 μm. No significant relation was found with the water-extractable arabinoxylan content or the relative arabinoxylan viscosity. The width and height of the Mixogram was related to the proportion of unaggregated proteins before the mixing peak and to polymeric proteins after the dough consistency reached a maximum. The Mixograph proved to be a powerful tool to investigate indices of bread making quality.     

Investigation by Confocal Raman Microspectroscopy of the Molecular Factors Responsible for Grain Cohesion in theTriticum aestivum Bread Wheat. Role of the Cell Walls in the Starchy Endosperm

Confocal Raman microspectroscopy has previously been employed to investigate the protein content and composition of the starchy endosperm of the wheat grain. With the same objective, that is to determine the molecular basis of grain cohesion and more specifically of kernel hardness, the contribution of endosperm cell walls in the kernel structure and cohesion was explored. The technique showed that endosperm cell walls consist not only of arabinoxylan chains with ramifications of ferulic esters, but also of others components such as proteins and lipids that could play some role in the mechanical properties of the endosperm cell walls. A new model of interaction between ferulic ramifications and a phospholipid component was proposed. The investigation of cell wall composition at successive stages of grain development revealed a decrease in the protein to arabinoxylan ratio and simultaneously an increase of the ferulic acid to arabinoxylan ratio that could be associated with a strengthening of the cell wall structure. The study confirms the effectiveness of confocal Raman microspectroscopy to approach the structure of wheat grain at the micrometer scale and to identify specific molecular factors responsible for grain cohesion and involved in the fracture modes generated during the milling process.     

Relative Activity of Endoxylanases Towards Water-extractable and Water-unextractable Arabinoxylan

Differences in endo-β-1,4-xylanase functionality result from the different ability of the enzymes to convert water-unextractable arabinoxylan (WU-AX) into solubilised AX (S-AX) and to degrade water-extractable arabinoxylan (WE-AX) and S-AX to lower molecular weight. These respective endoxylanase activities and their relative proportions were determined for two endoxylanases from Bacillus subtilis and Aspergillus aculeatus, according to a generally applicable procedure. Isolated wheat WE-AX and WU-AX containing wheat squeegee starch were prepared and incubated with different levels of endoxylanases. The supernatants were analysed for reducing xylose (RX) formed and WU-AX solubilised. Molecular weights of the products formed were assessed. Relative activities differed widely for the two endoxylanases. Based on the results and additional enzyme-substrate binding experiments, mechanisms possibly accounting for these differences are discussed.     

The influence of native and modified arabinoxylan preparations on baking properties of rye flour

Water soluble arabinoxylans, especially of rye origin, are known to considerably improve quality of bread. The aim of the current research was to check the changes in properties of rye dough and bread after the addition of different arabinoxylan preparations. Structure forming properties of these additives were given by the application of hydrolysis, and cross-linking, which influenced molar mass and reactivity of arabinoxylans.     

Study of grain cell wall structures by microscopic analysis with four different staining techniques

Four different grain cell wall staining techniques were compared. Two techniques specifically detected arabinoxylan (AX). The first technique used a xylanase probe, while the other one was based on immunolabeling of AX using monoclonal antibodies. The two other staining techniques, one based on Calcofluor and the other on immunolabeling using monoclonal antibodies, stained mixed-linkage β-glucan. Cell walls of wheat, barley, oat and rye grains, differing both in content and location of AX and β-glucan, were examined. The staining methods were complementary to each other in revealing the location and distribution of the major cereal dietary fiber components AX and β-glucan in the different grains. AX was mostly concentrated in nucellar epidermis and aleurone cells, whereas β-glucan was concentrated more in subaleurone cells. Furthermore, in the case of barley and rye, the endosperm cell walls also contained high amounts of β-glucan. Interestingly, β-glucan in rye and barley endosperm cell walls was located adjacent to the cell contents, suggesting that it is not evenly distributed in the endosperm cell walls. The results give new insight into the structure of the cereal dietary fiber complex. Further development of microscopic techniques will help in elucidating the cereal cell wall structure even in more detail.     

Quantities of phenolic compounds and their impacts on the perceived flavour attributes of rye grain

The use of whole grain rye products, beneficial to one's health, could be substantially extended if the typical intensively bitter flavour of rye could be modified without losing the characteristic rye-like flavour. The aim of the study was to evaluate the contribution of non-volatile phenolic compounds on the perceived flavour. Rye grain was milled into five milling fractions. The levels of phenolic compounds, i.e. the phenolic acids, alkylresorcinols and lignans, of the fractions were analysed and related statistically to sensory flavour profiles by partial least-squares (PLS) regression. The non-bound (free) phenolic acids are suggested to be most flavour-active. Cereal and intense flavour and aftertaste were related to vanillic and veratric acids, alkylresorcinol C_23:0, and other lignans except for pinoresinol. The perceived bitterness of the bran fractions was suggested to result from pinoresinol and syringic acid. Sinapic and ferulic acids, alkylresorcinols, except for alkylresorcinol C_23:0, and syringaresinol seemed to cause the germ-like flavour. Phenolic acids, alkylresorcinols and lignans distributed with a similar pattern in the rye grain as the flavour attributes: the phenolic compounds were clearly located in the outer bran fractions being intense in flavour, but not in the mild-tasting inner layers of the grain.     

Evidence for the presence of arabinoxylan hydrolysing enzymes in European wheat flours

Water extracts of flour samples prepared from six sound European wheat varieties hydrolyse p-nitrophenyl-β-d-xylopyranoside and p-nitrophenyl-α-l-arabino-furanoside and, in addition, release soluble, dyed fragments from azurine crosslinked xylan. Incubation of water soluble wheat arabinoxylan with water extracts of flour results in the release of low molecular weight fractions consisting mainly of arabinose and xylose and small proportions of oligosaccharides as detected by high performance anion exchange chromatography. Gel permeation profiles of the incubation mixtures show a clear breakdown of the arabinoxylan.     

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.     

Structural Characterisation of Water-extractable and Water-unextractable Arabinoxylans in Wheat Bran

Water-extractable arabinoxylan (WE-AX) in wheat bran, of which only a minority originates from adherent endosperm, amounted to 6% of the total arabinoxylan (AX) content in such bran. WE-AX had an arabinose to xylose (A/X)-ratio of 0·45. Graded ethanol precipitation (20–80% range) yielded AX with an A/X-ratio increasing from 0·31 to 0·85. A population of molecular weight (MW) of 50 kDa precipitated between 0 and 40% ethanol and one with much lower apparent MW precipitated at higher ethanol concentrations. Wheat bran unextractable cell wall material (UCM) was obtained as the residue withstanding thermostable α-amylase and protease treatments and consisted mainly of AX (ca 45%) and cellulose (30–35%). Two consecutive extractions of UCM with alkaline hydrogen peroxide (AHP; 2·0%, pH 11·5, 4 h, 60 °C) resulted in a cellulose rich residue (CRR) containing 33% of the AX originally present in UCM. The average A/X-ratio of AX in CRR was lower than that in UCM. The extracted AX polymers (ca 45% of the AX originally present in UCM) had a high A/X-ratio (0·82). Their elution profiles showed two polydisperse peaks with apparent MW of respectively 100–120 kDa and 5–10 kDa. Graded ethanol precipitation resulted in a lowly substituted fraction (A/X=0·40; 0–40% ethanol) of high MW and a fraction of highly substituted AX (A/X>0·95; 40–70% ethanol) containing both low molecular weight (LMW-) as well as high molecular weight (HMW-) AX. At an ethanol concentration of 70% or more, only LMW-AX precipitated.     

Effect of the addition of bran fractions on bread properties

High fibre breads were produced adding durum wheat bran fractions of different composition and particle size. Fresh products were characterized for texture, crumb grain, volume, colour, water status (water activity, moisture content, frozen water content, ¹H molecular mobility).     

硬质小麦籽粒阿拉伯木聚糖含量的遗传变异研究

为了更好地理解环境和基因型对小麦阿拉伯木聚糖含量的影响,为小麦品质改良提供科学依据,采用比色法测定了美国西部51个硬质小麦品种的阿拉伯木聚糖组分含量,这些小麦品种包括冬小麦和春小麦,分别被种植在3个不同的地点.结果表明,参试冬小麦品种水溶性和总阿拉伯木聚糖含量的变异范围分别为0.39%～0.81%和3.09%～4.04%,春小麦品种这两种木聚糖含量范围分别为0.48%～0.92%和3.94%～4.70%.小麦籽粒阿拉伯木聚糖含量的变异同时受品种、环境及其互作效应的显著影响,环境效应太于品种效应.小麦品种间阿拉伯木聚糖含量的变异主要表现在水溶性组分上.就不同组分而言,基因型对水溶性阿拉伯木聚糖组分的影响相对较大,环境则分别对冬小麦水不溶性、春小麦水溶性阿拉伯木聚糖组分有较大的影响.品种间和环境间阿拉伯木聚糖含量的变异,冬小麦大于春小麦;水溶性阿拉伯木聚糖含量的遗传型方差春小麦大于冬小麦.冬、春小麦的水溶性阿拉伯木聚糖含量的广义遗传力均大于95%,通过遗传育种的方法改良小麦阿拉伯木聚糖含量是可行的.冬小麦与春小麦阿拉伯木聚糖含量的平均值之间无明显差异.     

Variation in the degree of D-Xylose substitution in arabinoxylans extracted from a European wheat flour

Total water-extractable arabinoxylan from flour of the European bread making wheat cultivar Camp Remy was fractionated by ethanol precipitation. Both ¹H-NMR spectroscopy and methylation analysis of the isolated arabinoxylan fractions showed wide variation in the degree of xylose substitution. More highly substituted arabinoxylan fractions were precipitated at higher ethanol concentrations. At ethanol concentrations of 10–30% (v/v) (1→3),(1→4)-β-d-glucan co-precipitated with arabinoxylan. Re-precipitation of the polymer mixture resulted in a partial separation of the arabinoxylan from the mixture. Arabinoxylans with wide structural variation were also isolated by extraction of the flour with aqueous ethanol solutions of decreasing concentations. By this isolation technique, fractions with the highest degree of substitution were extracted with the most concentrated (50%) ethanolic solutions. The range of structural variation in the fractions was quite similar for both isolation methods. A decrease in the proportion of unsubstituted xylose residues occurred concomitantly with a decrease in the proportion of monosubstituted xylose units and with an increase in the proportion of disubstituted xylose units. An increase in the proportion of paired disubstituted xylose units as the arabinose to xylose ratio of the arabinoxylan fractions increased, and the presence of monosubstituted xylose residues next to disubstituted xylose residues in the highly substituted fractions, were illustrated by ¹H-NMR spectroscopy. Methylation analysis indicated an increase in traces of side chains of arabinose residues and in the levels of O-2 substituted xylose residues as the arabinose to xylose ratio increased.     

Endogenous arabinoxylans variability in refined wheat flour and its relationship with quality traits

Arabinoxylans (AXs) are the major dietary fiber (DF) component in wheat and their consumption has been associated with several health benefits. Genetic improvement of the AX in refined wheat flour could be a good solution to improve the DF daily consumption while maintaining the flour desirable quality. In this study, 193 common wheat lines were analyzed for their AX content in refined flour and end-use quality. Wide variation in both the total arabinoxylan (TOT-AX) (10.8–16.5 mg/g) and water-extractable arabinoxylan (WE-AX) (3.2–7.6 mg/g) was identified and, in both cases, the genotype had the greatest impact on the observed phenotypes. Variation in the endogenous AX fractions appeared to have a moderate effect on wheat quality. The WE-AX, specifically, were positively correlated with gluten strength (r = 0.11 to 0.32) and bread loaf volume (r = 0.16), whereas the TOT-AX were negatively correlated with dough extensibility (r = −0.11) and bread making quality (r = −0.11). Overall, results of this study show that the genetic improvement of grain AX is feasible and that the AXs present in refined flour do not dramatically alter wheat quality indicating that it is possible to select varieties with high AX endosperm content end desired end-use quality.     

The influence of nitrogen fertilizer on the yield and combustion quality of whole grain crops for solid fuel use

Whole grain crops can be suitable for the production of solid biofuels because they have a high biomass yield and can be harvested with a low water concentration. The concentrations of water, ash, nitrogen (N), sulphur (S), chlorine (Cl) and potassium (K) in solid biofuels should be as low as possible and calcium (Ca) concentrations high to avoid technical problems and environmentally harmful emissions during the combustion process. Since N fertilization can negatively influence the combustion quality of biomass, a conflict between yield and quality aims can arise. The aim of this study is to investigate the influence of the dosage of N fertilization on the yield and quality of the whole crop biomass of triticale, rye and wheat. In 1996 and 1997, field trials with winter triticale, winter rye and winter wheat were conducted at three locations in South-West Germany. N fertilizer doses were varied from 0 to 70 and 140 kg N ha⁻¹ a⁻¹. All N doses were applied between March and May. The whole crop biomass was harvested. The water concentrations and concentrations of ash, N, K, Cl and Ca in straw and grain were measured. A dose of 70 kg N significantly increased the yield of all cereal species, but yield increases at 140 kg N were not always significant when compared with 70 kg N. At 70 kg N the energy yields reached 137–249 GJ ha⁻¹, for wheat, 142–263 GJ ha⁻¹ for rye and 182–250 GJ ha⁻¹ for triticale. The water concentration of the biomass, mainly of the straw, was significantly increased by N fertilization when the harvest was performed early at comparatively high water concentrations. For all cereal species a significant increase of N concentrations, especially in the grain, was measured at increased N fertilizer levels. The K concentrations of the straw and the Ca concentrations of straw and grain of all cereal species were also increased by N fertilization. N fertilization had little or no effect on the ash and Cl concentrations, which slightly decreased with increased N fertilization. N fertilization can, therefore, be used as a tool to influence the concentrations of N and K in the biomass. When combining yield and quality aims, 70–100 kg ha⁻¹ a⁻¹ N fertilizer was the best dosage for the whole grain crops at the southwestern German locations tested here.     

Isolation and characterisation of friabilin genes in rye

The Hardness locus on chromosome 5D is the main determinant of grain texture in hexaploid wheat. Puroindoline-a (pin-a), puroindoline-b (pin-b) and Grain Softness Protein (GSP) genes are tightly linked at this locus and their products are the predominant components of friabilin, a 15 kDa endosperm protein complex. Differences in grain texture are mainly caused by specific puroindoline genes mutations and are known to play a large impact on the end-product quality of wheat, contributing to the distinction of well-suited market classes for specific end-uses. We investigated friabilin genes in rye (Secale cereale L.), aiming at an increase information on the friabilin molecular system in cereals and to further investigate their potential use for the genetic improvement of this crop. Using a PCR approach, puroindoline-b and GSP-like sequences were identified for the first time in several rye and triticale cultivars, sequenced and located on specific chromosomal arms. The primary structures of deduced proteins were determined and compared with those from other cereal species. In contrast with data from previous studies, secaloindoline-a was not found. Our results introduce new evidence for a discrete allelic variation at the secaloindoline loci in rye, indicating that future larger screenings may facilitate preliminary selection of germplasm for rye breeding purposes and triticale production.