Assessment of biochemical markers identified in wheat for monitoring barley grain tissue

The possible use of specific biochemical compounds identified in wheat grains was evaluated for monitoring barley grain tissues during fractionation. First barley grain anatomy was studied through microscopic observation and quantification of the relative proportion of each anatomical part in four distinct barley samples from both hulled and hulless genotypes. As expected from cereal phylogeny and irrespective of the possible presence of hull, common features were observed between barley and wheat grains, but the aleurone layer predominated in the outer layers. The specific location of the compounds identified in wheat was established. Phytic acid was specifically localized in the aleurone layer and alkylresorcinols in the composite layer containing the testa, even if their concentration differed from that observed in wheat grain tissues. Thus, these two markers identified in wheat can be used to monitor the corresponding barley tissues, independent of the presence of hulls. Conversely, phenolic compounds, either ferulic acid trimer or p-coumaric acid, cannot be used to monitor respectively the outer pericarp or the aleurone cell walls in barley grains. p-coumaric acid was identified as an efficient marker of the hull and could be used to distinguish hulled or hulless barley grains and to help monitor the dehulling process.     

Grain weight and grain number responsiveness to pre-anthesis temperature in wheat, barley and triticale

In temperate cereals are commonly accepted that determination of grain number (GN) and grain weight (GW) scarcely overlap during the crop cycle. However, the assumption that GW is determined exclusively after anthesis needs to be critically reviewed in the light of reports published over the few years where temperature treatments imposed before anthesis decreased GW of bread wheat. Although these evidences suggest that both GW and GN could be affected by environmental conditions before anthesis little is known about the effect of pre-anthesis temperature on these two main yield components in wheat, barley and triticale at field conditions. In addition, the effect of temperature on GW and GN at different stages prior to anthesis has been scarcely evaluated. The objectives of the current study were: (i) to evaluate the overall response, and specific differences, of GN and GW to pre-anthesis temperature, and (ii) to study the effect of different timings of high temperature at pre-anthesis on GN and GW in wheat, barley and triticale. Three fully irrigated field experiments were carried out in three successive seasons. At each season, a wheat, barley and triticale high yielding cultivar was evaluated at three temperature regimes: control, and two timings of heating before anthesis. During the first and second seasons, the timings of heating were booting-anthesis and heading-anthesis. In the thirst season, the timings were beginning of stem elongation-booting and booting-anthesis. Plots were arranged in a split-plot design with three replicates, where the main plot was assigned to thermal regime and the sub-plots to crop species. To apply heat, transparent chambers equipped with thermostatically controlled electric heaters were used. The thermal regime was controlled by sensors connected to a temperature regulator and recorded using data loggers. Temperature within the chambers was stable across developmental stages, crops, and seasons; it averaged 5.5 °C higher than air temperature. Thermal treatments consistently reduced grain yield (p < 0.05), the magnitude of the effect ranged between 5 and 52%. The highest effect was found when temperature increased during stem elongation (yield decrease: 46%), lowest when treatments were imposed during heading-anthesis (15%) and intermediate for treatments imposed during booting-anthesis (27%). Most effects of thermal treatments on yield were due to parallel effects on GN. However, thermal treatments significantly (p < 0.05) decreased GW during the three seasons. The most effecting treatment on GW was when the crops were heated during the B-A period, i.e. GW decreased up to 23%.     

Determinants of barley grain yield in a wide range of Mediterranean environments

Barley grain yield in rainfed Mediterranean regions can be largely influenced by terminal drought events. In this study the ecophysiological performance of the ‘Nure’ (winter) × ‘Tremois’ (spring) barley mapping population (118 Doubled Haploids, DHs) was evaluated in a multi-environment trial of eighteen site–year combinations across the Mediterranean Basin during two consecutive harvest years (2004 and 2005). Mean grain yield of sites ranged from 0.07 to 5.43 t ha⁻¹, clearly dependent upon both the total water input (rainfall plus irrigation) and the water stress index (WSI) accumulated during the growing season. All DHs were characterized for possessing molecular marker alleles tagging four genes that regulate barley cycle, i.e. Vrn-H1, Vrn-H2, Ppd-H2 and Eam6. Grain yield differences were initially interpreted in terms of mean differences between genotypes (G), environments (E), and for each combination of genotype and environment (GE) through a “full interaction” ANOVA model. Variance components estimates clearly showed the greater importance of GE over G, although both were much lower than E. Alternative linear and bilinear models of increasing complexity were used to describe GE. A linear model fitting allelic variation at the four genes explained genotype main effect and genotype × environment interaction much better than growth habit itself. Adaptation was primarily driven by the allelic constitution at three out of the four segregating major genes, i.e. Vrn-H1, Ppd-H2 and Eam6. In fact, the three genes together explained 47.2% of G and 26.3% of GE sum of squares. Grain yield performance was more determined by the number of grains per unit area than by the grain weight (phenotypic correlation across all genotypic values: r = 0.948 and 0.559, respectively). The inter-relationships among a series of characters defining grain yield and its components were also explored as a function of the length of the different barley developmental phases, i.e. vegetative, reproductive, and grain filling stages. In most environments, the best performing (adapted) genotypes were those with faster development until early occurrence of anthesis. This confirmed the crucial role of the period defining the number of grains per unit area in grain yield determination under Mediterranean environments.     

Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates

Antioxidative properties of proteins from barley and rice bran and their hydrolysates were examined. Three major hordein fractions of barley, B, C and D hordeins, were partially purified by gel filtration. Albumin, globulin, prolamin and glutelin fractions of rice bran were fractioned by the Osborne method. Hydrolysates of these protein fractions were prepared by digesting with pepsin followed by trypsin. Antioxidant properties in terms of antioxidative activity against linoleic acid peroxidation and reducing activity without the lipid adjuvant were investigated. The globulin fraction from rice bran protein revealed the strongest antioxidative activity throughout the incubation time of 7 days (p ≤ 0.05). The albumin fraction of rice bran protein showed the highest reducing activity (6964 μmol of Fe²⁺) followed by globulin, prolamin, glutelin and hordein fractions with activities of 2904, 2017, 1809 and 1333 μmol of Fe²⁺, respectively (p ≤ 0.05). Partially purified C hordein exhibited the highest reducing activity compared with B and D hordeins. Protein hydrolysates obtained after digestion with pepsin and trypsin exhibited much greater antioxidative, as well as reducing, activities than those from before digestion.     

Quality of rolled barley flakes as affected by batch of grain and processing technique

Rolled barley flakes were prepared from three different batches of grain by pearling, steaming and rolling. Autoclaved and malted barleys from the three batches were also processed in the same way. Analysis of the nine products showed that both batch of barley and process had significant effects on chemical composition and viscosity. Puddings were prepared from the products and mechanical consistency, juiciness and grain consistency were graded on both newly prepared and heated puddings by a sensory taste panel. Batch of barley had no effect on mechanical consistency but significant effects on juiciness and grain consistency. Type of processing had significant effect on all three parameters for both newly prepared and heated puddings.     

叶酸代谢途径folE基因表达载体的构建及在大豆中的转化

fol E基因是高叶酸调控基因,来源于细菌,已证明在西红柿等植物中,提高了叶酸含量。本试验采用农杆菌介导的大豆子叶节法,将fol E基因在大豆受体材料Williams82中表达。共侵染了200个外植体,得到34株转化苗。通过对转化苗进行除草剂(135 mg·L-1)涂抹鉴定,获得14株除草剂抗性转化苗。PCR检测结果显示其中12株为阳性。Southern杂交结果表明,fol E全长c DNA已成功插入到大豆基因组中,并得到2个单拷贝株系。     

富含叶酸水稻研究进展

叶酸缺乏是世界性的公共健康问题,严重影响人类健康。水稻是世界第一大主粮,也是我国大部分人日常能量摄入的主要来源。但是水稻籽粒中叶酸含量极低,因此,通过提高水稻的叶酸含量将有助于解决我国以及全球许多以水稻为主粮的国家和地区的叶酸缺乏问题。本文介绍了叶酸的结构、在生物中的生理功能及叶酸缺乏带来的影响,阐述了叶酸在植物体内生物合成的途径,以及目前水稻叶酸强化的研究进展。     

Differences in dry matter production,grain production,and photosynthetic rate in barley cultivars under long-term salinity

Soil salinity is a major environmental stress causing significant loss of crop productivity. Barley (Hordeum vulgare L.) is one of the few field crops that can grow in salt-affected fields and varietal differences in productivity under salinity conditions were known. To clarify the trait most responsible for grain production under salt stress, barley cultivars that were salt tolerant (OUE812) or salt sensitive (OUC613) were grown from seedling to harvest stage in vermiculite containing various concentrations of NaCl. Dry weight of aboveground parts and grain weight decreased significantly with increasing NaCl concentration. The dry weight of the aboveground parts and grain weight decreased more significantly in OUC613 than in OUE812 for plants treated with 150 mM and 200 mM NaCl. A marked reduction in ripening percentage caused significantly decreased grain production in OUC613 as compared with OUE812. In plants treated with 200 mM NaCl, the photosynthetic rate decreased three weeks after starting the NaCl treatment, but a significant difference between cultivars in photosynthetic rate did not appear until seven weeks of NaCl treatment. OUE812 kept a higher photosynthetic rate during ripening than did OUC613 and dry matter production during the period from ripening to harvest was significantly larger in OUE812 than in OUC613. Keeping a higher photosynthetic rate might have contributed to higher grain production in OUE812. Higher ripening percentage and higher rate of photosynthesis during ripening might be target traits in breeding to improve the tolerance of barley to long-term salt stress.     

Measurement of genetic and environmental variation in barley (Hordeum vulgare) grain hardness

In this study, we assessed a broad range of barley breeding lines and commercial varieties by three hardness methods (two particle size methods and one crush resistance method (SKCS—Single-Kernel Characterization System), grown at multiple sites to see if there was variation in barley hardness and if that variation was genetic or environmentally controlled. We also developed near-infrared reflectance (NIR) calibrations for these three hardness methods to ascertain if NIR technology was suitable for rapid screening of breeding lines or specific populations. In addition, we used this data to identify genetic regions that may be associated with hardness. There were significant (p<0.05) genetic effects for the three hardness methods. There were also environmental effects, possibly linked to the effect of protein on hardness, i.e. increasing protein resulted in harder grain. Heritability values were calculated at >85% for all methods. The NIR calibrations, with R² values of >90%, had Standard Error of Prediction values of 0.90, 72 and 4.0, respectively, for the three hardness methods. These equations were used to predict hardness values of a mapping population which resulted in genetic markers being identified on all chromosomes but chromosomes 2H, 3H, 5H, 6H and 7H had markers with significant LOD scores. The two regions on 5H were on the distal end of both the long and short arms. The region that showed significant LOD score was on the long arm. However, the region on the short arm associated with the hardness (hordoindoline) genes did not have significant LOD scores. The results indicate that barley hardness is influenced by both genotype and environment and that the trait is heritable, which would allow breeders to develop very hard or soft varieties if required. In addition, NIR was shown to be a reliable tool for screening for hardness. While the data set used in this study has a relatively low variation in hardness, the tools developed could be applied to breeding populations that have large variation in barley grain hardness.     

Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley

The effects on barley starch and grain properties of four starch synthesis mutations were studied during the introgression of the mutations from diverse backgrounds into an elite variety. The lys5f (ADPglucose transporter), wax (granule-bound starch synthase), isa1 (debranching enzyme isoamylase 1) and sex6 (starch synthase IIa) mutations were introgressed into NFC Tipple to give mutant and wild-type BC₂F₄ families with different genomic contributions of the donor parent. Comparison of starch and grain properties between the donor parents, the BC₂F₄ families and NFC Tipple allowed the effects of the mutations to be distinguished from genetic background effects. The wax and sex6 mutations had marked effects on starch properties regardless of genetic background. The sex6 mutation conditioned low grain weight and starch content, but the wax mutation did not. The lys5 mutation conditioned low grain weight and starch content, but exceptionally high β-glucan contents. The isa1 mutation promotes synthesis of soluble α-glucan (phytoglycogen). Its introgression into NFC Tipple increased grain weight and total α-glucan content relative to the donor parent, but reduced the ratio of phytoglycogen to starch. This study shows that introgression of mutations into a common, commercial background provides new insights that could not be gained from the donor parent.     

Improving spring and summer fungicide treatments for winter barley

The response of winter barley to broad-spectrum fungicides applied at first node stage [growth stage (GS) 31] or as two-spray programmes [first node stage and flag leaf to ear emergence (GS 39–59)] was examined at 22 sites over five years during 1986–1990. Single sprays were also applied at an intermediate timing [third node to flag leaf emerging (GS 33–37)] at all sites in 1989 and 1990. The mean response to the single spray at GS 30–31 was 0.86 t ha⁻¹ but ranged from 0.25 to 3.03 t ha⁻¹ (4–80%). With the addition of a late spray (GS 39–59) of propiconazole plus tridemorph to the spring treatment, mean response was 1.25 t ha⁻¹ [range 0.35–3.32 t ha⁻¹ (5–87%)]. There were some differences between fungicides and fungicide mixtures applied at GS 31 and a prochloraz component was needed for effective control of eyespot. Seasonal differences in the mean response to the GS 39–59 treatment reflected differences in disease pressure after GS 59. The GS 30–31 treatment failed to maintain disease control up to the milky ripe stage (GS 75) in years when disease pressure was high. Yield responses were correlated with disease severity on leaf 2 at the time of the spring (GS 31) application and with the degree of control of eyespot on the stem base and of foliar diseases on the flag leaf at GS 75. A single spray applied at GS 33–37 gave comparable yields to the two-spray programmes and has potential to improve profitability where disease levels are low at the first node stage.     

Spectroscopic and chemical fingerprints in malted barley

A unique “Matrix” of malted barley samples was produced to validate spectroscopic procedures for monitoring the malting process. Three critical factors that were examined in controlling the rate of germination were moisture content, temperature and germination time. Of interest to the malting industry, the analysis indicates the potential to identify new germplasm that, under optimized malting conditions, would produce suitably modified malt in three days of germination. It is also clear that the control of both moisture and temperature is essential for undertaking malting studies. The study suggests that Raman and FTIR could usefully complement NIR spectroscopy for monitoring grain during the malting process. For whole grain NIR measurements, the differences between test grain and control grain at optimal wavelengths of 1280 nm and 2224 nm were found to be valuable parameters for tracking progress during the malting process. The study showed the whole grain NIR most likely assessed changing properties of the periphery of the grain. This research suggested that specific calibration models using NIR for predicting malt quality attributes such as diastatic activity on whole malt are misleading and difficult to interpret because they are highly correlated with other carbohydrate/protein-related attributes of the malt.     

Microstructure of hard and soft kernels of barley

Kernel hardness, an important quality trait of cereal grains, is known to influence pearling properties and malting quality of barley. To understand the endosperm micro-structural features of kernels and their relationship to kernel hardness, endosperms of three hard and three soft hulled spring barley lines based on single kernel characterization system hardness index were observed under light (LM) and scanning electron (SEM) microscopy. Under LM, endosperm cell wall of the three hard kernel lines was significantly thicker than that of the three soft kernel lines. Hard and soft lines showed differences in the degree of starch-protein association and continuity of protein matrix under the SEM. Hard kernel lines with a continuous protein matrix exhibited greater starch-protein adhesion than the soft kernel lines, suggesting that starch-protein binding may be one of the factors influencing barley kernel hardness. SEM of flour particles of soft kernel lines showed numerous well defined individual A and B-type starch granules, while, flour of hard kernel lines mostly showed small flour aggregates with few individual starch granules.     

大麦条纹病侵染研究

针对大麦条纹病初侵染源和再侵染问题,进行了田间试验测定和大量的调查分析,明确了种子带菌是大麦条纹病的主要初侵染源。病指比较,种子带菌占整个初侵染源的90．8％,发病率比较,种子带菌占整个初侵染源的75％。种子消毒处理,能很好的控制大麦条纹病的发生发展。土壤中残存的大麦条纹病病菌,也是大麦条纹病的初侵染源,但在整个初侵染源中所占比例很小。大麦条纹病再侵染程度主要取决于初侵染量和气候条件。     

The potential of naked barley sourdough to improve the quality and dietary fibre content of barley enriched wheat bread

The possibility of using naked barley for food products is gaining popularity due to its dietary fibre content, especially β-glucans. The technological process (dough preparation, fermentation and baking) influences bread quality but also may contribute to degradation or preservation of valuable grain components. The aim of the study was to investigate the effects of different wholemeal barley flour share and bread production method on the quality of bread and non-starch polysaccharides content and solubility.Barley enriched bread contained more both soluble and insoluble dietary fibre and β-glucans, products of 40% barley share contained 67% more total dietary fibre and 160% more β-glucans than control. However, barley incorporation decreased the amount of soluble arabinoxylans. High barley contents contributed to the breads’ volume reduction by 14% and change in their crumb and crust colour. However, barley enriched breads gained higher ratings of taste than wheat bread. Barley sourdough fermentation improved breads’ volume, colour and sensory properties. Sourdough fermentation also resulted in higher concentration of dietary fibre, arabinoxylans and β-glucans. The beneficial effect of barley addition to wheat bread may be successfully enhanced by using barley wholemeal sourdough fermentation.     

不同类型大麦品种籽粒中酚酸类化合物含量的变异规律

为了探明不同类型大麦品种籽粒中酚酸类化合物含量的变异规律,采用超声辅助提取、高效液相色谱法测定的方法对15个大麦品种（系）籽粒中的13种酚酸进行了提取、测定和分析。结果表明,大麦籽粒中的酚酸类化合物主要是羟基苯甲酸衍生物,羟基肉桂酸衍生物含量较少。不同品种和不同类型大麦品种籽粒中酚酸类化合物的含量存在极显著的差异,黑皮大麦籽粒中总羟基苯甲酸衍生物（THBA）、总羟基肉桂酸衍生物（THCA）和总酚酸（TPA）的含量均高于非黑皮大麦;二棱大麦籽粒中THBA、THCA和TPA的含量均高于多棱大麦;皮大麦籽粒中THCA和TPA的含量高于裸大麦,但THBA的含量与裸大麦无显著差异。另外,与裸大麦相比,皮大麦籽粒中含有更为丰富的酚酸种类。     

Impact of field fungal contamination of barley on ensiling properties,nutritional quality and the microbiome of barley silage

Barley varieties of differing fungal disease resistance were grown in triplicate plots at Lacombe and Lethbridge, Alberta with the disease resistant variety (FR) sprayed with a foliar fungicide to maximize differences in field fungal disease. Both varieties were harvested at soft dough and ensiled in minisilos to assess differences in fungal contamination on ensiling properties, nutritional quality, aerobic stability and associated bacterial and fungal microbiomes. Data were analysed as repeated measures with the effect of treatment × time (duration of ensiling or aerobic exposure) included in the model. The percentage leaf area diseased by net form net blotch was higher (p < 0.05) in the untreated barley cv. Sundre (UN, 59.1% leaf area affected at Lacombe and 25.2% at Lethbridge) than in the FR barley cv. Chigwell (0.7% leaf area affected at Lacombe and 0.1% at Lethbridge). Fungal resistant barley had a lower (p < 0.01) acid and neutral detergent fibre content. Relative abundance of Xanthomonadales was higher (p = 0.02) for FR than UN, while Lactobacillales dominated the bacterial microbiome after 60 day of ensiling in both silages. Bacillales dominated both FR and UN after 21 day of aerobic exposure. Fungal resistant fresh barley forage had a tendency (p = 0.10) for a greater relative abundance of Pleosporales, while UN had higher (p < 0.01) Hypocreales. Mould counts were lower (p = 0.01) for FR than UN after 7 day of ensiling. Fungal resistant barley had minimal influence on the fungal community that contributed to the aerobic deterioration of barley silage.     

Localization of alkylresorcinols in wheat,rye and barley kernels

Cereal alkylresorcinols (AR), a group of phenolic lipids mainly found in the outer parts of wheat and rye kernels, are currently being studied for the possibility to use them as biomarkers for the intake of whole grain wheat and rye foods. In this work, AR were localised in grains by using light microscopy and gas chromatographic analysis of hand-dissected botanical and pearling fractions. GC-analysis of hand-dissected fractions showed that more than 99% of the total AR content was located in an intermediate layer of the caryopsis, including the hyaline layer, testa and inner pericarp. Microscopic examination showed that the outer cuticle of testa/inner cuticle of pericarp was the exact location, and that no AR were found in the endosperm or in the germ, suggesting that AR could be used as a selective marker of testa.     

Effects of sulphur on yield and malting quality of barley

Eight field experiments were conducted at four sites in the UK in 2003 and 2004 to investigate the effects of sulphur (S) application on yield and malting quality of barley. Significant yield responses to S additions were obtained in five out of the eight experiments, with yield increases ranging from 0.2 to 1.2 t/ha (4.7–22.5%). At the two most S-deficient sites, S application significantly increased malt diastatic power, alpha-amylase activity, friability and homogeneity, and decreased (1→3,1→4)-β-glucan concentration in the wort, indicating an improved endosperm modification during malting. Sulphur applications also significantly increased the concentration of S-methylmethionine (the precursor of dimethylsulphide) in kilned malt, which could impact on beer flavour. When the supply of N was limiting, S applications decreased grain N concentration due to a dilution effect as a result of increased grain yield. In some cases, S applications resulted in decreased grain size. At sites non-deficient or marginally deficient in S, applications of S had little effect on grain or malting quality parameters. The need to maintain an adequate S supply to barley for both yield and malting quality was demonstrated.