Genetic markers for CslF6 gene associated with (1,3;1,4)-β-glucan concentration in barley grain

The amount of (1,3;1,4)-β-D glucan accumulated in barley (Hordeum vulgare L.) cell walls is an important consideration for grain end-use. One of the major genes responsible for (1,3;1,4)-β-glucan biosynthesis is HvCslF6, which was analyzed in this study to determine the allelic variation between low (1,3;1,4)-β-glucan (∼3.3%) cultivar CDC Bold and high β-glucan (∼5.2%) line TR251. The CDC Bold HvCslF6 allele showed 16 single nucleotide polymorphisms (SNPs) and two indels when genomic region downstream of the ATG start codon was compared to TR251 allele. Both indels added 16 nucleotides to HvCslF6 first intron of CDC Bold and a single SNP in the third exon altered alanine 590 codon in the CDC Bold sequence to a threonine codon in TR251 allele. Genetic markers were developed for five polymorphic sites and confirmed useful to select low and high β-glucan lines in a previously characterized CDC Bold/TR251 mapping population and a novel F₅ recombinant inbred line (RIL) population derived from a Merit/H93174006 (4.8 and 5.3% (1,3;1,4)-β-glucan) cross. An analysis of parental lines of six populations segregating for β-glucan concentration validated association between the TR251 HvCslF6 haplotype and high (1,3;1,4)-β-glucan concentration in populations showing a (1,3;1,4)-β-glucan quantitative trait locus (QTL) on chromosome 7H.     

Effects of Cultivar and Environment on β-(1,3)-(1,4)-D-glucan Content and Acid Extract Viscosity of Spanish Barleys

The acid extract viscosities and β-glucan contents of ten two- and six-rowed barley cultivars grown at seven locations in three consecutive years in Spain were studied in the present work. The viscosities varied from 2·4 to 24·8 centistokes (cSt) and the mean value was 6·4 cSt. The average β-glucan content of barleys determined by HPLC was 3·5% with a range of 1·9–5·5%. Significant differences were found in both β-glucan content and acid extract viscosity between different cultivars, locations and years. The β-glucan contents and viscosities of winter cultivars were higher than those of spring. Cvs. Barbarrosa and Hatif de Grignon were the genotypes with the highest values for both parameters, while cv. Beka had the lowest viscosity and β-glucan content. Environmental factors influenced both parameters. The acid extract viscosities of barleys were correlated negatively with the amount of precipitation (r=−0·754;P<0·05). Barleys grown in wet and rainy areas (Girona and La Coruña) had lower viscosity values.     

Molecular weight and structure units of (1→3, 1→4)-β-glucans in dough and bread made from hull-less barley milling fractions

Milling fractions of hull-less barley, and dough and bread with hull-less barley flour (40%) and wheat flour (60%) were analysed in an investigation of how the properties of (1→3, 1→4)-β-glucan were affected by milling, dough formation and bread making. Calcofluor average molecular weight $({\overline{M}}_{\text{cf}})$ and molecular weight distribution and the cellotriosyl/cellotetraosyl ratio of the (1→3, 1→4)-β-glucan were determined. Four different hull-less barley samples were milled to produce straight-run white flours, shorts, bran and whole-meal flours. The molecular weight distributions were unimodal for all fractions, and the $({\overline{M}}_{\text{cf}})$ range was between 117×10⁴ and 188×10⁴. These parameters were similar for all barleys, although $({\overline{M}}_{\text{cf}})$ was somewhat lower in white flour and bran fractions and somewhat higher in shorts and whole-meal. The cellotriosyl/cellotetraosyl ratio (1.5–1.8) was also similar in all fractions. Doughs and breads were made to study how flour type (sifted or whole-meal barley flour), water content, yeast, mixing time and fermentation time affect (1→3, 1→4)-β-glucan. The molecular weight distribution of (1→3, 1→4)-β-glucan was polymodal with two or three populations for all doughs and breads, and the $({\overline{M}}_{\text{cf}})$ decreased with increasing mixing and fermentation time. These results indicated that (1→3, 1→4)-β-glucan was degraded by endogenous β-glucanases in the barley and/or wheat flour. The molecular weight was not significantly affected by bread-baking and other factors. After mixing and fermentation the cellotriosyl/cellotetraosyl ratio was about 1.7–1.8 and was thus not significantly different from that of the flour blends. Thus to retain high molecular weight (1→3, 1→4)-β-glucan, which is important for its cholesterol-lowering effect, it is thus important to keep the mixing and fermentation time as short as possible when baking hull-less barley bread.     

Adaptation of the standard enzymatic protocol (Megazyme method) to microplaque format for β-(1,3)(1,4)-d-glucan determination in cereal based samples with a wide range of β-glucan content

The growing interest in β-glucans and the dietary recommendations of an exact daily intake will require rapid and accurate quantification methods of β-glucans that can be used routinely by the food industry. The objective of the present study was to adapt the standard enzymatic procedure (Megazyme method) to quantify (1-3)(1-4)-β-D-glucans to micro-plate format and further application to analyze cereal based samples with a wide range of (1-3)(1-4)-β-D-glucan content (from 0.27–75%). The samples used in this study included two breads (wheat and barley/wheat), barley flours (4% and 8% β-glucans) and two samples of oat bran (28% and 75% β-glucans). Results showed that there was no significant differences in the quantification of β-(1,3)(1,4)-D-glucans in different samples by using the Megazyme method or the micro-method. The methodology developed was also compared in terms of sensitivity and reproducibility with the results obtained by the Megazyme kit method and no differences were observed. In conclusion, the developed method allows the β-glucan quantification (specifically for mixed-linkage [(1-3)(1-4)]-β-D-glucan) to be conducted rapidly and by an efficient and sensitive micro-method in a wide range of concentrations.     

In vitro protein digestibility of developing and mature sorghum grain in relation to α-, β-, and γ-kafirin disulfide crosslinking

Sorghum (P721N, 1992) was harvested at selected days after half-bloom (DAHB) and at maturity, and analyzed for protein and moisture contents, protein digestibility, α-, β-, and γ-kafirin contents, and unextractable disulfide-bound complexes. α-Kafirin synthesis began before 10 DAHB, and β- and γ-kafirin at 20 DAHB. All the kafirins were as abundant at 40 DAHB as at maturity. Protein digestibilities of uncooked flour were about 90%, and dropped to 73% at maturity. Digestibilities of cooked flour dropped markedly at 35 DAHB, 40 DAHB and maturity. A comparison of 1992 and 1987 data indicates that digestibility decreases with moisture content, not days of development. The amount of disulfide “crosslinked” β- and γ-kafirins was insignificant at the earlier stages, though increased as the grain matured. Since kafirin synthesis is complete by 40 DAHB, we suggest that the decrease in digestibility in maturing grain is due to the drying effect and formation of disulfide-bound complexes involving β- and γ-kafirins. Cooking the flour may further promote protein interactions, particularly after kafirin synthesis has ceased.     

Distribution of (1-3)(1-4)-β-d-glucans in kernels of selected cultivars of naked and hulled barley

The objective of this study was to determine the distribution of (1-3)(1-4)-β-d-glucans in the kernels of three high-glucan lines of naked barley (STH 4561, STH 4671 and STH 4676), a low-glucan line (STH 4572), and two low-glucan comparative cultivars (naked – Rastik, and hulled – Stratus). The content of (1-3)(1-4)-β-d-glucans assayed with the enzymatic method varied within the range from 4.04 to 5.71% dry matter basis. Microspectrophotometric analysis revealed significant differentiation in the structure of the kernels and in the distribution of (1-3)(1-4)-β-d-glucans; it also indicated a necessity of applying such processing methods as are appropriate to the distribution of functional components. The distribution of functional components in the high-glucan lines of naked barley makes them ideal for utilization both in the milling technology and in the production of grits and flakes.     

Synthesis of poly(norbornene ester)s by using (<Emphasis Type="Italic">η</Emphasis><Superscript>3</Superscript>-substituted allyl) palladium (NHC) complex as catalyst and investigation of their chemical structure - Glass transition temperature - Refractive index relationships

The synthesis of poly(norbornene ester)s by using a (η ³-substituted allyl) palladium (N-heterocyclic carbene (NHC)) complex as catalyst was performed and the relationship between chemical structure and glass transition temperature or refractive index of poly(norbornene ester)s was investigated. Norbornene ester monomers were synthesized via esterification of 5-norbornene-2-methyl alcohol and aromatic carboxylic acids. The polymerization catalyst, (η ³-substituted allyl) palladium (NHC) complex, was synthesized according to a published procedure. ¹H-NMR spectroscopy was performed to determine chemical structure of monomers and polymers. The molecular weight of the polymers was measured via gel permeation chromatography and the thermal properties were analyzed via thermogravimetric analysis and dynamic mechanical analysis. Refractive indices of polymer films were measured using a prism coupler. Polymers with the highest M _n (between 100 kg/mol and 300 kg/mol) were synthesized when the ratio of monomer to catalyst was 2000:1. The glass transition temperature of synthesized polymers was about 100 °C lower than that of conventional norbornene polymers. Among the six polymers of different chemical structures, four polymers exhibited a refractive index of 1.6 or more at a wavelength in the visible light region.     

Decomposition of (1–3,1–4)-β-Glucan and Expression of the (1–3,1–4) -β-Glucanase Gene in Rice Stems during Ripening

Abstract

To elucidate the possible participation of hemicellulose decomposition in lodging resistance, we studied the change of hemicellulose and cellulose content in the stems of rice during the ripening stage by methylation analysis and the expression of related genes by Northern blotting. In the rice stem in ripening stage, content of (1-3,1-4)-β-glucan, a component of hemicellulose, decreased markedly although the content of arabinoxylan, a major component of hemicellulose, and cellulose showed little change during the same growth period. On the other hand, expression of the Gns 1 gene, which may encode (1-3,1-4) -β-glucanase that catalyzes the degradation of (1-3,14) -β-glucan, increased sharply in the stem. The mechanism of decomposition of (1-3,1-4) -β-glucan in rice stem and the possible association with lodging resistance is discussed.     

Determination of (1→3),(1→4)-β-D-Glucanase Activity by a Calcofluor-Flow Injection Analysis Method

A new methodology for the determination of (1→3),(1→4)-β-d-glucanase activity has been developed. The concentration decay curves corresponding to the depolymerisation of high molecular weight barley (1→3),(1→4)-β-d-glucan by pure (1→3),(1→4)-β-d-glucanase (E.C. 3.2.1.73) fromBacillus subtilisand by crude (1→3),(1→4)-β-d-glucanase from different malts were monitored by the Calcofluor-FIA method. In all cases, the high molecular weight (17Instituto de Agroquı&#x0301;mica y Tecnologı&#x0301;a de Alimentos (CSIC)rarr;3),(1→4)-β-d-glucan decay curves fitted very well to an empirical formula describing the change in substrate concentration with time. The curves possess an inflexion point at which the depolymerisation rate of the substrate reaches a maximum. This maximum depolymerisation rate correlates with the initial concentrations of enzyme and substrate,E_oandS_o, and the enzyme kinetic constantsV_mandK_mthrough a hyperbola similar to that of Michaelis-Menten. TheK_mdetermined forB. subtilisβ-glucanase was rather low, about 0·99 g β-glucan/l, when compared with those corresponding to (1→3),(1→4)-β-d-glucanase from different malts, which were, in turn, practically identical at about 2·92 g β-glucan/l. Experiments with barley (1→3),(1→4)-β-d-glucans of different high initial molecular weights showed that initial molecular weight had no influence on the kinetics. Thus, this new methodology permits the determination of (1→3),(1→4)-β-d-glucanase activity in a direct way, i.e. the amount of (1→3),(1→4)-β-d-glucan degraded per amount of enzyme (or malt) per unit of time. Moreover, since it is insensitive to the initial molecular weight of the substrate, it seems to be well-suited for inter-laboratory comparisons of (1→3),(1→4)-β-d-glucanase activities.     

Management of Eumusae leaf spot disease of banana caused by Mycosphaerella eumusae with Zimmu (Allium sativum × Allium cepa) leaf extract

Among extracts of 33 plant species screened against Mycosphaerella eumusae, the causal agent of Eumusae leaf spot disease of banana, water extract of Cassia senna, Zimmu (Interspecific hybrid between Allium cepa × Allium sativum) and Rhincanthus nasutus provided 100% inhibition of spore germination and 1.7–2.0 cm zone of inhibition of mycelial growth under in vitro conditions. The maximum efficacy of mycelial inhibition was observed with Zimmu leaf extract. When Zimmu leaf extract was tested at different concentrations (5, 10, 25, 50 and 100% w/v), all tested concentrations provided complete inhibition of mycelial growth of the pathogen. The field evaluation of Zimmu leaf extract at different concentrations in cv. Grand Naine showed that the application of the water extract of Zimmu leaf at 50% concentration (w/v) provided 55% reduction of disease severity compared to the unsprayed control. Besides, the application of Zimmu leaf extract increased the value of youngest leaf spotted-0 (up to 60.5%) as well as increased the yield of banana (up to 46.8%) as compared to control. The effect of Zimmu in increasing the value of YLS-0 and the bunch yield was comparable with the chemical fungicide Propiconazole 25% EC (0.1%). Thin layer chromatography (TLC) analysis showed that among different major compounds, two lipid compounds (LP-B1 and LP-B2) extracted using methanol had the ability to inhibit M. eumusae growth (0.7–1.5 cm zone of inhibition). The gas chromatography–mass spectrometry (GC–MS) analysis of lipid bands revealed the presence of six different lipid compounds, which may be responsible for the growth inhibition of the pathogen. Since the application of water extract of Zimmu was found to be not only effective in controlling the leaf spot disease severity but also increased the number of green leaves and yield of banana fruits, the Zimmu extract can be used effectively in integrated disease management of Eumusae leaf spot disease for enhancing banana production in an ecologically sustainable manner.     

Accumulation of mixed linkage (1 → 3) (1 → 4)-β-d-glucan during grain filling in barley: A vibrational spectroscopy study

The accumulation of mixed linkage barley (1 → 3) (1 → 4)-β-d-glucan (BG) during grain filling at eight stages was studied using standard reference methods and infrared spectroscopy. Two mutant barley genotypes having higher (starch mutant lys5f) and lower (high lysine mutant lys3a) BG content than the normal control Cork were studied. The Cork and lys3a genotypes showed a linear BG accumulation throughout the grain filling to reach a maximum of approximately 6 and 4% BG (w/w) dry matter, respectively. However, lys5f mutant exhibited an exponential increase in BG synthesis to a maximum of approximately 18% BG (w/w) dry matter 30 days after flowering (DAF), seemingly compensating for a decreased synthesis of starch.     

Does relative time of emergence affect stand composition and yield in a grass–legume mixture? Kura clover (Trifolium ambiguum)–meadow bromegrass (Bromus biebersteinii) and Kura clover–orchardgrass (Dactylis glomerata) mixtures

Establishing Kura clover (Trifolium ambiguum) in mixtures with grass species is challenging, because slow growth of clover seedlings results in low competitive ability. This study examined establishment success by altering time of seeding of the grass component to reduce competition with Kura clover seedlings. Two trials, one of Kura clover–meadow bromegrass (Bromus biebersteinii) and the other Kura clover–orchardgrass (Dactylis glomerata) mixtures were planted in Edmonton, Alberta. Grasses were seeded at the same time as the clover, or introduced when the clover reached one true leaf or three true leaves, in the autumn of the planting year or the following spring. Species composition varied significantly between treatments. When sown at the same time, Kura clover contributed 31 and 14% of yield in the establishment year when sown with meadow bromegrass and orchard grass, respectively. Delaying grass sowing until Kura clover had one or three leaves gave a higher percentage of Kura clover compared with planting at the same time. Autumn and spring grass sowing resulted in stands of 78 and 80% clover with meadow bromegrass, and 74 and 67% clover with orchardgrass. Altering the competitive advantage of the grass species to produce a more balanced mixture was successfully achieved by delaying seeding of the grass relative to Kura clover. A long interval before introducing the grass (autumn or following spring), was not successful as established Kura clover seedlings have an increased competitive ability.     

Effects of microbial,organically acceptable,and reduced risk insecticides on Anthonomus signatus (Curculionidae: Coleoptera) in strawberries (Fragaria × ananassa)

Anthonomus signatus, the strawberry bud weevil, is active in early spring coinciding with the bloom period of host plants and when managed and unmanaged pollinators are active. Female A. signatus cause injury to the host plant during egg laying when they deposit a single egg inside an unopened flower bud and then girdle or “clip” the bud at the pedicel. Past efforts to control A. signatus populations have relied on the use of broad spectrum insecticides. Fields are either treated at the first signs of damage, or precautionary treatments may be applied prophylactically in fields with a history of damage. Because A. signatus damage occurs during bloom, there is concern about the potential harm to pollinators caused by these treatments. In order to identify materials more compatible for use during bloom, the efficacy of reduced risk pesticides against A. signatus was tested in semi-field bioassays over two years. Beauveria bassiana, cyantraniliprole, novaluron, and pyrethrins were assessed in the first year, and none caused significant A. signatus mortality. Acetamiprid, flonicamid, and spinosad were compared in the second year, acetamiprid and spinosad had higher A. signatus mortality than an untreated control, and exposure to spinosad resulted in similar mortality to bifenthrin, an industry standard material.     

Floating Photocatalyst Based on Poly(ε-caprolactone) Foam and TiO<Subscript>2</Subscript> Nanoparticles for Removal of Textile Dyes

This study discusses a novel approach for fabrication of floating photocatalyst which can be efficiently exploited for photodegradation of dyes in aqueous solutions. A fabrication of the floating photocatalyst consisted of two steps: transformation of the poly(ε-caprolactone) beads (PCL_b) into poly(ε-caprolactone) foam (PCL_f) with porous structure in supercritical carbon-dioxide and subsequent loading of PCL_f with TiO₂ nanoparticles (NPs). Morphological characterization of the PCL_f before and after TiO₂ NPs loading was carried out by FESEM. The presence of titanium on the surface and inside the PCL_f was detected by EDX. Photocatalytical activity of the floating photocatalyst was investigated in aqueous solution of textile dyes C.I. Acid Orange 7 (AO7) and C.I. Basic Yellow 28 (BY28) which were exposed to lamp that simulates the sun light. In addition to sustainable floatability for a long period of time, developed floating photocatalyst exhibited high rate of photodegradation since the complete discoloration of AO7 and BY28 solutions and photocatalysts alone occurred after 300 and 180 min of illumination, respectively. Its photocatalytic activity was preserved after three repeated photodegradation cycles with unchanged chemical structure that was confirmed by FTIR analysis.