Effect of particle size on kinetics of starch digestion in milled barley and sorghum grains by porcine alpha-amylase

The influence of milled grain particle size on the kinetics of enzymatic starch digestion was examined. Two types of cereals (barley and sorghum) were ground, and the resulting grounds separated by size using sieving, with sizes ranging from 0.1 to 3 mm. In vitro enzymatic digestion was performed, using pancreatic alpha-amylase, amyloglucosidase and protease, to determine fractional-digestion rates over 24 h. The resulting glucose production rate data were well fitted by simple first-order kinetics. For each sieve screen size, the digestion rate of barley was always higher than that of sorghum. The rate coefficients for digestion showed a decrease with increasing size, and could be well fitted by an inverse square relationship. This is consistent with the supposition that starch digestion in these systems is controlled by diffusion of enzyme through the grain fragment. Apparent diffusion coefficients of alpha-amylase obtained by fitting the size dependence were 0.76 (sorghum) and 1.7 (barley) × 10⁻⁷ cm² s⁻¹, 9 (sorghum) and 4 (barley) times slower than predicted for a molecule of the size of alpha-amylase in water.     

Similarities and differences in secondary structure of viscoelastic polymers of maize α-zein and wheat gluten proteins

The secondary structure of a dough-like zein polymer was compared to the structure present in a wheat viscoelastic system using FT-IR spectroscopy. When zein was mixed at 35 °C, which is above its glass transition temperature (T_g), changes in its secondary structure suggested that the protein loses its native structure, mainly composed of α-helices (68%), and a viscoelastic system is formed by a structural rearrangement that favors β-sheet structures. This rearrangement is very similar to the structural changes observed in gluten viscoelastic polymers. Upon removal of shear stress, the zein polymer showed a rapid decrease in the proportion of β-sheet structures (from 48% to 28% after the first 3 min) in favor of unordered structures. At the same time, the viscoelasticity of the polymer decreased rapidly. In contrast, gluten, in a similar viscoelastic system and held at the same temperature, showed a fairly constant high content of β-sheet structures (49%) coinciding with the slow relaxation time typical of gluten networks after the removal of shear. We speculate that the addition of a protein capable of causing extensive and stable β-sheet formation in the zein–starch viscoelastic polymer could increase the stability and relaxation time of the zein system and, thereby, create the possibility of a zein dough with similar functionality to a wheat viscoelastic system.     

Arabinoxylan and hydroxycinnamate content of wheat bran in relation to endoxylanase susceptibility

Hydroxycinnamate and protein contents and monosaccharide composition were determined for 11 industrial wheat (Triticum aestivum) brans and related to the susceptibility of their arabinoxylans (AX) to enzymatic degradation. There was significant variation in carbohydrate, A/X ratio, protein, hydroxycinnamic acid and diferulic acid (DiFA) content among the wheat brans. In addition, a strong correlation was found between AX and ferulic acid contents. Brans were extracted with water followed by a dry-thermal treatment to remove 90% starch and to inactivate endogenous enzymes and proteinaceous inhibitors. Treated brans were compared with respect to AX degradation by a family 11 xylanase. Digestion with xylanase had a strong impact on the chemical composition of the residual bran. The A/X ratio changed from 0.60 to 1.07. The solubilised AX had an A/X ratio of 0.32. The A/X ratio of enzyme-depleted bran was negatively correlated with the loss of AX. Total DiFA in destarched brans was negatively correlated with the amount of soluble AX. These relationships indicate that structural features of AX and the extent of its cross-linking in the cell walls of the bran tissues influence its susceptibility to xylanase treatment. Thus, the amount of enzyme-solubilised AX was not directly related to the content of AX in the destarched bran. However, brans differed in their susceptibility to xylanase attack.     

The identification of a barley haze active protein that influences beer haze stability: The genetic basis of a barley malt haze active protein

In bright beers, the formation of haze is a serious quality problem, which places limitations on the storage life of the product. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) immunoblot analysis using an antiserum that was raised against a silica eluate (SE) protein fraction (obtained from silica gel, used for the colloidal stabilisation of beer), detected a range of protein bands in barley, malt, beer and haze. A polymorphism was observed in which some barley varieties contained a molecular weight (MW) 12,000 band (SE +ve) while in other varieties this band was absent (SE −ve). A survey of 219 Australian and international barley varieties, including a comprehensive selection of current and past malting varieties, identified 181 varieties as SE +ve, and 38 varieties as SE −ve. Previous pilot brewing trials demonstrated that SE −ve varieties are desirable as the beer brewed from the malt of these varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The genetic basis for the absence of the SE protein was conferred by a recessive allele at a single locus. Interval mapping analysis showed that the MW 12,000 band mapped to the short arm of chromosome 3H.     

The effect of gluten on the retrogradation of wheat starch

The retrogradation of amylopectin in a wheat starch and a wheat starch/gluten (10:1) blend prepared by extrusion and containing 34% water (wet weight basis) was studied using X-ray diffraction, differential scanning calorimetry and NMR relaxometry during storage at constant water content and temperature (25 °C). For both samples, amylopectin ‘fully’ retrograded after 2–3 days storage, i.e. the different parameters monitored with time to follow the retrogradation had reached their maximum value, and crystallised predominantly into the A polymorph. Under the experimental conditions used, there was no evidence of any significant effects of the presence of gluten on the kinetics, extent or polymorphism of amylopectin retrogradation.     

Phenolic constituents of glandular trichomes onSolanum berthaultii andS. polyadenium

Phenolic compounds in Type A glandular trichomes of two insectresistant potato species were separated and identified using a combination of high pressure liquid chromatography (HPLC), UV-visible spectrometry, and¹H proton nuclear magnetic resonance (NMR) procedures. Type A trichomes ofSolanum berthaultii andS. polyadenium contained a phenolic compound tentatively identified as the glucose ester of p-hydroxyphenylpropionic acid. Its presence in trichomes of both species suggests that this compound may play a major role in formation of the brownish, insect entrapping exudate characteristic of trichome-mediated insect resistance in these species. Chlorogenic acid, a major phenolic constituent of Type A trichomes ofSolanum polyadenium was not detected in trichomes of 5.berthaultii. The exclusive presence of chlorogenic acid in trichome exudates ofS. polyadenium may be responsible, at least in part, for the greater levels of insect entrapment by this species, compared with that ofS. berthaultii.     

A modified enzymic browning assay for potato glandular trichomes

Tetralobulate (Type A) glandular trichomes are present on foliage of many wild as well as cultivated species of potatoes. These trichomes confer resistance to insect pests if their density, gland size and biochemical products are similar as those ofSolanum berthaultti Hawkes andS. polyadenium Greenm. Hybrids ofS. tuberosum L. ×S. berthaultii have been screened for resistance by measurement of phenolic oxidation activity in their Type A trichomes by use of an enzymic browning assay. However, variability in the procedure has prompted the development of an improved assay.     

Mechanisms of resistance to potato leafhopper,Empoasca fabae (Harris), in potato

Potato genotypes were evaluated for resistance to natural infestations of the potato leafhopper,Empoasca fabae (Harris), in the field. Characterization of Type A and B glandular trichomes, phenolic properties of Type A trichomes, glycoalkaloid content and composition,E. fabae adult and nymph infestation, leaf damage (hopperburn), yield, and yield reduction were determined on genotypes PI 473331 and PI 473334 (accessions ofSolatium berthaultii (Hawkes)), NY123 and Q174-2 (advanced hybrids), and Elba and Allegany (commercial cultivars). Regarding the Type A trichome, PI 473334 was most pubescent, followed by PI 473331, both had high levels of enzymatic browning; on PI 473334 no glycoalkaloids were detected, while on PI 473331 solamargine and solasonine were present. Those genotypes were the least infested by adults; moreover, nymphs were never found on their leaves. PI 473331 and Q174-2 possess Type B trichomes, the latter had levels of Type A trichome comparable to the first and to NY123, while displaying high levels of phenolic oxidation of Type A and very low levels of glycoalkaloids. Although Q174-2 had intermediate infestation ofE. fabae, it suffered a yield reduction comparable to the control cultivar Allegany. The density of Type A trichomes of NY123 was relatively low, even though the browning assay results were significantly elevated. Foliage of this genotype was found to contain the four types of glycoalkaloids studied at high levels.Empoasca fabae infestation on NY123 was intermediate, and it yielded well on infested plots, but suffered an intermediate yield reduction. Elba presented densities of Type A similar to NY123. It suffered the lowest yield reduction despite its high levels of leafhopper infestation and hopperburn. Its tolerance might be associated with its phenology, since it is a late-maturing cultivar. High scores of the modified enzymatic browning assay were correlated with high infestation and high leaf damage. Results indicated a significant correlation between leafhopper infestation and hopperburn.     

Identification of quantitative trait loci for β-glucan concentration in barley grain

The amount of (1 → 3),(1 → 4)-β-d glucan (β-glucan) accumulated in cell walls of barley (Hordeum vulgare L.) kernel is an important determinant for grain end-use. Grain β-glucan concentration is affected by environmental and genetic factors and usually varies from 3 to 6%. In this study, we have analyzed the β-glucan trait in a doubled-haploid (DH) population of 170 lines grown in three separate field trials. Most of the DH lines showed β-glucan values that ranged from that of the low β-glucan parent (cultivar CDC Bold; 3.3%) to that of the high β-glucan parent (breeding line TR251; 5.4%). Eighty-eight lines of the DH population were genotyped using simple sequence repeat (SSR), amplified fragment length polymorphism (AFLP) and diversity array technology (DArT) markers, which were subsequently integrated into a barley genetic map spanning 1059 cM. Interval mapping and multiple-QTL-mapping (MQM) of quantitative trait loci (QTL) from the three trials indicated seven genomic regions associated with low grain β-glucan concentration. For all putative QTLs, the low β-glucan concentration was contributed by alleles from CDC Bold except for two loci on chromosomes 5H that were derived from TR251. A major QTL located to the centromere region of chromosome 7H was identified by both mapping methods for all three trials. The 7H QTL explained up to 39% of the β-glucan concentration and genetic markers associated with the locus may be used to aid selection of high and low β-glucan barley lines.     

Inheritance of glandular trichomes in crosses withSolanum berthaultii

The inheritance of trichome density and droplet size was investigated in diploid and tetraploid crosses. In haploidTuberosum ×Solanum berthaultii and (S.phureja ×haploid Tuberosum) × S.berthaultii crosses, droplet size on type B trichomes was small or droplets were lacking on F₁ hybrids. Parental phenotypes were recovered in the F₂ and backcross generations. Trichome density was more variable within a clone that was droplet size. Density of both type A and type B trichomes appears to be under the control of small numbers of genes, as parental phenotypes were recovered in the F₂ and backcross generations. The variation observed for each of the three traits studied could not be explained by hypotheses based on the segregation of one or two loci. The insect resistance ofSolanum berthaultii appears to be due to the interaction of several chemical and physical factors, thus potato breeders should handle it as a quantitatively inherited trait.     

The identification of a barley haze active protein that influences beer haze stability: Cloning and characterisation of the barley SE protein as a barley trypsin inhibitor of the chloroform/methanol type

Previous pilot brewing trials have demonstrated that in the absence of a molecular weight (MW) 12,000 (barley silica eluate (SE) protein), the beer brewed from the malt of these SE −ve varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The previously described SE protein was characterised using comparative two-dimensional (2-D) gel electrophoresis immunoblots of barley seed extracts from both SE +ve and SE −ve varieties. The SE protein spot identified was excised and its partial sequence determined, after in-gel cleavage using trypsin and separation of the resulting fragments by reversed-phase high performance liquid chromatography (HPLC). N-terminal sequence analysis of the tryptic peptides from SE +ve and SE −ve varieties identified the SE protein as the barley trypsin inhibitor-CMe precursor (BTI-CMe). The mature BTI-CMe protein is 13.3 kDa and its functional gene is located on chromosome 3H. Cloning of the BTI-CMe protein demonstrated that both SE −ve and SE +ve barley varieties contain a BTI-CMe protein family member that is similar but has a consistently different sequence, primarily in the last 30 amino acid residues of their C-termini.     

Lipid oxidation and amylopectin molecular weight changes occurring during storage of extruded starch samples

Using thermomechanical extrusion, waxy maize starch and 4% (w/w) lipid were formed into strips. The lipids used were free fatty acids (mostly linoleic) and antioxidant stripped sunflower oil (either previously oxidised or fresh, with and without copper ions). By altering their water content it was possible to store, at the same temperature, sample strips in the glassy and rubbery states. Lipid oxidation was monitored by determining hexanal in the headspace above all stored samples. The molecular weights of the amylopectin in the samples of extruded products (dissolved in dimethylsulfoxide, precipitated and redissolved using a pressure cell) were determined by asymmetric flow-field-flow fractionation, analytical ultracentrifugation and light scattering. The initial rate of hexanal generation was higher in samples stored in the glassy state compared with those in the rubbery state. Waxy maize and extruded samples, at the start of the storage period and after 42 days, were used to establish the molecular weight of the amylopectin. A significant fall (a decrease of 40%) in molecular weight was found on storage of samples containing sunflower oil and copper ions and those containing free fatty acids, irrespective of the method used for molecular weight determination.     

Green peach aphid resistance by glandular trichomes inSolanum tuberosum xS. berthaultii hybrids

Three interspecific F₃ hybrid clones ofSolarium tuberosum L. x 5.berthaultii Hawkes bearing both Type A and Type B glandular trichomes were assessed for resistance to the green peach aphid,Myzus persicae (Sulzer). In glasshouse trials, aphid mortality and population development on hybrid clones were similar to those on the aphid-resistant parent,S. berthaultii. Trichome exudate of the hybrid, B227-63, was as effective in aphid immobilization as that ofS. berthaultii. Field populations of the green peach aphid on hybrids were intermediate between those onS. berthaultii and susceptibleS. tuberosum cultivars.     

Postharvest decay control and quality retention in litchi (cv. McLean's Red) by combined application of modified atmosphere packaging and antimicrobial agents

The effects of modified atmosphere packaging (MAP) [bioriented polypropylene (BOPP-1 or BOPP-2)] in combination with antimicrobial agents Bacillus subtilis, 10⁷ colony-forming units (cfu) ml⁻¹; ethylenediamine tetraacetic acid, calcium disodium salt hydrate (EDTA) (0.1%); or 4-hexylresorcinol (4-HR) (0.15%) on postharvest decay control and quality retention of litchi cv. McLean's Red were assessed as possible replacements for commercial SO₂ fumigation. Fruits dipped in B. subtilis, EDTA or 4-HR (5 min) separately, blow dried (25 °C, 3 min), packed in BOPP-1, held for 18 d at 2 °C, 95% RH, and 2 d at 14 °C, 75% RH were significantly less decayed. The antagonist–BOPP-1 combination also promoted the best bacterial survival during storage. B. subtilis was observed to survive effectively in BOPP-1 (16% O₂, 6% CO₂; 90% RH), but its survival was adversely affected in BOPP-2 (5% O₂, 8% CO₂; 93% RH). Alternaria alternata and Cladosporium spp. were the major decay-causing fungi in BOPP-1 treatments, and Candida, Cryptococcus and Zygosaccharomyces were the predominant yeasts in BOPP-2 treatments. Combination treatments EDTA, 4-HR or B. subtilis in BOPP-1 inhibited polyphenol oxidase (PPO) and significantly reduced pericarp browning and severity. Although the combination treatments EDTA, 4-HR or B. subtilis in BOPP were equally effective in controlling decay and browning, the EDTA and 4-HR affected the natural pinkish-red colour of the pericarp by showing higher h° values (orange–pink). Among the combination treatments, B. subtilis+BOPP-1 had the best potential to control decay, retain the colour and the overall litchi fruit quality during a marketing chain of 20 d.     

Relationship between the glass transition temperature and the melt flow behavior for gluten, casein and soya

The effects of moisture content (25–45% wwb) and temperature (75–120 °C) on the viscosity of gluten, soya and rennet casein systems was studied using a capillary rheometer. An attempt was made to relate the viscosities to the glass transition temperature measured by differential scanning calorimetry, dynamic mechanical thermal analysis and the phase transition analyzer. The temperature where the material flowed was also determined by the latter technique. All three-protein systems showed shear and extension thinning. Over the shear rate range investigated (1–10³ s⁻¹), gluten had a substantially lower viscosity than the other two proteins, although the difference was less pronounced at the highest temperature studied. This low viscosity is reflected by lower values of the glass transition temperature, the melt flow temperature and the dynamic moduli E′ and E″ in the rubbery state. The results are discussed in terms of the structure and heat induced changes for the three proteins and their relevance to food processing considered.     

Effect of different levels of nitrogen and farmyard manure on yield and quality of spinach (Spinacea oleracea L.)

The effect of different levels of nitrogen N₀(0kg/ha), N₁(30 kg/ha), N₂ (60 kg/ha), and N₃ (90 kg/ha) and farmyard manure F₀ (0 tonnes/ha), F₁ (10 tonnes/ha), and F₂ (20 tonnes/ha) on the yield and nutrient composition of spinach and its uptake was investigated on a sandy loam soil. Yield; phosphorus, iron, manganese, zinc, and copper uptakes; and ascorbic acid content increased with the application of both the inorganic nitrogen fertilizer and the farmyard manure, with a maximum at the N₃F₂ level, i.e. at 90 kg N/ha with 20 tonnes FYM/ha. However, the contents of protein, -carotene, and reducing sugars were maximum at the highest dose of nitrogen without the application of farmyard manure.     

Molecular Basis of the Gelatinisation and Swelling Characteristics of Waxy Rice Starches Grown in the Same Location During the Same Season

Six waxy rice samples (grown at the same site during the same season) were investigated to identify those aspects of amylopectin structure and granule composition which differentiate the starches into high (T_p 77·8 °C on average) or low (T_p 66·2 °C on average) gelatinisation temperature. All starches contained less than 2·5% amylose with negligible lipid. Granule dimensions were similar (mean diameter 4·9–5·7 μm) as was the β-amylolysis limit (59·9–63·0%). However, the structural elements of amylopectins within the two groups varied. The low gelatinisation temperature starches contained two major chain fractions upon debranching with iso-amylase, F3 (DP 16) and F1 (DP 51) on average while the high gelatinisation temperature starches contained three fractions, F3 (DP 16), F2 (DP 19) and F1 (DP 40). The proportion of F1/(F2+F3) for the low and high gelatinisation starches representing on average 20·2 and 10·5% respectively. Hence, the higher proportion of exterior chains and especially those with DP 19 appear to confer the higher gelatinisation temperature characteristic of these starches. Debranched β-limit dextrins were prepared from these starches. Regardless of their origin, (high or low gelatinisation temperature), they contained three fractions: F_3β (DP 5), F_2β (DP 13) and F_1β (DP 19). Hence, the high versus low gelatinisation characteristic was imparted by the exterior chains. Using 13-C CP-MAS/NMR it was confirmed that the number of double helices within the starches were approximately constant although small differences in crystallinity were identified by X-ray diffraction. Retrogradation of the solubilised native high and low gelatinisation temperature amylopectin molecules confirmed that the ‘high temperature gelatinisation amylopectin’ molecules form higher dissociation temperature and enthalpy crystalline domains. This confirms that the higher proportion of short and especially relatively longer short chains promotes crystallite formation.     

Influence of growth conditions on the value of crisphead lettuce

The marketable and nutritional quality of crisphead lettuce as affected by nitrogen supply, cultivar, and plant age at harvest was investigated in six plantings during 1989 and 1990. The optimum yield of marketable heads was obtained at a total nitrogen supply of 150 kg N per ha although only small differences were observed to the yield at 100 and 200 kg total N per ha. The total nitrogen supply included the amount of mineral nitrogen within the rhizosphere. The incidence of dry tipburn in older leaves was clearly decreased by an increased nitrogen supply, especially at late planting. The content of nitrate was increased and the content of dry matter and vitamin C decreased with increased nitrogen supply. The vitamin C content was higher for the cultivar Marius than for Saladin. As plants got older, the nutritional quality of crisphead lettuce decreased because the content of nutrients, especially vitamin C, decreased with increased plant age at harvest.     

Immunodetection of Polyphenol oxidase in glandular trichomes ofS. berthaultii,S. tuberosum and their hybrids

Type A glandular trichomes confer insect resistance to the wild potato,S. berthaultii Hawkes by oxidatively polymerizing their contents upon breakage, resulting in insect entrapment. Polyphenol oxidase (PPO) is responsible for the oxidative polymerization of Type A glandular trichome exudate.S. tuberosum L. trichomes possess low amounts of PPO and lack the ability to either polymerize their contents or entrap insects. Polyclonal antibodies raised against purifiedS. berthaultii glandular trichome PPO were used to develop an enzyme-linked immunosorbent assay (ELISA) to quantify PPO content ofS. berthaultii, S tuberosum, and hybrid type A glandular trichomes. The PPO ELISA and immunoblots were used to quantify PPO expression as a function of leaf development and genetic background. PPO accumulates to its maximal levels in trichomes prior to completion of leaflet expansion. In advanced breeding generations, trichome PPO is expressed at levels varying from that found inS. tuberosum to that found inS. berthaultii. By serving as an initial screen for trichome PPO, this ELISA may be useful to assist rapid introgression of this insect resistance trait into adaptedS. tuberosum backgrounds.