Normal Barley Research Articles

Effect of phytase supplementation on phosphorus digestibility in low-phytate barley fed to finishing pigs

Forty crossbred barrows (Camborough 15 Line female×Canabred sire) weighing an average of 79.6±8.0 kg were used in a factorial design experiment (5 barleys×2 enzyme levels) conducted to determine the effects of phytase supplementation on nutrient digestibility in low-phytate barleys fed to finishing pigs. The pigs were assigned to one of 10 dietary treatments comprised of a normal 2-rowed, hulled variety of barley (CDC Fleet, 0.26% phytate) or 2 low-phytate hulled genotypes designated as LP422 (0.14% phytate) and LP635 (0.09% phytate). A normal, hulless barley (CDC Dawn, 0.26% phytate) and a hulless genotype designated as LP422H (0.14% phytate) were also included. All barleys were fed with and without phytase (Natuphos 5000 FTU/kg). The diets fed contained 98% barley, 0.5% vitamin premix, 0.5% trace mineral premix, 0.5% NaCl and 0.5% chromic oxide but no supplemental phosphorus. The marked feed was provided for a 7-day acclimatization period, followed by a 3-day faecal collection. In the absence of phytase, phosphorus digestibility increased substantially (P<0.05) as the level of phytate in the barley declined. For the hulled varieties, phosphorus digestibility increased from 12.9% for the normal barley (0.26% phytate) to 35.3 and 39.8% for the two low-phytate genotypes (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 9.2% for the normal barley (0.26% phytate) to 34.7% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In contrast, when phytase was added to the diet, there was little difference in phosphorus digestibility between pigs fed normal barley and those fed the low-phytate genotypes (significant barley×enzyme interaction, P=0.01). For the hulled varieties, phosphorus digestibility was 50.1% for the barley with the normal level of phytate (0.26% phytate) compared with 51.1 and 52.4% for the varieties with 54 and 35% of the normal level of phytate (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 47.1% for the normal barley (0.26% phytate) to 54.4% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In conclusion, both supplementation with phytase and selection for low-phytate genotypes of barley were successful in increasing the digestibility of phosphorus for pigs. Unfortunately, the effects did not appear to be additive. Whether or not swine producers will choose low-phytate barley or supplementation with phytase as a means to improve phosphorus utilization, will likely depend on the yield potential of low-phytate barley and the additional costs associated with supplementation with phytase.

Distribution and Structural Variation of Nonstarch Polysaccharides in Milling Fractions of Hull‐less Barley with Variable Amylose Content

ABSTRACTThree hull‐less barley genotypes containing starches with variable amylose content (23.8% normal, 4.3% waxy, 41.8% high‐amylose barley) were pearled to 10% and then roller‐milled to produce pearling by‐products (PBP), flour, and fiber‐rich fractions (FRF). PBP were enriched in arabinoxylans, protein, and ash and contained small amounts of starch and β‐glucans. FRF were considerably enriched in β‐glucans and arabinoxylans. The solubility of β‐glucans was higher in PBP than in FRF. The solubility of arabinoxylans was higher in FRF than in PBP. Small amounts of arabinogalactans detected in barley were concentrated in the outer portion of the barley kernel. The content and solubility of nonstarch polysaccharides (NSP) in various milling fractions was also dependent on the type of barley. To obtain more detailed information about the content and molecular structure of NSP, each milling fraction was sequentially extracted with water, alkaline [Ba(OH)2], again with water, and finally with NaOH. These extractions resulted in four sub‐fractions: WE, Ba(OH)2, Ba(OH)2/H2O, and NaOH. β‐Glucans and arabinoxylans exhibited structural heterogeneity derived from differences in their location within the kernel as well as from the genetic origin of barley. The WE arabinoxylans from FRF and flour had a substantially lower degree of branching than those from PBP. The WE arabinoxylans from FRF of high‐amylose and normal barley contained more unsubstituted Xylp residues but fewer doubly‐substituted and singly‐substituted Xylp at O‐2 than their counterparts from PBP. The WE arabinoxylans from FRF of waxy barley had a relatively high content of doubly‐substituted, but very few singly‐substituted Xylp residues. In all three barley genotypes, the ratio of tri‐ to tetrasaccharides in β‐glucans from PBP was higher than from flour and FRF. Substantial differences in the molecular weight of NSP in different milling fractions were also observed.

A semidwarf phenotype of barley uzu results from a nucleotide substitution in the gene encoding a putative brassinosteroid receptor.

Brassinosteroids (BRs) play important roles throughout plant growth and development. Despite the importance of clarifying the mechanism of BR-related growth regulation in cereal crops, BR-related cereal mutants have been identified only in rice (Oryza sativa). We previously found that semidwarf barley (Hordeum vulgare) accessions carrying the "uzu" gene, called "uzu" barley in Japan, are non-responding for brassinolide (BL). We then performed chemical and molecular analyses to clarify the mechanisms of uzu dwarfism using isogenic line pairs of uzu gene. The response of the uzu line to BL was significantly lower than that of its corresponding normal line. Measurement of BRs showed that the uzu line accumulates BRs, similar to known BR-insensitive mutants. The marker synteny of rice and barley chromosomes suggests that the uzu gene may be homologous to rice D61, a rice homolog of Arabidopsis BR-insensitive 1 (BRI1), encoding a BR-receptor protein. A barley homolog of BRI1, HvBRI1, was isolated by using degenerate primers. A comparison of HvBRI1 sequences in uzu and normal barley varieties showed that the uzu phenotype is correlated with a single nucleotide substitution. This substitution results in an amino acid change at a highly conserved residue in the kinase domain of the BR-receptor protein. These results may indicate that uzu dwarfism is caused by the missense mutation in HvBRI1. The uzu gene is being introduced into all hull-less barley cultivars in Japan as an effective dwarf gene for practical use, and this is the first report about an agronomically important mutation related to BRs.

Supplements of transgenic malt or grain containing (1,3-1,4)-ß-glucanase increase the nutritive value of barley-based broiler diets to that of maize

1. A diet with addition to normal barley of malt from transgenic barley expressing a protein engineered, thermotolerant Bacillus (1,3-1,4)-β-glucanase during germination has previously been demonstrated to provide a broiler chicken weight gain comparable to maize diets. It also reduced dramatically the number of birds with adhering sticky droppings, but did not entirely eliminate sticky droppings. One of the objectives of the broiler chicken trials reported here was to determine if higher concentrations of transgenic malt could alleviate the sticky droppings. 2. Another aim was to investigate the feasibility of using mature transgenic grain containing the thermotolerant (1,3-1,4)-β-glucanase as feed addition and to compare diets containing transgenic grain to a diet with the recommended amount of a commercial β-glucanase-based product. 3. Inclusion of 75 or 151 g/kg transgenic malt containing 4·7 or 98 mg/kg thermotolerant (1,3-1,4)-β-glucanase with 545 or 469 g/kg non-transgenic barley instead of maize yielded a weight gain in Cornish Cross broiler chickens indistinguishable from presently used maize diets. The gene encoding the enzyme is expressed in the aleurone with a barley α-amylase gene promoter and the enzyme is synthesised with a signal peptide for secretion into the endosperm of the malting grain. 4. Equal weight gain was achieved, when the feed included 39 g/kg transgenic barley grain [containing 66 mg/kg thermotolerant (1,3-1,4)-β-glucanase] and 581 g/kg non-transgenic barley instead of maize. In this case, the gene encoding the enzyme has been expressed with the D-hordein gene (Hor3-1) promoter during grain maturation. The enzyme is synthesised as a precursor with a signal peptide for transport through the endoplasmic reticulum and targeted into the storage vacuoles. Deposition of the enzyme in the prolamin storage protein bodies of the endosperm protects it from degradation during the programmed cell death of the endosperm in the final stages of grain maturation and provides extraordinary heat stability. The large amount of highly active (1,3-1,4)-β-glucanase in the mature grain allowed the reduction of the transgenic grain ingredient to 0·2 g/kg diet, thus making the ingredient comparable to that of the trace minerals added to standard diets. 5. A direct comparison using transgenic grain supplement at the level of 1 g/kg of feed with the standard recommended addition of the commercial enzyme preparation Avizyme 1100® at 1 g/kg yielded equal weight gain, feed consumption and feed efficiency in birds fed a barley-based diet. 6. The production of sticky droppings characteristic of broilers fed on barley diets was avoided with all 9 experimental diets and reduced to the level observed with a standard maize diet by supplementation with transgenic barley. 7. The excellent growth and normal survival of the 400 broilers tested on barley diets supplemented with transgenic grain or malt showed the grain and malt not to be toxic. 8. The barley feed with added transgenic grain or malt containing thermotolerant (1,3-1,4)-β-glucanase provides an environmentally friendly alternative to enzyme additives, as it uses photosynthetic energy for production of the enzyme in the grain and thus avoids use of non-renewable energy for fermentation. The deposition of the enzyme in the protein bodies of the grain in the field makes coating procedures for stabilisation of enzyme activity superfluous. 9. Barley feed with the small amount of transgenic grain as additive to normal barley provides an alternative for broiler feed in areas where grain maize cannot be grown for climatic reasons or because of unsuitable soil and thus has to be imported.

Phosphorus digestibility in low-phytate barley fed to finishing pigs

Twenty-four crossbred barrows (Camborough 15 Line female × Canabred sire) weighing an average of 51.3 ± 5.1 kg were used in a trial conducted during two periods according to a completely randomized block design. The pigs were assigned to one of six dietary treatments comprised of a normal two-rowed, hulled variety of barley (CDC Bold) or three low-phytate genotypes selected to contain approximately 68, 41 or 3% of the normal level of phytate. A normal hulless barley (CDC Dawn) and a hulless genotype selected to contain 54% of the normal phytate content were also included. The diets fed contained 99.5% barley and 0.5% chromic oxide. The marked feed was provided for a 7-d acclimatization period, followed by a 3-d fecal collection. At the conclusion of the first replicate, the pigs were assigned to different treatments and the procedure was repeated resulting in eight observations per treatment. Selection for phytate content had no effects on the apparent fecal digestibility of dry matter, crude protein or gross energy (P &gt; 0.05). The digestibility coefficients for dry matter and energy were significantly higher for the hulless varieties of barley than the hulled genotypes. Phosphorus digestibility increased as the level of phytate in the barley declined. The linear relationship relating phosphorus digestibility (y) to phytate content (x) was: y = 67.7 − 12.54x (r2= 0.64; P &lt; 0.0001).

Influence of naked barley cultivar with normal, amylose-rich or amylopectin-rich starch and enzyme supplementation on digestibility and piglet performance

Nutrient utilization and performance were studied in two experiments with weaning piglets fed diets based on naked barley cultivars with normal, amylose-rich and amylopectin-rich starch (ST), with or without supplementation of an enzyme preparation with glucanase and xylanase activity. The coefficient of apparent digestibility (CAD) of components in all diets gradually increased from the small intestine to the rectum, although the extent of digestion in the small intestine and caecum was different for the barley cultivars. The diet based on naked normal barley showed a lower CAD (P<0.05) for organic matter (OM), crude protein (CP) and starch (ST) at the small intestine (0.25, 0.12 and 0.72) than the amylose-rich (0.61, 0.64 and 0.87) and amylopectin-rich (0.54, 0.49 and 0.87) barley cultivars. In contrast, at the caecum the naked normal barley showed higher CAD values (P<0.05) for OM, CP and ST (0.48, 0.49 and 0.26) than the amylose-rich (0.15, 0.08 and 0.10) and amylopectin-rich (0.23, 0.21 and 0.11) barley cultivars. There were no cultivar related differences in the extent of digestion at the colon and rectum. The coefficient of total tract apparent digestibility (CTTAD) of OM and CP was higher (P<0.05) for the amylose-rich (0.89 and 0.83) and amylopectin-rich (0.89 and 0.80) barley cultivars than for the normal barley cultivar (0.85 and 0.77). However, there were no differences in performance (P>0.05) between diets based the different barley cultivars. Enzyme supplementation of the diets improved the CTTAD of CP (P<0.05), while it had no effect (P>0.05) on the performance. In conclusion, the starch composition of barley appears to have a limited effect on its utilization in piglets. However, the impaired digestibility of the naked normal barley cultivar indicates that there are properties in the hulless grain that imposes limitation to its utilization and which could not be alleviated by enzyme supplementation. Overall, the present results support the contention that the potential for improvement of piglet growth performance by supplementing barley-based diets with cell wall degrading enzymes is limited.

Influence of naked barley cultivar with normal, amylose-rich or amylopectin-rich starch and enzyme supplementation on digestibility and piglet performance

Nutrient utilization and performance were studied in two experiments with weaning piglets fed diets based on naked barley cultivars with normal, amylose-rich and amylopectin-rich starch (ST), with or without supplementation of an enzyme preparation with glucanase and xylanase activity. The coefficient of apparent digestibility (CAD) of components in all diets gradually increased from the small intestine to the rectum, although the extent of digestion in the small intestine and caecum was different for the barley cultivars. The diet based on naked normal barley showed a lower CAD ( P<0.05) for organic matter (OM), crude protein (CP) and starch (ST) at the small intestine (0.25, 0.12 and 0.72) than the amylose-rich (0.61, 0.64 and 0.87) and amylopectin-rich (0.54, 0.49 and 0.87) barley cultivars. In contrast, at the caecum the naked normal barley showed higher CAD values ( P<0.05) for OM, CP and ST (0.48, 0.49 and 0.26) than the amylose-rich (0.15, 0.08 and 0.10) and amylopectin-rich (0.23, 0.21 and 0.11) barley cultivars. There were no cultivar related differences in the extent of digestion at the colon and rectum. The coefficient of total tract apparent digestibility (CTTAD) of OM and CP was higher ( P<0.05) for the amylose-rich (0.89 and 0.83) and amylopectin-rich (0.89 and 0.80) barley cultivars than for the normal barley cultivar (0.85 and 0.77). However, there were no differences in performance ( P>0.05) between diets based the different barley cultivars. Enzyme supplementation of the diets improved the CTTAD of CP ( P<0.05), while it had no effect ( P>0.05) on the performance. In conclusion, the starch composition of barley appears to have a limited effect on its utilization in piglets. However, the impaired digestibility of the naked normal barley cultivar indicates that there are properties in the hulless grain that imposes limitation to its utilization and which could not be alleviated by enzyme supplementation. Overall, the present results support the contention that the potential for improvement of piglet growth performance by supplementing barley-based diets with cell wall degrading enzymes is limited.

Low-phytic acid barley improves calcium and phosphorus utilization and growth performance in growing pigs1

Thirty-five crossbred barrows averaging 13.5 kg starting BW were used in a 35-d experiment to compare the availability of P and the nutritional value of two near-isogenic progeny of the barley cultivar 'Harrington'. Low-phytic acid barley (LPB, 0.35% total P, 0.14% phytic acid P) was homozygous for the low-phytic acid 1-1 allele, and the normal barley (NB, 0.35% total P, 0.24% phytic acid P) was homozygous for the normal allele of that gene. Pigs were fed individually twice daily in metabolism pens. Barley was the only source of phytate in semipurified diets, 1 to 3. Diet 1 contained 75% NB, 0.14% estimated available P (aP), and 0.50% Ca. Diet 2 contained 75% LPB, 0.22% aP, and 0.50% Ca. No inorganic P (iP) was added to Diets 1 and 2 in order to measure the animal response to the different concentrations of aP in these cultivars. Diet 3 was NB Diet 1 supplemented with iP to equal the concentration of aP in LPB Diet 2. Practical barley-soybean meal (SBM)-type diets, NB Diet 4 and LPB Diet 5, were formulated to meet all minimum nutrient requirements, and contained 0.30% aP and 0.65% Ca. For the semipurified diets, pigs fed LPB Diet 2 had higher (P < or = 0.05) bone ash weight, bone breaking strength, P absorption and retention, and Ca absorption and retention compared with pigs fed NB Diet 1, with a trend (P = 0.10) for pigs fed LPB Diet 2 to have a higher ADG and gain:feed ratio than pigs fed NB Diet 1. However, pigs fed LPB Diet 2 or NB Diet 3 were not different (P > or = 0.3) in growth performance, fresh bone weight, fat-free dry bone weight, bone ash, bone breaking strength, or N utilization. This indicates that LPB and NB were equal in nutritional value after supplementation of NB with iP to equal the estimated aP in LPB. For the practical barley-SBM diets, there were no differences (P > or = 0.4) between pigs fed NB Diet 4 or LPB Diet 5 for growth performance, fresh bone weight, bone breaking strength, the percentages of P and Ca utilization, or N, DE, and ME utilization. The use of LPB in pig diets reduced P excretion in swine waste by 55% and 16% in our semipurified and practical diets, respectively, compared with NB. Using our in vitro procedure designed to mimic the digestive system of the pig, the availability of P for pigs was estimated at 52% for LPB and 32% for NB.

Starch Properties of a Waxy Mutant Line of Hull-less Barley (Hordeum vulgare L.)

Endosperm starch isolated from an amylose-free waxy mutant hull-less barley line, Shikoku Hadaka 97, had an amylose content of 0.3% and higher swelling power than ordinary waxy barley cultivars/lines with amylose contents of 2.2-6.5%. A highly significant correlation was observed between amylose content and swelling power among waxy barley starches. No clear differences were detected in the chain-length distribution profiles or thermal properties between the amylose-free starch and ordinary waxy starch. The chain-length distribution profile of waxy barley starch was slightly different from that of normal barley starch. Gelatinization temperatures and gelatinization enthalpy of waxy barley starch were higher than those of normal barley starch. Significant correlations were observed between amylose content and thermal properties of starch samples analyzed. Waxy barley starch stained with a concentrated iodine-potassium iodide solution showed a ghost-like appearance.

Measuring Gene Flow in the Cultivation of Transgenic Barley

Genetic engineering is becoming a useful tool in the improvement of plants and plant-based raw materials. Varieties with value-added traits are developed for nonfood use in industrial and medical production, and different production lines must be kept separate. For good management practices, knowledge of relevant gene flow parameters is required. In the present study, pollen-mediated dispersal of transgenes via cross-fertilization was examined. A transgenic barley (Hordeum vulgare L.) line carrying a marker gene coding for neomycin phosphotransferase II (nptII) was used as a pollen donor. For maximum resolution, a cytoplasmically male-sterile barley line was utilized as recipient and the flow of nptII transgene was monitored at distances of 1, 2, 3, 6, 12, 25, 50, and 100 m from the donor plots of 225 and 2000 m2 Male-fertile plots at a distance of 1 m were included to measure the transgene flow in normal barley. The number of seeds obtained from male-sterile heads diminished rapidly with distance and only a few seeds were found at distances of 50 and 100 m. Molecular genetic analysis (polymerase chain reaction—PCR) revealed that all seeds obtained from male-sterile heads at a distance of 1 m were transgenic, as anticipated. However, only 3% of the distant seeds (50 m) actually carried the transgene, whereas most of them resulted from fertilization with nontransgenic background pollen. This background pollen was mainly due to pollen leakage in some male-sterile heads. In normal male-fertile barley, the cross-fertilization frequency with transgenic pollen varied from 0 to 7% at a distance of 1 m, depending on weather conditions on the heading day. We conclude that, because of competing self-produced and nontransgenic background pollen, the possibility of cross-pollination is very low between a transgenic barley field and an adjacent field cultivated with normal barley. However, adequate isolation distances and best management practices are needed for cultivation of transgenic barley.

Measuring Gene Flow in the Cultivation of Transgenic Barley

Genetic engineering is becoming a useful tool in the improvement of plants and plant-based raw materials. Varieties with value-added traits are developed for nonfood use in industrial and medical production, and different production lines must be kept separate. For good management practices, knowledge of relevant gene flow parameters is required. In the present study, pollen-mediated dispersal of transgenes via cross-fertilization was examined. A transgenic barley (Hordeum vulgare L.) line carrying a marker gene coding for neomycin phosphotransferase II (nptII) was used as a pollen donor. For maximum resolution, a cytoplasmically male-sterile barley line was utilized as recipient and the flow of nptII transgene was monitored at distances of 1, 2, 3, 6, 12, 25, 50, and 100 m from the donor plots of 225 and 2000 m(2). Male-fertile plots at a distance of 1 m were included to measure the transgene flow in normal barley. The number of seeds obtained from male-sterile heads diminished rapidly with distance and only a few seeds were found at distances of 50 and 100 m. Molecular genetic analysis (polymerase chain reaction-PCR) revealed that all seeds obtained from male-sterile heads at a distance of 1 m were transgenic, as anticipated. However, only 3% of the distant seeds (50 m) actually carried the transgene, whereas most of them resulted from fertilization with nontransgenic background pollen. This background pollen was mainly due to pollen leakage in some male-sterile heads. In normal male-fertile barley, the cross-fertilization frequency with transgenic pollen varied from 0 to 7% at a distance of 1 m, depending on weather conditions on the heading day. We conclude that, because of competing self-produced and nontransgenic background pollen, the possibility of cross-pollination is very low between a transgenic barley field and an adjacent field cultivated with normal barley. However, adequate isolation distances and best management practices are needed for cultivation of transgenic barley.

Low-phytic acid barley improves calcium and phosphorus utilization and growth performance in growing pigs

Thirty-five crossbred barrows averaging 13.5 kg starting BW were used in a 35-d experiment to compare the availability of P and the nutritional value of two near-isogenic progeny of the barley cultivar 'Harrington'. Low-phytic acid barley (LPB, 0.35% total P, 0.14% phytic acid P) was homozygous for the low-phytic acid 1-1 allele, and the normal barley (NB, 0.35% total P, 0.24% phytic acid P) was homozygous for the normal allele of that gene. Pigs were fed individually twice daily in metabolism pens. Barley was the only source of phytate in semipurified diets, 1 to 3. Diet 1 contained 75% NB, 0.14% estimated available P (aP), and 0.50% Ca. Diet 2 contained 75% LPB, 0.22% aP, and 0.50% Ca. No inorganic P (iP) was added to Diets 1 and 2 in order to measure the animal response to the different concentrations of aP in these cultivars. Diet 3 was NB Diet 1 supplemented with iP to equal the concentration of aP in LPB Diet 2. Practical barley-soybean meal (SBM)-type diets, NB Diet 4 and LPB Diet 5, were formulated to meet all minimum nutrient requirements, and contained 0.30% aP and 0.65% Ca. For the semipurified diets, pigs fed LPB Diet 2 had higher (P < or = 0.05) bone ash weight, bone breaking strength, P absorption and retention, and Ca absorption and retention compared with pigs fed NB Diet 1, with a trend (P = 0.10) for pigs fed LPB Diet 2 to have a higher ADG and gain:feed ratio than pigs fed NB Diet 1. However, pigs fed LPB Diet 2 or NB Diet 3 were not different (P > or = 0.3) in growth performance, fresh bone weight, fat-free dry bone weight, bone ash, bone breaking strength, or N utilization. This indicates that LPB and NB were equal in nutritional value after supplementation of NB with iP to equal the estimated aP in LPB. For the practical barley-SBM diets, there were no differences (P > or = 0.4) between pigs fed NB Diet 4 or LPB Diet 5 for growth performance, fresh bone weight, bone breaking strength, the percentages of P and Ca utilization, or N, DE, and ME utilization. The use of LPB in pig diets reduced P excretion in swine waste by 55% and 16% in our semipurified and practical diets, respectively, compared with NB. Using our in vitro procedure designed to mimic the digestive system of the pig, the availability of P for pigs was estimated at 52% for LPB and 32% for NB.

Improvement of malting quality of barley by complementing the malt enzyme spectrum

The processing quality of cereals can be modified by altering the structural grain constituents or the enzyme activities that mobilize storage reserves of the seeds. In order to complement the malt enzyme spectrum, a gene encoding for a thermotolerant fungal endo-(1,4)-β-glucanase was introduced into two barley cultivars, Kymppi and Golden Promise. The gene was expressed in the seeds during germination, thus providing a thermotolerant enzyme that is active under mashing conditions. The amount of thermotolerant β-glucanase produced by the seeds (ca. 0.025% soluble seed protein) has been shown to be sufficient to reduce wort viscosity by decreasing the soluble β-glucan content. For the safe commercial cultivation of transgenic plants risk assessment of their cultivation is needed. In our study experimental estimates of the transgene flow from transgenic barley by pollen dispersal were produced. Field trials were conducted during the summers of 1996 and 1997. A transgenic barley line homozygous for the gene encoding for neomycin phosphotransferase was used as a source of pollen and male-sterile barley lines as recipients. In order to be able to transform the cross-fertilization frequencies to corresponding values of normal male-fertile barley, plots of normal barley were also included in the experimental plan. On the basis of our study, cross-fertilization in male-sterile recipient barley is possible with very low frequency up to 50 meters from the donor area. However, the frequency dramatically decreases with distance and due to self-pollination the possibility of cross-fertilization remains very low in normal cultivated barley.

Exploring the phenotypic expression of a regulatory proteome-altering gene by spectroscopy and chemometrics

Evaluating gene effects on proteomes and the resulting indirect pleiotropic effects through the cell machinery on the chemical phenotype constitutes a formidable challenge to the analytical chemist. This paper demonstrates that near-infrared (NIR) spectroscopy and chemometrics on the level of the barley seed phenotype is able to differentiate between genetic and environmental effects in a PCA model involving normal barley lines and the gene regulator lys3a in different genetic backgrounds. The gene drastically changes the proteome quantitatively and qualitatively, as displayed in two-dimensional electrophoresis, resulting in a radically changed amino acid and chemical composition. A synergy interval partial least squares regression model (si-PLSR) is tested to select combinations of spectral segments which have a high correlation to defined chemical components indicative of the lys3a gene, such as direct effects of the changed proteome, for example, the amide content, or indirect effects due to changes in carbohydrate and fat composition. It is concluded that the redundancy of biological information on the DNA sequence level is also represented at the phenotypic level in the dataset read by the NIR spectroscopic sensor from the chemical physical fingerprint. The PLS algorithm chooses spectral intervals which combine both direct and indirect proteome effects. This explains the robustness of NIR spectral predictions by PLSR for a wide range of chemical components. The new option of using spectroscopy, analytical chemistry and chemometrics in modeling the genetically based covariance of physical/chemical fingerprints of the intact phenotype in plant breeding and biotechnology is discussed.

Physicochemical properties and structure of large, medium and small granule starches in fractions of normal barley endosperm

Normal barley grain was milled to flour with a machine used to polish brewers’ rice from the surface layer to the center. Large (18.4 μm, median size), medium (12.3 μm) and small (2.2 μm) granule starches were isolated from classified flours. Their physicochemical properties and fine structure were investigated. The percentage (w%) of large granules decreased from the surface layer to the center, while the amounts of medium and small granules increased. Although all the starch granules were an A-type crystal, the relative crystallinity varied from 22.0 to 27.4%. The DP n of the amyloses was around 1600 and similar for all the samples. But the amylose content of the starches varied from 21.9 to 26.4%. Also, the amylopectins showed differences in DP n (around 5700–7900) and chain-length distribution between granule size or fractions. The transition temperature ranges and the enthalpy values of the starch granules differed with granule size. The gelatinization properties showed no correlation with any of the parameters, except the enthalpy value and relative crystallinity ( γ=+0.73). The findings suggested that the structural characteristics of the starches in classified flours of normal barley differed essentially from those of waxy barley.

Molecular Structure and Pasting Properties of Normal Barley Starches.

Molecular Structure and Pasting Properties of Normal Barley Starches.

Variation in total and soluble beta-glucan content in hulless barley: effects of thermal, physical, and enzymic treatments.

Total and soluble beta-glucan content and effects of various treatments of barley grain on extractability and molecular characteristics of soluble beta-glucan were studied. Four types of hulless barley (normal, high amylose, waxy, and zero amylose waxy) from 29 registered and experimental genotypes were analyzed. For each, moisture, protein, amylose, 100 kernel weight, starch, beta-glucan (total and soluble), beta-glucanase activity, and slurry viscosity were determined. Significant differences in total beta-glucan were observed among the groups, with average values of 7. 49%, 6.86%, 6.30%, and 4.38% for high amylose, waxy, zero amylose waxy, and normal barley, respectively. The extractability of beta-glucan in high amylose barley was relatively low (20.6-29.7%) compared to that in normal (29.8-44.3%), zero amylose waxy (34.0-52. 5%), and waxy (36.7-52.7%) barley genotypes. Viscosity of barley flour slurries was affected by the content of soluble beta-glucans, beta-glucanase activity, and molecular weight of beta-glucans. Hydrothermal treatments (autoclaving and steaming) of barley had no effect on extractability of beta-glucans, but prevented enzymic hydrolysis of beta-glucans, and thereby substantially improved their molecular weight. The addition of enzymes (protease and esterase) during extraction and/or physical treatments (sonication) increased extractability of beta-glucans from barley.

Morphological Changes of Granules of Different Starches by Surface Gelatinization with Calcium Chloride

ABSTRACTNative starch granules of 11 selected cultivars (potato, waxy potato, sweet potato, normal maize, high‐amylose maize, waxy maize, wheat, normal barley, high‐amylose barley, waxy barley, and rice) were treated with a calcium chloride solution (4M) for surface gelatinization. The surface‐gelatinized starch granules were investigated using light microscopy and scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). In general, those starches with larger granule sizes required longer treatment time to complete the gelatinization. The salt solution treatment of starch was monitored by light microscopy and stopped when the outer layer of the granule was gelatinized. The surface gelatinized starch granules were studied using scanning electron microscopy. On the basis of the gelatinization pattern from calcium chloride treatments, the starches could be divided into three groups: 1) starches with evenly gelatinized granule surface, such as normal potato, waxy potato, sweet potato, maize, and high‐amylose maize; 2) starches with salt gelatinization concentrated on specific sites of the granule (i.e., equatorial groove), such as wheat, barley, and high‐amylose barley; and 3) starches that, after surface gelatinization, can no longer be separated to individual granules for SEM studies, such as waxy barley, waxy maize, and normal rice. The morphology of the surface gelatinized starch resembled that of enzyme‐hydrolyzed starch granules.

Long-term effects of several biochemical and physical factors on cellulose biosynthesis in callus and suspension cultures of normal and mutant barley (Hordeum vulgare L.) strains producing less cellulose

To clarify a low level of cellulose biosynthesis of thein vitro cultured cells, the effects of several biochemical factors such as carbon sources (sucrose, maltose, and UDPG), antioxidants (ascorbic acid and glutathione) and physical factors such as artificial pressure, high gravity, on the cellulose production in barley callus and suspension cultures were investigated. In the suspension culture of two barley strains, the supplement of different concentrations (0, 1.5, 3.0, and 4.5%) of sucrose or maltose into the medium for 30 days did not promote the cellulose production and 4.5% of sugar supplement was rather inhibitory in one strain. However, in the presence of sucrose at 3%, UDPG (3 or 10 mM) supplement, as a precursor for cellulose, promoted 1.2–13 fold of the production in two strains. A low concentration (3 mM) of ascorbic acid and glutathione promoted 1.5 and 1.2 fold of the production in two strains, respectively. These results suggest that low cellulose biosynthesis of thein vitro cultured cells is due to a decreased level of the UDPG in the cytosol, and that the oxidative condition of external medium impedes cellulose synthesis in some manners. Artificial pressure applied to the callus promoted 1.4 fold of the cellulose production. High gravity (5,000 or 10,000g) applied to the suspension-cultured cells by centrifugation did not cause a substantial change.

Hull-less barley for swine: ileal and faecal digestibility of proximate nutrients, amino acids and non-starch polysaccharides

The nutritive value of three hull-less barley cultivars (Condor, Falcon and CDC Buck) were evaluated. The samples were analysed for proximate composition, amino acids and non-starch polysaccharides (NSP). Wheat and normal barley were used for comparison. Apparent digestibilities were determined with eight cannulated pigs. The diets were formulated to have all energy and protein supplied by the cereals. Compared to normal barley, hull-less barley has higher contents of protein and amino-acids, and lower levels of fibre components. The levels of several essential amino acids in hull-less barley are higher than in wheat. No obvious differences in NSP composition were observed, but the hull-less barley contained considerably higher levels of β-glucan compared to hulled barley and wheat. In digestion trials the piglets did not accept the diet in which conventional barley was formulated as the only energy and protein source, whereas hull-less barleys were well consumed. The results showed that, in the order of Condor, Falcon, CDC Buck and wheat, the overall tract digestibilities of intake energy were 83·1, 80·3, 83·2 and 82·9%, and of protein were 72·6, 69·5, 76·5 and 77·6%, respectively. The averages of apparent ileal amino acid digestibility were 56·5, 58·4, 65·3 and 65·4%, respectively. Hull-less barleys showed 20–37% ileal digestibility of total NSP. This study provides digestibility coefficients of hull-less barleys, and it is concluded that the digestibility of hull-less barley is comparable to wheat and superior to hulled barley in terms of nutrient composition and acceptability. © 1998 SCI.