Session 6 Barley Protein, Feed and Food

Oral Presentation

Functional Genomics in the Productivity and End-Use Quality
of Barley

E. Newbigin1, A. Bacic1, P. Langridge2 and G. B. Fincher2

1School of Botany, University of Melbourne, Parkville, VIC 3052, Australia,
E-mail: edwardjn@unimelb.edu.au; 2Australian Centre for Plant Functional Genomics, University
of Adelaide, Glen Osmond, SA 5064, Australia

Functional genomics is a technology through which large sets of genes that influence a particular biological process are identified. Functional genomics comprises several sub-disciplines, including genomics, proteomics and metabolomics. The total number of genes in plant genomes lies somewhere in the range of 25,000 to 40,000. Thus, functional genomics technologies must be geared to high throughput data collection and high throughput analyses of gene structure and function. Additional specialised resources, including barley mutant libraries, barley DNA microarrays and high-density genetic maps of barley, represent critical support technologies. The technologies will be illustrated by reference to specific applications relating to malting and brewing performance, such as the biosynthesis of cell wall polysaccharides, and to productivity, where tolerance to a range of abiotic stresses is a key determinant of yield. Finally, the potential impacts and benefits of functional genomics research for the barley industry will be addressed. Where a gene controlling an important trait has been isolated, a diagnostic marker that is 100% accurate in predicting the phenotype of the plants for the target trait could be used in high throughput screening of germplasm in breeding programs.

Supported by grants from the Grains Research and Development Corporation and the Australian Research Council.

Barley Foods for Good Nutrition and Health

C. W. Newman and R. K. Newman

College of Agriculture, Montana State University, Bozeman, MT 59717, USA,
E-mail: cwn@montana.com

Barley furnishes a multitude of dietary nutrients including amino acids, minerals, vitamins, insoluble and soluble dietary fiber and physiologically active phytochemicals. Many genotypes of barley are available for processing into foods and some are more concentrated in various nutrients than others and differ in processing requirements. Concentration of nutrients in barley can be altered by standard breeding practices. New breeding techniques are being used to reduce phytate, which is considered to be an antinutrient, although some reports indicate beneficial effects of phytate. Barley can be used in numerous ways to prepare delicious, nutritious and health promoting foods, depending upon the innovation of a food processor. Traditional use of barley as a food has been pearl barley in soups, stews and as a replacement or extender for white rice. Processing techniques, such as milling, flaking and extrusion increase the variety of barley food products. Research has shown that the consumption of barley food products is effective in lowering blood cholesterol thus reducing risk of heart disease. Barley also has a very low glycemic index which helps to prevent high peaks in blood sugar and insulin after meals. This effect is beneficial in the prevention of type 2 diabetes and controlling blood sugar in both type 1 and type 2 diabetes. Because of the nutritional and health benefits of barley, its consumption in human foods should be actively promoted.

Genetic Studies of Barley Seed Phosphate and Phytic Acid

V. Raboy1, H. Saneoka2, J. A. Dorsch1, K. Peterson1, D. Bowen3, J. Fu3, M. Guttieri3 and E. Souza3

1USDA-ARS, Aberdeen, ID 83210-0530, USA, E-mail: vraboy@uidaho.edu;
2Hiroshima University, Higashi-Hiroshima, 739-8526 Japan; 3University of Idaho, Aberdeen,
ID 83210-0530, USA

Phytic acid represents about 65% of seed total P. More than 20 heritable barley low phytic acid (lpa) mutants have been isolated. These mutations result in reductions in seed phytic acid P ranging from 50% (lpa1-1) to > 90% (M 955), but have little effect on seed total P. Barley lpa1-1 appears to be aleurone specific. The first three barley lpa mutations that were mapped were found to represent three non-linked loci, termed lpa1 (chromosome 2H), lpa2 (chromosome 7H) and lpa3 (chromosome 1H; Roslinsky, Eckstein, Rossnagel & Scoles pers. com.), respectively. Studies are in progress to determine the alleleic relationship and map positions of the additional 15 as yet unmapped barley lpa mutations. The pathway to phytic acid begins with the synthesis of inositol (Ins), via the conversion of glucose-6-P to Ins(3)P1, catalyzed by the enzyme d-myo-inositol 3-phosphate synthase (MIPS, E.C. 5.5.1.4), and then hydrolysis of Ins(3)P1 to Ins, catalyzed by an Ins monophosphatase (EC 3.1.3.25). We isolated and mapped (to chromosome 4H) the barley genomes single copy MIPS gene, and are now working on Ins monophosphatase-encoding sequences. We plan to screen for mutations in these sequences using the TILLING reverse genetics approach. We are also studying the impact of lpa mutations on expression of genes outside those in the pathways of phytic acid synthesis. In separate studies we are pursuing candidate gene hypotheses concerning specific Ins phosphate kinases and lpa loci. Progress in breeding Low Phytate barley is also progressing, and agronomic and nutritional studies will be reviewed.

Investigation of Barley Proteome

J. Chmelik and P. Rehulka

Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, 611 42 Brno,
Czech Republic, E-mail: chmelik@iach.cz

In contrast to the static genome, the proteome is dynamic and reflexes the real state of life. The proteomic approach to the characterization of the metabolic and storage protein content in barley grains may give fundamental information on the relationships between protein composition/quality, changes of the protein composition during development and under different environmental conditions (location, season) or differences in the protein composition between varieties. Recent research advances at gene level are providing a lot of nucleotide sequences, genetic map and DNA markers for barley. However, knowledge of the genotype alone does not permit either to select better cultivars, or to determine the environmental effects on grain quality. To this aim, detailed elucidation of the protein composition is necessary in order to assess the quality barley. In addition, identification of individual proteins as markers of phenotype has great value at all stages of barley production. Detailed knowledge of the grain protein composition may produce further insight into the relationships between protein composition and the technological properties of barley. Identification of several proteins extracted with various solvents from barley grains by the proteomic strategy based on a combination of gel electrophoresis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and bioinformatics was performed. The mass spectrometry-based strategy designed for protein identification exhibits good sensitivity and rapid protein identification in comparison to other methods. In order to simplify the protein mixture extracted from grains, we used sequential extractions of grains with various solvents (based on modified Osborne system). Several proteins (e.g. beta-amylase, B3 hordein) were identified in this way. It seems that this approach can become an important tool for the investigation of proteome of cereals.

Poster Presentation

The Characteristics of Barley Germplasm with Increased
Crude Protein Content

I. Belicka, U. Briedis and A. Liepa

Department of Crop Production, Latvia University of Agriculture, Jelgava, LV 3001, Latvia,
E-mail: stende.selekcija@apollo lv.

Evaluation of barley germplasm with increased protein content (15.023.5%) received from USDA-ARS Small Grain Collection was carried out in Latvia University of Agriculture. There were hulled and hulless both two-row and six-row genotypes from different continents and states. There was wide variability in yield, coefficient of variation medium to high among the genotypes represented Europe, America, Australia, Asia and Africa. The mean yield of genotypes from Asia and Africa differed significantly by other groups. Two-row hulless barley had higher yield, if compared to six-row. The coefficient of variation of crude protein was 6.18.8% and genotypes of Asia and Africa differed significantly by this parameter from European and American genotypes. Starch content of barley genotypes with high protein content was less variable trait, and there were no significant differences among investigated accessions. With increased yield, the content of crude protein decreased, coefficient of correlation for hulled two-row barley r = 0.307 (r0.05 = 0.175), for six-row r = 0.541 (r0.05 = 0.253). There was significantly negative correlation between starch and crude protein content. Our experiments showed that barley germplasm comprises genotypes with increased crude protein content and good agronomic traits. It is possible to select the initial material for crossing and including them in barley breeding programme for feed.

Relationships between Some Hordein Components
and Quality Properties in Two Tunisian Barley Varieties
as Influenced by N Fertilization

L. Bettaieb Ben Kaab1, M. El Felah2, M. H. Ghorbal1 and M. Lauriere3

1Département de Biologie, Faculté des Sciences de Tunis, Campus Universitaire,1060 Tunis, Tunisie, E-mail: leila.bk@planet.tn;
2Département de Génétique, INRAT, 2049-Ariana, Tunis, Tunisie; 3Centre de Biotechnologie
Agro-Industrielle, INRA, Paris-Grignon, 78850 Thiverval-Grignon, France

Two Tunisian barley varieties, Martin and Rihane, differing in degrees of adaptation to environmental conditions and grain yield potential, were grown under different levels of nitrogen (N) fertilization. Effect of N supply on the hordein components (expressed in mg per albumen) was determined by Nu-PAGE electrophoresis system using the MOBS running buffer and densitometric analysis. Increasing N fertilization increased protein content by increasing the amount of C-hordeins. The subfraction BIII of high molecular weight B-hordeins appeared, in the present study, as the best criterion for distinguishing between the two barley varieties genetic adaptation and its quantitation could be recommended for understanding of quality properties of barley response to different environmental conditions particularly N fertilization which appear to be the main origin of differences in the protein aggregation mode of different B-hordeins sulphur-rich subfractions.

The Barley Starch Granule Proteome

M. Boren, H. Larsson, A. Falk and C. Jansson

Plant Biology and Forest Genetics, Swedish Agricultural University, 750 07 Uppsala, Sweden,
E-mail: mats.boren@ubsg.slu.se

We describe the initial characterization of the starch granule proteome of barley, containing the proteins firmly associated with the granule. Starch granule preparations from mature barley seeds were subjected to SDS washes and enzymatic shaving with thermolysin prior to protein extraction in boiling SDS buffer. Proteins were examined by two-dimensional and one dimensional electrophoresis. A typical granule proteome contained around 150 proteins spots. A total of 74 spots were identified by mass fingerprinting and sequencing using mass spectrometry with integrated NCBI database searches. Four of the central enzymes in starch synthesis, i.e. GBSSI, SSI, SSII and SBEIIb, together with hordeins B and D, serpin Z4, pyruvate orthophosphate dikinase and an enzyme involved in thiamine biosynthesis, were identified. Unexpectedly, of the identified spots on the two-dimensional gel, 49 were found to belong to GBSSI, representing variations in both relative molecular mass and isoelectric point. The GBSSI fragmentation was consistent between preparations and cultivars and is inferred to result from in vivo proteolysis. Based on the enzymatic shaving, we conclude that the starch synthesis enzymes, including the GBSSI fragments, are integral components of the granule, whereas the other granule-bound proteins are attached to the surface.

Protein Fractions and the Assessment of Their Genotype Variability

V. Curn1 and J. Barta2

1Biotechnological Centre and 2Department of Plant Production, Faculty of Agriculture, University of South Bohemia in Ceske Budejovice, 370 05 Ceske Budejovice, Czech Republic, E-mail: curn@zf.jcu.cz

Proteins represent an important part of barley grain, mainly from the point of view of the food industry application. In this presentation, 20 barley genotypes (cultivars and new breeding lines) of world-wide range were evaluated for their total protein content and relative percentage of protein solubility fraction. The modified method of extraction of protein fractions (Albumins + Globulins, Prolamins, Glutelins) based on the Osbornes principle was used. Total protein content and protein fraction contents were determined as total nitrogen content in dry matter (N × 6.25) by modified Dumas method on elementar analyser Flash EA 1112 (ThermoQuest, Italy). Genotype variability in protein fraction composition was evaluated by electrophoresis (SDS-PAGE). In total protein content, the statistically significant differences among analyzed genotypes were found. The values of total protein content were in range 9 to 21% of dry matter. The significant differences were also found in individual protein fraction mean proportions of albumins + globulins, prolamins and, glutelins represent 16.1%, 22.4%, and 38.2% of the crude protein, respectively. SDS-PAGE profiles of prolamin fraction may be considered as a suitable tool for barley genotype characterization.

Supported by the Ministry of Agriculture of the Czech Republic, Grant No. NAZV QD 1365.

Variability of Vitamin E Content in Caryopses of Spring Barley Lines of Food Type

J. Ehrenbergerova1, J. Pryma2, N. Belcrediova1 and K. Vaculova3

1Department of Crop Science and Plant Breeding, Mendel University of Agriculture and Forestry Brno,
613 00 Brno, Czech Republic, E-mail: ehren@mendelu.cz;
2Research Institute of Brewing and Malting, PCL, Malting Institute in Brno, 614 00 Brno, Czech Republic;
3Agricultural Research Institute Kromeriz, Ltd., 767 01 Kromeriz, Czech Republic

Concentration of vitamin E and its isomers (-, -, -, -tocopherols and tocotrienols, i.e. tocols generally) was followed (in caryopses F2 to F6 of generations of crosses and lines. For crossing the malting type varieties registered in the CR with the varieties of the waxy type (obtained from the USA from professor C. W. Newman) were used. Content of all isomers of vitamin E varied from 16 to 114 mg/kg and the main share was that of isomers - and -, -tocotrienols (50 and 32%). The parental waxy varieties of the American provenience Wanubet and Washonubet (73.9478.82 mg/kg), lines Kompakt × Wabet (72.32 mg/kg) and line Wabet × Krona (64.8 mg/kg) had statistically significantly higher average content of all tocols in comparison with the other lines and varieties. Activity of vitamin E, expressed in mg of -tocopherol-equivalent moved from 16.92 to 26.46 mg/kg in waxy varieties of the American provenience, which was significantly more than in varieties of the malting type (12.1520.81 mg/kg). We determined statistically higher: (1) -tocopherol content in the line Krona × Kompakt (5.92 mg/kg)
versus the variety Krona (4.80 mg/kg) (2) -, -tocopherol content in the lines Kompakt × Krona (4.16 mg/kg), Krona × Wanubet (3.93 mg/kg), Krona × Kompakt (3.78 mg/kg), and Wanubet × Krona (3.40 mg/kg) in comparison with the parental variety Krona (2.60 mg/kg) (3) -tocopherol content in the line Kompakt × Wabet (1.23 mg/kg) surpassing thus significantly the parental variety Kompakt (1.01 mg/kg) (4) -tocotrienol content versus the variety Krona (21.02 mg/kg) in the line Krona × Wanubet (23.07 mg/kg) and Wanubet × Krona (25.28 mg/kg). By hybridization of the malting type varieties with the varieties of the type waxy, significant increase of content of some effective isomers of vitamin E in the caryopses of some lines was achieved, which helps to raise antioxidant capacity and suitability of these lines for utilization in the food industry.

Hulled and Hull-Less Barley Genotypes for the Development of Functional Foods

A. Gianinetti, F. Finocchiaro, B. Ferrari and A. M. Stanca

Istituto Sperimentale per la Cerealicoltura, 29017 Fiorenzuola dArda (PC), Italy,
E-mail: kimhongs@rda.go.kr

Among cereals, barley is the main cereal grain for the development of functional foods, as it contains several classes of compounds of strong nutritional interest: tocols, -glucans and proanthocyanidins. Tocols are well recognised for their biological effects, including antioxidant and hypocholesterolemic activity. Barley is considered a good source of this class of compounds, because it contains both a high concentration of total tocols and a favourable distribution of the most biologically active isomers. In fact, our results indicate that different species of cereals can be distinguished from their tocol isomers distribution, with barley richest in isomers more biologically active (- and -tocotrienols). Beta-glucans have beneficial effects on blood cholesterol level and on glycemic response. They are a complex class of polysaccharide and can be divided into soluble and insoluble fractions. It has been suggested that these different fractions could have different physiological effects. For this reason we are evaluating different barley genotypes with the aim of investigating the concentration of soluble and insoluble
-glucans in the grain. The proanthocyanidins, a class of polyphenols with a strong antioxidant activity, are interesting from a nutritional point of view. Barley grains contain these compounds and we are evaluating different genotypes for high concentration of proanthocyanidins.

A Functional Genomics Approach for Improving Amino Acid Composition in the Developing Barley Grain

M. Hansen, E. Vincze and P. B. Holm

Department of Plant Biology, Danish Institute of Agricultural Sciences, DK-4200 Slagelse, Denmark,
E-mail: michael.hansen@agrsci.dk, vikingen@get2net.dk

Barley is extensively used as feed for pigs and chickens. However, since it has a suboptimal content of the essential amino acids lysine, methionine and threonine, massive supplementation with other protein sources or amino acids are required to meet the nutritional demands. The second problem is that prolamins, the major barley storage proteins, contain large amounts of glutamine and proline. These amino acids cannot be utilized completely by the animals. The excess is degraded and the nitrogen excreted in the urine contributing significantly to the environmental nitrogen load. The objective of our project is to get more insight into amino acid metabolism in the developing barley grain at the molecular level and eventually to alter the amino acid composition by molecular breeding techniques. Our primary experimental approach comprise the generation of pathway specific microarrays for developing barley grains where we have assembled a comprehensive set of genes involved in nitrogen mobilization, transport and amino acid metabolism. Genes of major significance for carbon and lipid metabolism are also included in order to assess the interconnections between the basic metabolic pathways in the grain. The collected target materials used for microarrays are barley grains grown under different nitrogen regimes in the field and the high lysine mutant, lys 3a that possess an improved amino acid composition.

Inheritance of (1-3) (1-4)--D-Glucan Content
in Barley (Hordeum vulgare L.)

H. S. Kim, K. G. Park, S. B. Baek, Y. G. Son, C. W. Lee, J. C. Kim, J. G. Kim and J. H. Nam

National Institute of Crop Science, R.D.A. Suwon, 441-857, Korea,
E-mail: kimhongs@rda.go.kr

Beta-glucan is the soluble dietary fiber component and occurs in highest amounts in the endosperm of barley. It has been shown to have health benefits such as cholesterol reducing activity as human nutrition. On the other hand, a high -glucan level has a bad effect on the grain quality for malting and feeding. Our objectives were to study the inheritance of -glucan content in barley and mapping quantitative trait loci (QTL) associated with this trait. F5-derived 107 lines from the cross of the six-rowed waxy hulless barley cultivar originating from Japan cv. Yonezawa Mochi and the six-rowed non-waxy hulless barley cultivar originating from Korea cv. Neulssalbori were measured for the agronomic traits and -glucan level. Eighty-eight lines of them were also analyzed for 88 genetic markers to map QTLs. The -glucan content of recombinant lines was normally distributed with a range of 3.06.65% across environments. By single point analysis, 12 significant markers were found. At least, three QTLs could be mapped on chromosome 1H and 4H with genetic markers explaining 5.821.4% of the variability for -glucan content. On average, 16.4% of the total phenotypic variance of -glucan content could be explained by the waxy gene on chromosome 1H. The effects of genotypes and environment on the -glucan level of barley are discussed.

Development of New Waxy Barley Cultivars for Healthy Diet Purpose in Korea

J. H. Nam, H. S. Kim, J. G. Kim, S. B. Baek, Y. G. Son, C. W. Lee, M. S. Kang and S. J. Suh

National Institute of Crop Science, R.D.A. Suwon, 441-857, Korea,
E-mail: namjhy@rda.go.kr

In Korea, barley has been used as one of the major food crops with a form of whole grain diets usually cooked with rice. Improvement of cooking quality and specific nutrition components of food barley is facing a new challenge with the growing awareness of the beneficial effects of a healthy diet. In our studies, various isogenic lines were developed with four genetic combinations of waxy/hulled, waxy/hulless, non-waxy/hulled, and non-waxy/hulless forms by backcrossing a waxy hulless Korean landrace, Masangwamaek and a waxy hulless Japanese cultivar, Yonezawa Mochi to the non-waxy hulled Korean cultivars, Kangbori and Olbori, respectively. The effects of these genes were analyzed on the agronomic traits, cooking quality, and texture properties. No typical differences were observed among isogenic lines for agronomic traits and yield components. The amylase content of waxy lines (2.12.9%) was lower than that of non-waxy lines (18.919.7%) regardless of hulled/hulless forms. The addition of waxy gene could affect hydrolysis pattern of grains with lowering temperature for gelatinization as well as for the highest viscosity level by 13oC and 15oC, respectively. The duration for gelatinization of waxy starch grains was shorter by 11 minutes than non-waxy starch grains. On the other hand, there was no significant difference between hulled and hulless lines for amylase content and cooking qualities.

Genotypic Variations in Phytate-Phosphorus of Barley
and Other Cereals

J. M. Nyachiro, J. L. Zantinge, J. H. Helm, P. E. Juskiw and D. Salmon

Field Crop Development Centre, Alberta Agriculture, Food and Rural Development, Lacombe,
AB T4L 1W8, Canada, E-mail: joseph.nyachiro@gov.ab.ca

Phytate is a salt of phytic acid (myo-inositol-1,2,3,4,5,6-hexakiphosphate) and it is the most abundant form of phosporus (P) in seeds where it is used as a phosphorus reservoir during seed germination. This study describes the range of phytate-P in cereal genotypes of barley, wheat, triticale, rye and oat. Phytate is indigestible by humans and non-ruminant livestock. Phytic acid in feed grain seeds can bind with micronutrients (e.g., iron, manganese and zinc) and macronutrients (e.g., calcium and magnesium). Excessive non-digested phytate from feed could end up in animal waste (manure). This could pollute the environment and cause eutrophication of waterways. Samples of 423 barley, 114 wheat, 38 triticale, 24 oat, and 8 rye genotypes samples were analyzed for phytate-P and total P on a wet and dry basis. The phytate-P percent of total P, on a dry matter basis, ranged between 2.6 and 70.2 for barley, 39.3 and 84.1 for wheat, 44.6 and 84.8 for triticale, 43.5 and 64.3 for oats, 52.1 and 63.5 for rye. The widest variability of phytate-P was determined in barley compared to other cereal species. Results of the current study show that there is variability in phytate-P content in the genotypes of cereals analyzed. This variability could be exploited for the benefit of livestock feed and human food.

Improving the Plumpness and Yield of Barley in Western Australia

B. H. Paynter1, M. Cakir2, P. J. Eckermann3 and C. Li4

1Department of Agriculture, Northam, WA 6401, Australia,
E-mail: bpaynter@agric.wa.gov.au;
2State Agricultural Biotechnology Centre, Mudroch University, Mudroch, WA 6150, Australia;
3Biometrics SA, Adelaide University, Glen Osmond, SA 5064, Australia;
4Western Australia Department of Agriculture, South Perth, WA 6151, Australia

Since its release in 1983, the malting barley variety Stirling has been a benchmark variety for its ability to produce plump grain under a wide range of conditions in Western Australia. In the higher rainfall areas (> 400 mm), improvements in grain yield have resulted from the release of later maturing, feed barley varieties such as Onslow, Skiff and Fitzgerald. Attempts to breed for improvements in malting quality have resulted in the release of Franklin, Baudin and Gairdner. These varieties lack the grain plumpness of Stirling and the ability to maintain grain plumpness under a range of environmental stresses, although they have a higher yield potential. The propensity of crosses using European or Canadian malting germplasm to produce small grain in Western Australia is a major hurdle for barley breeding in the long term. This study is examining whether or not a relationship between carbon isotope discrimination, stem carbohydrate levels and silicon concentration correlate with both grain yield and grain plumpness in the Western Australian environment. Markers associated with grain plumpness were identified from a VB9104/Dash mapping population that has been phenotyped over the last two years as part of a scoping study for the current research. VB9104 (Europa/IBON7.148) is a high yielding, plump grained crossbred from Victoria and Dash (Chad/Joline//Cask) is a narrow grained, high yielding variety from the United Kingdom.

Selection of Barley Lines with Waxy Endosperm and Hulless Grains: Genotyping and Phenotyping

K. Polakova1, K. Vaculova2 and L. Kucera1

1Research Institute of Crop Production, 161 06 Prague-Ruzyne, Czech Republic,
E-mail: polakova@vurv.cz;
2Agriculture Research Institute Kromeriz, Ltd., 767 01 Kromeriz, Czech Republic

Traditional barley breeding has taken many years of selection based on phenotypic expression and selfing to develop lines for a commercial distribution. It is necessary to bridge the gap between plant molecular biology and conventional plant breeding. DNA marker-assisted selection (MAS) accelerates the breeding process. We present here DNA-MAS of F2 lines with waxy endosperm and hulless grains. We used three types of co-dominant molecular markers; SNP (G/A) marker, which is in the third exon of the amylase gene Amy32b,
G allele is presented in waxy hulless cultivars, A allele is presented in covered cultivars; SSR marker, which we named Waxy06 (AG)6 and an insertion/deletion marker were used to detect the waxy recessive allele, which results in waxy phenotype. The high throughput DNA isolation system (2 × 96 samples within 2 hours, Qiagen) was used in this study. Genomic DNA was extracted from leave segments (50 mg) of seedling plants, so that we were able to directly characterise individual plants using molecular markers and immediately applied DNA-MAS. Therefore, selected plants can be used for further breeding purposes in the same vegetative season.

Supported by the Ministry of Agriculture of the Czech Republic, Project No. QD 1365.

Verification of Nutritive Value and Hypocholesterolemic Effect
of Spring Barley Lines

J. Prochazkova1, S. Pipalova1, J. Ehrenbergerova2 and K. Vaculova3

1Department of Animal Nutrition and 2Department of Crop Science and Plant Breeding,
Mendel University of Agriculture and Forestry Brno, 613 00 Brno, Czech Republic,
E-mail: xpipalov@node.mendelu.cz;
3Agricultural Research Institute Kromeriz, Ltd., 767 01 Kromeriz, Czech Republic

The nutritive value of the set of newly bred spring barley lines with different grain characteristics was checked in growth model experiment on laboratory rats. The set contained hulless lines bred in the ARI Kroměříž: feed type KM 1771 (1) and KM 1057 (2), food type KM 2082 (3), KM 2092 (4), KM 2062 (5) and lines formed on the basis of crossing with food donors of a waxy endosperm type hulless line Wabet × Washonubet (6), hulled line Wabet × Krona (7) and Wabet × Kompakt (8). The line Kompakt × Krona (9) and the malting variety Nordus (10) were used as a control. The best results of an average daily gain and feed conversion (9.19g; 2.29g/g) were achieved by feeding the line KM 1057 (2), conversely, the worst by feeding food barley with highest -glucan (7.1%) of the waxy type (6 average daily gain was 8.01g and feed conversion 2.54g/g). The lowest -glucan content was detected in sample 9 (3.88%). Polynomial dependence between -glucan content (x) and cholesterol content (y = 0.0061x2 0.419x + 8.9265, R2 = 0.5387, P < 0.01) was calculated from the statistical data. The highest values of digestibility coefficients of nitrogen substances NS (75.96%) and fibre (49.63%) were measured in animals in group 2, on the contrary the lowest digestibility of fibre (31.10%), fat (56.96%) and ash (45.65%) was exhibited by control group 10. Differences in digestibility of all followed nutrients after the 14th day and at the end (28th day) of the experiment were observed.

Molecular Marker Development in Low Phytate Barley
(Hordeum vulgare)

V. Roslinsky1, P. E. Eckstein1, V. Raboy2, B. G. Rossnagel1 and G. J. Scoles1

1Department of Plant Sciences/Crop Development Centre, University of Saskatchewan,
Saskatoon, S7N 5A8, Canada, E-mail: peter.eckstein@usask.ca; 2U.S.D.A. Agricultural Research Service, National Small Grains Research Facility, Aberdeen, ID 83210, USA

Barley grain contains significant levels of phosphorous, though most is bound in the form of phytic acid (phytate) and is nutritionally unavailable to monogastrics. Through mutation breeding, USDA researchers have recently developed several low phytate barley mutants which have significantly less phytate. Of the three low phytate mutants investigated, the lpa1-1 locus shows a 50% decrease in phytic acid and has been mapped to the long arm of chromosome 2H. A RFLP marker (ABC153) located in this region has been converted to a dominant SCAR marker. Segregation analysis in the doubled haploid population from the cross CDC McGwire by low phytate mutant Hvlpa1-1 has confirmed linkage between the SCAR marker and lpa1-1 with 15% recombination. The mutation in a second low phytate mutant, M 635 (75% phytate decrease), has been mapped to the long arm of chromosome 1H via linkage with ISSR marker LP75. Analysis with ABC153 confirms this locus is not allelic to lpa1-1, and the locus has been designated lpa3-1. Both markers are polymorphic in a cross involving a third mutant M 955 (95% phytate decrease). Initial results indicate that the mutation in M 955 and locus lpa3-1 are in the same region on chromosome 1H, and screening of a larger population is in progress to determine if the mutations are allelic.

High-Resolution Mapping of the Naked-Caryopsis Gene (nud) and the Origin of Naked Barley

S. Taketa1, S. Kikuchi1, T. Awayama1, S. Yamamoto1, M. Ichii1 and S. Kawasaki2

1Faculty of Agriculture, Kagawa University, Ikenobe, Miki-cho, Kita-gun, Kagawa, 761-0795, Japan,
E-mail: staketa@ag.kagawa-u.ac.jp;
2National Institute of Agrobiological Sciences, Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

The hulled or naked caryopsis character of barley is an important trait for edibility and to follow its domestication process. It is well established that a single recessive gene, nud, located on the long arm of chromosome 7H, controls the naked caryopsis character. With an ultimate goal of map-based cloning of this economically important gene, we initiated bulked segregant analysis of this trait in combination with amplified fragment length polymorphism (AFLP). From analyses of 2,742 primer combinations, 9 AFLP fragments were selected as linked markers and six of them were converted into easy-to-use PCR based markers. A fine map surrounding the nud locus was developed in an F2 population of 458 individuals derived from a cross between cv. Kobinkatagi (naked type) and cv. Triumph (hulled type). This map has one order higher resolution than those reported before. One marker sKT3, which is linked with the nud locus with a distance of 0.2 cM, were used to trace the origin of naked barley. A total of 253 barley accessions including 47 wild barleys, were examined for genetic polymorphisms in this marker. Our data suggest that naked barleys today were originated from a small number of mutation events that occurred in hulled type.

Relationship between Grain Hardness and Endosperm
Cell Wall Polysaccharides in Barley

T. Tohno-oka1, N. Kawada1,2 and T. Yoshioka1

1National Institute of Crop Science, Tsukuba, Ibaraki 305-8518, Japan,
E-mail: tohnooka@affrc.go.jp;
2National Agricultural Research Center for Kyushu Okinawa Region,
Chikugo, Fukuoka 833-0041, Japan

Barley has been revaluated for food and source of dietary fiber, so pearling quality of barley is one of important breeding objectives. Hardness and whiteness of endosperm are important characters for it. To clarify factors contributing to hardness of endosperm, we analyzed relationship between it and chemical components. Endosperm hardness was evaluated as the time to grind grain from 55% to 40% pearl yield in pearling test mill (Satake TM-05). Protein, starch, -(1-3,1-4)-glucan and arabinoxylan content in the bran generated during pearling were measured. Beta-glucan and arabinoxylan are polysaccharides contained in cell walls of grain and the former is a major component of cell walls in barley endosperm. Both protein and -glucan content affected pearling time and it had a higher correlation with content of -glucan than that of protein. The effect of arabinoxylan was low. Beta-glucan contained in barley endosperm is useful soluble dietary fiber, but its content influences grain hardness.

Ways of Hulless Barley Grain Processing and Nutritional Quality

K. Vaculova1, D. Gabrovska2, J. Prokes3, V. Erban2, J. Ouhrabkova2, K. Hoke2,
M. Houska2 and J. Rysova2

1Agricultural Research Institute Kromeriz, Ltd., 767 01 Kromeriz, Czech Republic,
E-mail: vaculova@vukrom.cz;
2Food Research Institute Prague, 102 31 Prague, Czech Republic;
3Research Institute of Brewing and Malting, PCL, Malting Institute in Brno, 614 00 Brno, Czech Republic

Different technological processing methods (separated milling, malting, puffing) were applied in two our own hulless breeding lines of spring barley, KM1910 and KM2283, with different genetic backgrounds of grain chemical composition. Based on nutritionally differentiated products (barley and malt flour, barley and barley malt flakes, scratch bran, malt honey, and others), recipes for preparation of salty and sweet bakery and pastry products (bread, pasta, bulgur, griddle-cakes, pancakes, fancy bread, gingerbread, etc.), products with beneficial health effects (Granola, a basis for milk fermented products, malt drink), children nutrition and products of Chalva and Nugeta types have been developed. Both sensory evaluations and chemical analyses of traditional and new types of products showed that hulless barley is a suitable raw material to enlarge the foodstuffs assortment in the Czech Republic for both wholesome preventive human nutrition of the population and consumers of specific dietary requirements.

Supported by the Ministry of Agriculture of the Czech Republic, Projects Nos. QD 0183 and QF 3291.