Session 5 Malting and Brewing Technological Parameters

 

Oral Presentation

 

 

Barley Varieties Suitable for the Production of a Czech Type Beer

K. Kosar, V. Psota and A. Mikyska

Research Institute for Malting and Brewing, PLC, Malting Institute in Brno, 614 00 Brno,
Czech Republic, E-mail: kosar@brno.beerresearch.cz

In the course of the 20th century the production of beer was substantially changed. Economic pressure led to concentration, modernization and automation of the beer and malt production. These changes were reflected in a new view of the malting barley varieties quality. In two last decades varieties with a strong enzymatic activity, high extract content and high final attenuation have been promoted. Due to historical reasons the traditional production of pale lager has been preserved in the Czech brewing. For the production of a Czech type beer, those varieties of malting spring barley are suitable that enable the production of beer with a higher level of residual extract, strong palatefulness, excellent foaming power and relatively lower alcohol content.

 

Are We Any Closer to Predicting How Well a Malt Will Behave
in the Brewhouse?

E. D. Baxter, C. D. Booer and A. Faulkner

Brewing Research International, Lyttel Hall, Nutfield, Surrey, RH1 4HY, UK
E-mail: d.baxter@brewingresearch.co.uk

An essential element in the breeding of new barleys for malting is the ability to predict the malting and brewing quality of potential new varieties, often when only a limited amount of seed is available. Recent years have seen the publication of a number of laboratory techniques which claim to predict brewhouse performance. These include measurements of kernel homogeneity, and aspects of wort filterability. This paper describes pilot brewing trials with a set of six commercial malts, all of which conformed to the same typical specification, in an effort to identify which of these new techniques is the most useful in terms of predicting processability in the brewery. The malts were tested for a wide range of standard and non-standard quality parameters and then brewed in the BRi pilot brewery, using standard brewing protocols designed to simulate common brewing practices. Each malt was scored for ease of lautering, fermentation behaviour and yield of ethanol, and the results compared with the laboratory tests. It was apparent that, although the six malts had very similar standard specifications, they differed significantly in their brewing behaviour. The results from suggested that, within this sample set, laboratory tests of wort filterability were the best predictors of processability.

 

 

 

Distilling Barley Bringing Certainty to the Future

J. M. Brosnan1, T. A. Bringhurst1, K. Denyer2, J. S. Swanston3 and W. T. B. Thomas3

1Scotch Whisky Research Institute, Research Park North, Riccarton Edinburgh, EH14 4AP, UK,
E-mail: james.brosnan@swri.co.uk;
2John Innes Institute, Norwich Research Park, Norwich, NR4 7UH, UK; 3Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK

The distilling industry is one of the most historic and economically important end users of barley. The Scotch Whisky industry alone earns over 3 billion in exports for the United Kingdom. The quality of malting barley for distilling in the United Kingdom has been greatly improved because of a rigorous varietal selection procedure endorsed by all those in the barley supply chain. From Golden Promise in the 1960s to Optic today distillers have seen a steady increase in the alcohol yield potential of barley varieties. However, the challenges which face distillers in the near future demand more than high alcohol yield from barley. Exciting opportunities do exist to significantly improve distilling barley by utilising non-GMO genetic techniques combined with an understanding of the phenotypic variation already available in the barley gene pool. Making the jump in technology transfer between knowledge of barley genes to innovation in a distillery requires partnership. In this paper the future needs of distillers will be described in the context of research work carried out by the distilling industry in Scotland in collaboration with other barley research groups.

 

 

 

Poster Presentation

 

DIStilling Gene Function in Barley: A Reverse Genetics Approach

D. G. Caldwell, N. McCallum, G. Muehlbauer and R. Waugh

Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK,
E-mail: rwaugh@scri.sari.ac.uk

Functional genomics in barley (Hordeum vulgare L.) is a relatively under-developed area of research due to the lack of structured reverse genetics populations and its inability to transform efficiently. Here, we describe the creation and implementation of a large-scale reverse genetics population derived from ethyl-methanesulfonate (EMS) mutagenised barley (cv. Optic). The population is comprised of ~10,000 plants from two different doses of EMS (20mM and 30mM). The ~10K M2 lines are currently arrayed into 1,152 pools of 8 plants. However, recent advances in SNP detection (Cel-Nuclease/HSX detector, Transgenomic) has shown the ability to detect one SNP in 32 alleles (16 plants) and in some cases 1:48 alleles (24 plants). These deeper pooling strategies will be implemented making the screening more time and cost effective. The population is now currently available for use by the research community either through collaboration or cost recovery. Implementation, description, and initial test screening will be described.

 

 

Understanding the Genetic Bases of Modification

H. M. Collins, S. J. Logue and J. K. Eglinton

School of Agriculture and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia,
E-mail: helen.collins@adelaide.edu.au

The proportion and degree of starch, protein and cell wall modification during malting are critical in determining the quality of the final product. While high levels of cell wall modification are desirable to prevent filtration problems, only partial degradation of protein is required. The relationship between cell wall and protein modification is complex with numerous genes likely to be involved. A number of commonly measured traits give an indication of the level of modification. Free amino nitrogen (FAN), Kolbach Index (KI) and soluble protein (SP) are used as indicators of the level of modification of the proteins, while wort -glucan (WBG) and wort viscosity indicate the level of modification of the cell walls. The Australian National Barley Molecular Marker Program (NBMMP) has investigated these traits using a number of Australian mapping populations. Due to the complex relationship between protein and carbohydrate modification this information will not be useful to barley breeders unless knowledge of how and why these regions are influencing modification is known. This paper investigates the regions found to be associated with modification related traits and in doing so aims to gain a greater understanding of the processes involved in modification.

 

 

 

An Investigation of the Relative Rates of Protein and Carbohydrate Modification of a Number of International Malting Varieties,
Grown in Three Countries

H. M. Collins1, J. S. Swanston2, B. G. Rossnagel3 and S. J. Logue1

1School of Agriculture and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia,
E-mail: helen.collins@adelaide.edu.au;
2Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK; 3Crop Development Centre, University of Saskatchewan, Saskatoon, S7N 5A8, Canada

During malting, barley grains are steeped in water and then allowed to germinate under controlled conditions. This process is generically termed modification. The overall degree and relative proportion of starch, protein and cell wall modification during malting are critical in determining the quality of the final product. The modification process is greatly influenced by both the variety investigated and the environment in which the variety is grown. Currently, there are a large number of high malting quality varieties being grown worldwide that differ dramatically in the way in which they modify. These varieties are adapted to vastly different growing environments and often perform poorly when removed from their ideal environment. In this study, twelve international malting varieties were grown in Scotland, Canada and Australia in 2002 and 2003, in order to compare their modification profiles. They were malted in a Phoenix micromalter using a standard micromalting regime from each country. These regimes varied greatly, with large differences in both the number and length of steeps and the length of time for germination. Both the country and micromalter schedule had large effects on the degree of modification of the grain. The implications of these results will be discussed.

 

 

 

Biochemical and Genetic Basis of Wort Fermentability

M. J. Edney1, J. K. Eglinton2, H. M. Collins2 and A. R. Barr3

1Grain Research Laboratory, Canadian Grain Commission, Winnipeg, R3C 3G8, Canada,
E-mail: medney@grainscanada.gc.ca;
2School of Agriculture and Wine, University of Adelaide,
Adelaide, SA 5064, Australia; 3Australian Grain Technologies, University of Adelaide,
Roseworthy, SA 5064, Australia

Factors that affect wort fermentability were investigated with 180 doubled haploid lines from an Arapiles/Franklin cross. Barley, grown in South Australia during 2001, was micromalted and analysed using standard methods. Samples were found to have a wide range in malting quality with lines both superior and inferior to the parents for all traits analysed. QTL and statistical analysis of data indicated, for this population, that poor malt modification, beta-glucan in particular, had the greatest negative effect on fermentability. Fermentability was also negatively correlated with the maltotetrose content of wort. A significant QTL for this sugar was coincident with the chromosome 4H locus for the heat-stable beta-amylase (sd2H). Lines with the heat stable enzyme, inherited from Arapiles, had improved fermentabilities of IOB extracts but not of EBC extracts. Glucose, possibly coming from beta-glucan hydrolysis, was found to have a positive effect on fermentability. A significant QTL for wort fructose was in the area of the sdw semi-dwarfing gene on chromosome 3H, highlighting the potential impact of agronomic traits on the genetic analysis of malt quality.

 

 

 

Malting Quality of a Hulless/Covered Doubled Haploid
Barley Population

M. J. Edney1, W. G. Legge2, B. G. Rossnagel3 and H. M. Collins4

1Grain Research Laboratory, Canadian Grain Commission, Winnipeg, R3C 3G8, Canada,
E-mail: medney@grainscanada.gc.ca; 2Brandon Research Station, Agriculture and Agri-Food Canada, Brandon, R7A 5Y3, Canada; 3Crop Development Centre, University of Saskatchewan, Saskatoon,
S7N 5A8, Canada; 4School of Agriculture and Wine, University of Adelaide, Adelaide, SA 5064, Australia

The potential to improve malting quality of hulless barley was investigated with a two-row doubled haploid population (TR251/HB345). TR251 is a covered, malt breeding line with good malting potential while HB345 is a hulless line with good agronomic traits and the gene for heat stable -amylase (sd2H). Barley grown in standard yield test plots at Brandon, Manitoba during 2002 was micromalted and analysed using standard conditions. The 62 hulless lines, on average, had significantly higher levels of malt extract than the 62 covered lines. A significantly lower percentage of the hulless extract was fermentable but the hulless lines still fermented a greater amount of total extract. Levels of diastatic power were similar among hulless and covered malts but -amylase was significantly lower in the hulless lines. Heat stable -amylase improved fermentability but not significantly. Samples showed an acceptable degree of malt modification but -glucan levels still had a negative effect on fermentability. Levels of total free amino acids were similar among hulless and covered worts; however, levels of several individual amino acids were significantly lower in the hulless worts which had a negative effect on fermentability. In summary, hulless malt had some superior attributes compared to covered malt but increased levels of some specific enzymes would further enhance its appeal.

 

 

Proteome Analysis of Barley Seeds and Malt

C. Finnie1, O. Oestergaard1, M. Christiansen2, G. Backes3, K. S. Bak-Jensen1, S. Laugesen4,
P. Roepstorff4, J. Larsen5
and B. Svensson1

1Department of Chemistry, Carlsberg Laboratory, DK-2500 Copenhagen, Denmark, E-mail: bis@crc.dk;
2Sejet Plantbreeding, DK-8700 Horsens, Denmark; 3Plant Research Department, Risoe National Laboratory, DK-4000 Roskilde, Denmark; 4Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense, Denmark; 5Carlsberg Research Laboratory, DK-2500 Copenhagen, Denmark

Barley proteomics, using 2D-gel electrophoresis and mass spectrometry for protein identification, provides biologically relevant information even without genome sequence. Proteins in over 450 spots in developing and germinating seeds and dissected tissues have been identified, including multiple forms of -amylase, -amylase, -amylase/subtilisin inhibitor, serpins, CM-proteins, peroxidases, enzymes involved in defence: PR-proteins, chitinase, thaumatin-like proteins, and oxidative or desiccation stress: ascorbate peroxidase, peroxiredoxins, sHSP, LEA-proteins. Previously uncharacterised proteins are identified by searching EST databases with peptide masses and post-translational modifications characterised by advanced mass spectrometry. The project aims to identify proteins important for malting. Sixteen cultivars differing in malting quality were grouped using independent methods: (a) clustering analysis of spot differences, (b) malt quality scoring from micromalting. The best malting cultivars were grouped identically by both methods, thus malting quality is reflected by the proteome. Analysis of additional spot differences and cultivars will explore the potential for predicting malting quality. Most spot differences are genetically determined, illustrated by spots containing allelic -amylase forms. Spot differences segregated in a doubled haploid population (Scarlett × Meltan) and thus were located on the genetic map. Micromalt analysis of this population enabled correlation of proteome, genetic information and malting quality.

 

 

 

Hull-Cracked Grain and Grain with Ventral Swelling
as Damaged Grains in Malting Barley

M. Furusho1, T. Baba2, H. Kai1, M. Tsukazaki1, Y. Uchimura1 and O. Yamaguchi1

1Fukuoka Agricultural Research Center, Chikushino, Fukuoka 818-8549, Japan,
E-mail: furusho@farc.pref.fukuoka.jp;
2National Agricultural Research Center, Hokuriku Research Center, Inada, Joetsu 943-0193, Japan

Hull-cracked grain (HCG) and grain with ventral swelling (GVS) are damaged grains, which constitute deteriorated grain qualities in malting barley. This study was carried out to solve this problem by developing evaluation methods for occurrence of these damaged grains. It was possible to select HCG-tolerant varieties by early seeding and cultivation under insufficient sunshine during the internodes elongation stage. When water was sprinkled on the barley materials in the early-ripening stage, the middle-ripening stage and the late-ripening stage, sprinkling in the late-ripening stage most notably reproduced the occurrence of GVS. A method to evaluate the occurrence of GVS by intermittent, artificial water sprinkling in the late-ripening stage was established. To establish a Marker Assisted Selection (MAS) system for lines with a low occurrence of HCG and GVS, we screened about 1,000 DNA markers to detect polymorphisms between Kinuyutaka (low occurrence of HCG and high of GVS) and Yoshikei 15 (high occurrence of HCG and low of GVS). Then, we constructed a genetic map for Kinuyutaka × Yoshikei 15 using 150 doubled haploid lines. The QTL analyses revealed three QTLs for HCG on 2H, 3H and 6H and one for GVS on 1H of barley chromosomes, respectively.

 

 

Possibility of Matrix Assisted Laser Desorption Ionisation Time-Of-Flight Mass Spectrometry for Evaluation of Barley Quality

D. Gajdosova1, J. Havel1, K. Novotna1, O. Sedo1, P. Havlova2 and J. Ehrenbergerova3

1Department of Analytical Chemistry, Faculty of Science, Masaryk University, 611 37 Brno,
Czech Republic, E-mail: havel@chemi.muni.cz; 2Research Institute for Brewing and Malting, PCL, Malting Institute in Brno, 614 00 Brno, Czech Republic; E-mail: havlova@brno.beerresearch.cz; 3Department of Crop Science and Plant Breeding, Mendel University of Agriculture and Forestry Brno, Brno, Czech Republic; E-mail: ehren@mendelu.cz

Barley and consequently of malt quality is of fundamental importance to reach good quality of beer. There are several factors, genetics, cultivation conditions, use of fungicides, weather conditions, stress, etc. Barley can be e.g. attacked by fusaria (Aspergillus, Rhizopus, Penicillium or Nigrospora) while Fusarium head blight (FHB) illness seems to be the main reason for beer over-gushing, for example. Also the content of various metabolites, pathogenesis-related protein, etc. is important for barley quality. Detection of mycotoxins and other metabolites requires detailed and often tedious and laborious analysis. Matrix Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI TOF MS) is rather novel technique able to ionize and detect with excellent sensitivity various compounds from low up to quite high molecular weight proteins. This work deals with examination of MALDI TOF MS possibilities for detection, identification or determination of various toxins or metabolites in barley. It was found that selective identification of different types of metabolites is possible, while some of them can be detected directly from barley, others only after suitable extraction procedure. Advantages and disadvantages of MALDI TOF analyses are demonstrated and evaluated. Comparison of results obtained via this technology and classical analytical approaches is discussed.

 

 

 

Beta-Glucan Content of Old and Modern Spring Barleys

H. Grausgruber1, R. Schoenlechner2, V. Zahlner2, E. Berghofer2 and P. Ruckenbauer1

1BOKU, Institute of Agronomy and Plant Breeding, 1180 Vienna, Austria,
E-mail: grausgruber@ipp.boku.ac.at;
2BOKU, Institute of Food Technology, 1190 Vienna, Austria

Numerous studies have verified the cholesterol-lowering benefits of barley foods, due to -glucan, the major soluble fiber component. The health benefit of barley, therefore, depends on the direct relationship between blood cholesterol and the risk of heart attacks. Hence, the interest in barley as food source is currently increasing. We studied the -glucan content of spring barley landraces and cultivated varieties from a time period of more than 150 years, beginning with the Nuremberg barleys of 1832 up to recent malting barleys. Hypothesis was that probably old barleys exhibit higher -glucan contents because of a less intensive selection for malting quality. The latter trait, namely, is negatively correlated with the -glucan content. We analysed 30 genotypes from the harvests of 2000, 2002 and 2003. In 2003 also a few hulless varieties were available for analysis. Moreover, in 2003 a comparison between organic and conventional grown barleys was made using 10 selected genotypes. A significant variation in -glucan was estimated both for genotypes, as well as for years. Generally, no clear trend in -glucan content over time of registration/cultivation of the varieties was observed, however, the most recent malting barleys exhibited as supposed the lowest contents. The highest contents were observed for some hulless barleys.

 

Investigation of the Molecular Characteristics of Barley Hordeins
by HPLC and MALDI-TOF MS Techniques

M. S. Izydorczyk1, S. Bazin1, H. Liu1,3, K. Preston1, Y. Qian2, W. Ens2, E. Lefol3 and B. L. Harvey3

1Grain Research Laboratory, CGC, Winnipeg, R3C 3G8, Canada,
E-mail: mizydorczyk@grainscanada.gc.ca;
2Department of Physics and Astronomy, University of Manitoba, Winnipeg, R3T 2N2, Canada; 3Department of Plant Science, University of Saskatchewan, Saskatoon, S7N 5A8, Canada

The effect of barley proteins on malt and beer quality characteristics is complex because of the large number of proteins present in barley grain, their diverse characteristics, and their extensive modification during the malting and mashing processes. A better understanding of the protein-related phenomena associated with beer quality will assist in developing improved malting barley. In this study, ground barley samples were extracted sequentially with NaCl, 50% (v/v) propanol, and 50% (v/v) propanol containing 1% dithiothreitol. The monomeric and polymeric proteins were separated by selective precipitation. The extracts were subjected to HPSEC combined with RI, UV, and multiangle light scattering (MALS) detectors. The average molecular weight of intact monomeric and polymeric fractions was determined. The reduced polymeric fraction was separated by RP-HPLC and peaks were collected for subsequent peptide mapping by MALDI-TOF MS. The masses of polypeptides in the B-C-, and D-hordeins were determined. Tryptic peptide maps of the major polypetides in the B-subgroup were also obtained. Wort and beer polypeptide fractions, obtained by sequential precipitation with ammonium sulphate and purification by ion-exchange chromatography, were also characterized by the HPLC and MS techniques.

 

 

 

Impact of Chromosome 5HL QTL on the Malting Quality
and Proteinase Activity in Malt

M. Kihara1, Y. Okada1, W. Saito1, N. Kawada2,3, T. Kaneko1, T. Asakura1 and K. Ito1

1Plant Bioengineering Research Laboratories, Sapporo Breweries Ltd., Gunma 370-0393, Japan,
E-mail: makoto.kihara@sapporobeer.co.jp;
2National Agricultural Research Center for Kyushu Okinawa Region, Fukuoka 833-0041, Japan

We investigated the QTLs for the malting quality and proteinase activity in the doubled-haploid lines derived from a cross between Mikamo Golden, a Japanese malting barley, and Harrington, a North American malting barley. Our results clearly indicated that one QTL located on chromosome 5HL had a large influence on Kolbach Index and proteinase activity in malt. In order to clarify the effects of chromosome 5HL QTL, we analyzed the malting quality and proteinase activity of 27 malting barley cultivars and breeding lines with various origins, and also examined RFLP pattern of the QTL region. Kolbach Index and proteinase activity of the malt varied among the lines tested. The RFLP pattern showed polymorphism of the QTL region and was classified into several groups. One of the groups including Klages, a North American malting barley, showed the higher Kolbach Index and proteinase activity than other groups, and the RFLP pattern closely related with these characteristics. These results suggest that the QTL located on chromosome 5HL has a considerable effect on the malting quality.


 

Improvement of Barley Malting Quality
Using a Gene from Hordeum spontaneum

C. D. Li1, R. Lance1, A. N. Tarr1, S. Broughton1, S. Harasymow1, R. Appels1 and M. Jones2

1Western Australia Department of Agriculture, South Perth, WA 6151, Australia,
E-mail: cli@agric.wa.gov.au;
2State Agricultural Biotechnology Centre, Murdoch University, South Perth, WA 6150, Australia

The gene pool of Hordeum spontaneum has great potential to improve barley yield and quality. Utilization of this gene pool is frequently limited by the genetic drags. Molecular markers provided a tool to target transferring the unique genes. A special Sd3 -amylase from Hordeum spontaneum has high enzyme activity and thermostability. SNP, SSR and CAP molecular markers were developed to distinguish different -amylase genes (alleles) or isoenzymes and target transfer the desirable -amylase gene into the elite genetic background. Molecular markers by combining with doubled haploid technique were used to facilitate transferring the Sd3 gene into local barley variety Gairdner. Four backcrosses were completed with the recurrent parent. Three hundred lines with the Sd3 -amylase were selected using molecular markers and 380 DH lines were generated from the third backcross. Quality analysis showed that the Sd3 gene has significantly enhanced the diastatic power by increasing the -amylase enzyme activity and thermostability. There was little affect on malt extract and wort viscosity but a slight increase in wort soluble nitrogen content was observed. The malt beta-amylase activity and thermostability increased by an average 30% and 77% by introducing the Sd3 gene.

 

 

Can Anonymous QTLs Be Introgressed Successfully
into Another Genetic Background?
Results from a Barley Malting Quality Parameter

R. C. Meyer1, J. S. Swanston1, J. M. Brosnan2, M. Field3, R. Waugh1, W. Powell1
and W. T. B. Thomas1

1Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK, E-mail: wthoma@scri.sari.ac.uk;
2Scotch Whisky Research Institute, Riccarton, Edinburgh, EH14 4AP, UK; 3Advanta Seeds UK,
Docking, Kings Lynn, Norfolk, PE31 8LS, UK

Spirit yield, the product of hot water extract and it fermentability, is an important production parameter for Scotch Whisky. We utilised a barley mapping population to identify a QTL associated with an increase in fermentability and used a backcrossing programme to transfer it into cv. Landlord in an attempt to improve its predicted spirit yield (PSY). We genotyped and phenotyped the resultant BC1DH population and to demonstrate that the introgressed region was associated with an increased fermentability. The QTL region was also associated with a decrease in hot water extract with the net effect that there was a resultant decrease in predicted spirit yield. Fine mapping in the region is required to establish whether or not the observed effects are due to closely linked loci or the pleiotropic effects of a single locus. Alternatively, it is possible that a gene detected in a relatively poor malting quality background may not be expressed to the same extent in a better background. Analysis using all the marker data indicated that other loci in the BC1DH population had a greater effect upon PSY.

MAFF Agro/Food LINK funded this project (CSAA4306) with the Home Grown Cereals Authority (Project No. 1573), the Scotch Whisky Research Institute, Advanta Seeds UK and Mylnefield Research Services Ltd. as industrial partners.

 

 

A New Two Dimensional Germinative Classification for Malting Barley Quality Based on Separate Estimates for Vigour and Viability

B. Moeller and L. Munck

Department of Dairy and Food Science, The Royal Veterinary and Agricultural University,
DK-1958 Frederiksberg C, Denmark, E-mail: lmu@kvl.dk; www.model.kvl.dk

Optimal germination performance is with no doubt the most important quality criterion for malting barley. The industry and trade are dependent on reproducible and representive analyses as expressed in EBC Analyticas analyses for germinative energy (GE) % and capacity (GC) %. Quality classification indices based on elaborate pilot malting analyses and expert evaluation without the germinative analyses has been developed by Molina-Cano (1987) for EBC and recently by us (Nielsen et al. 2002) using fuzzy logic analysis. Monnez (1987) found it difficult to embrace the quality complex in one figure and suggested a hierarchical classification with on 23 principal components obtained by multivariate data analysis. Based on multivariate data analyses of germination profiles we have suggested that the percentage values of one day and three days germination obtained from current EBC analyses for germinative energy and capacity are used in a two dimensional classification plot for barley samples representing vigour x and viability y. The 7 barley classes obtained visualises the quality differences in a consistent and instructive way clearly differencing and ordering malting barleys with falling extract % and increasing wort -glucan (mg/l) according to a subsequent validation analysis (Mller 2003). The classification can in principle by obtained within 24 h combining 1-day germination with the tetrazolium germinative capacity test (EBC 3.5.2) for viability. The vigour and viability classification test maybe used in micro and pilot malting and in full-scale trials as an essential description of the germination quality trait of fundamental importance in malting barley breeding.

 

 

 

Genetic Analysis of Seed Dormancy in Barley

D. Prada1, I. Romagosa1, S. E. Ullrich2, L. Cistue3, J. A. Clancy2 and J. L. Molina-Cano1

1Centre UdL-IRTA, 25198 Lleida, Spain, E-mail: joseluis.molina@irta.es; 2Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164-6420, USA; 3Estación Experimental de Aula Dei, CSIC, 50080 Zaragoza, Spain

Moderate seed dormancy in barley (Hordeum vulgare L.) is a very important target for many breeding programs when seed is intended for the malting industry. Seed dormancy is quantitatively inherited and variously influenced by the environment. The advent of quantitative trait locus (QTL) analysis provided a powerful tool to dissect complex traits into their genetic components. Although some molecular markers for dormancy QTL have been identified, the corresponding genes involved in the regulation of the process have not been cloned. Induced barley mutants may constitute useful material to study the physiology and genetics of seed dormancy. Overlapping positions between mapped monogenic mutants involved in the germination process and known dormancy QTLs could provide important clues to possible candidate genes underlying the expression of seed dormancy in barley. In the present study two different approaches have been evaluated to identify chromosomal regions associated with the expression of seed dormancy: (1) QTL analyses in a barley doubled haploid line (DHL) population derived from the cross between cv. Triumph, a major two-row malting cultivar in Europe that is very prone to dormancy, and cv. Morex, a six-row non-dormant malting quality standard cultivar in North America; and (2) identification of a mutant genomic region induced in TL43 (sodium azide-induced mutant obtained from cv. Triumph) that affects its level of seed dormancy. QTL analysis showed a major and consistent dormancy QTL near the centromere on chromosome 7(5H) associated with the establishment of dormancy during seed development; and a minor QTL near the long arm telomere on chromosome 7(5H) for the rate of dormancy release during after-ripening.

 

 

Genotypic and Environmental Variation of Some Chemical Compounds Related to Quality of Malting Barley

J. c. Qi1,2, K. Wei1, J. X. Chen1, J. M. Wang1, F. B. Wu1 and G. P. Zhang 1

1Agronomy Department, Zhejiang University, Hangzhou, 310029, P. R. China,
E-mail: zhanggp@zju.edu.cn;
2Department of Agricultural Science, Shihezi University, Shihezi, 832003, P. R. China

The main parameters involved in malt barley quality include malt extract, Kolbach Index, diastatic power and viscosity, which are in turn dependent on the carbohydrate and protein content and their components as well as activities of the hydrolysis enzymes associated with these chemicals. It has been well documented that protein and -glucan content, -glucanase and -amylase activities are the chemical compounds related closely to quality of malt barley. In the present review on the genotypic and environmental effect of these compounds and their relationships with malt quality, genetic variation of Chinese barley germplasm in these compounds and the characterization of main malt barleys planted on the different ecological locations are comprehensively illustrated. The influence of climatic and agronomic factors on these chemical compounds is discussed. Moreover, the future approaches of improving malt barley quality are proposed in reduction of -glucan content and increase of diastatic power through enhancing the activities of both -glucanase and -amylase.

 

 

 

Genotype-Environment Interaction for Malt Characters in a Series
of Experiments with Two- and Six-Rowed Barley Doubled Haploids

M. Rebarz, K. Krystkowiak, A. Kuczynska, T. Adamski, Z. Kaczmarek, M. Surma
and S. Jezowski

Institute of Plant Genetics, Polish Academy of Sciences, 60-479 Poznań, Poland,
E-mail: mreb@igr.poznan.pl

Genotype-environment interaction of barley doubled haploids was studied in relation to malting quality characters. Material for the studies covered two- and six-rowed barley doubled haploids lines derived from F1 hybrids of the malting cultivar Maresi (two-rowed) and non-malting cultivar Pomo (six-rowed). Doubled haploids were produced by the bulbosum method using the standard crossing procedure. Experiment was carried out in 6 environments (three years and 2 locations). In each environment the experiment was performed in a complete block design with three replications. Malt protein content, Kolbach index, extract yieldand malt fine-coarse difference were measured. Samples of plump kernels were malted in Automatic Micromalting System (Phoenix Biosystems, Australia). Malt analyses were performed using standard methods according to European Brewery Convention.Genotype-environment (GE) interaction was studied using SERGEN software. Main effect for each genotype was evaluated andits GE interaction was tested. On the basis of F-statistic for GE interaction genotypes were regarded as stable (non-significant GE interaction) and unstable (significant GE interaction). Unstable lines were classified as extensive when their regression coefficient was significant and negative, and intensive if their regression coefficient was significant and positive. On the basis of obtained resultslines with stable malt characters were distinguished.

 

 

Transgenic Barley in Brewing Detection of Transgene
and Heterologous Protein Levels

A. Ritala1, T. H. Teeri2, S. Marttila3, A. Wilhelmson1, S. Kaur2, S. Rasmussen4
and A. M. Nuutila1

1VTT Biotechnology, Espoo, FIN-020440, Finland, E-mail: anneli.ritala@vtt.fi; 2Department of Applied Biology, University of Helsinki, FIN-0014 Helsinki, Finland; 3Swedish University of Agricultural Sciences, 23053 Alnarp, Sweden; 4Risoe National Laboratory, DK-4000 Roskilde, Denmark

The aim of the project was through a model case to find out wether the threshold of 1% genetically modified material in foods is detectable with the present analytical methods. The transgenic barley material was homozygotized through microspore culture resulting in doubled haploid plants. The transgenic and non-transgenic barley samples were malted, mashed and fermented, and the malts, worts and beers were analyzed in respect to the transgene and the heterologous protein. The detection of the heterologous protein from the malted grains was successful only by immunomicroscopy. The heterologous enzyme was localized in the aleurone layer and scutellum of the transgenic malt, but no enzyme was detected in the starchy endosperm. This suggests that the heterologous protein was expressed in seeds but only in very low levels. The real-time PCR proved to be repeatable and extremely sensitive for the detection of the transgene. The threshold of 1:1,000 (0.1%) which is in the range of the legal requirements was obtainable with high confidence. Detectable DNA was also obtained from wort, but repeated DNA extractions from beer were not successsful.

 

 

 

The Genetic Basis of Improved Beer Haze Stability

L. H. Robinson1, C. M. Ford1, P. Healy2, C. E. Gibson3, A. R. Barr4 and D. E. Evans5

1School of Agriculture and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia,
E-mail: louise.robinson@adelaide.edu.au; 2Lion Nathan, GPO, Milton, Queensland, 4001, Australia; 3Joe White Maltings Ltd, AusMalt Technical Centre, Cavan, SA 5094, Australia; 4Australian Grain Technologies, University of Adelaide, Roseworthy, SA 5371, Australia; 5Tasmanian Institute of Agricultural Research, University of Tasmania, Hobart, Tasmania 7001, Australia

Formation of permanent haze in beer is a serious quality problem, which limits the storage life of the product. From silica gel, used for beer colloidal stabilisation, a silica eluent (SE) protein fraction was isolated and an antibody raised against this fraction. SDS-PAGE immunoblot analysis using the SE antiserum detected a range of protein bands in barley, malt, beer and haze. Interestingly, a polymorphism was observed in some barley varieties which contained a ~12 kDa band (SE +ve) while in other varieties this band was absent (SE ve). The genetic basis for the presence/absence of the SE protein was determined by QTL analysis which found that the ~12 kDa band mapped to the short arm of chromosome 3H. Two-dimensional gel electrophoresis and sequence analysis was used to identify the SE protein as the barley trypsin inhibitor CMe precursor (BTI-CMe). This protein is 13.3 kDa and its gene is located on chromosome 3H, consistent with information available on the SE protein. Pilot brewing trials found that beer brewed from varieties that did not contain the ~12kDa SE band, SE ve varieties, were more resistant to haze force testing than beer produced from SE +ve varieties. These results demonstrate that the selection of SE ve varieties will provide an opportunity to improve the colloidal stability of beer.

 

 

Polymorphism of the Intron IIIExon IV Region of the -Amylase Gene Bmy1 in North European Barley Varieties

T. G. Sjakste1 and M. S. Roeder2

1Laboratory of Plant Genetics, Institute of Biology, LV 2169, Salaspils, Latvia,
E-mail: tanjaskste@email.lubi.edu.lv;
2Gene and Genome Mapping, Institute of Plant Genetics and Crop Research (IPK),
D-06466 Gatersleben, Germany

Allelic diversity and inheritance of the C698T polymorphism encoding alleles of -amylase with different thermostability, a deletion/insertion event and a microsatellite (TG)n(G)n in the 3region of intron III of the seed specific -amylase gene Bmy1 were studied in a set of barley accessions composed of 21 Latvian, 1 Indian and 29 European old and modern commercial barley varieties, the Himalaya landrace Brachytic and 3 Hordeum spontaneum lines from Israel. The data obtained indicated a low frequency of the C698 genotype encoding an enzyme of higher thermostability in Europe. Results are discussed in the frame of genetic diversity and inheritance of the polymorphic loci genotyped and of their availability for selection of barley -amylase alleles.

 

 

Comparison of Malting Quality Parameters of Spring
and Winter Barley Genotypes in the Czech Republic

J. Spunar, M. Spunarova, Z. Nesvadba and K. Vaculova

Agricultural Reearch Institute Kromeriz, Ltd., 767 01 Kromeriz, Czech Republic,
E-mail: spunar@vukrom.cz

The Czech Republic has a long tradition in spring barley breeding which was started in 1872 by Proskowetz who created the first spring barley variety Proskowetz Hana Pedigree. During the 100-year period of systematical breeding, parameters of malting quality have improved. The progress in malting quality manifested in increased extract content manifested in higher relative extract, Kolbach Index, diastatic power, friability, etc. In winter barley the particular progress was reached in winterhardiness which enables to profit from the shorter vegetation period in comparison with spring barley. In spite of the fact that Czech malting industry prefers strongly spring barley varieties two-row winter barley still remains the reserve for malt production in the years when the production of spring barley is insufficient for malt production due to both the amount and quality. Low area of cultivation due to economical reasons, low yield, infestation by Fusarium head blight manifested in the lack spring barley with acceptable malting quality parameters in the year 2000. On the contrary winter barley varieties, particularly the German variety Tiffany provided raw material with required parameters of malting quality. In the poster the relationships between spring and winter barley crop and particularly correlation between individual parameters of malting quality will be analyzed.

Supported by the Ministry of Agriculture of the Czech Republic, Project No. QE 1093.

 

 

Do Components of Barley Variety Mixtures Converge
for Malting Quality Attributes?

J. S. Swanston and A. C. Newton

Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK,
E-mail: jswans@scri.sari.ac.uk

Barley plants grown in a variety mixture may be subject to competitive and/or complementary interactions. These may alter how the plants respond to their environment and, in some situations, mixture components appear to converge for characters such as height and maturity. To date, no investigation of whether such effects also occur for factors that influence quality has been made, although convergence could clearly reduce the variation that is perceived to be a major obstacle to the commercial utilisation of mixtures. To investigate possible convergence, three diverse spring barley varieties were grown as pure stands and also as equal proportion, three-component mixtures in replicated trial. Mixtures were sown with random distribution of grain from the components and also in a format where the components comprised alternate rows within a plot. Harvesting was carried out manually to preserve the identity of individual plants. Quality attributes were assessed, where possible, on individual plants, to enable the variation within and between mixture components to be determined and the performance of the three varieties, from the three different plot arrangements was compared. Implications for the choice and optimal spatial distribution of mixture components and the potential for malting are discussed.

 

 

 

Genotypic Differences of Translocated Dry Matter at Spring Barley

M. Uzik and A. Zofajova

Department of Breeding Methods, Research Institute of Plant Production, 921 01 Piestany, Slovakia, E-mail: zofajova@vurv.sk

Sets of 7 parental varieties and 135 F2.3 lines in the year 2000 and 5 parental varieties and 42 F2.4 lines in the year 2001 were evaluated for biomass yield at heading and at maturity, SPAD index, grain yield and N content in straw and in grain (only in 2001). Portion of dry matter at heading from total at maturity was lower in the year 2000 (43.0% at varieties and 52.3% at lines) by reason of short period from emergence to heading than in the year 2001 (51.8% at varieties and 56.9% at lines) when the same vegetative period was longer by 14 days. Portion of translocated dry matter in grain was higher in the year 2001 when was higher sink capacity for grain (or biomass at heading) (649.2 g/m2 at varieties and 641.4 g/m2 at lines) than in the year 2000 (467 g/m2 at varieties and 527 g/m2 at lines). Portion of translocated N in grain was 82.02% at varieties and 93.07% at lines. Genotypes with higher translocated dry matter had lower grain yield. For the most characteristics portion of variability for crossing was higher than for lines within crossing.