17^th North American Barley Researchers Workshop (NABRW)

September 22-25, 2002

Ramada Plaza Suites and Conference Center

Fargo, North Dakota, USA

Host Organizing Committee

Richard Horsley, Co-Chair

Michael Edwards, Co-Chair

Paul Schwarz

Lynn Dahleen

Marcia McMullen

Stephen Neate

Jerome Franckowiak

Barley and it's potential as a forage crop in dairy production in Mexico. Jorge A. Correa¹, Flavio Capettini² and Sanjaya Rajaram³, ¹Semillas Correa Mexicana, S.A. de C.V., Celaya, Guanajuato; ²ICARDA/CIMMYT Barley Breeding Program, Apdo Postal 6-641, Mexico DF, México; ³CIMMYT Wheat Program, Apdo Postal 6-641, Mexico DF, Mexico

The development of the Mexican dairy industry created a great increase in demand of high quality forage, during the summer-spring as well as in fall-winter seasons. The 10-year drought in Mexico has generated the need to produce forage for the short-term demand and in highly technified areas such as in Laguna, Northern Mexico. It is important to mention that this demand has also brought the need to grow forage in two cycles during fall and winter, including oat, wheat, triticale and more recently, barley. Semillas Correa Mexicana, a national seed producing company, has been working intensively in the last two years (two cycles per season) in order to obtain new varieties of oat, triticale and awnless barley. It is important to underline that during fall-winter, barley results were superior to all other crops, exceeding the expected. Therefore, we will focus our presentation in the results obtained in three different states in México, and will also see the future importance that barley will play in our country.

Jorge A. Correa, semillas@scorrea.com, (01461) 61 15135

Root Disease Suppression In Small Grains In A Low Rainfall And Low Soil Fertility Environment. Stephen M. Neate¹ and Vadakattu Gupta², ¹Department of Plant Pathology, North Dakota State University, Fargo, ND,;²CSIRO Land and Water, SA Australia.

Traditionally, ecological research into mechanisms which lead to disease suppressive soils has involved study of interactions between the pathogen and only one or a few microorganisms or physical or chemical factors in the soil. As the temporal and spatial growth of a pathogenic organism is influenced by ongoing interactions with different trophic groups of soil biota we propose that an ecological approach may be better suited to understand the mechanisms behind the development of a disease suppressive soil. We systematically tested the effect on disease of the potential factors that influence the survival of disease inoculum, both within and between seasons. The often transient nature of the interactions we detected confirmed the need for a sequential sampling approach. Spatial variations also required that that factor be taken into account when taking measurements. The complex biotic interactions measured confirmed the need for an integrated ecological approach i.e. combination of functional and trophic groups, and utilizing the food-web model. We compare the results we obtained in a low rainfall low soil fertility environment with other mechanistic studies of suppression in higher rainfall and higher fertility environments.

Stephen M. Neate, stephen.neate@ndsu.nodak.edu; 701 231-7078

Host-Parasite Genetics in the Hordeum vulgare:Cochliobolus sativus pathosystem. Brian Steffenson and Shaobin Zhong, Department of Plant Pathology, University of Minnesota, St. Paul, MN 55108

Spot blotch, caused by Cochliobolus sativus, is an important disease of barley in the United States. Six-rowed malting cultivars in the upper Midwest possess durable resistance to spot blotch, which was derived from the breeding line ND B112. Genetic analysis of six-rowed barley cultivars revealed that spot blotch resistance at the seedling stage is conferred by a single major gene (Rcs5) on chromosome 1(7H). Durable adult plant resistance in the field is controlled by two quantitative trait loci: one on chromosome 5(1H) explaining 63% of the phenotypic variance and a second one on chromosome 1(7H) explaining 9% of the variance. Three pathotypes (0, 1, and 2) of C. sativus were identified on three differential barley genotypes. A cross was made between the pathotype 2 isolate ND90Pr and the pathotype 0 isolate ND93-1. Virulence on cultivar Bowman was controlled by a single locus (VHv1) in isolate ND90Pr. A genetic map of the ND90Pr/ND93-1 cross was constructed using molecular markers, and six co-segregating AFLP markers were identified for VHv1 on one of the major linkage groups. Fifteen chromosomes were resolved from CHEF gel electrophoresis of C. sativus isolates ND90Pr and ND93-1. Hybridization of CHEF-separated DNA with a cloned AFLP marker co-segregating with VHv1 localized the barley virulence locus to a well-separated chromosome of 2.8 Mbp. Research is underway to clone both Rcs5 in barley and VHv1 in C. sativus. The successful cloning of these genes will allow us to develop a model system for studying host-parasite interactions at the molecular level.

Brian Steffenson, bsteffen@umn.edu, (612) 625-4735

Genetically engineered stem rust resistance in barley using the Rpg1 gene. Horvath, H., Rostoks, N., Brueggeman, R., Steffenson, B., von Wettstein, D. and Kleinhofs, A. Department of Crop and Soil Sciences, School of Molecular Biosciences, Washington State University.

Stem rust, caused by Puccinia graminis f. sp. tritici, was among the most devastating diseases of barley in the northern GreatPlains of the U.S. and Canada before the deployment of the stemrust-resistance gene Rpg1 in the 1940s. Since then, Rpg1 has provideddurable protection against severe stem rust losses, except for a few minor outbreaks. Recently, the Rpg1 gene was cloned by a map-based approach. Sequencing of Rpg1 alleles revealed a functional gene structure only in resistant cultivars and a defective gene structure or lack of Rpg1 in susceptiblecultivars. Here we present the first case of stable transformation of a susceptible cultivar with a cloned barley disease resistance gene. Using Agrobacterium-mediated transformation, the Rpg1 gene isolated from cv. Morex (including 864 bp upstream of the start codon and 651 bp downstream of the stop codon) was transferred into the susceptible cultivar Golden Promise. Forty-two transgenic T₀ plants containing the stem rust resistance gene were obtained. The T₁ generation of 12 transformants was analyzed for resistance to stem rust pathotype MCC. Resistance was observed in 10 transgenic barley lines, while two lines gave rise to susceptible progeny. Interestingly, the majority of transgenic stem rust resistant plants exhibited a level of resistance that was higher than Morex, the original source of the cloned resistance gene. The reason for this apparent anomaly as well as stability of transgenic plants is being investigated.

Henriette Horvath, PhD, henny@mail.wsu.edu, 509-335-5933

Barley production and the impact of seedbed utilization, row spacing, and fungicide. T.K. Turkington¹ , H.R. Kutcher², G.W. Clayton¹, J.D. O'Donovan³, A.M. Johnston⁴, K.N. Harker¹, J.H. Helm⁵, and F.C. Stevenson⁶. ¹Lacombe Res. Centre, Agric. and Agri-Food Canada (AAFC), Lacombe, AB, Canada; ²Melfort Research Farm, AAFC, Melfort, SK, Canada; ³Beaverlodge Res. Farm, AAFC, Beaverlodge, AB, Canada; ³Potash and Phosphate Institute of Canada, Saskatoon, SK, Canada; ⁵Field Crop Dev. Centre, Alberta Agric., Food and Rural Dev., Lacombe, AB, Canada; ⁶142 Rogers Rd., Saskatoon, SK, Canada.

Seeding systems that have openers spreading seed rather than placing it in distinct rows may enhance foliar cereal disease pressure by restricting canopy air movement, thus producing favourable microenvironmental conditions. In addition, narrow row spacing may also enhance disease development and negatively impact productivity. A field experiment was conducted at Lacombe and Beaverlodge, AB, and Melfort, SK, from 1999 to 2000 to evaluate the effect of seedbed utilization (SBU) using three seed placements (distinct row: 23 cm and 30 cm with a hoe opener; and spread band: a 20 cm spread using a 28 cm sweep on 23 cm row spacing) and fungicide (propiconazole) applications (untreated check, and applications at the 2–3 leaf stage, flag leaf stage, heading stage, 2–3 leaf plus flag leaf stage, and flag leaf plus heading stage). Net blotch severity and yield varied with location and year, with disease highest and yields lowest for those treatments that did not include a fungicide application at either the flag leaf stage or heading. Foliar disease severity and yield were influenced by the SBU treatments, but varied with location and year, with a trend of higher disease and lower yields for the two hoe opener treatments, which were similar. The increase in disease and reduction in grain yield, when no fungicide was applied, tended to be smaller for the spread band versus hoe treatments. Planting in distinct rows may have resulted in higher disease and lower yields by helping to facilitate spore dispersal and subsequent disease development compared with the spread band treatment.

T.K. Turkington, turkingtonk@agr.gc.ca, (403) 782-8138.

Efficacy of Fungicides in Controlling Barley Fusarim Head Blight in Lines With Partial Resistance. J.D. Pederson¹, R.D. Horsley¹, M. McMullen^2,3, and K. McKay³. ¹Dep. of Plant Sciences, North Dakota State Univ., ²Dep of Plant Pathology, North Dakota State Univ.; and ³North Dakota Extension Service.

Research to test the efficacy of fungicides in controlling Fusarium head blight (FHB) and deoxynivalenol (DON) levels in barley was previously conducted using cultivars (i.e. Robust, Foster, and Stander) that are susceptible to FHB. Results indicate that fungicides had little to no effect in reducing DON concentration to levels acceptable to the malting and brewing industry. Minimal information is available on the efficacy of fungicides in controlling FHB and DON levels on genotypes with partial FHB resistance. The objective of this study is to determine if the integrated use of fungicides and barley cultivars with partial resistance to FHB will control FHB severity and accumulation of DON. Experiments were conducted in the field in North Dakota since 2000 and included genotypes resistant, partially resistant, and susceptible to FHB. Fungicides used were Folicur in 2000, 2001, and 2002; and AMS21619 in 2001 and 2002. Folicur did not significantly reduce FHB severity or DON accumulation in resistant, moderately resistant, or susceptible genotypes. However, genotypes sprayed with Folicur generally had greater yield due to control of septoria speckled leaf blotch (SSLB), incited by Septoria passerinii. Yield gains due to control of SSLB tended to be sufficient to cover the cost of Folicur and its application on cultivars developed and released by upper Midwest barley breeding programs. Preliminary data indicates that efficacy of AMS21619 was slightly better than Folicur in reducing FHB and DON.

R. Horsley, Richard.Horsley@ndsu.nodak.edu, 701-231-8142

A Historical Perspective on Barley Breeding and Related Research at Aberdeen, Idaho. Darrell M. Wesenberg*, J. Michael Bonman, and Donald E. Obert

USDA-ARS ret.* and USDA-ARS, Aberdeen, Idaho 83210

The Aberdeen Research and Extension Center was established in 1911. Initially the Idaho Agricultural Experiment Station and the USDA Office of Cereal Investigations jointly operated the Center. Early objectives included the improvement of cereals in the intermountain region by introducing better varieties. Spring barley variety testing began in 1912. Winter barley, hulless barley, hooded, and beardless barley were studied. Composite cross populations were reported as early as 1923. The first reference to malting barley was the inclusion of ‘Canadian Malting No. 2’ in trials in 1925. Over time varieties resulted from reselections from introduced varieties, variety introductions, selection from composite crosses, and traditional pedigree breeding. G.A Wiebe played a key role in barley research at Aberdeen from the early 1920s. With his appointment as Junior Plant Breeder by the USDA in May 1922, he began a long career with the USDA, culminating in the national role of USDA-ARS Barley Investigations Leader. Other individuals who provided the basis for barley breeding and research accomplishments at Aberdeen include H.V. Harlan, M.L. Martini, Harland Stevens, Ralph Hayes, and Allen Dickson. Varieties introduced or developed and released at Aberdeen have been important commercial varieties as well as being widely used parents that have contributed to other significant varieties. Today basic project objectives are similar, but goals are more diverse and although traditional pedigree breeding remains an important approach, new breeding targets and new technologies involving tissue culture, transgenic germplasm, and molecular markers are creating greater opportunities for barley improvement.

D.M. Wesenberg, dwesen4285@aol.com, 208-226-2638

Principles Learned about Quantitative Traits in Barley from QTL Analysis.

Steven E. Ullrich, Washington State University, Pullman, WA 99164-6420

Quantitative traits (QTs) have always been difficult to study and understand and have greatly baffled geneticists. Conventional quantitative genetic analyses have been unsatisfactory and unsatisfying in terms of actual usable knowledge gained, and the assumptions underlying such analyses are generally unrealistic and unacceptable. The following is a quote about QTs from a well known plant breeding textbook published within the last decade: “Each multiple gene expresses a small effect on the phenotype relative to the total variation; normally it is not possible to identify individual gene effects.” Quantitative trait locus (QTL) analysis using advanced molecular genetic and computer tools has opened a floodgate of knowledge about quantitative traits not available through conventional genetic analyses. The term QTL was invented to describe a genetic determinant for a QT, which is not quite at the level of gene or locus in the conventional sense. This paper is intended to be a reflection of perspectives based on experience and observation.

Considerable new and usable information has been gained over the past 10 years about economically important QTs in barley starting with but not limited to QTL analysis. Much of this information has been made possible through the North American Barley Genome (Mapping) Project and based on key comprehensive molecular maps. First of all, QTL analyses have identified the number and approximate chromosome location of QTLs for many traits (7, 11, 12, 13, 16). These results in themselves are very exciting and satisfying. QTL analyses have given us the first real genetic glimpses of important QTs. In addition QTL analysis has allowed for the calculation of QTL effects (r²values or % variation explained), heritability, and environmental (E) and QTL x E, and QTL x QTL interaction effects.

QTLs that map coincidently with other QTLs of other traits have confirmed relationships among traits and have raised questions about the occurrence and role of pleiotropy and gene clusters. For example, malt extract content QTLs usually map coincidently with other QTLs, such as for a-amylase activity, diastatic power, malt b-glucanase activity, and/or malt b-glucan content, which are all sub-traits or contributing traits of malt extract (5, 18). Grain yield QTLs often overlap with sub-trait QTLs, such as heading date, plant height, lodging resistance, and/or shatter resistance (7, 12, 16). Fusarium head blight resistance QTLs may overlap with QTLs for plant height and inflorescence morphology (20). Chromosome regions that are rich in major related QTLs are logical candidates for marker-assisted selection (MAS) for trait maintenance or improvement in applied breeding.

QTL analysis has opened the door for fine mapping, further genetic study, map-based cloning, and MAS. Identified QTLs have been “isolated” by selection within doubled haploid line (DHL) mapping populations or by marker-assisted backcross isoline development. Such lines have been used for QTL verification and study of gene action and epistasis, e.g., for seed dormancy (3, 4). Marker-assisted selection has been used for QTL verification and for breeding applications. Barley stripe rust (BSR) resistance QTLs were mapped (1) and the information used for breeding BSR resistance cultivars (6,17). This involved a two gene-plus model. Verification of QTLs via tests of MAS has met with mixed results. Using DHLs form ‘Steptoe’/’Morex’ crosses not used in the mapping efforts, a major malting quality QTL region on chromosome 1 (7H) was verified and MAS improved malting quality compared with conventional phenotypic selection (2). However, another QTL region on chromosome 4 seemed to disappear! A similar study, using ‘Harrington’/TR306 DHLs not used in the mapping effort, produced similar results. Two major malting quality QTL regions on chromosome 7 (5H) were verified with MAS improving malting quality, but QTL regions on chromosomes 3 and 6 could not be definitively confirmed (8). Moving a grain yield QTL on chromosome 3 from Steptoe into Morex via MAS seemed not to adversely affect malting quality of progeny, but did lower test weight and apparently did not increase grain yield (10). Again, using a Steptoe/Morex DHL population not used in mapping, confirmed with consistent positive MAS results grain yield QTL regions on chromosome 3 and 6. MAS for chromosome 2 and 7 (5H) yield QTLs gave inconsistent results due apparently to QTL x E interactions (15). Introgressing three yield QTL regions from Steptoe into the Morex background via MAS resulted in little effect on malting quality, improvements in yield related traits (reduced plant height, lodging, shattering), but no improvement on grain yield itself (9). On the other hand a MAS study involving backcrossing ‘Baronesse’ grain yield QTLs into the Harrington background has been preliminarily successful in the development of lines with Harrington-like malting quality and Baronesse-like grain yield (Schmierer et al., 2002, this proceedings). However, there is one backcross line that has improved yield and reduced malting quality, and it does not contain any of the targeted Baronesse yield QTLs!

Experience in fine mapping studies using isolines developed by MAS backcrossing and in various MAS breeding schemes for improving or maintaining complex “mega traits”, such as malt extract and grain yield, indicates that the background genotype is of great importance. It appears isogenic lines are not usually isogenic even with the aid of molecular marker checks. Furthermore, behavior of QTLs is not always as expected in different backgrounds. Ramage’s (14 ) concepts of the role of background genotype and “happy homes” appear to be as important for QTLs as they are for qualitative genes. Lack of ‘purity” of genetic background also allows for epistatic effects not expected because of previously undetected “genes”. The effects of E and QTL x E interactions also complicate understanding. Crossover interactions in which a QTL expresses alternative favorable alleles (19) are also perplexing and complicate MAS. However, refinement of techniques and the ability to observe the genome more comprehensively, with for example micro-arrays, will continue to advance our understanding of QTs. But at present, even with QTL analysis and subsequent molecular genetic study, QTs can still be baffling.

References

1. Chen, F.Q. D. Prehn, P.M. Hayes, D. Mulrooney, A. Corey, and H. Vivar. 1994 Mapping genes for resistance to barley stripe rust (Puccinia striiformis f.sp. hordei). Theor. Appl. Genet. 88:215-219.

2. Han, F., I. Romagosa, S.E. Ullrich, B. Jones, P.M. Hayes, and D.M. Wesenberg. 1997. Molecular marker assisted selection for malting quality traits in barley. Molec. Breed. 3:427-437.

3. Han, F., S.E. Ullrich, J.A.. Clancy, V. Jitkov, A. Kilian, and I. Romagosa. 1996. Verification of barley seed dormancy loci via linked molecular markers. Theor. Appl. Genet. 92:87-91.

4. Han, F., S.E. Ullrich, J.A. Clancy, and I. Romagosa.1999. Inheritance and fine mapping of a major barley seed dormancy QTL. Plant Sci. 143:113-118.

5. Han, F., S.E. Ullrich, A. Kleinhofs, B.L. Jones, P.M. Hayes, and D.M. Wesenberg. 1997. Fine structure mapping of the barley chromosome 1 centromere region containing malting quality QTL. Theor. Appl. Genet. 95:903-910.

6. Hayes, P.M., A.E. Corey, R. Dovell, R. Karow, C. Undi, K. Rhinart, and H. Vivar. 2000. Registration of Orca barley. Crop Sci. 40:849.

7. Hayes, P. M., B. H. Liu, S. J. Knapp, F. Chen, B. Jones, T. Blake, J. Franckowiak, D Rasmusson, M. Sorrells, S. E. Ullrich, D. Wesenberg and A. Kleinhofs. 1993. Quantitative trait locus effects and environmental interaction in a sample of North American barley germplasm. Theor. Appl. Genet. 87: 392-401.

8. Igartua E., M. Edney, B.G. Rossnagel, D. Spaner, W.G. Legge, G.J. Scoles, P.E. Eckstein, G.A. Penner, N.A. Tinker, K.G. Briggs, and D.E. Falk. 2000. Marker-based selection of QTL affecting grain and malt quality in two-row barley. Crop Sci. 40:1426-1433.

9. Kandemir, N., B.L. Jones, D.M. Wesenberg, S.E. Ullrich, and A. Kleinhofs. 2000. Marker assisted analysis of three grain yield QTL in barley (Hordeum vulgare L.) using near isogenic lines. Molec. Breed. 6:157-167.

10. Larson, S.R., D.K. Habernicht, T.K. Blake, and M. Adamson. 1997. Backcross gains for six-rowed grain and malt qualities with introgression of a feed barley yield QTL. J. Am. Soc. Brew. Chem. 55:52-57.

11. Marquez-Cedillo, L.A., P.M. Hayes, B.L. Jones, A. Kleinhofs, W.G. Legge, B.G. Rossnagel, K. Sato, S.E. Ullrich, D.M. Wesenberg, and the NABGMP. 2000. QTL analysis of malting quality in barley based on the doubled haploid progeny of two elite North American varieties representing different germplasm groups. Theor. Appl. Genet. 101:173-184.

12. Marquez-Cedillo, L.A., P.M. Hayes, A. Kleinhofs, W.G. Legge, B.G. Rossnagel, K. Sato, S.E. Ullrich, D.M. Wesenberg, and the NABGMP. 2001. QTL analysis of agronomic traits in barley based on the doubled-haploid progeny of two elite North American varieties representing different germplasm groups. Theor. Appl. Genet. 103:625-637.

13. Mather, D.E., N.A. Tinker, D.E. LaBerge, M. Edney, B.L. Jones, B.G. Rossnagel, W.G. Legge, K.G. Briggs, R.B. Irvine, D.E. Falk, and K.J. Kasha. 1997. Regions of the genome that affect grain and malt quality in a North American two-row barley cross. Crop Sci. 37:544-554.

14. Ramage, R.T. 1977. Male sterile facilitated recurrent selection and happy homes. p. 92-98. in S. Barghout et al., (ed.). Proc. 4^th reg. winter cereals workshop, vol. 2. barley. Amman, Jordan.

15. Romagosa, I., F. Han, S.E. Ullrich, P.M. Hayes, and D.M. Wesenberg. 1999. Verification of yield QTL through realized molecular marker assisted selection responses in a barley cross. Molec. Breed. 5:143-152.

16. Tinker, N.A., D.E. Mather, B.G. Rossnagel, K.J. Kasha, A. Kleinhofs, P.M. Hayes, D.E. Falk, Ferguson, L.P. Shugar, W.G. Legg, R.B. Irvine, T.M. Choo, K.G. Briggs, S.E. Ullrich, J.D. Franckowiak, T. Blake, R.J. Graf, S.M. Dofing, M.A. Saghai Maroof G.J. Scoles, D. Hoffman, L.S. Dahleen, A. Kilian, F. Chen, R.M. Biyashev, D.A. Kudrna, and B.J. Steffenson. 1996. Regions of the genome that affect agronomic performance in two-row barley. Crop Sci. 36:1053-1062.

17. Toojinda, T., E. Baird, A. Booth, L. Broers, P. Hayes, W. Powell, W. Thomas, H. Vivar, and G. Young. 1998. Introgression of qualitative trait loci (QTLs) determining stripe rust resistance in barley: an example of marker-assisted line development. Theor. Appl. Genet. 96:123-131.

18. Ullrich S.E., F. Han, and B.L. Jones. 1997. Genetic complexiy of the malt extract trait in six-row spring barley suggested by QTL analysis. J. Am. Soc. Brew. Chem. 55:1-4.

19. Zhu, H., G. Briceno, R. Dovel, P.M. Hayes, B.H. Liu, C.T. Liu, and S.E. Ullrich. 1999. Molecular breeding for grain yield in barley: An evaluation of QTL effects in a spring barley cross. Theor. Appl. Genet. 98:772-779.

20. Zhu, H., L. Golchrist, P. Hayes, A. Kleinhofs, D. Kudrna, Z. Liu, L. Prom, B. Steffenson, T. Toojinda, and H. Vivar. 1999. Does function follow form? Principle QTLs for Fusarium head blight (FHB) resistance are coincident with QTLs for inflorescence traits and plant height in a doubled-haploid population of barley. Theor. Appl. Genet. 99:1221-1232.

S.E. Ullrich, Ullrich@wsu,edu, 509-335-4936

Digital Plant Breeding. Mario C. Therrien AAFC Brandon Research Centre, Brandon, MB. Canada

In recent years, electronic devices for measuring plant parameters, such as crop density, and gathering digital information, such as digital cameras, have become robust and relatively inexpensive. Their ability to rapidly measure physical and optical properties has made these devices a potentially useful set of tools in a breeding program by measuring or predicting useful agronomic traits. Five different digital devices were assessed for their practicality and utility in identifying high (grain and forage) yielding genotypes of barley for potential use in a breeding program. These included 1) a digital camera, 2) a crop canopy analyzer, 3) a chlorophyll fluorometer, 4) a SPAD meter, and 5) a multi-spectral radiometer. Nine barley cultivars were chosen, widely differing in genetic background and agronomic performance, and were planted at two locations over two years. Each device was used throughout the growing season at emergence, tillering, shot blade, boot, anthesis, and grainfill stages (except the block harvested as silage). Values obtained were correlated with grain and biomass yield. The digital camera and SPAD meter were also used to examine disease load and incidence at anthesis and grainfill. As well, the digital camera also measured harvested grain parameters. Each device was also assessed for ease of use and cost/benefit. Results show that all the devices were able to predict grain or biomass yield, or both, except for the fluorometer. The digital camera and SPAD meter were also useful for measuring other traits. The SPAD meter gave the best overall results.

Mario C. Therrien, Mtherrien@em.agr.ca, 204-726-7650

Breeding for lower DON response in six-row malting barley using conventional methods and existing germplasm. Dr. D.B. Cooper Busch Agricultural Resources Inc. 3515 E. Co. Rd. 52, Ft. Collins, CO 81024

Fusarium head blight (FHB) on malting barley has become an important limitation on the stable supply of acceptable malt barley in the upper mid-western United States. Considerable effort has been made to screen germplasm for sources of resistance to both the pathogen itself and for sources of reduced deoxynivalenol (DON) content in the grain. The relatively large environmental effects for both resistance and DON content have made it difficult to identify lines with better response. After several location x years of screening for DON content in our breeding program we tentatively identified several lines with lower DON content. We inter-crossed these lines in a partial diallel mating design. The resulting progeny were made available to all interested public malt barley breeding programs in the region and with their assistance we measured the DON content over several generations and over a wide range of environments. Combining ability analysis of DON content confirmed that the principle mode of gene action was additive. Some specific combinations had lower DON than predicted on an additive basis only, indicating limited broad-sense specific combining ability was also present. We recently solicited lines with better response to FHB from the public programs and hand crossed them with our best lines in the greenhouse. We obtained seed set on 322 of the possible 410 half-diallel combinations. The F₁ generation of these new crosses will be increased in Arizona during the fall of 2002 and returned to Ft. Collins for a first cycle of random mating in the spring of 2003 using Genesis^TM hybridizing agent. We expect cycle-1 F₂ seed should be available to interested parties for planting and selection in the spring of 2004.

Dr. D.B Cooper, blake.cooper@anheuser-busch.com, (970) 472-2327.

GrainGenes and Other Public Databases for Barley Genomics. Victoria Carollo¹, David Matthews², Gerard Lazo¹, Olin Anderson¹,¹USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710; ²Cornell University, Dept. of Plant Breeding, Ithaca NY 14853

Sequencing efforts in the last few years have yielded an enormous wealth of data for Triticeae genomics. GrainGenes (http://www.graingenes.org), the USDA-ARS sponsored Triticeae database holds records for over 150,000 barley gene EST sequences. Although sequences are deposited in to the GenBank and are available to the public, further efforts by the bioinformaticists at the USDA-ARS have improved the accessibility and offer unprecedented ways to view and manipulate these data. GrainGenes curators have worked with curators at Gramene (www.gramene.org) and NCBI (www.ncbi.nlm.nih.gov) to provide data and link to comparative mapping tools for barley. Sequence and locus records in GrainGenes are also linked to external databases including GenBank and the UniGene clusters for the Triticeae at NCBI, TIGR tentative consensus sequences (http://www.tigr.org/tdb/tgi/) and the “Genome View” at Gramene, the USDA-ARS sponsored resource for comparative mapping in grains. Gramene users can align the North American Barley Genome Mapping Project (NABGMP) Steptoe x Morex maps with selected maps from rice, wheat, oat, maize and sorghum. Another new comparative mapping tool is available at the Plant Genomes Central, a new site at NCBI dedicated to plant genomics (www.ncbi.nlm.nih.gov/PMGifs/Genomes/PlantList.html). Barley maps can be compared to each other and include two consensus maps, the NABGMP Steptoe x Morex and Harrington x TR306.

Victoria Carollo, vcarollo@pw.usda.gov, 510-559-5944

Genetic and physical mapping towards map-based cloning in barley. Kleinhofs, Andris, Dept Crop & Soil Sciences and School of Molecular Biosciences, Washington State University, Pullman, WA 99164-6420

Map-based cloning in large genome organisms such as barley is tedious and expensive. It should be avoided if the homologous gene can be obtained from the small genome monocot model rice. However, rice does not have all the genes we might be interested in i.e. rust resistance genes. Within the Triticeae barley is the organism of choice for map-based cloning due to the extensive genetic maps and an excellent publicly available BAC library. Map-based cloning in barley has already been successful. The mlo, Rar1, Mla, Rpg1 genes have been cloned using this approach and a BAC contig including the rpg4 gene has been constructed. Construction of the BAC contigs including the stem rust resistance genes Rpg1 and rpg4 and the cloning of Rpg1 will be presented. This work was accomplished by chromosome walking. In order to avoid this tedious approach, we developed a system of targeted map saturation based on the rice genome sequence data and Triticeae EST database. In this approach we identify Triticeae ESTs with homology to rice genomic sequence with synteny to the region of interest. The Triticeae ESTs are mapped at low resolution, BAC clones identified and contigs established. This approach was used to target the Rcs5 gene. Progress will be reported.

Andris Kleinhofs, andyk@wsu.edu, 509-335-4389

HarvEST Triticeae: a portable EST database viewer for barley researchers and others. Steve Wanamaker and Timothy J. Close, Department of Botany & Plant Sciences, University of California, Riverside, CA, 92521-0124, USA

HarvEST Triticeae is a portable EST database-viewer that emphasizes gene function and is oriented to the design of oligonucleotides, supporting microarray content design, BAC library screening, and PCR-based genetic mapping. The software is downloadable from http://harvest.ucr.edu and will run on any PC that uses Windows and meets the minimum hardware configuration. Version 0.86 (released July 2002) contains more than 258,000 barley EST sequences representing about 40,000 unigenes. All EST sequences have been quality trimmed, cleaned of vector, and assembled using CAP3. For key features of HarvEST, no internet connection is required. Key features include: unigene output from archived contig assemblies, Boolean searching with user-defined quantitative settings, archived BLAST hit information, and various other searching, reporting and export functions. The unigene feature can be applied to entire assemblies, individual libraries, or any combination of libraries. For Boolean searches, a user sets search parameters according to their interest. For example, the choice could be genes associated with a particular stress (e.g. low temperature, heat, drought, pathogen, nitrogen deprivation), developmental stage (e.g. malted seed, 20 DAP spike), or tissue (e.g. endosperm, pericarp, rachis, root tip). The user also decides what percentage representation is relevant. Once the choices have been made, the search is fully executed and a browsable output is displayed. The output can be viewed on-screen or exported as summary tables or sequence files. HarvEST also facilitates connection to NCBI for live BLAST searches of the public EST database.

Timothy J. Close, timothy.close@ucr.edu, 909-787-3318.

Using EST databases for functional and comparative bioinformatics. Warren M. Kruger and Gary J. Muehlbauer University of Minnesota, St. Paul MN 55108

High throughput sequencing capacity, comprehensive sequence databases and algorithms for finding sequence similarity has facilitated the use of expressed sequence tags (ESTs) for gene discovery. Functional and comparative analysis of ESTs was used to identify genes expressed when barley is infected by Fusarium graminearum (Fg) and Blumeria graminis f.sp. hordei (Bgh). A database of 8,400 non-redundant sequences was created from almost 20,000 ESTs, generated from a Fusarium- and three powdery mildew-infected cDNA libraries. Functional annotations were derived from BLASTX hits (P≤10^-5) to proteins in Genbank’s non-redundant protein database. We found 982 genes expressed in response to Fg and Bgh of which 29 were not found amongst 130,000 barley ESTs generated from various uninfected tissues. 72% (21/29) of these genes encode unknown proteins and 14% (4/29) encode amino acid metabolic proteins. Genes encoding structural, metabolic and stress-related proteins, e.g. PR proteins, glutathione S-transferase and aluminum-induced proteins, were the most abundantly expressed in response to Fg and Bgh. 143 Fg and Bgh genes were not found in over 8,000 Fg and 4,700 Bgh ESTs generated from non-infective fungal tissue. A subset of 13 genes did not have homologs in Saccharomyces cerevisiae and Neurospora crassa and may represent fungal virulence factors. We identified 70 genes specifically expressed in at least two plant-microbe interactions, suggesting that there are similarities between all plant-microbe interactions.

Warren M. Kruger, kruge002@umn.edu, (612) 625-9701

Parallel Expression Analysis using Barley Microarrays. Roger Wise¹, Rico Caldo¹, Stacy Turner², Dan Ashlock³, and Julie Dickerson⁴. ¹USDA-ARS & Plant Pathology, ²Bioinformatics and Computational Biology, ³Mathematics, and ⁴Computer & Electrical Engineering, Iowa State University, Ames, IA 50011-1020

In small grain Triticeae crops, the molecular characterization of genes coincident with disease, response to biotic or abiotic stresses, or cellular development has traditionally followed a “one-gene-at-a-time” approach. However, recent advances in microarray technology now allow the parallel investigation of up to 22,500 genes in a single experiment. Early next year, a USDA-IFAFS-funded barley GeneChip will be available from Affymetrix. We will utilize our microarray to identify new, undiscovered genes that up- or down-regulated in response to biotic and abiotic stresses. In order to effectively access, compare, and manipulate up to 500,000 data points per experiment, this project will create an on-line interactive database, BarleyBase, and develop a set of web-accessible tools for the analysis of Affymetrix GeneChip data. BarleyBase will feature “click through” integration of the data on the web and it will be interoperable with the Gramene comparative mapping resource for grains (http://www.gramene.org/). The web-based analysis tools will enable database users to identify subsets of genes that change expression in response to drought, cold stress, disease, or other treatments. These tools will accelerate agronomic and quality research in cereals, one of the world’s most important food sources.

Roger Wise, rpwise@iastate.edu, 515-294-9756

Effectiveness of in vitro selection in barley for Fusarium Head Blight Resistance.

M. Banik¹, W.G. Legge¹, B. Bizimungu², J. Tucker¹, M.C. Therrien¹, A. Tekauz², F. Edes³and M. Savard⁴. Agriculture and Agri-Food Canada- ¹ Brandon Research Centre, Brandon, MB, ²Cereal Research Centre, Winnipeg, MB, ³ Lethbridge Research Centre, Lethbridge, AB, ⁴Eastern Cereal and Oilseed Research Centre, Ottawa, ON, Canada

Deoxynivalenol (DON), the most prevalent mycotoxin produced by Fusarium graminearum Schwab, plays an important role in Fusarium Head Blight (FHB) pathogenesis. DON or a mixture of DON with other mycotoxins, 15-acetyldeoxynivalenol and T2, were added to anther culture media to evaluate feasibility of in vitro selection for increasing the level of Fusarium resistance in barley. Three barley genotypes and 7 F₁ hybrids with varying levels of FHB resistance were in vitro selected in the presence of toxin. All fertile, selected and control double haploids were tested in the FHB nursery at Brandon, MB, in 2000-2001 under artificial infection and DON content was determined by ELISA technique. Although not statistically significant (P>0.5), several lines showed a lower DON content (AC Metcalfe 99/30-2, Excel 99/12-3, Robust DT00-14) in comparison with their respective parental genotype. In vitro selected Excel lines generally had lower DON content and responded the best. Among 7 different crosses, only in vitro selected segregating lines of two row types from Chevron/CDC Fleet had significantly lower DON content than the two-row and six-row control lines. Thus, in vitro selection appeared to be effective in two row types of Chevron/CDC Fleet cross. Further testing with more replication over years will be required to confirm the results.

Mitali Banik, mbanik@agr.gc.ca, 204-726-7650

Characterization of the barley stem rust resistance gene Rpg1 family. R. Brueggeman^*, N. Rostoks^*, D. Kudrna^*, A. Druka^*, A. Kleinhofs^*‡.^*Department of Crop and Soil Sciences and ^‡School of Molecular Biosciences, Washington State University, Pullman, WA 99164-6420, USA

The barley stem rust resistance gene Rpg1 encodes a protein with homology to receptor like kinases (RLKs) with two tandem protein kinase domains, a novel structure for a plant disease resistance gene. The Rpg1 gene family was identified by hybridization of 10 arrayed different tissue cDNA libraries containing approximately 500,000 clones. Using an Rpg1 probe, one cDNA clone was identified from a very similar member of the gene family (ABC1037) and another from a more distantly related member (ABC1036). Hybridization of the more diverged family member ABC1036 back to the cDNA libraries identified a closely related member (ABC1040) and a more distant member ABC1041. A BLASTn search of the Triticeae EST database using the ABC1041 sequence revealed another gene (ABC1063) which is closely related to ABC 1041 , but further diverged from Rpg1. These five cDNA representatives of the Rpg1 gene family and their genomic counterparts were completely sequenced and mapped. The closely related member ABC1037 is linked to Rpg1 (about 40kb) and probably represents a recent duplication event. ABC1036 and ABC1040 mapped to chromosome 7(5H) and are very closely linked. ABC1041 mapped very close to the stem rust resistance gene rpg4 on chromosome 7(5H), but is not rpg4. ABC1063 has not been mapped. Characterization of this gene family will provide insight into function and evolution of this novel resistance gene family in barley and other Triticeae.

Robert Brueggeman, bigbass@wsu.edu, 509-523-5302

Multivariate Analysis of Malting Quality. Allen D. Budde, Lauri Herrin and Berne L. Jones. USDA/ARS, Cereal Crops Research Unit, Madison, WI 53726

Around 4500 barleys are malted and their malting qualities analyzed at the Cereal Crops Research Unit each year. For several years, we have utilized a computerized scoring system to assist us in selecting those lines having the best malting quality. For the 2000 and 2001 crop year submissions, Principal Component Analyses (PCA) were performed on the malting quality values to determine whether the best performing lines could be distinguished from the less desirable ones. The data from the two- and six-rowed varieties were analyzed separately because their ideal malt quality parameters, as listed by the American Malting Barley Association in their ‘Analytical Guidelines to Barley Breeders’, differ. The analytical results were normalized, subjected to PCA, and the results were plotted and compared with the computer generated ‘quality scores’ for each line. The scores of the best performing lines, as calculated by our quality scoring method, clustered together near the site that represented an ‘ideal’ malting barley. Principal component analyses of the barley submissions will provide information that can nicely supplement, or perhaps even replace, our current malt quality scoring system for identifying the best performing barleys.

Allen D. Budde, adbudde@wisc.edu, (608) 262-4483

Breeding BYDV Resistant Barley With an Agronomically Improved Background.

Flavio Capettini¹, Monique Henry² and Hugo Vivar². ¹ICARDA/CIMMYT,²CIMMYT, ^1&2Apdo Postal 6-641, 06600 México D.F., México

Barley Yellow Dwarf is the most important viral disease of barley, having the potential to destroy entire crops in some Latin American countries and worldwide. Breeding for this devastating disease is one of the most important objectives of the ICARDA/CIMMYT barley breeding program, which has been successful to produce resistant or tolerant germplasm in a agronomically enhanced genetic background. Selection against susceptible genotypes has been carried out for more than 20 years in the Toluca Experiment Station of CIMMYT in Mexico, where symptoms are frequent under natural conditions. Artificial inoculation with greenhouse-reared aphids is also done in screening nurseries under field conditions to assure uniform infection, to differentiate biotype reaction and reduce the risk of escapes. The three serotypes most commonly found in the Americas, PAV, MAV and RPV, are used. Plots inoculated with one biotype each and a fourth plot is a check kept free of aphids by insecticide applications. The most common and effective source of tolerance to BYDV in barley has been identified in Ethiopian barleys and found to be associated to the major semi dominant geneYd2. This gene has been reported to be associated with reduction in virus concentration with PAV and MAV but not with RPV. An assay based on a protein showing allelic variation that correlates with Yd2 and a PCR marker for the YD2 associated allele Ylp has been used in the program screened genotypes, finding that the Yd2 gene was present in the genotypes showing field tolerance. Elite lines tested showed field tolerance to PAV and MAV as well as to RPV, indicating that additional gene(s) conferring resistance to this serotype should also be present in the germplasm.

Flavio Capettini, f.capettini@cgiar.org, +52 55 804-7558.

GrainGenes: The Triticeae Genome Database. Victoria Carollo¹, David Matthews², Gerard Lazo¹, Olin Anderson¹;¹USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710; ²Cornell University, Dept. of Plant Breeding, Ithaca NY 14853

GrainGenes, the Triticeae genome database (http://www.graingenes.org), is the most comprehensive source of information on barley, wheat, rye and oats. GrainGenes is maintained at the USDA/ARS/WRRC in Albany, CA and is a central repository for information on gene and expressed sequence tag (EST) sequences, genetic and physical maps, DNA probes, germplasm, pathology, and other data types. GrainGenes may also be accessed through a mirror site at NRC-PBI, Saskatoon, Canada (gene.pbi.nrc.ca/graingenes/). Recent barley related additions to GrainGenes include an expanded image collection from the Oregon Wolfe Barleys, several interactive maps including two with SSR markers and the barley bin maps from Washington State University. These bin maps include markers from several map studies and are available via Gbrowser, a new map-viewing tool developed by the Generic Model Organism Database Project that has been implemented on GrainGenes. Other collaborative efforts are available via GrainGenes including TREP, a Triticeae repeat sequence database (wheat.pw.usda.gov/ITMI/Repeats/) and a Triticeae EST-SSR Coordination project (wheat.pw.usda.gov/ggpages/ITMI/2002/EST-SSR).

Victoria Carollo, vcarollo@pw.usda.gov, 510-559-5944

A proposal for the use of pedigree and historical data for marker trait association in barley. Federico Condon¹, Kevin P. Smith¹ Brian Steffenson², ¹Department of Agronomy and Plant Genetics, University of Minnesota, 1991 Upper Buford Circle, St. Paul, MN 55108. ²Plant Pathology Department, 1991 Upper Buford Circle, St. Paul MN 55108

The identification of associations between phenotypes and molecular markers in the core germplasm of a breeding program could be very useful in crop improvement. This information would eventually allow the identification of potential QTLs without having to develop mapping populations, avoid the presence of undesirable traits in the population and represent the true genetic diversity available to a breeder (Jannink et al, 2002). Since the assumptions of normal distribution and independence is not valid in a non-structured population, this approach should take into account population structure. With this objective, the co-ancestry coefficients between pairs of genotypes is used as a factor in a linear mixed model to asses the expected covariance among genotypes. A set of historical genotypes comprised of variety candidates from the University of Minnesota barley breeding program, and their ancestors were genotyped with 60 SSR primers, and evaluated in the field at three locations in 2002. We observed significant variation in plant height (54 to 97cm), heading date (up to 20 difference), and disease severity for F. graminearum (23 - 93%) and spot blotch (Cochliobolus sativus) (3.5 –65%). The preliminary results of association mapping between the molecular markers and this set of phenotypes will be presented.

Kevin Smith, smith376@tc.umn.edu, (612) 624-1211

Evaluation of Bowman Backcross-derived Barley Genetic Stocks. Jerome D. Franckowiakand An Hang. Department of Plant Sciences, North Dakota State University, Fargo, ND 58105 and USDA-ARS, National Small Grains Germplasm Research Facility, Aberdeen, ID 83210.

Morphological mutants are a valuable genetic resource in barley, Hordeum vulgare. Placing visually identifiable mutants into a common genetic background, represented by the spring, two-rowed cultivar Bowman, can help determine their pleiotropic effects and locate closely linked genetic traits. For example, linkage drag with traits such as spike type (vrs1 and Int-c), smooth awn (raw1 and raw2), anthocyanin pigmentation (Rst1), and brittle rachis (Btr) has helped to associate mutants with specific chromosome regions (Franckowiak, 1995). (See the attached maps.) Agronomic data were collected on over 650 Bowman backcross-derived genetic stocks in nurseries grown near Christchurch, New Zealand and Aberdeen, ID. These sites were chosen to optimize survival of weak lines and to provide seed for the USDA-ARS Barley Genetic Stock Collection at Aberdeen. Based on the data collected, a number of general observations can be made. Many mutants modify the expression of several agronomic traits. Chlorophyll mutants frequently showed maturity delays and had smaller grain. Lines with early heading or reduced tillering genes tended to have large kernels. Resistance to leaf rust, incited by Puccinia hordei, was associated with larger grain and higher yields in New Zealand. Lines having a dominant trait that is found in wild barley often had higher yields. Most semidwarf lines yielded less than Bowman. Based on loci position and agronomic data, about 190 of the Bowman backcross-derived lines were chosen for inclusion in the genetic stocks portion of the barley core collection.

Jerome D. Franckowiak, j.franckowiak@ndsu.nodak.edu (701) 231-7540

Evaluation of Hulless Barley Germplasm at Aberdeen, ID. A. Hang, K. Satterfield, and C. S. Burton. USDA-ARS, National Small Grains Germplasm Research Facility, Aberdeen, ID.

Over 95% of barley grain produced in North America is for animal feed and for malting while the rest is used for seed or for human consumption. Although most barley cultivars are “hulled”, researchers recently have indicated that hulless grain has a higher feed value than hulled barley. In human nutrition studies, beta-glucan and tocotrienols in barley grain are reported to have cholesterol-lowering properties. Beta-glucan in barley also reduces glucose intolerance and insulin resistance, which could improve glucose metabolism and delay or reduce the risk of developing type 2 diabetes. Development of hulless barley with increased beta-glucan and protein contents could help meet the demand for healthy food and could provide new opportunities for growers and the food industry. Forty hulless barley lines from the USDA National Small Grains Collection were planted at various locations in Idaho; agronomic data including heading date, plant height, seed yield, and test weight were recorded. Eight-gram seed samples from each line also were sent to the USDA Cereal Research Unit at Madison, WI for analysis of protein, beta-glucan, and lipid contents. Seed yield and test weight of several hulless lines were comparable with the check cultivars. Eleven lines had protein content varying from 17.12 to 18.28% and three lines had beta-glucan levels ranging from 7.60 to 9.47%. These lines will be used as germplasm for further improvement of hulless barley.

An Hang, anhang@uidaho.edu (208) 397-4162. Ext. 119

A New Look at Location Yield Data. James H. Helm, Patricia Juskiw and Tim Duggan, Field Crop Development Centre, Lacombe, AB, Canada.

When multi-year and multi-location data are analyzed, they are most often summarized on a geographic basis over years. This combines sites and years that are in a geographic area despite production levels. The fallacy of this becomes evident when drought , soil fertility or heavy rainfall makes a site or year significantly different than the norm. The Field Crop Development Centre has developed a SAS based program called the FCDC Data Miner that allows the search of all data bases across years and locations and has the power to present the summary of the data in either geographic zonal format or by yield level of the test that the selected varieties are in. The presentation of data according to yield level of the trial has the advantage of comparing lines in low, medium low, medium high and high yielding environments. The yield response curves can be graphed in relation to a specific check variety or to the mean of the test. This type of analysis shows the best varieties for low yielding environments as well as those varieties that respond well to higher yielding environments. Also built into the software is the potential to screen by protein content so that malting barleys grown in tests with selectable protein content can be compared together while those tests that deliver higher or lower protein content can be evaluated for yield potential in a feed environment. Beside yield and protein, we can also look at kernel weight, test weight, maturity, height, whole plant biomass and other agronomic characteristics.

James H. Helm, James.helm@gov.ab.ca, 403-782-8696

The Stability of Diastatic Power QTL Expression in Western Six-rowed Barley Backgrounds and Environments: The Initial Phases. David Hoffman and An Hang, USDA-ARS Aberdeen, Idaho

Malting barley is an important crop in Idaho and other western US states. Emphasis at Aberdeen has been on developing two-rowed spring malting cultivars although some two- and six-rowed winter lines look promising. Several spring two-row malting spring cultivars approved for malting are available to western growers, but six-rowed barleys have greater yield potential. Currently, there are no western spring six-rowed cultivars recommended for malting. Midwestern malting six-rowed barleys have been used in the western US, but, in the west, they are usually tall and weak-strawed, and often lodge, especially under irrigation. In contrast, western-adapted six-rowed barleys such as Colter, have stiff straw, high yield potential, and adequate malt extract, but lack the sufficient diastatic power (DP) levels required for commercial malting. We have initiated a study to evaluate three major DP QTL regions in various western six-rowed genetic backgrounds when grown in various production environments. Following selection of the recipient cultivars, the next step will be to develop the appropriate near-isogenic lines for multi-environment DP testing. We wish to learn which DP QTL are the most effective in various six-rowed backgrounds and how environment influences DP expression and if genotype by environment interactions exist. The information gathered from this project will help barley geneticists perform marker-assisted DP enhancements of high-yielding western six-rowed barleys and will serve as a guide for other malting traits that may need adjustment.

David Hoffman, dhoffman@uidaho.edu, 208-397-4162 ext. 125

Non-Starch Polysaccharides in Barley. M. S. Izydorczyk and S. Bazin, Canadian Grain Commission, Grain Research Laboratory, 1404-303 Main St., Winnipeg, MB, Canada

Non-starch polysaccharides in barley –β-glucans, arabinoxylans and arabinogalactans – together with the enzymes responsible for their modification, play an important role in barley processing and quality attributes of barley-derived products. Clear understanding of the molecular basis of functionality of these polysaccharides is necessary to gain control over mechanisms responsible for their behaviour during the malting and brewing processes. In the present study, non-starch polysaccharides were extracted from barley, malt, and wort to monitor the changes these polymers undergo during processing. Both water (at various temperature) as well as alkali were used during the extraction to fully elucidate the structure and properties of water soluble and insoluble polymers. Both polysaccharides, β$-glucans and arabinoxylans, exhibited a high degree of polydispersity and very high molecular weights. Both polymers have a tendency for intermolecular aggregation and even formation of three-dimensional networks, although the mechanisms of gelation differ substantially between these polysaccharides. The molecular structure of β$-glucans and arabinoxylans will be discussed in terms of monosaccharides and linkage composition, degree of branching and substitution patterns. Arabinogalactans constitute the third group of barley non-starch polysaccharides. Our results indicate that the outer tissues of barley grain contain more arabinogalactans than the endosperm and that the molecular structure of arabinogalactans changes depending on their location in the kernel.

M.S. Izydorczyk, (204)983-1300

Proteinases and Proteinase Inhibitors of Barley and Malt. Berne L. Jones, USDA-ARS, Cereal Crops Research Unit, Madison, WI 53726

For several years, we have been studying the biochemistry behind the protein degradation that must occur during malting and brewing in order to produce good beer. Two protein groups apparently control this solubilization of barley proteins; the endoproteinases and their proteinaceous inhibitors. The cysteine class proteinases probably play the biggest role in protein solubilization, but representatives of all four protease classes are present in green malt. We previously identified, purified and studied some of the cysteine protease inhibitors. Recently, we purified and characterized some of the metalloproteinases and what appears to be the main serine proteinase (which we have called SEP-1) of green malt. The metalloproteinases seem to play a role in solubilizing storage proteins during malting and mashing, whereas the serine proteinase apparently neither hydrolyzes storage proteins nor affects the mash soluble protein levels. Its role in germination is still not known. We have now identified, purified and characterized compounds in barley and malt extracts that inhibit the SEP-1 activity. Some of these were proteins that belonged to the ‘CM’ (chloroform-methanol soluble) proteins group, but not all CM proteins inhibited the SEP-1. Although we have neither looked for nor detected any proteinaceous inhibitors that affect the metalloproteinases, their existence has been reported. It thus appears that it is common for inhibitors of the various proteinase classes to occur in malted barley. It should, therefore, be possible to manipulate the concentrations and activities of both the enzymes and their inhibitors to improve the malting quality of future barleys.

Berne L. Jones, bljones@facstaff.wisc.edu, 608-262-4480

Fast neutron induced barley mutants. Kleinhofs, A., and Kudrna, D., Dept. Crop and Soil Sciences, Washington State University, Pullman, WA 99164-6420

Fast neutron irradiation is assumed to induce deletions and perhaps other chromosome rearrangements as opposed to the strictly base substitution mutations induced by azide. In order to facilitate gene cloning and identification, we started a fast neutron induced mutant collection. Seeds were irradiated at 3.5 and 4.0 Gy at the FAO/IAEA Seibersdorf SNIF facility starting in 1992 and continuing every year until the facility was closed. M₁ seeds were grown at Pullman and M₂ spikes as well as bulk seed collected. M₂ populations were grown in the field and interesting visual mutants selected. Over 500 mutants have been cataloged with some phenotype information. A few mutants have been mapped. Information along with pictures of some of the more interesting mutants will be presented.

Andris Kleinhofs, andyk@wsu.edu, 509-335-4389

Spike morphology and FHB reaction in barley. Nadejda Krasheninnik and Jerome D. Franckowiak; Department of Plant Sciences, North Dakota State University, Fargo, ND 58102

Spike morphology in barley, controlled by the six-rowed spike 1 (vrs1) locus located in 2HL, is associated with Fusarium head blight (FHB) severity, which is generally lower for most two-rowed genotypes than for six-rowed genotypes. The vrs1.a allele is present in most six-rowed cultivars and produces well-developed lateral spikelets, the Vrs1.b allele in two-rowed cultivars reduces lateral spikelets to sterile bracts with a rounded tip, and the Vrs1.t (deficiens) allele causes an extreme reduction in the size of lateral spikelets. Disease occurrence and severity were thought to be determined in part by the spike morphology, because reduced air-movement and higher humidity in case of six-rowed spikes were considered favorable for FHB onset. FHB resistance QTLs were mapped to the centromeric region of chromosome 2, close to the vrs1 locus, suggesting that a genetic association between spike morphology and FHB reaction might exist. To test this hypothesis thirty-six cultivars and experimental lines, containing spike morphology variants Vrs1.t, Vrs1.b, and vrs1.a, were inoculated and screened for FHB resistance. The results suggest that barley spike morphology and FHB reaction may be independent traits associated only by linkage.

Nadejda Krasheninnik, Nadejda.Krasheninnik@ndsu.nodak.edu, (701) 231-7825

Mapping Genes Conferring Fusarium Head Blight Resistance in C93-3230-24.

K.E. Lamb¹, M.J. Green¹, R.D. Horsley¹, and Zhang Bingxing²; ¹Dep. of Plant Sciences, North Dakota State Univ. and ²Dep. of Plant Protection, Zhejiang Univ.

The six-rowed line C93-3230-24, from the cross B2912/Hietpas 5, was identified by researches at Busch Agricultural Resources, Inc. (BARI) to have (FHB) resistance similar to Chevron, and better FHB resistance than either of its parent in a greenhouse test. The genetic background of C93-3230-24 appears to be completely different than that of any of the FBH resistant accessions identified. Thus, this line may have alleles for FHB resistance and DON accumulation not currently identified. The objectives of this study are: 1) to construct skeletal maps that includes RFLP and SSR markers for an F₁-dervived DH mapping population developed from the cross Foster/C93-3230-24 and 2) determine the position of QTL controlling FHB resistance, DON accumulation, days to heading and maturity, plant height, and spike nodding angle. Field experiments were conducted in mist-irrigated FHB nurseries in 2001 and 2002 in North Dakota and Zhejiang Province China using 118 DH lines and parents. Single locus analysis using available marker data identified six regions in five chromosomes associated with FHB resistance. The regions are located in chromosomes 2H, 4H, 5H, 6H, and 7H. The region with the largest effect on FHB resistance appears to be in chromosome 2H. Associations between the markers and maturity and/or plant height were found in the same regions as FHB resistance. Results in this study are similar to those obtained in studies using the resistant six-rowed cultivar ‘Chevron’ and the ICARDA/CIMMYT cultivar ‘Gobernadora’. Thus, preliminary results suggest that C93-3230, Chevron, and Gobernadora may have similar alleles for FHB resistance.

R. Horsley, Richard.Horsley@ndsu.nodak.edu, 701-231-8142

Transformation of Barley cv. Conlon with Genes for Resistance to Fusarium Head Blight. M. Manoharan¹, L.S. Dahleen², T. Hohn³, S.P. McCormick⁴, N.A. Alexander⁴, P. Schwarz⁵, S. Neate⁶ and R.D. Horsley¹. ¹Dept. of Plant Science, North Dakota State University, Fargo, ND; ²USDA-ARS Fargo, ND; ³Syngenta, Inc., Research Triangle Park, NC; ⁴USDA-ARS, Peoria, IL; ⁵Dept. of Cereal Science, ⁶Dept. of Plant Pathology, NDSU, Fargo, ND.

Fusarium head blight, incited primarily by Fusarium graminearum, has caused devastating losses to barley since the 1990’s. Production of the mycotoxin deoxynivalenol (DON) by F. graminearum is harmful to humans and livestock. Expressing certain anti-toxin genes such as TRI101 and PDR5 could improve resistance to fungal infection and reduce DON levels. TRI101 encodes a 3-OH trichothecene acetyltransferase that converts DON to a less toxic acetylated form. PDR5, an ATP-binding cassette, acts as an efflux transporter, shunting DON across the plasma membrane from the interior of the cell. We have transformed the commercial malting barley cultivar Conlon with these genes to reduce DON levels in infected grain. Ten-day old calli derived from immature embryos were co-bombarded with the herbicide-resistance gene bar as the selectable marker. Putative transgenic plants were confirmed by Southern analysis. A total of seven independent events with TRI101 and six with PDR5 were recovered. Northern analysis indicated the expression of TRI101 and PDR5. Expression of TRI101 was further confirmed by detecting acetyltransferase activity in seeds of the transgenic plants. T₂ lines of three events with TRI101 and two events with PDR5 were tested in the field and the T₃in thegreenhouse for disease and toxin level. Both genes appeared to reduce FHB infection and DON accumulation in some of the transgenic plants.

L.S. Dahleen, dahleenl@fargo.ars.usda.gov, 701-239-1384

Using the Re-steep Function for Hydrating Grain in the Joe White Micromalter.

Christopher H. Martens, Allen D. Budde, and Berne L. Jones. USDA/ARS, Cereal Crops Research Unit, Madison, WI 53726

Can the re-steep function of the Joe White Micromalter be used to replace the manual hydration of germinating grain during the malting process? To find out, four different barleys were steeped for 30 hours (19°C), germinated for 24 hours, and re-steeped for five minutes. The imbibition of water and subsequent malting losses of the re-steeped samples were significantly greater than those of manually hydrated controls. The malt from the re-steeped grain was over-modified, resulting in higher wort color, soluble protein, S/T, alpha-amylase, and free amino nitrogen values, and lower beta-glucan levels than those of the well-modified, manually hydrated controls. Even a one-minute re-steep, after 30 hours of steep and six hours of germination yielded grain that was too highly hydrated. Although our initial question has not been answered, it appears that briefly re-steeping early in germination, before the rootlets have forked, might be a useful way to increase the modification of grain that has been under-hydrated at steep out.

Christopher H. Martens, chmarten@wisc.edu, (608) 262-4483

Fungicide Application Timings of Folicur and AMS 21619 for Fusarium Head Blight Control in Six-Row and Two-Row Malting Barley. Kent McKay and Kristie Clark, North Central Research Extension Center, Minot, North Dakota.

A six-row barley fungicide trial was conducted at Minot, ND and a two-row barley fungicide trial was conducted at Mohall, ND in 2002. The primary objectives of these studies were to evaluate a single fungicide application with two split fungicide applications of Folicur and AMS 21619 for Fusarium head blight control. A six-row malting barley, ‘Robust’, was planted on May 16 at Minot and a two-row Malting barley, ‘Conlon’, was planted on May 18 at Mohall. Fungicide treatments at both locations were applied with a 10 foot hand-held CO₂ sprayer delivering 20 GPA water at 30 psi using twin-jet nozzles. At both locations, the single fungicide application and the first split fungicide application were applied when the barley was 100% headed. The second split application was applied four days after the first application. Weather conditions during heading and kernel formation stages were very dry at Minot and Mohall. At both locations, the untreated plots had very low incidence of Fusarium head blight (under 5%). Yield, test weight, and percent DON will be evaluated with all treatments and will be presented at the poster session.

Kent McKay, Area Agronomy Specialist, kmckay@ndsuext.nodak.edu, 701-857-7682

Fungicide Studies for Control of Fusarium Head Blight of Barley in North Dakota, 1998-2002. McMullen, M¹., Meyer, S¹., Jordahl, J¹., Pederson, J²., and Halley, S³. ¹ Dept. of Plant Pathology, ²Dept. of Plant Sciences, North Dakota State University, Fargo, ND 58105; ³ Langdon Research Extension Center, Langdon, ND 58249

Ten fungicides, applied alone or in combination, were evaluated for control of Fusarium head blight (FHB) in barley field plots at Fargo, North Dakota from 1998-2002. Fungicides evaluated included Folicur (tebuconazole), Tilt (propiconazole), Benlate (benomyl), Penncozeb (mancozeb), Quadris (azoxystrobin), Stratego (Tilt + trifloxystrobin), BAS 500 (pyrachlostrobin), Caramba (metconazole), BAS 505 (experimental from BASF) and AMS 21619 (experimental from Bayer CropScience). Of products tested, only mancozeb had federal registration allowing application to barley heads, and Folicur had a Section 18 emergency exemption since 1998 in North Dakota for use on barley for suppression of FHB. Fungicides were applied in 18 gpa at 40 psi to six-row malting barley cultivars at early full head emergence, using a hand-held boom equipped with forward/backward facing XR8001 flat fan nozzles oriented 30⁰ from horizontal. Of the ten fungicides tested, only Folicur at 4 fl oz/acre was included in the field tests each year, as it had shown good efficacy in early tests. Over the five years, Folicur reduced FHB field severity (incidence x head severity) an average of 53.4% compared to the untreated check. DON levels were > 20 ppm in two of the five years and were reduced by only 12.9% with Folicur treatment. Two fungicides, BAS 505 and AMS 21619, available for testing in 2001 and 2002, provided slightly better reduction of FHB field severity than Folicur. Further evaluation of experimental fungicides for reduction of FHB and DON levels in barley is needed.

Marcia McMullen, mmcmulle@ndsuext.nodak.edu, 701-231-7627

Mapping of QTL associated with nitrogen storage and remobilization in barley (Hordeum vulgare L.) leaves. Suzanne Mickelson¹, Deven See², Fletcher D. Meyer¹, John P. Garner¹, Curt R. Foster¹, Tom K. Blake³ and Andreas M. Fischer¹. ¹Montana State University, Bozeman, MT 59717; ²Kansas State University, Manhattan, KS 66506; ³International Center for Agricultural Research in the Dry Areas ICARDA, Aleppo, Syrian Arab Republic.

Nitrogen uptake and metabolism are central for vegetative and reproductive plant growth. This is reflected by the fact that N can be remobilized and reused within a plant, and this process is crucial for yield in most annual crops. A population of 146 barley RILs, derived from a cross between two varieties differing markedly in grain protein concentration, was used to compare the location of QTL associated with N uptake, storage and remobilization in flag leaves relative to QTL controlling developmental parameters and grain protein accumulation. Overlaps of support intervals for such QTL were found on several chromosomes, with chromosomes 3 and 6 being especially important. For QTL on these chromosomes, alleles associated with inefficient N remobilization were associated with depressed yield and higher levels of total or soluble organic N during grain filling and vice versa; therefore, genes directly involved in or regulating N recycling may be located on these chromosomes. Interestingly, the most prominent QTL for grain protein concentration (on chromosome 6) did not co-localize with QTL for N remobilization. However, QTL peaks for nitrate and soluble organic N were detected at this locus for plants grown in one of two years in the study. For these, alleles associated with low grain protein concentration were associated with higher soluble N levels in leaves during grain filling; therefore, gene(s) found at this locus might influence the N sink strength of developing barley grains.

Suzanne Mickelson, smickelson@montana.edu, (406) 994-5055

Status of RWA-resistant barley germplasm. D.W. Mornhinweg, USDA-ARS, Stillwater, OK, P.P. Bregitzer, Darrell Wesenberg USDA-ARS, Aberdeen, ID, Bob Hammon, Frank Peairs, Colorado State U.

Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), is a devastating pest on barley in the western U.S. First identified in the US in 1986 it spread throughout the intermountain regions of the western US and into Canada by 1988. All currently grown barley cultivars were highly susceptible to RWA and losses were high. Screening of the entire National Small Grains Collection of Hordeum vulgare by the USDA-ARS in Stillwater, identified 109 accessions with some level of RWA resistance. Unadapted RWA-resistant germplasm lines were developed from each of these accessions and two were released to breeders, STARS-9301B and STARS-9577B. Although highly resistant, these lines were very unadapted to US barley production and were quite poor in terms of grain yield and malting quality. A prebreeding program was initiated cooperatively by the USDA-ARS in Stillwater, OK and Aberdeen, ID to bring RWA resistance into adapted backgrounds that could be utilized more effectively for cultivar development by breeders in all barley growing areas of the U.S After 2 years of preliminary yield testing, 56 RWA-resistant germplasm lines are possible for release involving 14 susceptible backgrounds (spring, winter, feed, malt) and 32 resistant sources.

D.W. Mornhinweg, dmornhinweg@pswcrl.ars.usda.gov, (405) 624-4141 ext. 237

Evaluation of near-isogenic lines for Fusarium Head Blight resistance QTL in barley. L. M. Nduulu, A. Mesfin, G. J. Muehlbauer, and K. P. Smith*.

Dept. of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108.

In a mapping population created with the FHB resistant cultivar Chevron and elite susceptible breeding line M69, we identified two candidate quantitative trait loci (QTL) for marker-assisted-selection (MAS). However, to employ MAS effectively, these QTL must be validated and more precisely mapped to identify markers that will be useful in breeding. In the current study, we have used two methods to develop near-isogenic lines (NILs) for two QTL regions (one on chromosome 2H and another on chromosome 6H) associated with FHB resistance. In the first method, we used molecular marker-assisted backcrossing to develop BC₃ NILs using markers flanking the two regions of interest. In the second method, we used molecular markers to identify NILs from heterogeneous inbred families (HIFs) that are segregating for the regions of interest. A total of 30 NILs isogenic for these two regions were evaluated for percent of FHB infection, heading date, and plant height in two locations in 2002.The mean FHB severity of the BC derived NILs carrying the Chevron allele for the chromosome 2H QTL was 18.4% compared to 42.9% for M69. On the contrary, the mean FHB severity of the BC derived NIL carrying the Chevron allele for the chromosome 6H QTL was 44.0% (not significantly different from M69). These preliminary results indicate that it is more efficient to use markers flanking the chromosome 2H QTL region for MAS as compared to using markers flanking the chromosome 6H QTL region. Further analyses of all traits evaluated for the BC derived NILs will be presented. Additionally, we are in the process of analyzing data from the HIF derived NILs. Results of those analyses will also be presented.

Kevin Smith, smith376@tc.umn.edu, (612) 624-1211

Predicting Relative Maturity in Barley Using Spikes. Joseph Nyachiro*, James Helm, Patricia Juskiw, Donald Salmon, Kequan Xi and Jennifer Zantinge, Field Crop Development Centre, Lacombe, AB Canada.

Relative physiological maturity in barley is an important genetic trait especially in temperate regions where the thriving of barley cultivars could be influenced by the number of frost-free days. We studied relative physiological maturity, with the objective of determining the effect of genotype, environment (location-by-years), and genotype-by-environment interactions on maturity. A wide range of barley genotypes was grown in non-replicated field tests in multiple locations in Alberta, Canada in three consecutive years 1998, 1999 and 2000. Spikes were harvested and used to determine relative maturity of each genotype. The results showed relative spike maturity trends in genotypes were similar among years although the magnitude differed from year to year. There was significant (P < 0.05) location x year interaction but non-significant location x genotype and genotype x year interactions. Barley spikes could be used to predict relative maturity of barley genotypes.

Joseph Nyachiro, joseph.nyachiro@gov.ab.ca, 403-782-8692

Potential NIRS Application for Plant Breeding & Production Research. Lori Oatway and Jim Helm, Field Crop Development Centre, Lacombe, Alberta, Canada.

Near Infrared Spectroscopy (NIRS) has been used in the Cereal Breeding Program at the Field Crop Development Centre (FCDC) for over 20 years but recent advances in NIRS technology and computers has further increased its potential. Over the past eight years the FCDC has taken advantage of the new NIRS advances and developed equations for all areas of the barley breeding program. In the feed quality area these equations include Protein, Protein Digestibility, Energy, Energy Digestibility, Digestible Energy Content and Lysine. NIR equations developed to determine malting quality include Fine Extract, Diastatic Power, Alpha-Amylase, Total Malt Protein, Soluble Malt Protein, Malt B-Glucan, Friability, Homogeneity and Viscosity. Food quality characteristics that are analyzed by NIRS include Starch, B-Glucan, Pentosans, Ash, Lipid, Soluble Fiber, Total Fibre and color. All of these equations were developed using clean, whole grain samples and have R² values ranging between 0.89 to 0.98. By creating robust equations using the correct samples, it is possible to have accurate and repeatable equations that can stand up over time. The future of NIRS is promising. NIRS has shown potential in the areas of barley phytate, water sensitivity and even genetic ‘fingerprinting’. The limits of this technology are yet to be determined.

Lori Oatway, lori.oatway@gov.ab.ca, 403-782-8048

Analysis of the barley stem rust resistance gene Rpg1 messenger RNA.

Nils Rostoks, Brian Steffenson, David Kudrna, Andris Kleinhofs. Washington State University, Dept. of Crop and Soil Sciences, Pullman WA 99164-6420, USA; Dept. of Plant Pathology, 495 Borlaug Hall, 1991 Upper Buford Circle, St. Paul, MN 55108-6030, USA

Recently, we (Brueggeman et al. PNAS 2002, 99:9328-33) cloned the barley stem rust resistance gene Rpg1 by a map-based approach (GenBank accession AF509748). Expression of the gene appeared to be low with only one partial cDNA clone found among 500,000 colonies searched. To obtain a better understanding of the Rpg1 gene expression, we initiated a qualitative and quantitative analysis of the mRNA. Different tissue types, such as green and etiolated seedling, mature leaf, root and immature inflorescence all appeared to express the gene as judged by RT-PCR and Northern blot techniques. Quantification of the Rpg1 mRNA upon infection with Puccinia graminis f. sp. tritici and in different tissues using real time RT-PCR is in progress. A single transcription start site 132 bp upstream of the Rpg1 gene translation start was detected by 5’ RACE technique in the root and the mature leaf tissues. Thus the total length of the Rpg1 mRNA is 2964 n. Analyses of the promoter-containing upstream region will be presented.

Nils Rostoks, nrostoks@wsu.edu, 509-335-9692

Genetic Linkage Map of cv. Foster x Fusarium Resistant Line CI4196.

Deric Schmierer¹, David Kudrna², Thomas Drader¹, and Andris Kleinhofs³.¹Dept. of Crop & Soil Sciences, Washington State University, Pullman, WA 99164-6420, USA; ²Plant Genomics Institute, Dept. of Plant Sciences, University of Arizona, Tucson, AZ 85721-0036, USA; ³School of Molecular Biosciences and Dept. of Crop & Soil Sciences, Washington State University, Pullman, WA 99164-6420, USA

A population of 144 recombinant inbred lines (RILs) from a cross between barley cv. Foster and a Fusarium resistant line CI4196 was used to construct a genetic linkage map. Restriction Fragment Length Polymorphic markers were used primarily and include: known function genes, genomic DNA, resistance gene analogs (RGA), and cDNA and EST clones. A skeletal map consisting of 106 loci covering 1,124 cM has been completed. This map still contains 12 gaps over 20 cM. The goal is to create a skeletal map with a marker approximately every 10cM. The map will be used to map Fusarium resistance QTL.

Andris Kleinhofs, andyk@wsu.edu (509)335-4389

Effects of deoxynivalenol on detached barley leaf segments. Seeland, T.M., Bushnell, W.R., and D.E. Krueger. USDA-ARS Cereal Disease Laboratory and Department of Plant Pathology, University of Minnesota, St. Paul MN 55108.

Deoxynivalenol (DON) is known to be a potent inhibitor of protein synthesis and has been implicated as a virulence factor for development of Fusarium graminearum in Fusarium head blight (FHB). From histological observations of diseased tissues, we postulate that DON contributes to the necrotrophic phase of FHB development. However, the physiological role of DON in pathogenesis is largely unknown. As a first step toward understanding effects of DON, we are using uninfected barley leaf segments to assess cytological and physiological effects of the toxin on green tissues. We partially stripped the abaxial epidermis from detached Robust leaf segments (1.2 cm long) and floated them with stripped mesophyll in contact with aqueous DON solutions at 30-200 ppm. When incubated in light (150-450 μmol m^-2 s^-1 ), tissues on DON (at all tested concentrations) usually turned white within 48-72 hr from loss of chlorophyll and carotenoid pigments. In darkness, DON-treated tissues remained green. As viewed by transmission electron microscopy, DON induced a characteristic increase in spacing of thylakoid chloroplast membranes in light, but not in darkness. In both light and dark, however, DON markedly increased loss of electrolytes from tissues as measured by electrical conductivity of incubating solutions. This indicates that DON had a detrimental effect on plant cell plasma membranes, causing loss of electrolytes. In summary, DON in leaves mimicked effects of Fusarium head blight in heads by causing loss of chloroplast pigments (although only in the light) and by causing other degenerative changes (in light and dark). The results support the hypothesis that DON toxicity has a role in pathogenesis.

W. R. Bushnell, billb@cdl.umn.edu, 612-625-7781

Can Molecular Breeding Overcome Yield Ceilings in Western Two-Row Malting Barley? D.A. Schmierer, A. Kleinhofs, S.E. Ullrich, D.A. Kudrna, V.A. Jitkov, B.L. Jones*; Washington State Univ., Pullman, WA 99164-6420; * USDA-ARS, Madison, WI 53705-2334

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Evaluation of the National Small Grains Collection of barley for resistance to Fusarium head blight and deoxynivalenol accumulation. L.G. Skoglund and J.L. Menert. Busch Agricultural Resources, Inc. Fort Collins, CO. 80524

A total of 7,475 barley accessions from the USDA-ARS, National Small Grains Collection, Aberdeen, Idaho, were evaluated in 1998 for resistance to Fusarium head blight (FHB) and deoxynivalenol (DON) accumulation. Selections were made and advanced for further testing in 1999, 2000 and 2001. Disease evaluations were carried out in the field with natural inoculum at locations in North Dakota and Minnesota. Additional screening was carried out in greenhouses in Colorado. Accessions were evaluated for percent FHB and ppm DON. The six-rowed barley accession Chevron (CIho 1111) and the two-rowed barley accessions Svanhals (CIho 2274) and Svansota (CIho 1907), which had previously been identified as resistant, continued to show good levels of resistance to both FHB and DON accumulation. Other accessions were identified for use by barley breeding programs.

Linnea G. Skoglund, linnea.Skoglund@anheuser-busch.com, 970 472-2332

Reactions of Barley Cultivars to Fusarium Head Blight (Fusarium graminearum) in Western Canada. J. R. Tucker¹, W. G. Legge¹, M. Savard², M. C. Therrien¹, A. Tekauz³, B. G. Rossnagel⁴, B. L. Harvey⁵, E. Lefol⁴, D. Voth⁴ and T. Zatorski⁴.¹Agriculture and Agri-Food Canada (AAFC), Research Centre, Brandon, MB; ²AAFC Eastern Cereal and Oilseed Research Centre, Ottawa, ON; ³AAFC Cereal Research Centre, Winnipeg, MB; ⁴Crop Development Centre, University of Saskatchewan, Saskatoon, SK; and ⁵Dept. Plant Sciences, University of Saskatchewan, Saskatoon, SK

Fusarium head blight (FHB) has recently emerged as the most devastating barley disease in western Canada. The objective of the present study was to evaluate disease reactions on a set of 82 new and current western Canadian barley cultivars in a FHB disease nursery at Brandon, MB, for 2000 and 2001. Barley rows were artificially inoculated with corn infected with Fusarium graminearum, and visually scored for disease reaction 2-3 weeks following heading. DON (deoxynivalenol) levels were assessed for each cultivar using the ELISA method. FHB reactions on the cultivar set were also visually assessed in a disease nursery at Hangzhou, China. On average, two-row barley showed more resistance than six-row barley for both FHB rating and DON content. Hulless cultivars showed a decrease in DON content in comparison with covered cultivars. All correlations between measured disease parameters were statistically significant, with the strongest relationship observed between DON content in 2000 and 2001 (r = 0.86). All cultivars tested showed FHB symptoms and accumulated DON, but a broad range of reactions was observed. Several moderately resistant cultivars were identified.

J. R. Tucker jtucker@agr.gc.ca (204) 726-7650

An Alberta perspective on Fusarium head blight. T.K. Turkington¹, R.M. Clear², J. Calpas³, and J.P. Tewari⁴. ¹Lacombe Res. Centre, Agric. and Agri-Food Canada, Lacombe, AB, Canada; ²Canadian Grain Commission, Grain Res. Laboratory, Winnipeg, MB, Canada, ³Alberta Agric., Food and Rural Dev., Pest Risk Management Unit, Edmonton, AB, Canada; ⁴Univ. of Alberta, Faculty of Agric., Forestry & Home Econ., Edmonton, AB, Canada.

The continued westward appearance of Fusarium graminearum (F.g.) in the Canadian Prairie Provinces has occurred over the last several years, with this pathogen becoming well established in southeastern Saskatchewan (SK), causing cereal yield and grade losses. Awareness of Fusarium head blight (FHB) caused by F.g. in Alberta (AB) has increased as a result of popular farm press articles, chemical advertisements, and extension efforts. The risk of FHB in AB has been looked at from the perspective of the disease triangle. Currently, all cereal varieties are susceptible to FHB, but vary in their level of susceptibility, while many of the same varieties are grown across the three Prairie Provinces. Surveys conducted by several organizations since 1994 have indicated that F.g. is not a common pathogen in cereal grain and residue, and grass residue. Concerns have been expressed surrounding further introduction and spread of this pathogen via infected seed and ascospore production from spilled infected feed grain. Although July temperatures in central and northern areas of AB can be 3-4^oC lower compared with epidemic areas in Manitoba (MB), these differences decrease for the central regions of western MB and eastern SK. Moreover, several reports indicate the potential for FHB at lower temperatures. In contrast, central and northern AB tend to have similar or even higher rainfall compared with locations in MB and SK. Efforts are underway to gain a better understanding of the potential of FHB caused by F.g. in AB, create awareness, and recommend management strategies to slow down FHB caused by F.g. until more effective control options become available.

T.K. Turkington, turkingtonk@agr.gc.ca, (403) 782-8138.

Mapping Quantitative Stripe Rust Resistance in a Large Doubled Haploid Population of Barley. Vales, M. Isabel¹; Hayes, Patrick M¹; Castro, Ariel¹; Corey, Ann¹; Mundt, Chris²; Capettini, Flavio³; Vivar, Hugo³; Sandoval-Islas, Sergio⁴; Schoen, Chris⁵.¹Dept. of Crop and Soil Sciences, Oregon State University, Corvallis, OR;²Dept. of Botany and Plant Pathology, Oregon State University, Corvallis, OR;³ICARDA/CIMMYT Barley Program, Apdo 6-641, Mexico, DF Mexico;⁴Instituto de Fitosanidad, Colegio de Postgraduados, Montecillo, Texcoco, Mexico;⁵University of Hohenheim, State Plant Breeding Institute, Stuttgart, Germany.

Genetic resistance to barley stripe rust can be qualitatively or quantitatively inherited. The use of qualitative sources of resistance alone is risky because there is evidence that pathogen virulence can evolve more quickly than plant breeders can deploy single resistance genes in new varieties. Quantitative resistance, alone or in combination with qualitative resistance, is in general more durable and is a key consideration in disease resistance breeding. A large (n=409) barley doubled haploid population was developed from the cross: BCD47 x Baronesse. BCD47 is a doubled haploid selection developed by molecular marker-assisted pyramiding of stripe rust resistant alleles at quantitative trait loci (QTL) on chromosomes 4 and 7. Baronesse is the most widely grown barley variety in the Pacific Northwest and it is quite susceptible to barley stripe rust. The goal of this project is to identify genetic factors that influence stripe rust resistance. So far, QTL analysis allowed to identify genetic factors affecting quantitative stripe rust resistance on several chromosomes, the main QTL are on chromosomes 3, 4, 6 and 7. Further genotypic and phenotypic studies will allow to better estimate the effect of population size on the estimation of QTL number, effects, and its interactions with each other and with the environment.

M. Isabel Vales Isabel.Vales@orst.edu (541) 737-3539

Summary of QTL Analyses of the Seed Dormancy Trait in Barley S.E. Ullrich, F. Han, W. Gao, D. Prada*, J. Clancy, A. Kleinhofs, I. Romagosa*, J.L. Molina-Cano*, D.A. Schmierer. Washington State Univ., Pullman, WA 99164-6420; * Univ. de Lleida-IRTA, 25198 Lleida, Spain

Moderate seed dormancy is desirable in barley, but the trait is difficult to manipulate in breeding because of complex inheritance and large environmental effects. Quantitative trait locus (QTL) mapping has shed light on genes involved and has opened the way for further genetic study and marker-assisted breeding. QTL analyses [MapMaker-QTL (3) or MQTL (10)] for the seed dormancy trait in barley have been performed on the North American Barley Genome Project doubled haploid line (DHL) mapping populations of ‘Steptoe’ (dormant) / ‘Morex’ (S/M) and ‘Harrington’ / TR306 (dormant) (H/TR), as well as, a DHL population from ‘Triumph’ (dormant) / Morex (T/M). Germination percentage after various after-ripening periods (0 – 30 d) was used to measure the expression of dormancy.

Major dormancy QTL regions attributed to Steptoe from the S/M cross were identified near the centromere (SD1, 50% variation explained) and long arm telomere (SD2, 15% variation explained) on chromosome 7(5H), and minor QTLs attributed to Steptoe (5% variation explained each) were identified on chromosomes 1(7H) and 4 (4H), Table 1 (1, 6, 11, 12). Fine mapping efforts using a substitution mapping method with molecular marker assisted backcross generated isogenic lines have resolved SD1 into a 4.4 from 8.7 cM region (2) and SD2 into a 0.8 from 9.0 cM region (Gao, Kleinhofs, Ullrich, unpublished). From the H/TR cross, putatively, a major QTL from TR306 was identified near the chromosome 7 (5H) long arm telomere (58% variation explained) and a minor QTL from Harrington was found on chromosome 5 (1H) (9% variation explained), Table 1 (9). Takeda (9) measured germinations of 86 and 40% for Harrington and TR306, respectively. Repeat of this analysis based on growthroom grown material verified these QTL results. Although, Harrington and TR306 had 0 d after-ripening germinations of 78 and 85%, respectively, with a range of 19-100% for the mapping population (Clancy and Ullrich, unpublished). From the T/M cross, a major dormancy QTL from Triumph was identified near the centromere of chromosome 7 (5H) (24% variation explained) and minor QTLs were indicated near the centromere, on the short arm, and long arm of chromosomes 2 (2H), 5 (1H), and 7 (5H), respectively (6-9% variation explained), Table 1 (Prada, Romagosa, Ullrich un-published).

Several of the QTL identified in the various different mapping populations could be the same or closely linked. The major dormancy QTL attributed to Triumph near the centromere of chromosome 7 (5H) is likely the same as SD1, the major QTL attributed to Steptoe based on common markers on the maps from the two crosses. Potentially the chromosome 5 (1H) QTLs identified from the H/TR and T/M crosses are the same. However, the chromosome 7 (5H) QTLs near the long arm telomere identified from S/M (SD2) and T/M are probably not the same, but linked, since the dormancy allele is from Steptoe in the S/M cross, and from Morex in the T/M cross. On the other hand, the dormancy QTL from TR306, which is also near the telomere of chromosome 7 (5H), could be same as either one of the Steptoe or Morex QTLs identified in that region.

Table 1. Summary of barley seed dormancy QTLs detected from three crosses.

Cross/Chrom. Map Interval¹ Test % Var. Parent w/ Express. in Refer.

Stat. Dor. Allele Environ.

Step./Morex

7 (5H) Ale – ABC324 23^a50 Steptoe All 2, 6, 11, 12

7 (5H) MWG851D – 5^a15 Steptoe All 6, 11, 12

MWG851B

1 (7H) Amy2 – Ubi1 3^a 5 Steptoe Some 6, 11, 12

4 (4H) MG622 – BCD402B 2^a 5 Steptoe Some 6, 11, 12

Harr./TR306

7 (5H) ABC622-ABC718 28^a 58 TR306 One 9

5 (1H) N I 3^a 9 Harrington One 9

Trium./Morex

7 (5H) E32M49e - 131^b 24 Triumph All Unpub.

Bmac96

7 (5H) E32M49o - 46^b 9 Morex Some Unpub.

E39M49b

5 (1H) GM521 – 28^b 6 Morex Some Unpub.

E32M59d

2 (2H) Vrs1 – Bmag125 27^b 6 Morex Some Unpub.

¹ Distance between markers in table order: 4.4, 0.8, 16.0, 29.1, 3.0, 9.3, 18.0, 11.0 cM.

N I Not indicated.

^a LOD score; MapMaker-QTL (3); LOD of 2.0 sig. @ p< 0.001.

^b SIM; MQTL (10); threshold significance at p< 0.05. Only QTLs with coincident peaks from SIM and sCIM analyses are included.

Further study of the S/M dormancy QTLs indicate that additivity, epistasis (QTL x QTL interaction), QTL x environment interaction, and potentially cytoplasmic effects occur among the QTLs (1, 2, 3, 5, 6, 9). Based on an F₂backcross population, SD1 appears to segregate as a single Mendelian gene, but with environmental and potentially modifier gene effects (2). Based on time-course studies from anthesis through grain-filling, maturity and after-ripening, the S/M QTLs seem to act to maintain/release dormancy after maturity and during after-ripening and are not to be involved in the inception of dormancy (6).

Another potential QTL or major gene that conditions dormancy may be located on chromosome 6 (6H). A set of F₅ recombinant inbred lines (RILs) were developed from a cross of the near isogenic lines; Triumph (dormant) and TL43, a non-dormant sodium azide induced mutant of Triumph (4). Subsets of dormant and non-dormant lines were used for bulk segregant analysis with 60 RFLPs, SSRs, and STSs distributed across the seven barley chromosomes . Differences between the parents and between the respective dormant and non-dormant RILs were only detected in the centromere region of chromosome 6 (6H). Since the TL43 mutant also shows reduced sensitivity to ABA (7), it is hypothesized that the putative chromosome 6 gene is involved in hormonal regulation of germination. Study is underway to further characterize this mutant gene.

References

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29. Takeda, K. 1996. Varietal variation and inheritance of seed dormancy in barley. p. 205-212. In K. Noda and D.J. Mares (ed.). Pre-harvest sprouting in cereals 1995. Center for Academic Societies, Osaka, Japan.

30. Tinker, N.A. and D.E. Mather. 1995. MQTL: Software for simplified composite interval mapping of QTL in multiple environments. J. QTL available: J. Agric. Genomics : http://www.ncgr.org/jag/papers95/paper295/index295.html.

31. Ullrich, S.E., F. Han, T.K. Blake, L.E. Oberthur, W.E. Dyer, and J.A. Clancy. 1996. Seed dormancy in barley. Genetic resolution and relationship to other traits. p. 157-163. In K. Noda and D.J. Mares (ed.). Pre-harvest sprouting in cereals 1995. Center for Academic Societies, Osaka, Japan.

32. Ullrich, S. E., P. M. Hayes, W. E. Dyer, T. K. Blake, and J. A. Clancy. 1993. Quantitative trait locus analysis of seed dormancy in "Steptoe" barley. p. 136-145. In: M. K. Walker-Simmons and J. L. Reid (eds.) Preharvest sprouting in cereals 1992. Amer.Assoc. Cereal Chemist, St. Paul.

S.E. Ullrich, Ullrich@wsu,edu, 509-335-4936

Phenotypic Associative Microsatellite (SSR) Marker Assisted Selection. L. J. Wright, D. B. Cooper, BARI Ft. Collins CO.; P. Hayes, OSU, Corvallis, OR.

In a joint project between Busch Agricultural Resources, Inc. and Oregon State University, seventy-seven SSR markers were assayed on 192 6-rowed and two-rowed BARI malting barley lines using either an ABI 377 or ABI 3700 Automated Sequencer. The lines tested included elite BARI experimentals and North American malting varieties. These lines were ranked from 1 – 9 (low to high) for thirty traits, including malting quality and DON. Marker data were then sorted according to the phenotypic ranking for each trait. High and low rankings for the phenotypic trait were then compared to the SSR database to determine if an association could be made for each SSR marker. Many phenotypic associative SSR markers were made for both agronomic and malting traits. When associative markers for five malting barley characteristics were compared to malting QTLs there was good agreement between the two.

Les Wright, Les.Wright@Anheuser-Busch.com, (970) 473-2326

Cultivar resistance to scald of barley in Alberta from 1997 to 2001. K. Xi¹, T.K. Turkington², M. Cortez¹, J. Helm¹, P. Juskiw¹ and J. Nyachiro¹. ¹Alberta Agriculture, Food and Rural Development, Field Crop Development Centre. Lacombe, AB. T4L 1W1; ²Agriculture and Agri-Food Canada, Lacombe Research Centre, Lacombe, AB. T4L 1W1

Cultivar resistance to scald caused by Rhynchosporium secalis (Oud) J.J. Davis was evaluated in field hill plots across Alberta, Canada, from 1997 to 1999 
as the first period, and from 2000 to 2001as the second period.  The barley (Hordeum vulgare L.) used consisted of commercial cultivars with different levels of
resistance or susceptibility and cultivars with major resistance genes that have been used as differentials worldwide.  Consistently highly
 to intermediate resistant reactions in the differentials suggest that R. secalis pathotypes were similar for the differentials during the two test periods.  
Resistance in few commercial cultivars including AC Stacey, CDC Dolly, Kasota, Mahigan and Seebe, held up at all locations during the entire test period.
Cultivars previously considered as resistant were found to be intermediate in reaction in the first period and became susceptible in the second period,
indicating the resistance was completely overcome by the occurrence of virulent pathotypes.  In addition, most commercial cultivars tended to display relatively
more susceptible reactions in the second period compared with that in the first one.  Scald reaction of commercial cultivars was significantly different among site
and season, and pathotype distribution was considerably different among sites.  Consequently, scald management via the selection of cultivar will be dependent
on location.

K. Xi, kequan.xi@gov.ab.ca,  (403) 782-8100 ext 260,

Genetic structure of Rhynchosporium secalis in Alberta. J. Zantinge, J. Hillson and K. Xi. Alberta Agriculture, Field Crop Development Centre, 6000 C & E Trail, Lacombe, AB T4L 1W1, Canada

Amplified fragment length polymorphism (AFLP) markers were used to determine the genetic diversity and structure of Alberta field populations of Rhynchosporium secalis. In the first study, 14 representative isolates generally segregated according to cultivar origin in the resulting UPMGA dendrogram based on AFLP markers. The second study using 110 isolates revealed a highly divergent R. secalis population with many clads and similarity coefficients ranging from 0.56 to 1.00. The population subdivided weakly according to location and pathotype. The Westlock and Beaverlodge sites that had less virulent scald races shared greater genotypic similarity while the Trochu site with more virulent scald races had greater genotypic variation. The highly divergent fingerprints and clustering from some moderately resistant varieties support the theory that “new” resistant varieties select for increased variation and virulence in the R. secalis populations.

J. Zantinge Jennifer.zantinge@gov.ab.ca, 403-782-8100.

Host Organizing Committee

Sponsors

Monday September 23

Tuesday September 24

Pathology/Entomology, Marcia McMullen, Moderator

Breeding/Germplasm/Agronomy, Jerome Franckowiak, Moderator

Wednesday September 25

Biotechnology/Genomics, Lynn Dahleen, Moderator

Root Disease Suppression In Small Grains In A Low Rainfall And Low Soil Fertility Environment. Stephen M. Neate¹ and Vadakattu Gupta², ¹Department of Plant Pathology, North Dakota State University, Fargo, ND,;²CSIRO Land and Water, SA Australia.

References

Step./Morex

Harr./TR306

Trium./Morex

References

Host Organizing Committee

Sponsors

Monday September 23

Tuesday September 24

Pathology/Entomology, Marcia McMullen, Moderator

Breeding/Germplasm/Agronomy, Jerome Franckowiak, Moderator

Wednesday September 25

Biotechnology/Genomics, Lynn Dahleen, Moderator

Root Disease Suppression In Small Grains In A Low Rainfall And Low Soil Fertility Environment. Stephen M. Neate1 and Vadakattu Gupta2, 1Department of Plant Pathology, North Dakota State University, Fargo, ND,;2CSIRO Land and Water, SA Australia.

References

Step./Morex

Harr./TR306

Trium./Morex

References

Root Disease Suppression In Small Grains In A Low Rainfall And Low Soil Fertility Environment. Stephen M. Neate¹ and Vadakattu Gupta², ¹Department of Plant Pathology, North Dakota State University, Fargo, ND,;²CSIRO Land and Water, SA Australia.