Wiley Acta Medica Okayama 1936-5209 19 3 2025 Oregon Wolfe barley genetic stocks – Research and teaching tools for next generation scientists e70004 EN Margaret R. Krause Department of Crop and Soil Science, Oregon State University Juan David Arbelaez Department of Crop Sciences, University of Illinois at Urbana-Champaign Åsmund Asdal Nordic Genetic Resource Centre Ramzi Belkodja CIHEAM-Zaragoza Nancy Boury Department of Plant Pathology, Entomology, and Microbiology, Iowa State University Victoria C. Blake Department of Plant Sciences and Plant Pathology, Montana State University Patrick J. Brown Department of Plant Sciences, University of California-Davis Ana Casas Departamento de Genética y Producción Vegetal, Estación Experimental Aula Dei–CSIC Luis Cistué Departamento de Genética y Producción Vegetal, Estación Experimental Aula Dei–CSIC Alba Farré‐Martínez AGROTECNIO-CERCA Center, Universidad de Lleida Scott Fisk Department of Crop and Soil Science, Oregon State University Gregory S. Fuerst U.S. Department of Agriculture-Agricultural Research Service, Corn Insects and Crop Genetics Research Unit, Iowa State University Estela Giménez Department of Biotechnology-Plant Biology, School of Agricultural, Food and Biosystems Engineering, Universidad Politécnica de Madrid Carla Guijarro‐Real Department of Biotechnology-Plant Biology, School of Agricultural, Food and Biosystems Engineering, Universidad Politécnica de Madrid Katy Guthrie Department of Agronomy and Plant Genetics, University of Minnesota Margaret Halstead Aardevo North America Laura Helgerson Department of Crop and Soil Science, Oregon State University Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Ernesto Igartua Departamento de Genética y Producción Vegetal, Estación Experimental Aula Dei–CSIC Morten Lillemo Department of Plant Sciences, Norwegian University of Life Sciences Marina Martínez‐García Department of Biotechnology-Plant Biology, School of Agricultural, Food and Biosystems Engineering, Universidad Politécnica de Madrid Mariona Martínez‐Subirà AGROTECNIO-CERCA Center, Universidad de Lleida Susan McCouch Plant Breeding and Genetics Section, School of Integrative Plant Science, Cornell University Laurie McGhee Colfax-Mingo Community High School Travis Nickols Department of Crop and Soil Science, Oregon State University Nick Peters Department of Plant Pathology, Entomology, and Microbiology, Iowa State University Raymond Porter Haupert Institute for Agricultural Studies, Huntington University Ignacio Romagosa AGROTECNIO-CERCA Center, Universidad de Lleida Anja Karine Ruud Department of Plant Sciences, Norwegian University of Life Sciences Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Silvio Salvi Department of Agricultural and Food Sciences, University of Bologna Giuseppe Sangiorgi Department of Agricultural and Food Sciences, University of Bologna Rebekka Schüller Department of Crop Sciences, University of Illinois at Urbana-Champaign Taner Z. Sen Crop Improvement and Genetics Research Unit, U.S. Department of Agriculture-Agricultural Research Service José Miguel Soriano AGROTECNIO-CERCA Center, Universidad de Lleida Robert M. Stupar Department of Agronomy and Plant Genetics, University of Minnesota To‐Chia Ting Agronomy Department, Purdue University Kelly Vining Department of Crop and Soil Science, Oregon State University Maria von Korff Institute of Plant Genetics, Heinrich-Heine-Universität Düsseldorf Agatha Walla Institute of Plant Genetics, Heinrich-Heine-Universität Düsseldorf Diane R. Wang Agronomy Department, Purdue University Robbie Waugh Division of Plant Sciences, School of Life Sciences, University of Dundee Roger P. Wise Department of Plant Pathology, Entomology, and Microbiology, Iowa State University Robert Wolfe Agriculture and Agri-Food Canada Eric Yao Crop Improvement and Genetics Research Unit, U.S. Department of Agriculture-Agricultural Research Service Patrick M. Hayes Department of Crop and Soil Science, Oregon State University The Oregon Wolfe Barley (OWB) mapping population (Reg. no. MP-4, NSL 554937 MAP) is a resource for genetics research and instruction. The OWBs are a set of doubled haploid barley (Hordeum vulgare L.) lines developed at Oregon State University from the F1 of a cross between Dr. Robert Wolfe's dominant and recessive marker stocks. Exhibiting a high level of genetic and phenotypic diversity, the OWBs are used throughout the world as a research tool for barley genetics. To date, these endeavors have led to 56 peer-reviewed publications, as well as three reports in the Barley Genetics Newsletter. At the same time, the OWBs are widely used as an instructor resource at the K–12, undergraduate, graduate, and professional levels. They are currently used at universities and/or institutes in German, Italy, Norway, Spain, and the United States and are currently being developed further for educational use in other countries. Genotype and phenotype data, lesson plans, and seed availability information are available herein and online. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0040-5752 137 9 2024 Mutations in starch BRANCHING ENZYME 2a suppress the traits caused by the loss of ISOAMYLASE1 in barley 212 EN Ryo Matsushima Institute of Plant Science and Resources, Okayama University Hiroshi Hisano Institute of Plant Science and Resources, Okayama University June-Sik Kim Institute of Plant Science and Resources, Okayama University Rose McNelly John Innes Centre, Norwich Research Park Naoko F. Oitome Department of Biological Production, Akita Prefectural University David Seung John Innes Centre, Norwich Research Park Naoko Fujita Department of Biological Production, Akita Prefectural University Kazuhiro Sato Institute of Plant Science and Resources, Okayama University The genetic interactions among starch biosynthesis genes can be exploited to alter starch properties, but they remain poorly understood due to the various combinations of mutations to be tested. Here, we isolated two novel barley mutants defective in starch BRANCHING ENZYME 2a (hvbe2a-1 and hvbe2a-2) based on the starch granule (SG) morphology. Both hvbe2a mutants showed elongated SGs in the endosperm and increased resistant starch content. hvbe2a-1 had a base change in HvBE2a gene, substituting the amino acid essential for its enzyme activity, while hvbe2a-2 is completely missing HvBE2a due to a chromosomal deletion. Further genetic crosses with barley isoamylase1 mutants (hvisa1) revealed that both hvbe2a mutations could suppress defects in endosperm caused by hvisa1, such as reduction in starch, increase in phytoglycogen, and changes in the glucan chain length distribution. Remarkably, hvbe2a mutations also transformed the endosperm SG morphology from the compound SG caused by hvisa1 to bimodal simple SGs, resembling that of wild-type barley. The suppressive impact was in competition with floury endosperm 6 mutation (hvflo6), which could enhance the phenotype of hvisa1 in the endosperm. In contrast, the compound SG formation induced by the hvflo6 hvisa1 mutation in pollen was not suppressed by hvbe2a mutations. Our findings provide new insights into genetic interactions in the starch biosynthetic pathway, demonstrating how specific genetic alterations can influence starch properties and SG morphology, with potential applications in cereal breeding for desired starch properties. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0040-5752 136 4 2023 FLOURY ENDOSPERM 6 mutations enhance the sugary phenotype caused by the loss of ISOAMYLASE1 in barley 94 EN Ryo Matsushima Institute of Plant Science and Resources, Okayama University Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Ivan Galis Institute of Plant Science and Resources, Okayama University Satoko Miura Department of Biological Production, Akita Prefectural University Naoko Crofts Department of Biological Production, Akita Prefectural University Yuto Takenaka College of Life Sciences, Ritsumeikan University Naoko F. Oitome Department of Biological Production, Akita Prefectural University Takeshi Ishimizu College of Life Sciences, Ritsumeikan University Naoko Fujita Department of Biological Production, Akita Prefectural University Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Starch is a biologically and commercially important glucose polymer synthesized by plants as semicrystalline starch granules (SGs). Because SG morphology affects starch properties, mutants with altered SG morphology may be useful in breeding crops with desirable starch properties, including potentially novel properties. In this study, we employed a simple screen for mutants with altered SG morphology in barley (Hordeum vulgare). We isolated mutants that formed compound SGs together with the normal simple SGs in the endosperm and found that they were allelic mutants of the starch biosynthesis genes ISOAMYLASE1 (HvISA1) and FLOURY ENDOSPERM 6 (HvFLO6), encoding starch debranching enzyme and CARBOHYDRATE-BINDING MODULE 48-containing protein, respectively. We generated the hvflo6 hvisa1 double mutant and showed that it had significantly reduced starch biosynthesis and developed shrunken grains. In contrast to starch, soluble α-glucan, phytoglycogen, and sugars accumulated to higher levels in the double mutant than in the single mutants. In addition, the double mutants showed defects in SG morphology in the endosperm and in the pollen. This novel genetic interaction suggests that hvflo6 acts as an enhancer of the sugary phenotype caused by hvisa1 mutation. No potential conflict of interest relevant to this article was reported.

Oxford University Press Acta Medica Okayama 1340-2838 29 1 2022 Chromosome-scale assembly of barley cv. 'Haruna Nijo' as a resource for barley genetics dsac001 EN Areej Sakkour Institute of Plant Science and Resources, Okayama University Martin Mascher Department Genebank, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Axel Himmelbach Department Genebank, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Georg Haberer Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Thomas Lux Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Manuel Spannagl Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Nils Stein Department Genebank, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Shoko Kawamoto Department of Informatics, National Institute of Genetics Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Cultivated barley (Hordeum vulgare ssp. vulgare) is used for food, animal feed, and alcoholic beverages and is widely grown in temperate regions. Both barley and its wild progenitor (H. vulgare ssp. spontaneum) have large 5.1-Gb genomes. High-quality chromosome-scale assemblies for several representative barley genotypes, both wild and domesticated, have been constructed recently to populate the nascent barley pan-genome infrastructure. Here, we release a chromosome-scale assembly of the Japanese elite malting barley cultivar 'Haruna Nijo' using a similar methodology as in the barley pan-genome project. The 4.28-Gb assembly had a scaffold N50 size of 18.9 Mb. The assembly showed high collinearity with the barley reference genome 'Morex' cultivar, with some inversions. The pseudomolecule assembly was characterized using transcript evidence of gene projection derived from the reference genome and de novo gene annotation achieved using published full-length cDNA sequences and RNA-Seq data for 'Haruna Nijo'. We found good concordance between our whole-genome assembly and the publicly available BAC clone sequence of 'Haruna Nijo'. Interesting phenotypes have since been identified in Haruna Nijo; its genome sequence assembly will facilitate the identification of the underlying genes. No potential conflict of interest relevant to this article was reported.

Elsevier Ltd. Acta Medica Okayama 07335210 101 2021 Germplasm evaluation for crop improvement: Analysis of grain quality and cadmium accumulation in barley 103297 EN Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Kazuyoshi Takeda Institute of Plant Science and Resources, Okayama University Jian Feng Ma Institute of Plant Science and Resources, Okayama University Evaluating genetic variation in barley (Hordeum vulgare) germplasm, combined with genome-wide genotyping, is vital for identifying genes controlling important grain-quality traits. For example, in addition to traditional grain quality properties such as starch and protein contents, grain safety parameters such as heavy metal content, are important in the use of barley for human food and animal feed. A number of genes affecting grain quality have been identified by map-based cloning strategies and functionally analyzed by genetic transformation experiments. Moreover, germplasm evaluation yielded information that enabled the introgression of a key gene controlling grain cadmium accumulation into an elite barley cultivar, reducing the content of this heavy metal in grain. Genotyping of molecular markers and resequencing of germplasm accessions may provide information about how grain quality–related loci evolved and how the current allelic diversity was established. In this review, we describe germplasm resources for barley grain quality–related traits and the methods used to analyze the functions of genes controlling these traits, illustrating cadmium accumulation as an example. We also discuss future directions for the efficient identification of grain quality–related genes. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 1467-7644 2021 Regulation of germination by targeted mutagenesis of grain dormancy genes in barley 1 10 EN Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Robert E. Hoffie Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Fumitaka Abe Institute of Crop Science, NARO Hiromi Munemori Institute of Plant Science and Resources, Okayama University Takakazu Matsuura Institute of Plant Science and Resources, Okayama University Masaki Endo Institute of Agrobiological Sciences, NARO Masafumi Mikami Institute of Agrobiological Sciences, NARO Shingo Nakamura Institute of Crop Science, NARO Jochen Kumlehn Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Kazuhiro Sato Institute of Plant Science and Resources, Okayama University High humidity during harvest season often causes pre-harvest sprouting in barley (Hordeum vulgare). Prolonged grain dormancy prevents pre-harvest sprouting; however, extended dormancy can interfere with malt production and uniform germination upon sowing. In this study, we used Cas9-induced targeted mutagenesis to create single and double mutants in QTL FOR SEED DORMANCY 1 (Qsd1) and Qsd2 in the same genetic background. We performed germination assays in independent qsd1 and qsd2 single mutants, as well as in two double mutants, which revealed a strong repression of germination in the mutants. These results demonstrated that normal early grain germination requires both Qsd1 and Qsd2 function. However, germination of qsd1 was promoted by treatment with 3% hydrogen peroxide, supporting the notion that the mutants exhibit delayed germination. Likewise, exposure to cold temperatures largely alleviated the block of germination in the single and double mutants. Notably, qsd1 mutants partially suppress the long dormancy phenotype of qsd2, while qsd2 mutant grains failed to germinate in the light, but not in the dark. Consistent with the delay in germination, abscisic acid accumulated in all mutants relative to the wild type, but abscisic acid levels cannot maintain long-term dormancy and only delay germination. Elucidation of mutant allele interactions, such as those shown in this study, are important for fine-tuning traits that will lead to the design of grain dormancy through combinations of mutant alleles. Thus, these mutants will provide the necessary germplasm to study grain dormancy and germination in barley. No potential conflict of interest relevant to this article was reported.

Oxford University Press (OUP) Acta Medica Okayama 1340-2838 28 3 2021 Chromosome-scale genome assembly of the transformation-amenable common wheat cultivar ‘Fielder’ dsab008 EN Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Fumitaka Abe Institute of Crop Science, NARO Martin Mascher Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Georg Haberer Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Heidrun Gundlach Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Manuel Spannagl Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Kenta Shirasawa Kazusa DNA Research Institute Sachiko Isobe Kazusa DNA Research Institute We have established a high-quality, chromosome-level genome assembly for the hexaploid common wheat cultivar ‘Fielder’, an American, soft, white, pastry-type wheat released in 1974 and known for its amenability to Agrobacterium tumefaciens-mediated transformation and genome editing. Accurate, long-read sequences were obtained using PacBio circular consensus sequencing with the HiFi approach. Sequence reads from 16 SMRT cells assembled using the hifiasm assembler produced assemblies with N50 greater than 20 Mb. We used the Omni-C chromosome conformation capture technique to order contigs into chromosome-level assemblies, resulting in 21 pseudomolecules with a cumulative size of 14.7 and 0.3 Gb of unanchored contigs. Mapping of published short reads from a transgenic wheat plant with an edited seed-dormancy gene, TaQsd1, identified four positions of transgene insertion into wheat chromosomes. Detection of guide RNA sequences in pseudomolecules provided candidates for off-target mutation induction. These results demonstrate the efficiency of chromosome-scale assembly using PacBio HiFi reads and their application in wheat genome-editing studies. No potential conflict of interest relevant to this article was reported.

Oxford University Press (OUP) Acta Medica Okayama 2160-1836 2021 Chromosome-scale assembly of wild barley accession “OUH602” jkab244 EN Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Martin Mascher Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Axel Himmelbach Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Georg Haberer Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Manuel Spannagl Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health Nils Stein Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Barley (Hordeum vulgare) was domesticated from its wild ancestral form ca. 10,000 years ago in the Fertile Crescent and is widely cultivated throughout the world, except for in tropical areas. The genome size of both cultivated barley and its conspecific wild ancestor is approximately 5 Gb. High-quality chromosome-level assemblies of 19 cultivated and one wild barley genotype were recently established by pan-genome analysis. Here, we release another equivalent short-read assembly of the wild barley accession “OUH602.” A series of genetic and genomic resources were developed for this genotype in prior studies. Our assembly contains more than 4.4 Gb of sequence, with a scaffold N50 value of over 10 Mb. The haplotype shows high collinearity with the most recently updated barley reference genome, “Morex” V3, with some inversions. Gene projections based on “Morex” gene models revealed 46,807 protein-coding sequences and 43,375 protein-coding genes. Alignments to publicly available sequences of bacterial artificial chromosome (BAC) clones of “OUH602” confirm the high accuracy of the assembly. Since more loci of interest have been identified in “OUH602,” the release of this assembly, with detailed genomic information, should accelerate gene identification and the utilization of this key wild barley accession. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1438-793X 2021 RNA-Seq-based DNA marker analysis of the genetics and molecular evolution of Triticeae species EN Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Kentaro Yoshida Graduate School of Agricultural Science, Kobe University Shigeo Takumi Graduate School of Agricultural Science, Kobe University The release of high-quality chromosome-level genome sequences of members of the Triticeae tribe has greatly facilitated genetic and genomic analyses of important crops such as wheat (Triticum aestivum) and barley (Hordeum vulgare). Due to the large diploid genome size of Triticeae plants (ca. 5 Gbp), transcript analysis is an important method for identifying genetic and genomic differences among Triticeae species. In this review, we summarize our results of RNA-Seq analyses of diploid wheat accessions belonging to the genera Aegilops and Triticum. We also describe studies of the molecular relationships among these accessions and provide insight into the evolution of common hexaploid wheat. DNA markers based on polymorphisms within species can be used to map loci of interest. Even though the genome sequence of diploid Aegilops tauschii, the D-genome donor of common wheat, has been released, the diploid barley genome continues to provide key information about the physical structures of diploid wheat genomes. We describe how a series of RNA-Seq analyses of wheat relatives has helped uncover the structural and evolutionary features of genomic and genetic systems in wild and cultivated Triticeae species. No potential conflict of interest relevant to this article was reported.

Oxford University Press Acta Medica Okayama 1340-2838 27 4 2020 History and future perspectives of barley genomics dsaa023 EN Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Barley (Hordeum vulgare), one of the most widely cultivated cereal crops, possesses a large genome of 5.1Gbp. Through various international collaborations, the genome has recently been sequenced and assembled at the chromosome-scale by exploiting available genetic and genomic resources. Many wild and cultivated barley accessions have been collected and preserved around the world. These accessions are crucial to obtain diverse natural and induced barley variants. The barley bioresource project aims to investigate the diversity of this crop based on purified seed and DNA samples of a large number of collected accessions. The long-term goal of this project is to analyse the genome sequences of major barley accessions worldwide. In view of technical limitations, a strategy has been employed to establish the exome structure of a selected number of accessions and to perform high-quality chromosome-scale assembly of the genomes of several major representative accessions. For the future project, an efficient annotation pipeline is essential for establishing the function of genomes and genes as well as for using this information for sequence-based digital barley breeding. In this article, the author reviews the existing barley resources along with their applications and discuss possible future directions of research in barley genomics. No potential conflict of interest relevant to this article was reported.

Cell Press Acta Medica Okayama 2666-1667 1 2 2020 Protocol for Genome Editing to Produce Multiple Mutants in Wheat 100053 EN Fumitaka Abe Division of Basic Research, Institute of Crop Science, NARO Yuji Ishida Plant Innovation Center, Japan Tobacco Inc. Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Masaki Endo Division of Applied Genetics, Institute of Agrobiological Sciences, NARO Toshihiko Komari Plant Innovation Center, Japan Tobacco Inc. Seiichi Toki Division of Applied Genetics, Institute of Agrobiological Sciences, NARO Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mutants can be generated. The technique for acceleration of generation advancement with embryo culture reduces time for mutant production. The mutants produced by this protocol can be used for the analysis of gene function and crop improvement. For complete details on the use and execution of this protocol, please refer to Abe et al. (2019). No potential conflict of interest relevant to this article was reported.

Frontiers Research Foundation Acta Medica Okayama 1664-462X 577 2019 Development of Genome-Wide SNP Markers for Barley via Reference- Based RNA-Seq Analysis 577 EN Tsuyoshi Tanaka Breeding Informatics Research Unit, Division of Basic Research, Institute of Crop Science, National Agriculture and Food Research Organization (NARO), Goro Ishikawa Breeding Strategies Research Unit, Division of Basic Research, Institute of Crop Science, National Agriculture and Food Research Organization (NARO) Eri Ogiso-Tanaka Soybean and Field Crop Applied Genomics Research Unit, Division of Field Crop Research, Institute of Crop Science, National Agriculture and Food Research Organization (NARO) Takashi Yanagisawa Wheat and Barley Breeding Unit, Division of Wheat and Barley Research, Institute of Crop Science, National Agriculture and Food Research Organization (NARO) Kazuhiro Sato Group of Genome Diversity, Institute of Plant Science and Resources, Okayama University Marker-assisted selection of crop plants requires DNA markers that can distinguish between the closely related strains often used in breeding. The availability of reference genome sequence facilitates the generation of markers, by elucidating the genomic positions of new markers as well as of their neighboring sequences. In 2017, a high quality genome sequence was released for the six-row barley (Hordeum vulgare) cultivar Morex. Here, we developed a de novo RNA-Seq-based genotyping procedure for barley strains used in Japanese breeding programs. Using RNA samples from the seedling shoot, seedling root, and immature flower spike, we mapped next-generation sequencing reads onto the transcribed regions, which correspond to ∼590 Mb of the whole ∼4.8-Gbp reference genome sequence. Using 150 samples from 108 strains, we detected 181,567 SNPs and 45,135 indels located in the 28,939 transcribed regions distributed throughout the Morex genome. We evaluated the quality of this polymorphism detection approach by analyzing 387 RNA-Seq-derived SNPs using amplicon sequencing. More than 85% of the RNA-Seq SNPs were validated using the highly redundant reads from the amplicon sequencing, although half of the indels and multiple-allele loci showed different polymorphisms between the platforms. These results demonstrated that our RNA-Seq-based de novo polymorphism detection system generates genome-wide markers, even in the closely related barley genotypes used in breeding programs. No potential conflict of interest relevant to this article was reported.

Cell Press Acta Medica Okayama 22111247 28 5 2019 Genome-Edited Triple-Recessive Mutation AltersSeed Dormancy in Wheat 1362 1369 EN Fumitaka Abe Division of Wheat and Barley Research, Institute of Crop Science, NARO Emdadul Haque Division of Wheat and Barley Research, Institute of Crop Science, NARO Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Tsuyoshi Tanaka Division of Basic Research, Institute of Crop Science, NARO Yoko Kamiya Kihara Institute for Biological Research, Yokohama City University Masafumi Mikami Graduate School of Nanobioscience, Yokohama City University Kanako Kawaura Kihara Institute for Biological Research, Yokohama City University Masaki Endo Division of Applied Genetics, Institute of Agrobiological Sciences, NARO Kazumitsu Onishi Department of Agro-Environmental Science, Obihiro University of Agriculture and Veterinary Medicine Takeshi Hayashi Division of Basic Research, Institute of Crop Science, NARO Kazuhiro Sato Institute of Plant Science and Resources, Okayama University 1Common wheat has three sets of sub-genomes, making mutations difficult to observe, especially for traits controlled by recessive genes. Here, we produced hexaploid wheat lines with loss of function of homeoalleles of Qsd1, which controls seed dormancy in barley, by Agrobacterium-mediated CRISPR/Cas9. Of the eight transformed wheat events produced, three independent events carrying multiple mutations in wheat Qsd1 homeoalleles were obtained. Notably, one line had mutations in every homeoallele. We crossed this plant with wild-type cultivar Fielder to generate a transgene-free triple-recessive mutant, as revealed by Mendelian segregation. The mutant showed a significantly longer seed dormancy period than wild-type, which may result in reduced pre-harvest sprouting of grains on spikes. PCR, southern blotting, and whole-genome shotgun sequencing revealed that this segregant lacked transgenes in its genomic sequence. This technique serves as a model for trait improvement in wheat, particularly for genetically recessive traits, based on locus information from diploid barley. No potential conflict of interest relevant to this article was reported.

Springer Acta Medica Okayama 0721-7714 36 4 2017 Selection of transformation-efficient barley genotypes based on TFA (transformation amenability) haplotype and higher resolution mapping of the TFA loci 611 620 EN Hiroshi Hisano Institute of Plant Science and Resources, Okayama University Brigid Meints Department Crop and Soil Sciences, Washington State University Matthew J. Moscou The Sainsbury Laboratory Luis Cistue Department Genetica y Produccion Vegetal Begoña Echávarri Kazuhiro Sato Institute of Plant Science and Resources, Okayama University Patrick M. Hayes Department Crop and Soil Science, Oregon State University Key message: The genetic substitution of transformation amenability alleles from ‘Golden Promise’ can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars. Abstract: Barley (Hordeum vulgare) cv. ‘Golden Promise’ is one of the most useful and well-studied cultivars for genetic manipulation. In a previous report, we identified several transformation amenability (TFA) loci responsible for Agrobacterium-mediated transformation using the F2 generation of immature embryos, derived from ‘Haruna Nijo’ × ‘Golden Promise,’ as explants. In this report, we describe higher density mapping of these TFA regions with additional SNP markers using the same transgenic plants. To demonstrate the robustness of transformability alleles at the TFA loci, we genotyped 202 doubled haploid progeny from the cross ‘Golden Promise’ × ‘Full Pint.’ Based on SNP genotype, we selected lines having ‘Golden Promise’ alleles at TFA loci and used them for transformation. Of the successfully transformed lines, DH120366 came the closest to achieving a level of transformation efficiency comparable to ‘Golden Promise.’ The results validate that the genetic substitution of TFA alleles from ‘Golden Promise’ can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars. No potential conflict of interest relevant to this article was reported.

Biomed Central Ltd Acta Medica Okayama 1471-2164 12 2011 454 sequencing of pooled BAC clones on chromosome 3H of barley EN Kazuhiro Sato Yuka Motoi Nami Yamaji Hideya Yoshida Background: Genome sequencing of barley has been delayed due to its large genome size (ca. 5,000Mbp). Among the fast sequencing systems, 454 liquid phase pyrosequencing provides the longest reads and is the most promising method for BAC clones. Here we report the results of pooled sequencing of BAC clones selected with ESTs genetically mapped to chromosome 3H. Results: We sequenced pooled barley BAC clones using a 454 parallel genome sequencer. A PCR screening system based on primer sets derived from genetically mapped ESTs on chromosome 3H was used for clone selection in a BAC library developed from cultivar "Haruna Nijo". The DNA samples of 10 or 20 BAC clones were pooled and used for shotgun library development. The homology between contig sequences generated in each pooled library and mapped EST sequences was studied. The number of contigs assigned on chromosome 3H was 372. Their lengths ranged from 1,230 bp to 58,322 bp with an average 14,891 bp. Of these contigs, 240 showed homology and colinearity with the genome sequence of rice chromosome 1. A contig annotation browser supplemented with query search by unique sequence or genetic map position was developed. The identified contigs can be annotated with barley cDNAs and reference sequences on the browser. Homology analysis of these contigs with rice genes indicated that 1,239 rice genes can be assigned to barley contigs by the simple comparison of sequence lengths in both species. Of these genes, 492 are assigned to rice chromosome 1. Conclusions: We demonstrate the efficiency of sequencing gene rich regions from barley chromosome 3H, with special reference to syntenic relationships with rice chromosome 1. No potential conflict of interest relevant to this article was reported.

Oxford University Press Acta Medica Okayama 0305-1048 38 2010 NBRP databases: databases of biological resources in Japan D26 D32 EN Yukiko Yamazaki Ryo Akashi Yutaka Banno Takashi Endo Hiroshi Ezura Kaoru Fukami-Kobayashi Kazuo Inaba Tadashi Isa Katsuhiko Kamei Fumie Kasai Masatomo Kobayashi Nori Kurata Makoto Kusaba Tetsuro Matuzawa Shohei Mitani Taro Nakamura Yukio Nakamura Norio Nakatsuji Kiyoshi Naruse Hironori Niki Eiji Nitasaka Yuichi Obata Hitoshi Okamoto Moriya Okuma Kazuhiro Sato Tadao Serikawa Toshihiko Shiroishi Hideaki Sugawara Hideko Urushibara Masatoshi Yamamoto Yoshio Yaoita Atsushi Yoshiki Yuji Kohara The National BioResource Project (NBRP) is a Japanese project that aims to establish a system for collecting, preserving and providing bioresources for use as experimental materials for life science research. It is promoted by 27 core resource facilities, each concerned with a particular group of organisms, and by one information center. The NBRP database is a product of this project. Thirty databases and an integrated database-retrieval system (BioResource World: BRW) have been created and made available through the NBRP home page (http://www.nbrp.jp). The 30 independent databases have individual features which directly reflect the data maintained by each resource facility. The BRW is designed for users who need to search across several resources without moving from one database to another. BRW provides access to a collection of 4.5-million records on bioresources including wild species, inbred lines, mutants, genetically engineered lines, DNA clones and so on. BRW supports summary browsing, keyword searching, and searching by DNA sequences or gene ontology. The results of searches provide links to online requests for distribution of research materials. A circulation system allows users to submit details of papers published on research conducted using NBRP resources. No potential conflict of interest relevant to this article was reported.

Biomed Central Ltd Acta Medica Okayama 1471-2164 10 2009 Development and implementation of high-throughput SNP genotyping in barley EN Timothy J. Close Prasanna R. Bhat Stefano Lonardi Yonghui Wu Nils Rostoks Luke Ramsay Arnis Druka Nils Stein Jan T. Svensson Steve Wanamaker Serdar Bozdag Mikeal L. Roose Matthew J. Moscou Shiaoman Chao Rajeev K. Varshney Peter Szuecs Kazuhiro Sato Patrick M. Hayes David E. Matthews Andris Kleinhofs Gary J. Muehlbauer Joseph DeYoung David F. Marshall Kavitha Madishetty Raymond D. Fenton Pascal Condamine Andreas Graner Robbie Waugh Background: High density genetic maps of plants have, nearly without exception, made use of marker datasets containing missing or questionable genotype calls derived from a variety of genic and non-genic or anonymous markers, and been presented as a single linear order of genetic loci for each linkage group. The consequences of missing or erroneous data include falsely separated markers, expansion of cM distances and incorrect marker order. These imperfections are amplified in consensus maps and problematic when fine resolution is critical including comparative genome analyses and map-based cloning. Here we provide a new paradigm, a high-density consensus genetic map of barley based only on complete and error-free datasets and genic markers, represented accurately by graphs and approximately by a best-fit linear order, and supported by a readily available SNP genotyping resource. Results: Approximately 22,000 SNPs were identified from barley ESTs and sequenced amplicons; 4,596 of them were tested for performance in three pilot phase Illumina GoldenGate assays. Data from three barley doubled haploid mapping populations supported the production of an initial consensus map. Over 200 germplasm selections, principally European and US breeding material, were used to estimate minor allele frequency (MAF) for each SNP. We selected 3,072 of these tested SNPs based on technical performance, map location, MAF and biological interest to fill two 1536-SNP "production" assays (BOPA1 and BOPA2), which were made available to the barley genetics community. Data were added using BOPA1 from a fourth mapping population to yield a consensus map containing 2,943 SNP loci in 975 marker bins covering a genetic distance of 1099 cM. Conclusion: The unprecedented density of genic markers and marker bins enabled a high resolution comparison of the genomes of barley and rice. Low recombination in pericentric regions is evident from bins containing many more than the average number of markers, meaning that a large number of genes are recombinationally locked into the genetic centromeric regions of several barley chromosomes. Examination of US breeding germplasm illustrated the usefulness of BOPA1 and BOPA2 in that they provide excellent marker density and sensitivity for detection of minor alleles in this genetically narrow material. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 61 1 2011 Expression and functional analysis of the barley Nud gene using transgenic rice 35 42 EN Katsuyuki Kakeda Mie Univ, Grad Sch Bioresources Norimitsu Ishihara Mie Univ, Grad Sch Bioresources Yohei Izumi Okayama Univ, Inst Plant Sci & Resources Kazuhiro Sato Okayama Univ, Inst Plant Sci & Resources Shin Taketa Okayama Univ, Inst Plant Sci & Resources Most cereal crops have hulless grains (naked caryopses) with a free-threshing trait, whereas the majority of barley cultivars show hulled (covered) caryopses. The naked caryopsis in barley is genetically controlled by a single locus, nod. The Nud gene (the covered caryopsis allele) encodes an ethylene response factor (ERF) family transcription factor that regulates a lipid biosynthetic pathway. For functional analysis of the barley Nud gene, we produced transgenic rice expressing Nod in the developing caryopses. All transgenic lines had caryopses that were easily dehulled at maturity, indicating that the naked caryopsis phenotype remained in spite of expression of the Nod transgene. Histochemical and lipid analyses of the transgenic rice caryopses did not show increased lipid accumulation on the surface of developing caryopses, suggesting that the Nud-mediated lipid pathway may not function in rice caryopses. The predicted rice ortholog of Nod, Os06ERF was expressed specifically in the developing caryopses. However, expression of Os06ERF ceased at an earlier developmental stage than that of the native Nod gene in barley caryopses, which was also the case for expression of the Nod transgene. This raises the alternative hypothesis that the timing of Nod expression may be critical for activating the pathway for hull-caryopsis adhesion. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 60 5 2010 NBRP, National Bioresource Project of Japan and plant bioresource management 461 468 EN Nori Kurata Hikaru Satoh Hidemi Kitano Yasuo Nagato Takashi Endo Kazuhiro Sato Ryo Akashi Hiroshi Ezura Makoto Kusaba Masatomo Kobayashi Eiji Nitasaka Fumie Kasai Yukiko Yamazaki Atsushi Yoshimura The National BioResource Project has been organized and established to promote research activities using valuable bioresources. A total of twenty-eight bioresources for ten animals, nine plants and nine microorganisms/cell lines developed or collected in Japan were selected for the project. Resources are categorized into several different groups in the project; genetic resources, germplasm, genome resources and their information. Choices of how many resources must be preserved and maintained and in which categories are dependent on the status of the research community of each organism. These resources, if utilized systematically and intelligently, are powerful means for leading new scientific discoveries. Some examples can be seen in this paper. This paper reviews plant bioresources with the main focus on rice resource activities within the project. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 59 4 2009 Mapping of QTL for intermedium spike on barley chromosome 4H using EST-based markers 383 390 EN Fahimeh Shahinnia Badraldin Ebrahim Sayed-Tabatabaei Kazuhiro Sato Mohammad Pourkheirandish Takao Komatsuda The lateral spikelets of two-rowed barley are reduced in size and sterile, but in six-rowed barley all three spikelets are fully fertile. The trait is largely controlled by alleles at the vrs1 locus on chromosome arm 2HL, as modified by the allele present at the I locus on chromosome arm 4HS. Molecular markers were developed to saturate the 4HS region by exploiting expressed sequence-tags, either previously mapped in barley to this region, or present in the syntenic region of rice chromosome 3. Collinearity between rice and barley was strong in the 4.8 cM interval BJ468164-AV933435 and the 10 cM interval AV942364-BJ455560. A major QTL for lateral spikelet fertility (the I locus) explained 44% of phenotypic variance, and was located in the interval CB873567-BJ473916. The genotyping of near-isogenic lines for I placed the locus in a region between CB873567 and EBmac635, and therefore the most likely position of the I locus was proximal to CB873567 in a 5.3 cM interval between CB873567-BJ473916. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 59 5 2009 Genetic analysis of seed dormancy QTL in barley 645 650 EN Kazuhiro Sato Takashi Matsumoto Natsuko Ooe Kazuyoshi Takeda Seed dormancy in wild barley enables drought escape by preventing germination during the hot summer in and environments. Dormancy in cultivated barley has different effects: it can delay the malting process and/or it can prevent pre-harvest sprouting. Thus, cloning dormancy genes in barley will contribute to understanding the domestication process and it will facilitate optimizing the trait for efficient agronomic and industrial uses. Rates of seed germination were used to evaluate dormancy on physiologically matured grain samples that were dried and stored frozen until use. With this phenotypic scoring procedure, many genetic factors controlling seed dormancy has been reported as quantitative trait loci (QTL). Of these QTL, one at the centrometic region of chromosome 5H (Qsd1) has been most frequently identified and shows the largest effect across mapping populations. We also identified this QTL using the EST map based on Haruna Nijo (H. vulgare ssp. vulgare) crossed with wild barley H602 (H. vulgare ssp. spontaneum). We have derived both doubled haploid and recombinant chromosome substitution lines (RCSLs) from this cross. At least four QTLs are segregating in this germplasm. RCSLs having only the Qsd1 segment of wild barley in a Haruna Nijo genetic background were identified and 910 BC(3)F(2) plants were scored for dormancy. In these lines, segregation for dormancy fit a mono-factorial ratio. These germplasm resources are appropriate for map based cloning of Qsd1. Strategies for cloning Qsd1 with these resources are discussed. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 59 4 2009 Validation of rice blast resistance genes in barley using a QTL mapping population and near-isolines 341 349 EN Phinyarat Kongprakhon Alfonso Cuesta-Marcos Patrick M. Hayes Kelley L. Richardson Pattama Sirithunya Kazuhiro Sato Brian Steffenson Theerayuth Toojinda There are prior reports of Pyricularia grisea-the causal agent of blast of rice-causing disease in barley. In order to determine the specificity of this resistance in barley, we extended our previous mapping efforts to include blast isolates from barley and rice grown in Thailand and we assessed two resistance phenotypes: leaf blast (LB) and neck blast (NB). The largest-effect resistance QTL, on chromosome I H, was associated with NB and LB and is located in a region rich in resistance genes, including QTL conferring resistance to stripe rust (incited by Puccinia striiformis f. sp. hordei) and the mildew (Blumeria graminis f. sp. hordei) resistance gene Mla. The LB, NB and mildew resistance alleles trace to one parent (Baronesse) whereas the stripe rust resistance allele traces to the other parent (BCD47) of the mapping population. Baronesse is the susceptible recurrent parent of a set of near-isogenic lines (NILs) for three stripe rust resistance QTL, including one on 1H. Unigene (EST) derived single nucleotide polymorphism haplotypes of these NILs were aligned with the blast mapping population QTL using Mla as an anchor. Baronesse and all NILs without the 1H introgression were resistant to LB and NB. However, two NILs with the I H introgression were resistant to LB and NB. Both are resistant to stripe rust. Therefore, the QTL conferring resistance to stripe rust is separable by recombination from the blast resistance QTL. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1344-7610 59 1 2009 Mapping of the eibi1 gene responsible for the drought hypersensitive cuticle in wild barley (Hordeum spontaneum) 21 26 EN Guoxiong Chen Takao Komatsudu Mohammad Pourkheirandish Mohammad Sameri Kazuhiro Sato Tamar Krugman Tzion Fahima Abraham B. Korol Eviatar Nevo Segregation analysis showed that eibi1, a drought hypersensitive Cuticle wild barley mutant, was monogenic and recessive, and mapped in two F, Populations, one made from a cross between the mutant and a Cultivated barley (cv. Morex), and the other between the mutant and another wild barley. A microsatellite marker screen showed that the gene was located oil barley chromosome 3H, and a set of markers already assigned to this chromosome, including both microsatellites and ESTs, was used to construct a genetic map. eibi1 co-segregated with barley EST AV918546, and was located to bin 6. The synteny between barley and rice ill this region is incomplete, with a large discrepancy in map distances, and the presence Of Multiple inversions. No potential conflict of interest relevant to this article was reported.

Oxford University Press Acta Medica Okayama 1340-2838 19 6 2012 Discovery of High-Confidence Single Nucleotide Polymorphisms from Large-Scale De Novo Analysis of Leaf Transcripts of Aegilops tauschii, A Wild Wheat Progenitor 487 497 EN Julio Cesar Masaru Iehisa Akifumi Shimizu Kazuhiro Sato Shuhei Nasuda Shigeo Takumi Construction of high-resolution genetic maps is important for genetic and genomic research, as well as for molecular breeding. Single nucleotide polymorphisms (SNPs) are the,predominant class of genetic variation and can be used as molecular markers. Aegilops tauschii, the D-genome donor of common wheat, is considered a valuable genetic resource for wheat improvement. Our previous study implied that Ae. tauschii accessions can be genealogically divided into two major lineages. In this study, the transcriptome of two Ae. tauschii accessions from each lineage, lineage 1 (L1) and 2 (L2), was sequenced, yielding 9435 SNPs and 739 insertion/deletion polymorphisms (indels) after de novo assembly of the reads. Based on 36 contig sequences, 31 SNPs and six indels were validated on 20 diverse Ae. tauschii accessions. Because almost all of the SNP markers were polymorphic between L1 and L2, and the D-genome donor of common wheat is presumed to belong to L2, these markers are available for D-genome typing in crosses between common wheat varieties and L1-derived synthetic wheat. Due to the conserved synteny between wheat and barley chromosomes, the high-density expressed sequence tag barley map and the hypothetical gene order in barley can be applied to develop markers on target chromosomal regions in wheat. No potential conflict of interest relevant to this article was reported.

Oxford University Press Acta Medica Okayama 1340-2838 16 2 2009 Development of 5006 Full-Length CDNAs in Barley: A Tool for Accessing Cereal Genomics Resources 81 89 EN Kazuhiro Sato Tadasu Shin-I Motoaki Seki Kazuo Shinozaki Hideya Yoshida Kazuyoshi Takeda Yukiko Yamazaki Matthieu Conte Yuji Kohara A collection of 5006 full-length (FL) cDNA sequences was developed in barley. Fifteen mRNA samples from various organs and treatments were pooled to develop a cDNA library using the CAP trapper method. More than 60% of the clones were confirmed to have complete coding sequences, based on comparison with rice amino acid and UniProt sequences. Blastn homologies (E < 1E-5) to rice genes and Arabidopsis genes were 89 and 47%, respectively. Of the 5028 possible amino acid sequences derived from the 5006 FLcDNAs, 4032 (80.2%) were classified into 1678 GreenPhyl multigenic families. There were 555 cDNAs showing low homology to both rice and Arabidopsis. Gene ontology annotation by InterProScan indicated that many of these cDNAs (71%) have no known molecular functions and may be unique to barley. The cDNAs showed high homology to Barley 1 GeneChip oligo probes (81%) and the wheat gene index (84%). The high homology between FLcDNAs (27%) and mapped barley expressed sequence tag enabled assigning linkage map positions to 151-233 FLcDNAs on each of the seven barley chromosomes. These comprehensive barley FLcDNAs provide strong platform to connect preexisting genomic and genetic resources and accelerate gene identification and genome analysis in barley and related species. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1040-4651 23 4 2011 Unlocking the Barley Genome by Chromosomal and Comparative Genomics 1249 1263 EN Klaus F. X. Mayer Mihaela Martis Pete E. Hedley Hana Simkova Hui Liu Jenny A. Morris Burkhard Steuernagel Stefan Taudien Stephan Roessner Heidrun Gundlach Marie Kubalakova Pavla Suchankova Florent Murat Marius Felder Thomas Nussbaumer Andreas Graner Jerome Salse Takashi Endo Hiroaki Sakai Tsuyoshi Tanaka Takeshi Itoh Kazuhiro Sato Matthias Platzer Takashi Matsumoto Uwe Scholz Jaroslav Dolezel Robbie Waugh Nils Stein We used a novel approach that incorporated chromosome sorting, next-generation sequencing, array hybridization, and systematic exploitation of conserved synteny with model grasses to assign; similar to 86% of the estimated; similar to 32,000 barley (Hordeum vulgare) genes to individual chromosome arms. Using a series of bioinformatically constructed genome zippers that integrate gene indices of rice (Oryza sativa), sorghum (Sorghum bicolor), and Brachypodium distachyon in a conserved synteny model, we were able to assemble 21,766 barley genes in a putative linear order. We show that the barley (H) genome displays a mosaic of structural similarity to hexaploid bread wheat (Triticum aestivum) A, B, and D subgenomes and that orthologous genes in different grasses exhibit signatures of positive selection in different lineages. We present an ordered, information-rich scaffold of the barley genome that provides a valuable and robust framework for the development of novel strategies in cereal breeding. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0032-0889 156 1 2011 Comprehensive Sequence Analysis of 24,783 Barley Full-Length cDNAs Derived from 12 Clone Libraries 20 28 EN Takashi Matsumoto Tsuyoshi Tanaka Hiroaki Sakai Naoki Amano Hiroyuki Kanamori Kanako Kurita Ari Kikuta Kozue Kamiya Mayu Yamamoto Hiroshi Ikawa Nobuyuki Fujii Kiyosumi Hori Takeshi Itoh Kazuhiro Sato Full-length cDNA (FLcDNA) libraries consisting of 172,000 clones were constructed from a two-row malting barley cultivar (Hordeum vulgare 'Haruna Nijo') under normal and stressed conditions. After sequencing the clones from both ends and clustering the sequences, a total of 24,783 complete sequences were produced. By removing duplicates between these and publicly available sequences, 22,651 representative sequences were obtained: 17,773 were novel barley FLcDNAs, and 1,699 were barley specific. Highly conserved genes were found in the barley FLcDNA sequences for 721 of 881 rice (Oryza sativa) trait genes with 50% or greater identity. These FLcDNA resources from our Haruna Nijo cDNA libraries and the full-length sequences of representative clones will improve our understanding of the biological functions of genes in barley, which is the cereal crop with the fourth highest production in the world, and will provide a powerful tool for annotating the barley genome sequences that will become available in the near future. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 3 1 1995 Selection Effectiveness for the Resistance to Net Blotch in Barley 43 53 EN Kazuhiro Sato Kazuyoshi Takeda Selection effectiveness for the resistance to net blotch was estimated by using two sets of F2 and F3 populations derived from the crosses between resistant and susceptible parents. In every F2 and F3 population, disease ratings showed a continuous distribution. As many F3 lines with intermediate resistance had a smaller variance and homozygous genotype, the resistance might be controlled by a few genes. The heritabilities of the disease rating were estimated　by correlation coefficients and regression coefficients between each F2 plant and the descended F3 lines. Another estimate for heritability was calculated by the selection differential in the F2 plants and genetic gain in the F3 lines. Despite the different level of resistance in the resistant parents of the two crosses, the three kinds of heritabilities estimated were similar and ranged from 0.6 to 0.8. Because of the fewer number of genes controlling the disease resistance and the higher heritabilities, selection in a early generation may be effective for net blotch resistance in barlcy. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 2 1 1994 Numerical Taxonomic Study in Annual Cyperaceous Weeds 123 134 EN Chozin Muhamad Ahmad Kazuhiro Satou Shozo Yasuda Three species of Cyperaceous weeds, Cyperus iria (12 strains), C.　microiria(12 strains) and C. amuricus (6 strains), were collected from different sites of Okayama, Tottori and Tokyo prefectures, and various morphological characters, biomass and seed production were observed on the plants which were cultivated at Kurashiki. The analysis of variance showed a significant difference among the strains in each character. However, the species overlapped with each other in most morphological characters. Prinipal component analysis on the 21 characters showed that 83% of the total variation could be explained by the first three components: the first component (37%) was regarded as factors concerning spikelet and seed production; the second component (28%) was regarded as factors concerned the size of vegetative parts; the third component (18%) was largely affected by seed weight and floret density. Scatter diagram on the first and third principal components showed that the 30 strains of three species divided into three groups, and strains in each group correspond to the three species without exception. Based on the second and third principal components, strains of C. microiria were further divided into three sub-groups according to size of vegetative parts. Using the cluster analysis, 30 strains of these species were divided into four large clusters; the first was composed of C. amuricus strains, the second was of three strains of C. microiria, the third included the remaining strains of C. microiria, and the last cluster was composed of C. iria strains. It may be concluded that C. microiria is composed of two or three ecotypes which are different in morphological and reproductive traits. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 2 1 1994 オオムギの野生種（Hordeum spontaneum C. Koch)と在来種におけるうどんこ病抵抗性の比較 111 122 EN Toshinori Fukuyama Hideo Heta Kazuhiro Sato Kazuyoshi Takeda A total of 162 strains of wild barley, Hordeum spontaneum C. Koch originating from Iran, Iraq, Turkey and Central Asia, were tested for resistance to powdery mildew. Then, the variation of resistance was compared with that of 145 local varieties of cultivated barley (Hordeum vulgare L.) originating from the same region of the wild barley collection. Ten different isolates of the parasite with Japanese origin were separately inoculated onto the first leaves of the host plants. The infection types were classified into the following: i, immunelike; R, highly resistant; M, moderately resistant; and S, highly susceptible. Resistant strains with i, R or M infection type were more frequent among wild barleys as compared with the cultivated forms. It is noteworthy that among these three resistant reactions,the M type was most frequent in the wild barley. To compare the degree of resistance to a total of 10 isolates, the resistance score was calculated in each of the wild and cultivated strains as the following: Scores 1,2,3 and 4 were given to the infection types of i, R, M and S, respectively, and the mean score for 10 isolates was calculated. Wild barley showed significantly low resistance scores as compared with those of cultivated barley. This was also confirmed by the cluster analysis; the cluster with more resistance to 10 isolates consisted of many strains of wild barley. Next, the resistance of wild barley was characterized by their broader effective ranges to different isolates. According to the x2 test for independence of reactions to two different isolates, the resistant factor(s) involved in wild barley was confirmed to be rather non-specific to the parasite. It was concluded that H. spontaneum may be useful genetic resources for the breeding of resistance to powdery mildew as well as local varieties. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 2 1 1994 大麦網斑病における抵抗性遺伝資源 91 102 EN Kazuhiro Sato Kazuyoshi Takeda Net blotch caused by a fungus Pyrenophora teres Drechs. is a common disease in barley. Its source of resistance has been screened by many researchers by field evaluations or seedling tests inoculating a single isolatc. Since the pathogcnic variation of isolates has been reported in net blotch, resistance of the varieties to the disease may be different among the isolates with different pathogenicities. In this study, the pathogenic variation was examined and the varietal variation of the resistance was evaluated by inoculating with four P. teres isolates collected from Japan and Canada to more than 2,200 barley varieties of the world collection preserved at the Barley Germplasm Center of Okayama University. A preliminary inoculation test showed that the disease rating was affected little by the inoculation seasons. Disease ratings of varieties showed a continuous variation with a single mode in the resistant range in each of the four isolates. However, the correlation coefficient between Japanese isolate K105 and Canadian isolate WRS102 was as low as 0.55, indicating a slight pathogenic differentiation between these isolates. Significant correlation coefficients (r=.55~.78) among the ratings of isolates indicated that the pathogenicity to the varieties was rather similar and that the pathogenic differentiation was small among the four isolates tested. In general, varieties from Ethiopia, North Africa and Korea were more resistant than those from other regions. Varieties from Turkey and Europe were susceptible to Japanese isolates, while Nepalese varieties were susceptible to Canadian isolates. Twenty of 25 varieties which were resistant to the isolate K105 but susceptible to the isolate WR102 were from Nepal and most of those were Oriental-type (Bt bt2) in brittleness of rachis. These findings revealed an example of regional concentration of resistant gene in net blotch. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 2 1 1994 オオムギの葉の剛毛に関する遺伝子分析 63 68 EN Kazuhiro Sato Kazuyoshi Takeda The inheritance and linkage relationship of a new hairiness trait "large trichome" was investigated in barley. Although the size of large trichome is about four times that of normal one, the character can not be recognized with the naked eye. However, it is easily identified by the roughness of leaf touch. The large trichomes develop on both sides of the leaf blades. The direction of trichome is both acropetal and basipetal. It is clearly distinguished from the extremely long trichome controlled by Pub gene. About 2,300 varieties of our Barley Germplasm Center were screened by the leaf touch to find nine varieties with large trichome. Two of them were six-rowed local variety from Pakistan, and other seven were two-rowed varieties from Europe and Japan. All of them were hulled type. Crosses of six large trichome varieties with a normal Japanese variety resulted in the large trichome type F1s, suggesting the dominant nature of the trait. The large trichome line Hokuiku 17 was crossed with various linkage testres to study the mode of inheritance and the linkage relationship of the gene. In the F2 populations, the large trichome was controlled by a single dominant gene named Ltr (large trichome), which was independentiy inherited from the following marker genes; br and gl-5 on chromosome 1; li and ν on chromosome 2; uz on chromosome 3; K and gl-3 on chromosome 4; trd on chromosome 5; ο on chromosome 6. On the other hand, from the cross between Hokuiku 17 and OUL166, Ltr was found to be linked with s and fs on chromosome 7. Although the allelism test has not been completed, the very low frequency of the large trichome type (9/2,300) indicates that the variant resulted from a recent mutation event, or the fitness of the variant is low in the natural and/or artificial selection. No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 1 1 1992 大麦網斑病における幼苗検定法の確立と抵抗性品種の検索 75 90 EN Kazihiro Sato Kazuyoshi Takeda 大麦網斑病は糸状菌の1種であるPyrenophora teres Drechs.の感染によって葉身、葉鞘等に網目状の病斑を生じ、子実の登熟低下によって減収する共に、ビールオオムギにおいては醸造品質であるエキス分が低下する重要病害である。本病害は世界各地のオオムギ栽培地帯のうち主として温暖・湿潤な地域に分布しており（Shipton　et　al.1973）、近年、連作や灌漑によって被害が増大しつつある（Mathre 1982）。我国においては従来からその存在が確認されていたものの、登熟後期の活性の衰えた葉に生じる病害として重要性は認識されていなかった。しかし、最近、北海道、鳥取県、鹿児島県などのビールオオムギ栽培地帯で局所的な激発事例が確認されている。（佐藤、未発表）。本病害に対する防除法としては種子消毒ならびに殺菌剤の茎葉撒布が有効であるが、その効果は完全ではない。また、茎葉撒布はコストが高く、環境汚染の問題もあるので、最も有効で経済的かつ安全な防除法は抵抗性品種を栽培することと言っても良い。従来、本病害の積極的な抵抗性育種は行われていなかったが、最近は抵抗性を有する品種も育成されている（Metcalfe 1987）。抵抗性品種を育成するためには、遺伝資源ならびに雑種後代を効率よく評価、選抜するための検定方法を確立しなければならない。本病抵抗性の検定方法としては幼苗検定法、圃場検定法が考案されて広く用いられているが（Buchannon and McDonald 1965, Holtmeyer and Webster 1981）、環境条件の変化によって抵抗性が変動する事例が報告されているので（Khan and Boyd 1970, Tekauz 1986）、抵抗性を確実に評価するための安定した検定条件を設定する必要がある。抵抗性に関する遺伝資源についてはSchaller and Wiebe （1952）、Dessouki et al.（1965）およびBuchannon and McDonald （1965）等がそれぞれ数千品種を評価し、中国東北部、トルコおよびエチオピアなどに抵抗性の遺伝資源が豊富であること報告している。それらの品種のいくつかについては、抵抗性の遺伝子分析が行われており（Bockelman et al. 1977, Davis et al. 1990）、本病抵抗性育種の交配親として使用されている（Tekazu and Buchannon 1977, Moseman and Smith 1985）。岡山大学資源生物科学研究所大麦系統保存施設は世界的にも貴重なと東アジアの遺伝資源をはじめ五千余の保存品種を有するが、著者らは大麦網斑病の幼苗検定法を確立し、これらの品種の抵抗性を評価したので報告する。 No potential conflict of interest relevant to this article was reported.

岡山大学資源生物科学研究所 Acta Medica Okayama 0916-930X 1 2 1993 Pathogenic Variation of Pyrenophora teres Isolates Collected from Japanese and Canadian Spring Barley 147 158 EN Kazuhiro Sato Kazuyoshi Takeda Twenty-two isolates of Pyrenophora teres Drechs. collected from Japanese and Canadian spring barleys were inoculated to 38 barley varieties having various genetic backgrounds. The analysis of variance for the discase ratings showed that there were significant differences both in the virulence of isolates and the resistance of varieties. However, the interaction among isolates and varieties was not statistically significant. Both Finlay-Wilkinson regression analysis and principal component analysis by Additive Main effects and Multiplicative Interaction effects(AMMI)model classified the isolates into three groups,which were different in origins and sympton types. A spot tyte isolate was distinguished from net type isolates by its generally high virulence. A slight pathogenic differentiation was suggested between Japanese and Canadian net type isolates. No potential conflict of interest relevant to this article was reported.