Updated on 2024/06/19

写真a

 
HISANO Hiroshi
 
Organization
Institute of Plant Science and Resources Professor
Position
Professor
Profile

I studied about fructan biosynthesis and freezing tolerance in forage grass species at Hokkaido University. After graduation, I worked for development of DNA markers for breeding in legume and grass species and for genetic modification of lignin biosynthesis in bioenergy crop. Now I perform transformation and genome editing in barley.

External link

Degree

  • PhD ( 2005.3   Hokkaido University )

Research Interests

  • Plant Molecular Breeding

  • transformation

  • barley

  • genome editing

  • 植物遺伝資源学

  • Plant tissue culture

Research Areas

  • Environmental Science/Agriculture Science / Science in plant genetics and breeding

Education

  • Hokkaido University   大学院 農学研究科   応用生命科学専攻 博士課程

    2002.4 - 2005.3

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    Country: Japan

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  • Hokkaido University   大学院 農学研究科   応用生命科学専攻 修士課程

    2000.4 - 2002.3

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    Country: Japan

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  • Hokkaido University   農学部   生物資源科学科

    1996.4 - 2000.3

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    Country: Japan

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Research History

  • Okayama University   Institute of Plant Science and Resources   Professor

    2024.4

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    Country:Japan

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  • IPSR, Okayama University   Associate Professor

    2018.2 - 2024.3

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  • Okayama University   Institute of Plant Science and Resources   Assistant Professor

    2012.2 - 2018.1

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  • The Samuel Roberts Noble Foundation, Inc   Postdoctoral fellow

    2008.3 - 2012.1

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    Country:United States

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  • Kazusa DNA Research Institute   Postdoctoral fellow

    2005.4 - 2008.3

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Professional Memberships

Committee Memberships

  • 農林水産省   農業資材審議会 委員  

    2024.6   

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    Committee type:Government

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  • 農林水産省   農業資材審議会 臨時委員  

    2021.4 - 2024.6   

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    Committee type:Government

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  • 日本育種学会   和文誌編集事務局  

    2018.4 - 2020.3   

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    Committee type:Academic society

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  • ナショナルバイオリソースプロジェクト「オオムギ」   運営委員  

    2012.4   

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Papers

  • FLOURY ENDOSPERM 6 mutations enhance the sugary phenotype caused by the loss of ISOAMYLASE1 in barley Reviewed

    Ryo Matsushima, Hiroshi Hisano, Ivan Galis, Satoko Miura, Naoko Crofts, Yuto Takenaka, Naoko F. Oitome, Takeshi Ishimizu, Naoko Fujita, Kazuhiro Sato

    Theoretical and Applied Genetics   136 ( 4 )   2023.4

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    Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Key message

    Barley double mutants in two genes involved in starch granule morphology, HvFLO6 and HvISA1, had impaired starch accumulation and higher grain sugar levels than either single mutant.

    Abstract

    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.

    DOI: 10.1007/s00122-023-04339-5

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    Other Link: https://link.springer.com/article/10.1007/s00122-023-04339-5/fulltext.html

  • Discovery and Genome Characterization of a Closterovirus from Wheat Plants with Yellowing Leaf Symptoms in Japan Invited Reviewed

    Hideki Kondo, Hitomi Sugahara, Miki Fujita, Kiwamu Hyodo, Ida Bagus Andika, Hiroshi Hisano, Nobuhiro Suzuki

    Pathogens   12 ( 3 )   358 - 358   2023.2

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    Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    Many aphid-borne viruses are important pathogens that affect wheat crops worldwide. An aphid-transmitted closterovirus named wheat yellow leaf virus (WYLV) was found to have infected wheat plants in Japan in the 1970s; however, since then, its viral genome sequence and occurrence in the field have not been investigated. We observed yellowing leaves in the 2018/2019 winter wheat-growing season in an experimental field in Japan where WYLV was detected five decades ago. A virome analysis of those yellow leaf samples lead to the discovery of a closterovirus together with a luteovirus (barley yellow dwarf virus PAV variant IIIa). The complete genomic sequence of this closterovirus, named wheat closterovirus 1 isolate WL19a (WhCV1-WL19a), consisted of 15,452 nucleotides harboring nine open reading frames. Additionally, we identified another WhCV1 isolate, WL20, in a wheat sample from the winter wheat-growing season of 2019/2020. A transmission test indicated that WhCV1-WL20 was able to form typical filamentous particles and transmissible by oat bird-cherry aphid (Rhopalosiphum pad). Sequence and phylogenetic analyses showed that WhCV1 was distantly related to members of the genus Closterovirus (family Closteroviridae), suggesting that the virus represents a novel species in the genus. Furthermore, the characterization of WhCV1-WL19a-derived small RNAs using high-throughput sequencing revealed highly abundant 22-nt-class small RNAs potentially derived from the 3′-terminal end of the WhCV1 negative-strand genomic RNA, indicating that this terminal end of the WhCV1 genome is likely particularly targeted for the synthesis of viral small RNAs in wheat plants. Our results provide further knowledge on closterovirus diversity and pathogenicity and suggest that the impact of WhCV1 on wheat production warrants further investigations.

    DOI: 10.3390/pathogens12030358

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  • CRISPR/Cas9-based generation of mlo mutants for allelic complementation experiments to elucidate MLO function in barley Reviewed

    Hina Koide, Hiroshi Hisano, Takashi Yaeno

    Journal of General Plant Pathology   2023

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    Publishing type:Research paper (scientific journal)  

    Barley (Hordeum vulgare) mildew locus o (mlo) mutants exhibit strong resistance to penetration by the powdery mildew fungus Blumeria graminis f. sp. hordei. MLO, a seven-transmembrane protein localized at the plasma membrane is thought to be involved in intracellular calcium signaling. However, its molecular function and the mechanism by which mlo mutations confer resistance to penetration by the fungus remain poorly understood. A large number of mlo alleles with different amino acid substitutions at each intracellular loop have been found in various cultivars. However, it is difficult to analyze how each amino acid is involved in penetration resistance by comparing these cultivars because they differ substantially in their genetic background and in the presence or absence of resistance genes recognizing avirulence factors from the pathogen. In this study, we used a CRISPR/Cas9-mediated genetic modification system to generate mlo mutants in the transformable cultivar Golden Promise to enable complementation experiments with the aim of elucidating the molecular function of MLO. An mlo mutant with a thymine insertion in the second exon and penetration resistance to B. graminis f. sp. hordei was obtained. Susceptibility was restored in cells in which the MLO-mCherry gene was introduced using particle bombardment, indicating that this mlo mutant could be a useful genetic tool for complementation experiments using transgenes expressing a variety of mlo alleles.

    DOI: 10.1007/s10327-023-01120-w

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  • Optimizing genome editing efficiency in wheat: Effects of heat treatments and different promoters for single guide RNA expression

    Mitsuko Kishi-Kaboshi, Fumitaka Abe, Yoko Kamiya, Kanako Kawaura, Hiroshi Hisano, Kazuhiro Sato

    Plant Biotechnology   40 ( 3 )   237 - 245   2023

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    Publishing type:Research paper (scientific journal)   Publisher:Japanese Society for Plant Cell and Molecular Biology  

    DOI: 10.5511/plantbiotechnology.23.0717a

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  • A bifurcated palea mutant infers functional differentiation of WOX3 genes in flower and leaf morphogenesis of barley Reviewed

    Takanori Yoshikawa, Hiroshi Hisano, Ken-Ichiro Hibara, Jilu Nie, Yuki Tanaka, Jun-Ichi Itoh, Shin Taketa

    AoB PLANTS   14 ( 3 )   2022.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

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    Barley (Hordeum vulgare) is the fourth most highly produced cereal in the world after wheat, rice and maize and is mainly utilized as malts and for animal feed. Barley, a model crop of the tribe Triticeae, is important in comparative analyses of Poaceae. However, molecular understanding about the developmental processes is limited in barley. Our previous work characterized one of two WUSCHEL-RELATED HOMEOBOX 3 (WOX3) genes present in the barley genome: NARROW LEAFED DWARF1 (NLD1). We demonstrated that NLD1 plays a pivotal role in the development of lateral organs. In the present study, we describe a bifurcated palea (bip) mutant of barley focusing on flower and leaf phenotypes. The palea in the bip mutant was split into two and develop towards inside the lemma surrounding the carpels and anthers. The bip mutant is devoid of lodicules, which develop in a pair at the base of the stamen within the lemma in normal barley. bip also exhibited malformations in leaves, such as narrow leaf due to underdeveloped leaf-blade width, and reduced trichome density. Map-based cloning and expression analysis indicated that BIP is identical to another barley WOX3 gene, named HvWOX3. The bip nld1 double mutant presented a more severe reduction in leaf-blade width and number of trichomes. By comparing the phenotypes and gene expression patterns of various WOX3 mutants, we concluded that leaf bilateral outgrowth and trichome development are promoted by both NLD1 and HvWOX3, but that HvWOX3 serves unique and pivotal functions in barley development that differ from those of NLD1.

    DOI: 10.1093/aobpla/plac019

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    Other Link: https://academic.oup.com/aobpla/article-pdf/14/3/plac019/43934060/plac019.pdf

  • A crucial role for a node‐localized transporter, HvSPDT, in loading phosphorus into barley grains

    Mian Gu, Hengliang Huang, Hiroshi Hisano, Guangda Ding, Sheng Huang, Namiki Mitani‐Ueno, Kengo Yokosho, Kazuhiro Sato, Naoki Yamaji, Jian Feng Ma

    New Phytologist   234 ( 4 )   1249 - 1261   2022.3

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    Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1111/nph.18057

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/nph.18057

  • Regulation of germination by targeted mutagenesis of grain dormancy genes in barley. Reviewed International journal

    Hiroshi Hisano, Robert E Hoffie, Fumitaka Abe, Hiromi Munemori, Takakazu Matsuura, Masaki Endo, Masafumi Mikami, Shingo Nakamura, Jochen Kumlehn, Kazuhiro Sato

    Plant biotechnology journal   20 ( 1 )   37 - 46   2022.1

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    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.

    DOI: 10.1111/pbi.13692

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pbi.13692

  • Identification of a Novel Quinvirus in the Family Betaflexiviridae That Infects Winter Wheat. Reviewed International journal

    Hideki Kondo, Naoto Yoshida, Miki Fujita, Kazuyuki Maruyama, Kiwamu Hyodo, Hiroshi Hisano, Tetsuo Tamada, Ida Bagus Andika, Nobuhiro Suzuki

    Frontiers in microbiology   12   715545 - 715545   2021.8

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    Yellow mosaic disease in winter wheat is usually attributed to the infection by bymoviruses or furoviruses; however, there is still limited information on whether other viral agents are also associated with this disease. To investigate the wheat viromes associated with yellow mosaic disease, we carried out de novo RNA sequencing (RNA-seq) analyses of symptomatic and asymptomatic wheat-leaf samples obtained from a field in Hokkaido, Japan, in 2018 and 2019. The analyses revealed the infection by a novel betaflexivirus, which tentatively named wheat virus Q (WVQ), together with wheat yellow mosaic virus (WYMV, a bymovirus) and northern cereal mosaic virus (a cytorhabdovirus). Basic local alignment search tool (BLAST) analyses showed that the WVQ strains (of which there are at least three) were related to the members of the genus Foveavirus in the subfamily Quinvirinae (family Betaflexiviridae). In the phylogenetic tree, they form a clade distant from that of the foveaviruses, suggesting that WVQ is a member of a novel genus in the Quinvirinae. Laboratory tests confirmed that WVQ, like WYMV, is potentially transmitted through the soil to wheat plants. WVQ was also found to infect rye plants grown in the same field. Moreover, WVQ-derived small interfering RNAs accumulated in the infected wheat plants, indicating that WVQ infection induces antiviral RNA silencing responses. Given its common coexistence with WYMV, the impact of WVQ infection on yellow mosaic disease in the field warrants detailed investigation.

    DOI: 10.3389/fmicb.2021.715545

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  • In planta Genome Editing in Commercial Wheat Varieties Reviewed

    Yuelin Liu, Weifeng Luo, Qianyan Linghu, Fumitaka Abe, Hiroshi Hisano, Kazuhiro Sato, Yoko Kamiya, Kanako Kawaura, Kazumitsu Onishi, Masaki Endo, Seiichi Toki, Haruyasu Hamada, Yozo Nagira, Naoaki Taoka, Ryozo Imai

    Frontiers in Plant Science   12   2021.3

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    Publishing type:Research paper (scientific journal)   Publisher:Frontiers Media SA  

    Limitations for the application of genome editing technologies on elite wheat (<italic>Triticum aestivum</italic> L.) varieties are mainly due to the dependency on <italic>in vitro</italic> culture and regeneration capabilities. Recently, we developed an <italic>in planta</italic> particle bombardment (iPB) method which has increased process efficiency since no culture steps are required to create stably genome-edited wheat plants. Here, we report the application of the iPB method to commercially relevant Japanese elite wheat varieties. The biolistic delivery of gold particles coated with plasmids expressing CRISPR/Cas9 components designed to target <italic>TaQsd1</italic> were bombarded into the embryos of imbibed seeds with their shoot apical meristem (SAM) exposed. Mutations in the target gene were subsequently analyzed within flag leaf tissue by using cleaved amplified polymorphic sequence (CAPS) analysis. A total of 9/358 (2.51%) of the bombarded plants (cv. “Haruyokoi,” spring type) carried mutant alleles in the tissue. Due to the chimeric nature of the T0 plants, only six of them were inherited to the next (T1) generation. Genotypic analysis of the T2 plants revealed a single triple-recessive homozygous mutant of the <italic>TaQsd1</italic> gene. Compared to wild type, the homozygous mutant exhibited a 7 days delay in the time required for 50% seed germination. The iPB method was also applied to two elite winter cultivars, “Yumechikara” and “Kitanokaori,” which resulted in successful genome editing at slightly lower efficiencies as compared to “Haruyokoi.” Taken together, this report demonstrates that the <italic>in planta</italic> genome editing method through SAM bombardment can be applicable to elite wheat varieties that are otherwise reluctant to callus culture.

    DOI: 10.3389/fpls.2021.648841

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  • Targeted genome modifications in cereal crops Invited Reviewed International coauthorship International journal

    Hiroshi Hisano, Fumitaka Abe, Robert E. Hoffie, Jochen Kumlehn

    Breeding Science   71 ( 4 )   405 - 416   2021

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japanese Society of Breeding  

    The recent advent of customizable endonucleases has led to remarkable advances in genetic engineering, as these molecular scissors allow for the targeted introduction of mutations or even precisely predefined genetic modifications into virtually any genomic target site of choice. Thanks to its unprecedented precision, efficiency, and functional versatility, this technology, commonly referred to as genome editing, has become an effective force not only in basic research devoted to the elucidation of gene function, but also for knowledge-based improvement of crop traits. Among the different platforms currently available for site-directed genome modifications, RNA-guided clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) endonucleases have proven to be the most powerful. This review provides an application-oriented overview of the development of customizable endonucleases, current approaches to cereal crop breeding, and future opportunities in this field.

    DOI: 10.1270/jsbbs.21019

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  • RACE1, a Japanese Blumeria graminis f. sp. hordei isolate, is capable of overcoming partially mlo-mediated penetration resistance in barley in an allele-specific manner. Reviewed International journal

    Takashi Yaeno, Miki Wahara, Mai Nagano, Hikaru Wanezaki, Hirotaka Toda, Hiroshi Inoue, Ayaka Eishima, Masamichi Nishiguchi, Hiroshi Hisano, Kappei Kobayashi, Kazuhiro Sato, Naoto Yamaoka

    PloS one   16 ( 8 )   e0256574   2021

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    Loss-of-function mutation of the MILDEW RESISTANCE LOCUS O (Mlo) gene confers durable and broad-spectrum resistance to powdery mildew fungi in various plants, including barley. In combination with the intracellular nucleotide-binding domain and leucine-rich repeat receptor (NLR) genes, which confer the race-specific resistance, the mlo alleles have long been used in barley breeding as genetic resources that confer robust non-race-specific resistance. However, a Japanese Blumeria graminis f. sp. hordei isolate, RACE1, has been reported to have the potential to overcome partially the mlo-mediated penetration resistance, although this is yet uncertain because the putative effects of NLR genes in the tested accessions have not been ruled out. In this study, we examined the reproducibility of the earlier report and found that the infectious ability of RACE1, which partially overcomes the mlo-mediated resistance, is only exerted in the absence of NLR genes recognizing RACE1. Furthermore, using the transient-induced gene silencing technique, we demonstrated that RACE1 can partially overcome the resistance in the host cells with suppressed MLO expression but not in plants possessing the null mutant allele mlo-5.

    DOI: 10.1371/journal.pone.0256574

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  • Site-Directed Mutagenesis in Barley Using RNA-Guided Cas Endonucleases During Microspore-Derived Generation of Doubled Haploids

    Hoffie, R.E., Otto, I., Hisano, H., Kumlehn, J.

    Methods in Molecular Biology   2287   2021

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    Publishing type:Research paper (scientific journal)  

    DOI: 10.1007/978-1-0716-1315-3_9

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  • Antagonistic regulation of the gibberellic acid response during stem growth in rice. Reviewed International journal

    Keisuke Nagai, Yoshinao Mori, Shin Ishikawa, Tomoyuki Furuta, Rico Gamuyao, Yoko Niimi, Tokunori Hobo, Moyuri Fukuda, Mikiko Kojima, Yumiko Takebayashi, Atsushi Fukushima, Yasuyo Himuro, Masatomo Kobayashi, Wataru Ackley, Hiroshi Hisano, Kazuhiro Sato, Aya Yoshida, Jianzhong Wu, Hitoshi Sakakibara, Yutaka Sato, Hiroyuki Tsuji, Takashi Akagi, Motoyuki Ashikari

    Nature   584 ( 7819 )   109 - 114   2020.8

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    The size of plants is largely determined by growth of the stem. Stem elongation is stimulated by gibberellic acid1-3. Here we show that internode stem elongation in rice is regulated antagonistically by an 'accelerator' and a 'decelerator' in concert with gibberellic acid. Expression of a gene we name ACCELERATOR OF INTERNODE ELONGATION 1 (ACE1), which encodes a protein of unknown function, confers cells of the intercalary meristematic region with the competence for cell division, leading to internode elongation in the presence of gibberellic acid. By contrast, upregulation of DECELERATOR OF INTERNODE ELONGATION 1 (DEC1), which encodes a zinc-finger transcription factor, suppresses internode elongation, whereas downregulation of DEC1 allows internode elongation. We also show that the mechanism of internode elongation that is mediated by ACE1 and DEC1 is conserved in the Gramineae family. Furthermore, an analysis of genetic diversity suggests that mutations in ACE1 and DEC1 have historically contributed to the selection of shorter plants in domesticated populations of rice to increase their resistance to lodging, and of taller plants in wild species of rice for adaptation to growth in deep water. Our identification of these antagonistic regulatory factors enhances our understanding of the gibberellic acid response as an additional mechanism that regulates internode elongation and environmental fitness, beyond biosynthesis and gibberellic acid signal transduction.

    DOI: 10.1038/s41586-020-2501-8

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  • Breeding for low cadmium barley by introgression of a Sukkula-like transposable element Reviewed

    Gui Jie Lei, Miho Fujii-Kashino, De Zhi Wu, Hiroshi Hisano, Daisuke Saisho, Fenglin Deng, Naoki Yamaji, Kazuhiro Sato, Fang-Jie Zhao, Jian Feng Ma

    Nature Food   1 ( 8 )   489 - 499   2020.8

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    Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media {LLC}  

    DOI: 10.1038/s43016-020-0130-x

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    Other Link: http://www.nature.com/articles/s43016-020-0130-x

  • Protocol for Genome Editing to Produce Multiple Mutants in Wheat Reviewed International journal

    Fumitaka Abe, Yuji Ishida, Hiroshi Hisano, Masaki Endo, Toshihiko Komari, Seiichi Toki, Kazuhiro Sato

    STAR Protocols   1 ( 2 )   100053 - 100053   2020.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    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).

    DOI: 10.1016/j.xpro.2020.100053

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  • Virome Analysis of Aphid Populations That Infest the Barley Field: The Discovery of Two Novel Groups of Nege/Kita-Like Viruses and Other Novel RNA Viruses. Reviewed International journal

    Hideki Kondo, Miki Fujita, Hiroshi Hisano, Kiwamu Hyodo, Ida Bagus Andika, Nobuhiro Suzuki

    Frontiers in microbiology   11   509 - 509   2020

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    Aphids (order Hemiptera) are important insect pests of crops and are also vectors of many plant viruses. However, little is known about aphid-infecting viruses, particularly their diversity and relationship to plant viruses. To investigate the aphid viromes, we performed deep sequencing analyses of the aphid transcriptomes from infested barley plants in a field in Japan. We discovered virus-like sequences related to nege/kita-, flavi-, tombus-, phenui-, mononega-, narna-, chryso-, partiti-, and luteoviruses. Using RT-PCR and sequence analyses, we determined almost complete sequences of seven nege/kitavirus-like virus genomes; one of which was a variant of the Wuhan house centipede virus (WHCV-1). The other six seem to belong to four novel viruses distantly related to Wuhan insect virus 9 (WhIV-9) or Hubei nege-like virus 4 (HVLV-4). We designated the four viruses as barley aphid RNA virus 1 to 4 (BARV-1 to -4). Moreover, some nege/kitavirus-like sequences were found by searches on the transcriptome shotgun assembly (TSA) libraries of arthropods and plants. Phylogenetic analyses showed that BARV-1 forms a clade with WHCV-1 and HVLV-4, whereas BARV-2 to -4 clustered with WhIV-9 and an aphid virus, Aphis glycines virus 3. Both virus groups (tentatively designated as Centivirus and Aphiglyvirus, respectively), together with arthropod virus-like TSAs, fill the phylogenetic gaps between the negeviruses and kitaviruses lineages. We also characterized the flavi/jingmen-like and tombus-like virus sequences as well as other RNA viruses, including six putative novel viruses, designated as barley aphid RNA viruses 5 to 10. Interestingly, we also discovered that some aphid-associated viruses, including nege/kita-like viruses, were present in different aphid species, raising a speculation that these viruses might be distributed across different aphid species with plants being the reservoirs. This study provides novel information on the diversity and spread of nege/kitavirus-related viruses and other RNA viruses that are associated with aphids.

    DOI: 10.3389/fmicb.2020.00509

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  • Genome-Edited Triple-Recessive Mutation Alters Seed Dormancy in Wheat. Reviewed International journal

    Fumitaka Abe, Emdadul Haque, Hiroshi Hisano, Tsuyoshi Tanaka, Yoko Kamiya, Masafumi Mikami, Kanako Kawaura, Masaki Endo, Kazumitsu Onishi, Takeshi Hayashi, Kazuhiro Sato

    Cell reports   28 ( 5 )   1362 - 1369   2019.7

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    Common 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.

    DOI: 10.1016/j.celrep.2019.06.090

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  • Imaging Amyloplasts in the Developing Endosperm of Barley and Rice Reviewed

    Ryo Matsushima, Hiroshi Hisano

    Scientific Reports   9   3745   2019.3

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    Authorship:Last author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/s41598-019-40424-w

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  • Agrobacterium tumefaciens Enhances Biosynthesis of Two Distinct Auxins in the Formation of Crown Galls. Reviewed

    Kiyoshi Mashiguchi, Hiroshi Hisano, Noriko Takeda-Kamiya, Yumiko Takebayashi, Tohru Ariizumi, Yangbin Gao, Hiroshi Ezura, Kazuhiro Sato, Yunde Zhao, Ken-Ichiro Hayashi, Hiroyuki Kasahara

    Plant & cell physiology   60 ( 1 )   29 - 37   2019.1

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    The plant pathogen Agrobacterium tumefaciens infects plants and introduces the transferred-DNA (T-DNA) region of the Ti-plasmid into nuclear DNA of host plants to induce the formation of tumors (crown galls). The T-DNA region carries iaaM and iaaH genes for synthesis of the plant hormone auxin, indole-3-acetic acid (IAA). It has been demonstrated that the iaaM gene encodes a tryptophan 2-monooxygenase which catalyzes the conversion of tryptophan to indole-3-acetamide (IAM), and the iaaH gene encodes an amidase for subsequent conversion of IAM to IAA. In this article, we demonstrate that A. tumefaciens enhances the production of both IAA and phenylacetic acid (PAA), another auxin which does not show polar transport characteristics, in the formation of crown galls. Using liquid chromatography-tandem mass spectroscopy, we found that the endogenous levels of phenylacetamide (PAM) and PAA metabolites, as well as IAM and IAA metabolites, are remarkably increased in crown galls formed on the stem of tomato plants, implying that two distinct auxins are simultaneously synthesized via the IaaM-IaaH pathway. Moreover, we found that the induction of the iaaM gene dramatically elevated the levels of PAM, PAA and its metabolites, along with IAM, IAA and its metabolites, in Arabidopsis and barley. From these results, we conclude that A. tumefaciens enhances biosynthesis of two distinct auxins in the formation of crown galls.

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  • QTLs maintaining grain fertility under salt stress detected by exome QTL-seq and interval mapping in barley. Reviewed

    Asuka Kodama, Ryouhei Narita, Makoto Yamaguchi, Hiroshi Hisano, Shunsuke Adachi, Hiroki Takagi, Taiichiro Ookawa, Kazuhiro Sato, Tadashi Hirasawa

    Breeding science   68 ( 5 )   561 - 570   2018.12

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    Enhancing salt stress tolerance is a key strategy for increasing global food production. We previously found that long-term salinity stress significantly reduced grain fertility in the salt-sensitive barley (Hordeum vulgare) accession, 'OUC613', but not in the salt-tolerant accession, 'OUE812', resulting in large differences in grain yield. Here, we examined the underlying causes of the difference in grain fertility between these accessions under long-term treatment with 150 or 200 mM NaCl from the seedling stage to harvest and identified quantitative trait loci (QTLs) for maintaining grain fertility. In an artificial pollination experiment of the two accessions, grain fertility was significantly reduced only in OUC613 plants produced using pollen from plants grown under NaCl stress, suggesting that the low grain fertility of OUC613 was mainly due to reduced pollen fertility. Using QTL-seq combined with exome-capture sequencing and composite interval mapping of recombinant inbred lines derived from a cross between OUE812 and OUC613, we identified a QTL (qRP-2Hb) for grain fertility on chromosome 2H. The QTL region includes two genes encoding an F-box protein and a TIFY protein that are associated with male sterility, highlighting the importance of this region for maintaining grain fertility under salt stress.

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  • Simultaneous regulation of F5H in COMT-RNAi transgenic switchgrass alters effects of COMT suppression on syringyl lignin biosynthesis. Reviewed

    Wu Z, Wang N, Hisano H, Cao Y, Wu F, Liu W, Bao Y, Wang ZY, Fu C

    Plant biotechnology journal   17   836 - 845   2018.10

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  • Detection of QTLs controlling alpha-amylase activity in a diversity panel of 343 barley accessions Reviewed

    Kazuhiro Sato, Hiroshi Hisano, Satoko Matsumoto, Tian-Su Zhou, Makoto Kihara

    Molecular Breeding   38 ( 1 )   14   2018.1

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    The α–amylase activity of cultivated barley is critically important to the brewing industry. Here, we surveyed variation in malt α–amylase activity in 343 cultivated barley accessions from around the world. Population structure analysis based on genotype data at 1536 SNPs clustered these accessions into two groups, one comprising South-East Asian and Ethiopian accessions and one group containing the other accessions. A genome-wide association study identified significant quantitative trait loci (QTLs) for α–amylase activity on all seven chromosomes of barley. Accessions showing high and low α–amylase activity were crossed with the high-quality Japanese malting barley cv. Harun Nijo to develop F2 mapping populations. We identified two QTLs on chromosome 6H in a cross between Haruna Nijo (high activity) × Weal (highest activity). Single QTLs were identified each on 3H, 4H, and 5H from a cross between Haruna Nijo (high activity) × VLB-1 (low activity), indicating that the high α–amylase activity in Haruna Nijo might be derived from loci on these chromosomes. The addition of the high α–amylase activity QTL alleles from chromosome 6H in cv. Weal further increased the α–amylase activity conferred by alleles of Haruna Nijo. These results demonstrate that a target haplotype can be successfully improved using a strategy comprising diversity analysis of ex situ collections followed by introducing effective new alleles.

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  • Development and use of a switchgrass (Panicum virgatum L.) transformation pipeline by the BioEnergy Science Center to evaluate plants for reduced cell wall recalcitrance Reviewed

    Richard S. Nelson, C. Neal Stewart, Jiqing Gou, Susan Holladay, Lina Gallego-Giraldo, Amy Flanagan, David G. J. Mann, Hiroshi Hisano, Wegi A. Wuddineh, Charleson R. Poovaiah, Avinash Srivastava, Ajaya K. Biswal, Hui Shen, Luis L. Escamilla-Trevino, Jiading Yang, C. Frank Hardin, Rangaraj Nandakumar, Chunxiang Fu, Jiyi Zhang, Xirong Xiao, Ryan Percifield, Fang Chen, Jeffrey L. Bennetzen, Michael Udvardi, Mitra Mazarei, Richard A. Dixon, Zeng-Yu Wang, Yuhong Tang, Debra Mohnen, Brian H. Davison

    BIOTECHNOLOGY FOR BIOFUELS   10   309   2017.12

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    Background: The mission of the BioEnergy Science Center (BESC) was to enable efficient lignocellulosic-based biofuel production. One BESC goal was to decrease poplar and switchgrass biomass recalcitrance to biofuel conversion while not affecting plant growth. A transformation pipeline (TP), to express transgenes or transgene fragments (constructs) in these feedstocks with the goal of understanding and decreasing recalcitrance, was considered essential for this goal. Centralized data storage for access by BESC members and later the public also was essential.
    Results: A BESC committee was established to codify procedures to evaluate and accept genes into the TP. A laboratory information management system (LIMS) was organized to catalog constructs, plant lines and results from their analyses. One hundred twenty-eight constructs were accepted into the TP for expression in switchgrass in the first 5 years of BESC. Here we provide information on 53 of these constructs and the BESC TP process. Eleven of the constructs could not be cloned into an expression vector for transformation. Of the remaining constructs, 22 modified expression of the gene target. Transgenic lines representing some constructs displayed decreased recalcitrance in the field and publications describing these results are tabulated here. Transcript levels of target genes and detailed wall analyses from transgenic lines expressing six additional tabulated constructs aimed toward modifying expression of genes associated with wall structure (xyloglucan and lignin components) are provided. Altered expression of xyloglucan endotransglucosylase/hydrolases did not modify lignin content in transgenic plants. Simultaneous silencing of two hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferases was necessary to decrease G and S lignin monomer and total lignin contents, but this reduced plant growth.
    Conclusions: A TP to produce plants with decreased recalcitrance and a LIMS for data compilation from these plants were created. While many genes accepted into the TP resulted in transgenic switchgrass without modified lignin or biomass content, a group of genes with potential to improve lignocellulosic biofuel yields was identified. Results from transgenic lines targeting xyloglucan and lignin structure provide examples of the types of information available on switchgrass lines produced within BESC. This report supplies useful information when developing coordinated, largescale, multi-institutional reverse genetic pipelines to improve crop traits.

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  • Global profiling of phytohormone dynamics during combined drought and pathogen stress in Arabidopsis thaliana reveals ABA and JA as major regulators Reviewed

    Aarti Gupta, Hiroshi Hisano, Yuko Hojo, Takakazu Matsuura, Yoko Ikeda, Izumi C. Mori, Muthappa Senthil-Kumar

    SCIENTIFIC REPORTS   7 ( 4017 )   2017.6

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    Global transcriptome studies demonstrated the existence of unique plant responses under combined stress which are otherwise not seen during individual stresses. In order to combat combined stress plants use signaling pathways and 'cross talk' mediated by hormones involved in stress and growth related processes. However, interactions among hormones' pathways in combined stressed plants are not yet known. Here we studied dynamics of different hormones under individual and combined drought and pathogen infection in Arabidopsis thaliana by liquid chromatography-mass spectrometry (LC-MS) based profiling. Our results revealed abscisic acid (ABA) and salicylic acid (SA) as key regulators under individual drought and pathogen stress respectively. Under combined drought and host pathogen stress (DH) we observed non-induced levels of ABA with an upsurge in SA and jasmonic acid (JA) concentrations, underscoring their role in basal tolerance against host pathogen. Under a non-host pathogen interaction with drought (DNH) stressed plants, ABA, SA and JA profiles were similar to those under DH or non-host pathogen alone. We propose that plants use SA/JA dependent signaling during DH stress which antagonize ABA biosynthesis and signaling pathways during early stage of stress. The study provides insights into hormone modulation at different time points during combined stress.

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  • Selection of transformation-efficient barley genotypes based on TFA (transformation amenability) haplotype and higher resolution mapping of the TFA loci. Reviewed International journal

    Hiroshi Hisano, Brigid Meints, Matthew J Moscou, Luis Cistue, Begoña Echávarri, Kazuhiro Sato, Patrick M Hayes

    Plant cell reports   36 ( 4 )   611 - 620   2017.4

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    KEY MESSAGE: The genetic substitution of transformation amenability alleles from 'Golden Promise' can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars. 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.

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  • Exome QTL-seq maps monogenic locus and QTLs in barley. Reviewed International journal

    Hiroshi Hisano, Kazuki Sakamoto, Hiroki Takagi, Ryohei Terauchi, Kazuhiro Sato

    BMC genomics   18 ( 1 )   125 - 125   2017.2

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    BACKGROUND: QTL-seq, in combination with bulked segregant analysis and next-generation sequencing (NGS), is used to identify loci in small plant genomes, but is technically challenging to perform in species with large genomes, such as barley. A combination of exome sequencing and QTL-seq (exome QTL-seq) was used to map the mono-factorial Mendelian locus black lemma and pericarp (Blp) and QTLs for resistance to net blotch disease, a common disease of barley caused by the fungus Pyrenophora teres, which segregated in a population of 100 doubled haploid barley lines. METHODS: The provisional exome sequences were prepared by ordering the loci of expressed genes based on the genome information and concatenating genes with intervals of 200-bp spacer "N" for each chromosome. The QTL-seq pipeline was used to analyze short reads from the exome-captured library. RESULTS: In this study, short NGS reads of bulked total DNA samples from segregants with extreme trait values were subjected to exome capture, and the resulting exome sequences were aligned to the reference genome. SNP allele frequencies were compared to identify the locations of genes/QTLs responsible for the trait value differences between lines. For both objective traits examined, exome QTL-seq identified the monogenic Mendelian locus and associated QTLs. These findings were validated using conventional mapping approaches. CONCLUSIONS: Exome QTL-seq broadens the utility of NGS-based gene/QTL mapping in organisms with large genomes.

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  • Genomic regions responsible for amenability to Agrobacterium-mediated transformation in barley. Reviewed International journal

    Hiroshi Hisano, Kazuhiro Sato

    Scientific reports   6   37505 - 37505   2016.11

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    Different plant cultivars of the same genus and species can exhibit vastly different genetic transformation efficiencies. However, the genetic factors underlying these differences in transformation rate remain largely unknown. In barley, 'Golden Promise' is one of a few cultivars reliable for Agrobacterium-mediated transformation. By contrast, cultivar 'Haruna Nijo' is recalcitrant to genetic transformation. We identified genomic regions of barley important for successful transformation with Agrobacterium, utilizing the 'Haruna Nijo' × 'Golden Promise' F2 generation and genotyping by 124 genome-wide SNP markers. We observed significant segregation distortions of these markers from the expected 1:2:1 ratio toward the 'Golden Promise'-type in regions of chromosomes 2H and 3H, indicating that the alleles of 'Golden Promise' in these regions might contribute to transformation efficiency. The same regions, which we termed Transformation Amenability (TFA) regions, were also conserved in transgenic F2 plants generated from a 'Morex' × 'Golden Promise' cross. The genomic regions identified herein likely include necessary factors for Agrobacterium-mediated transformation in barley. The potential to introduce these loci into any haplotype of barley opens the door to increasing the efficiency of transformation for target alleles into any haplotype of barley by the TFA-based methods proposed in this report.

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  • Genetic analysis of the resistance of barley to cryptic species of Pyricularia Reviewed

    Analiza Grubanzo Tagle, Izumi Chuma, Hiroshi Hisano, Kazuhiro Sato, Yukio Tosa

    JOURNAL OF GENERAL PLANT PATHOLOGY   82 ( 6 )   302 - 306   2016.11

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    The resistance of barley to Pyricularia oryzae isolates is controlled by the Rmo2 locus irrespective of their original hosts. The resistance of barley cultivar H.E.S.4 to isolate tO-7 of P. pennisetigena (a cryptic species in the P. oryzae/grisea species complex) cosegregated with the resistance to P. oryzae isolate GFSI-1-7-2 controlled by Rmo2.a. On the other hand, its resistance to isolate NI919, belonging to another cryptic species of Pyricularia, was controlled by another gene inherited independently from Rmo2.a. These results suggest that gene-for-gene interactions underlie the resistance of barley to cryptic species of Pyricularia.

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  • Mitochondrial genome sequences from wild and cultivated barley (Hordeum vulgare). Reviewed International journal

    Hiroshi Hisano, Mai Tsujimura, Hideya Yoshida, Toru Terachi, Kazuhiro Sato

    BMC genomics   17 ( 1 )   824 - 824   2016.10

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    BACKGROUND: Sequencing analysis of mitochondrial genomes is important for understanding the evolution and genome structures of various plant species. Barley is a self-pollinated diploid plant with seven chromosomes comprising a large haploid genome of 5.1 Gbp. Wild barley (Hordeum vulgare ssp. spontaneum) and cultivated barley (H. vulgare ssp. vulgare) have cross compatibility and closely related genomes, although a significant number of nucleotide polymorphisms have been reported between their genomes. RESULTS: We determined the complete nucleotide sequences of the mitochondrial genomes of wild and cultivated barley. Two independent circular maps of the 525,599 bp barley mitochondrial genome were constructed by de novo assembly of high-throughput sequencing reads of barley lines H602 and Haruna Nijo, with only three SNPs detected between haplotypes. These mitochondrial genomes contained 33 protein-coding genes, three ribosomal RNAs, 16 transfer RNAs, 188 new ORFs, six major repeat sequences and several types of transposable elements. Of the barley mitochondrial genome-encoded proteins, NAD6, NAD9 and RPS4 had unique structures among grass species. CONCLUSIONS: The mitochondrial genome of barley was similar to those of other grass species in terms of gene content, but the configuration of the genes was highly differentiated from that of other grass species. Mitochondrial genome sequencing is essential for annotating the barley nuclear genome; our mitochondrial sequencing identified a significant number of fragmented mitochondrial sequences in the reported nuclear genome sequences. Little polymorphism was detected in the barley mitochondrial genome sequences, which should be explored further to elucidate the evolution of barley.

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  • Endogenous hormone levels affect the regeneration ability of callus derived from different organs in barley. Reviewed International journal

    Hiroshi Hisano, Takakazu Matsuura, Izumi C Mori, Miki Yamane, Kazuhiro Sato

    Plant physiology and biochemistry : PPB   99   66 - 72   2016.2

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    Hordeum vulgare (barley) is an important agricultural crop worldwide. A simple and efficient transformation system is needed to analyze the functions of barley genes and generate lines with improved agronomic traits. Currently, Golden Promise and Igri are the most amenable barley cultivars for stable transformation. Here we evaluated the regeneration ratios and endogenous hormone levels of calli derived from various malting barley cultivars, including Golden Promise, Haruna Nijo, and Morex. We harvested samples not only from immature embryos, but also from different explants of juvenile plants, cotyledons, coleoptiles, and roots. The callus properties differed among genotypes and explant types. Calli derived from the immature embryos of Golden Promise, which showed the highest ratio of regeneration of green shoots, had the highest contents of indoleacetic acid, trans-zeatin, and cis-zeatin. By contrast, calli derived from the cotyledons of Morex and the immature embryos of Haruna Nijo had elevated levels of salicylic acid and abscisic acid, respectively. We thus propose that the former phytohormones are positively associated with the regeneration ability of callus but the later phytohormones are negatively associated.

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  • A Genomics Approach to Deciphering Lignin Biosynthesis in Switchgrass Reviewed

    Hui Shen, Mitra Mazarei, Hiroshi Hisano, Luis Escamilla-Trevino, Chunxiang Fu, Yunqiao Pu, Mary R. Rudis, Yuhong Tang, Xirong Xiao, Lisa Jackson, Guifen Li, Tim Hernandez, Fang Chen, Arthur J. Ragauskas, C. Neal Stewart, Zeng-Yu Wang, Richard A. Dixon

    PLANT CELL   25 ( 11 )   4342 - 4361   2013.11

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    It is necessary to overcome recalcitrance of the biomass to saccharification (sugar release) to make switchgrass (Panicum virgatum) economically viable as a feedstock for liquid biofuels. Lignin content correlates negatively with sugar release efficiency in switchgrass, but selecting the right gene candidates for engineering lignin biosynthesis in this tetraploid outcrossing species is not straightforward. To assist this endeavor, we have used an inducible switchgrass cell suspension system for studying lignin biosynthesis in response to exogenous brassinolide. By applying a combination of protein sequence phylogeny with whole-genome microarray analyses of induced cell cultures and developing stem internode sections, we have generated a list of candidate monolignol biosynthetic genes for switchgrass. Several genes that were strongly supported through our bioinformatics analysis as involved in lignin biosynthesis were confirmed by gene silencing studies, in which lignin levels were reduced as a result of targeting a single gene. However, candidate genes encoding enzymes involved in the early steps of the currently accepted monolignol biosynthesis pathway in dicots may have functionally redundant paralogues in switchgrass and therefore require further evaluation. This work provides a blueprint and resources for the systematic genome-wide study of the monolignol pathway in switchgrass, as well as other C4 monocot species.

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  • Standardization of Switchgrass Sample Collection for Cell Wall and Biomass Trait Analysis Reviewed

    C. Frank Hardin, Chunxiang Fu, Hiroshi Hisano, Xirong Xiao, Hui Shen, C. Neal Stewart, Wayne Parrott, Richard A. Dixon, Zeng-Yu Wang

    BIOENERGY RESEARCH   6 ( 2 )   755 - 762   2013.6

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    As a native, low-input crop with high biomass production, switchgrass (Panicum virgatum) has become a favorable feedstock for the production of cellulosic biofuels in the United States. Many efforts are being made to improve the production of cellulosic biofuels from switchgrass. Protocols regarding analysis of switchgrass biomass have been established; however, the developmental stage of the materials being analyzed has varied depending on researchers' discretion, and no standardized harvesting procedure has been defined. Developmental stages have a large impact on the results of biochemical analyses. We propose a standardized procedure for switchgrass sample collection for cell wall and biomass analyses by describing various developmental stages of switchgrass, defining the R1 stage as the stage at which tillers should be collected, and providing a detailed description of how and what material should be analyzed. Such a standardized procedure will help to maintain consistency in switchgrass evaluation methods, enable comparisons of data obtained from different approaches and studies, and facilitate efforts towards improving switchgrass as a bioenergy crop.

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  • Comparative Genetic Mapping and Discovery of Linkage Disequilibrium Across Linkage Groups in White Clover (Trifolium repens L.) Reviewed

    Sachiko N. Isobe, Hiroshi Hisano, Shusei Sato, Hideki Hirakawa, Kenji Okumura, Kenta Shirasawa, Shigemi Sasamoto, Akiko Watanabe, Tsuyuko Wada, Yoshie Kishida, Hisano Tsuruoka, Tsunakazu Fujishiro, Manabu Yamada, Mistuyo Kohara, Satoshi Tabata

    G3-GENES GENOMES GENETICS   2 ( 5 )   607 - 617   2012.5

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    White clover (Trifolium repens L.) is an allotetraploid species (2n - 4X - 32) that is widely distributed in temperate regions and cultivated as a forage legume. In this study, we developed expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers, constructed linkage maps, and performed comparative mapping with other legume species. A total of 7982 ESTs that could be assembled into 5400 contigs and 2582 singletons were generated. Using the EST sequences that were obtained, 1973 primer pairs to amplify EST-derived SSR markers were designed and used for linkage analysis of 188 F-1 progenies, which were generated by a cross between two Japanese plants, '273-7' and 'T17-349,' with previously published SSR markers. An integrated linkage map was constructed by combining parental-specific maps, which consisted of 1743 SSR loci on 16 homeologous linkage groups with a total length of 2511 cM. The primer sequences of the developed EST-SSR markers and their map positions are available on http://clovergarden.jp/. Linkage disequilibrium (LD) was observed on 9 of 16 linkage groups of a parental-specific map. The genome structures were compared among white clover, red clover (T. pratense L.), Medicago truncatula, and Lotus japonicus. Macrosynteny was observed across the four legume species. Surprisingly, the comparative genome structure between white clover and M. truncatula had a higher degree of conservation than that of the two clover species.

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  • Agrobacterium-Mediated Transformation of Switchgrass and Inheritance of the Transgenes Reviewed

    Yajun Xi, Chunxiang Fu, Yaxin Ge, Rangaraj Nandakumar, Hiroshi Hisano, Joseph Bouton, Zeng-Yu Wang

    BIOENERGY RESEARCH   2 ( 4 )   275 - 283   2009.12

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    Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.

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  • High-density Integrated Linkage Map Based on SSR Markers in Soybean Reviewed

    Tae-Young Hwang, Takashi Sayama, Masakazu Takahashi, Yoshitake Takada, Yumi Nakamoto, Hideyuki Funatsuki, Hiroshi Hisano, Shigemi Sasamoto, Shusei Sato, Satoshi Tabata, Izumi Kono, Masako Hoshi, Masayoshi Hanawa, Chizuru Yano, Zhengjun Xia, Kyuya Harada, Keisuke Kitamura, Masao Ishimoto

    DNA RESEARCH   16 ( 4 )   213 - 225   2009.8

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    A well-saturated molecular linkage map is a prerequisite for modern plant breeding. Several genetic maps have been developed for soybean with various types of molecular markers. Simple sequence repeats (SSRs) are single-locus markers with high allelic variation and are widely applicable to different genotypes. We have now mapped 1810 SSR or sequence-tagged site markers in one or more of three recombinant inbred populations of soybean (the US cultivar &apos;Jack&apos; x the Japanese cultivar &apos;Fukuyutaka&apos;, the Chinese cultivar &apos;Peking&apos; x the Japanese cultivar &apos;Akita&apos;, and the Japanese cultivar &apos;Misuzudaizu&apos; x the Chinese breeding line &apos;Moshidou Gong 503&apos;) and have aligned these markers with the 20 consensus linkage groups (LGs). The total length of the integrated linkage map was 2442.9 cM, and the average number of molecular markers was 90.5 (range of 70-114) for the 20 LGs. We examined allelic diversity for 1238 of the SSR markers among 23 soybean cultivars or lines and a wild accession. The number of alleles per locus ranged from 2 to 7, with an average of 2.8. Our high-density linkage map should facilitate ongoing and future genomic research such as analysis of quantitative trait loci and positional cloning in addition to marker-assisted selection in soybean breeding.

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  • Genetic modification of lignin biosynthesis for improved biofuel production Invited Reviewed

    Hiroshi Hisano, Rangaraj Nandakumar, Zeng-Yu Wang

    In Vitro Cellular & Developmental Biology - Plant   45 ( 3 )   306 - 313   2009.6

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  • Construction of a consensus linkage map for red clover (Trifolium pratense L.) Reviewed

    Sachiko Isobe, Roland Koelliker, Hiroshi Hisano, Shigemi Sasamoto, Tshyuko Wada, Irina Klimenko, Kenji Okumura, Satoshi Tabata

    BMC PLANT BIOLOGY   9 ( 57 )   2009.5

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    Background: Red clover (Trifolium pratense L.) is a major forage legume that has a strong self-incompatibility system and exhibits high genetic diversity within populations. For several crop species, integrated consensus linkage maps that combine information from multiple mapping populations have been developed. For red clover, three genetic linkage maps have been published, but the information in these existing maps has not been integrated.
    Results: A consensus linkage map was constructed using six mapping populations originating from eight parental accessions. Three of the six mapping populations were established for this study. The integrated red clover map was composed of 1804 loci, including 1414 microsatellite loci, 181 amplified fragment length polymorphism (AFLP) loci and 204 restriction fragment length polymorphism (RFLP) loci, in seven linkage groups. The average distance between loci and the total length of the consensus map were 0.46 cM and 836.6 cM, respectively. The locus order on the consensus map correlated highly with that of accession-specific maps. Segregation distortion was observed across linkage groups. We investigated genome-wide allele frequency in 1144 red clover individuals using 462 microsatellite loci randomly chosen from the consensus map. The average number of alleles and polymorphism information content (PIC) were 9.17 and 0.69, respectively.
    Conclusion: A consensus genetic linkage map for red clover was constructed for the first time based on six mapping populations. The locus order on the consensus map was highly conserved among linkage maps and was sufficiently reliable for use as a reference for genetic analysis of random red clover germplasms.

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  • Development of intron-flanking EST markers for the Lolium/Festuca complex using rice genomic information Reviewed

    Ken-ichi Tamura, Jun-ichi Yonemaru, Hiroshi Hisano, Hiroyuki Kanamori, Julie King, Ian P. King, Kazuhiro Tase, Yasuharu Sanada, Toshinori Komatsu, Toshihiko Yamada

    THEORETICAL AND APPLIED GENETICS   118 ( 8 )   1549 - 1560   2009.5

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    DNA markers able to distinguish species or genera with high specificity are valuable in the identification of introgressed regions in interspecific or intergeneric hybrids. Intergeneric hybridization between the genera of Lolium and Festuca, leading to the reciprocal introgression of chromosomal segments, can produce novel forage grasses with unique combinations of characteristics. To characterize Lolium/Festuca introgressions, novel PCR-based expression sequence tag (EST) markers were developed. These markers were designed around intronic regions which show higher polymorphism than exonic regions. Intronic regions of the grass genes were predicted from the sequenced rice genome. Two hundred and nine primer sets were designed from Lolium/Festuca ESTs that showed high similarity to unique rice genes dispersed uniformly throughout the rice genome. We selected 61 of these primer sets as insertion-deletion (indel)-type markers and 82 primer sets as cleaved amplified polymorphic sequence (CAPS) markers to distinguish between Lolium perenne and Festuca pratensis. Specificity of these markers to each species was evaluated by the genotyping of four cultivars and accessions (32 individuals) of L. perenne and F. pratensis, respectively. Evaluation using specificity indices proposed in this study suggested that many indel-type markers had high species specificity to L. perenne and F. pratensis, including 15 markers completely specific to both species. Forty-nine of the CAPS markers completely distinguish between the two species at bulk level. Chromosome mapping of these markers using a Lolium/Festuca substitution line revealed syntenic relationships between Lolium/Festuca and rice largely consistent with previous reports. This intron-based marker system that shows a high level of polymorphisms between species in combination with high species specificity will consequently be a valuable tool in Festulolium breeding.

    DOI: 10.1007/s00122-009-1003-8

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  • Transformation of androgenic-derived Festulolium plants (Lolium perenne L. x Festuca pratensis Huds.) by Agrobacterium tumefaciens Reviewed

    Yang-Dong Guo, Hiroshi Hisano, Yoshiya Shimamoto, Toshihiko Yamada

    PLANT CELL TISSUE AND ORGAN CULTURE   96 ( 2 )   219 - 227   2009.2

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    Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. x Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium "Bx351" were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and beta-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.

    DOI: 10.1007/s11240-008-9478-6

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  • Coordinated expression of functionally diverse fructosyltransferase genes is associated with fructan accumulation in response to low temperature in perennial ryegrass Reviewed

    Hiroshi Hisano, Akira Kanazawa, Midori Yoshida, Mervyn O. Humphreys, Masaru Iizuka, Keisuke Kitamura, Toshihiko Yamada

    NEW PHYTOLOGIST   178 ( 4 )   766 - 780   2008

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    Fructan is the major nonstructural carbohydrate reserve in temperate grasses. To understand regulatory mechanisms in fructan synthesis and adaptation to cold environments, the isolation, functional characterization and genetic mapping of fructosyltransferase (FT) genes in perennial ryegrass (Lolium perenne) are described.
    Six cDNAs (prft1-prft6) encoding FTs were isolated from cold-treated ryegrass plants, and three were positioned on a perennial ryegrass linkage map. Recombinant proteins were produced in Pichia pastoris and enzymatic activity was characterized. Changes in carbohydrate levels and mRNA levels of FT genes during cold treatment were also analysed.
    One gene encodes sucrose-sucrose 1-fructosyltransferase (1-SST), and two gene encode fructan-fructan 6G-fructosyltransferase (6G-FFT). Protein sequences for the other genes (prfts 1, 2 and 6) were similar to sucrose-fructan 6-fructosyltransferase (6-SFT). The 1-SST and prft1 genes were colocalized with an invertase gene on the ryegrass linkage map. The mRNA levels of prft1 and prft2 increased gradually during cold treatment, while those of the 1-SST and 6G-FFT genes first increased, but then decreased before increasing again during a longer period of cold treatment.
    Thus at least two different patterns of gene expression have developed during the evolution of functionally diverse FT genes, which are associated in a coordinated way with fructan synthesis in a cold environment.

    DOI: 10.1111/j.1469-8137.2008.02409.x

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  • Characterization of the soybean genome using EST-derived microsatellite markers. Reviewed International journal

    Hiroshi Hisano, Shusei Sato, Sachiko Isobe, Shigemi Sasamoto, Tsuyuko Wada, Ai Matsuno, Tsunakazu Fujishiro, Manabu Yamada, Shinobu Nakayama, Yasukazu Nakamura, Satoshi Watanabe, Kyuya Harada, Satoshi Tabata

    DNA research : an international journal for rapid publication of reports on genes and genomes   14 ( 6 )   271 - 81   2007.12

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    We generated a high-density genetic linkage map of soybean using expressed sequence tag (EST)-derived microsatellite markers. A total of 6920 primer pairs (10.9%) were designed to amplify simple sequence repeats (SSRs) from 63,676 publicly available non-redundant soybean ESTs. The polymorphism of two parent plants, the Japanese cultivar 'Misuzudaizu' and the Chinese line 'Moshidou Gong 503', were examined using 10% polyacrylamide gel electrophoresis. Primer pairs showing polymorphism were then used for genotyping 94 recombinant inbred lines (RILs) derived from a cross between the parents. In addition to previously reported markers, 680 EST-derived microsatellite markers were selected and subjected to linkage analysis. As a result, 935 marker loci were mapped successfully onto 20 linkage groups, which totaled 2700.3 cM in length; 693 loci were detected using the 668 EST-derived microsatellite markers developed in this study, the other 242 loci were detected with 105 RFLP markers, 136 genome-derived microsatellite markers, and one phenotypic marker. We examined allelic variation among 23 soybean cultivars/lines and a wild soybean line using 668 mapped EST-derived microsatellite markers (corresponding to 686 marker loci), in order to determine the transferability of the markers among soybean germplasms. A limited degree of macrosynteny was observed at the segmental level between the genomes of soybean and the model legume Lotus japonicus, which suggests that considerable genome shuffling occurred after separation of the species and during establishment of the paleopolyploid soybean genome.

    DOI: 10.1093/dnares/dsm025

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  • Gene expression and genetic mapping analyses of a perennial ryegrass glycine-rich RNA-binding protein gene suggest a role in cold adaptation Reviewed

    H Shinozuka, H Hisano, S Yoneyama, Y Shimamoto, ES Jones, JW Forster, T Yamada, A Kanazawa

    MOLECULAR GENETICS AND GENOMICS   275 ( 4 )   399 - 408   2006.4

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    A perennial ryegrass cDNA clone encoding a putative glycine-rich RNA binding protein (LpGRP1) was isolated from a cDNA library constructed from crown tissues of cold-treated plants. The deduced polypeptide sequence consists of 107 amino acids with a single N-terminal RNA recognition motif (RRM) and a single C-terminal glycine-rich domain. The sequence showed extensive homology to glycine-rich RNA binding proteins previously identified in other plant species. LpGRP1-specific genomic DNA sequence was isolated by an inverse PCR amplification. A single intron which shows conserved locations in plant genes was detected between the sequence motifs encoding RNP-1 and RNP-2 consensus protein domains. A significant increase in the mRNA level of LpGRP1 was detected in root, crown and leaf tissues during the treatment of plants at 4 degrees C, through which freezing tolerance is attained. The increase in the mRNA level was prominent at least 2 h after the commencement of the cold treatment, and persisted for at least 1 week. Changes in mRNA level induced by cold treatment were more obvious than those due to treatments with abscisic acid (ABA) and drought. The LpGRP1 protein was found to localise in the nucleus in onion epidermal cells, suggesting that it may be involved in pre-mRNA processing. The LpGRP1 gene locus was mapped to linkage group 2. Possible roles for the LpGRP1 protein in adaptation to cold environments are discussed.

    DOI: 10.1007/s00438-005-0095-3

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  • Molecular cloning and genetic mapping of perennial ryegrass casein protein kinase 2 alpha-subunit genes Reviewed

    H Shinozuka, H Hisano, RC Ponting, NOI Cogan, ES Jones, JW Forster, T Yamada

    THEORETICAL AND APPLIED GENETICS   112 ( 1 )   167 - 177   2005.12

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    The alpha-subunit of the casein protein kinase CK2 has been implicated in both light-regulated and circadian rhythm-controlled plant gene expression, including control of the flowering time. Two putative CK2 alpha genes of perennial ryegrass (Lolium perenne L.) have been obtained from a cDNA library constructed with mRNA isolated from cold-acclimated crown tissue. The genomic organisation of the two genes was determined by Southern hybridisation analysis. Primer designs to the Lpck2a-1 and Lpck2a-2 cDNA sequences permitted the amplification of genomic products containing large intron sequences. Amplicon sequence analysis detected single nucleotide polymorphisms (SNPs) within the p150/112 reference mapping population. Validated SNPs, within diagnostic restriction enzyme sites, were used to design cleaved amplified polymorphic sequence (CAPS) assays. The Lpck2a-1 CAPS marker was assigned to perennial ryegrass linkage group (LG) 4 and the Lpck2a-2 CAPS marker was assigned to LG2. The location of the Lpck2a-1 gene locus supports the previous conclusion of conserved synteny between perennial ryegrass LG4, the Triticeae homoeologous group 5L chromosomes and the corresponding segment of rice chromosome 3. Allelic variation at the Lpck2a-1 and Lpck2a-2 gene loci was correlated with phenotypic variation for heading date and winter survival, respectively. SNP polymorphism may be used for the further study of the role of CK2 alpha genes in the initiation of reproductive development and winter hardiness in grasses.

    DOI: 10.1007/s00122-005-0119-8

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  • Transgenic perennial ryegrass plants expressing wheat fructosyltransferase genes accumulate increased amounts of fructan and acquire increased tolerance on a cellular level to freezing Reviewed

    H Hisano, A Kanazawa, A Kawakami, M Yoshida, Y Shimamoto, T Yamada

    PLANT SCIENCE   167 ( 4 )   861 - 868   2004.10

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    The accumulation of fructan in grasses during autumn is linked to winter hardiness. Genetic manipulation of the accumulation of fructan could be an important molecular breeding strategy for the improvement of winter hardiness in grasses. We produced transgenic perennial ryegrass (Lolium perenne) plants that overexpress wheat fructosyltransferase genes, wft1 and wft2, which encode sucrose-fructan 6-fructosyltransferase (6-SFT) and sucrose-sucrose 1-fructosyltransferase (1-SST), respectively, under the control of CaMV 35S promoter using a particle bombardment-mediated method of transformation. Significant increases in fructan content were detected in the transgenic perennial ryegrass plants. A freezing test using the electrical conductivity method indicated that transgenic plants that accumulated a greater amount of fructan than non-transgenic plants have increased tolerance on a cellular level to freezing. The results suggest that the overexpression of the genes involved in fructan synthesis serves as a novel strategy to produce freezing-tolerant grasses. (C) 2004 Elsevier Ireland Ltd. All rights reserved.

    DOI: 10.1016/j.plantsci.2004.05.037

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  • High frequency Agrobacterium-mediated transformation and plant regeneration via direct shoot formation from leaf explants in Beta vulgaris and Beta maritima Reviewed

    H Hisano, Y Kimoto, H Hayakawa, J Takeichi, T Domae, R Hashimoto, J Abe, S Asano, A Kanazawa, Y Shimamoto

    PLANT CELL REPORTS   22 ( 12 )   910 - 918   2004.7

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    We have developed a new procedure for Agrobacterium-mediated transformation of plants in the genus Beta using shoot-base as the material for Agrobacterium infection. The frequency of regeneration from shoot bases was analyzed in seven accessions of sugarbeet (Beta vulgaris) and two accessions of B. maritima to select materials suitable for obtaining transformed plants. The frequency of transformation of the chosen accessions using Agrobacterium strain LBA4404 and selection on 150-mg/l kanamycin was found to be higher than that in previously published methods. Genomic DNA analysis and beta-glucuronidase reporter assays showed that the transgene was inherited and expressed in subsequent generations. In our method, shoot bases are prepared by a simple procedure, and transformation does not involve the callus phase, thus minimizing the occurrence of somaclonal variations.

    DOI: 10.1007/s00299-004-0773-3

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  • QTL analysis of morphological, developmental, and winter hardiness-associated traits in perennial ryegrass Reviewed

    T Yamada, ES Jones, NOI Cogan, AC Vecchies, T Nomura, H Hisano, Y Shimamoto, KF Smith, MD Hayward, JW Forster

    CROP SCIENCE   44 ( 3 )   925 - 935   2004.5

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    Quantitative trait loci (QTLs) for a number of agronomically important traits of perennial ryegrass (Lolium perenne L.) were identified by means of a reference molecular marker-based genetic map. Replicated phenotypic data was obtained for a number of field-assessed morphological and developmental traits as well as the winter hardiness-associated characters of winter survival and electrical conductivity. Marker-trait association analysis was performed by a number of methods, and a high degree of congruence was observed between the respective results. QTLs were detected for morphological traits such as plant height. tiller size, leaf length, leaf width, fresh weight at harvest, plant type, spikelet number per spike and spike length, as well as the developmental traits of heading date and degree of aftermath heading. A number of traits were significantly correlated, and coincident QTL locations were identified. No significant QTLs for winter survival in the field were identified. However, a QTL for electrical conductivity corresponding to frost tolerance was located close to it heading date QTL in a region that may show conserved synteny with chromosomal regions associated with both winter hardiness and flowering time variation in cereals. The QTL analysis of multiple phenotypic traits provides the basis for marker assisted selection (MAS) of important agronomic characters, allowing genetic improvement of yield, quality and adaptation in perennial ryegrass breeding.

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Books

  • ゲノム編集によるムギ類種子休眠性の改良 (ゲノム編集技術)

    久野裕, 加星光子, 安倍史高, 佐藤和広( Role: Contributor ,  pp183-192)

    株式会社 情報機構  2023.1  ( ISBN:9784865022421

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  • オオムギの標的変異 (植物のゲノム編集 実験プロトコール)

    久野 裕( Role: Contributor)

    化学同人  2022.12  ( ISBN:9784759820881

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  • オオムギの形質転換およびゲノム編集 (ひとりではじめる植物バイオテクノロジー入門 : 組織培養からゲノム編集まで)

    久野裕( Role: Contributor)

    国際文献社  2022.3  ( ISBN:9784910603032

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    Total pages:i, 408p   Language:Japanese

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  • Doubled Haploid Technology

    Robert Eric Hoffie, Ingrid Otto, Hiroshi Hisano, Jochen Kumlehn( Role: Contributor ,  Site-Directed Mutagenesis in Barley Using RNA-Guided Cas Endonucleases During Microspore-Derived Generation of Doubled Haploids.)

    Springer  2021.7 

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  • オオムギのゲノム編集

    久野裕( Role: Contributor)

    アグリバイオ  2017.1 

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  • Biofuels

    Hisano H, Nandakumar R, Wang ZY( Role: Contributor ,  Genetic modification of lignin biosynthesis for improved biofuel production)

    Springer  2010 

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  • マメ科植物の比較ゲノム(ゲノムから読み解く生命システム)

    佐藤 修正, 久野 裕, 田畑 哲之( Role: Contributor)

    細胞工学別冊  2007 

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MISC

  • Perspectives on the use of bioresources in breeding sciences: Lessons from successful studies

    Nasuda Shuhei, Ito Emi, Sato Yutaka, Hisano Hiroshi, Sato Kazuhiro, Komatsuda Takao, Ishikawa Ryo, Hashiguchi Masatsugu, Suzuki Akihiro, Hoshikawa Ken

    Breeding Research   21 ( 1 )   81 - 85   2019

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    Language:Japanese   Publisher:日本育種学会  

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  • Developing naked barley for brewing through site-directed mutagenesis

    久野裕, 坂井寛章, 濱岡美香, 宗森広美, 安倍史高, 佐藤和広, HAYES Patrick M.

    育種学研究   25   2023

  • Effects of promoters for gRNA expression and cultivation temperatures on genome editing efficiency in wheat

    加星光子, 安倍史高, 神谷容子, 川浦香奈子, 久野裕, 佐藤和広

    育種学研究   24   2022

  • Identification of a gene involved in loading phosphorus into barley grains

    黄衡亮, 久野裕, 黄勝, 三谷奈見季, 佐藤和広, 山地直樹, 馬建鋒

    日本土壌肥料学会講演要旨集(Web)   68   2022

  • Cloning of Rwt7, a wheat resistance gene corresponding to PWT7 of Pyricularia oryzae

    足助聡一郎, 庭本大輔, 堀江晶子, TAGLE Analiza Grubanzo, 久野裕, 佐藤和広, 土佐幸雄

    日本植物病理学会大会プログラム・講演要旨予稿集   2022   2022

  • Virome analysis of aphid populations that infest the barley field

    近藤秀樹, 藤田美貴, 久野裕, 兵頭究, 鈴木信弘

    日本植物病理学会大会プログラム・講演要旨予稿集   2021   2021

  • Regulation of germination by targeted mutagenesis of grain dormancy genes in barley

    久野裕, HOFFIE Robert, 安倍史高, 宗森広美, 松浦恭和, 遠藤真咲, 三上雅史, 中村信吾, KUMLEHN Jochen, 佐藤和広

    育種学研究   23   2021

  • 塩ストレス下でのオオムギの花粉稔性低下に関わる遺伝子の探索

    児玉明日香, 成田亮平, 山口真功, 久野裕, 安達俊輔, 高木宏樹, 平沢正, 佐藤和広, 大川泰一郎

    育種学研究   21   2019

  • ゲノム編集技術による穂発芽耐性の改良されたコムギ育種素材

    安倍史高, ハク エムダドウル, 久野裕, 田中剛, 神谷容子, 三上雅史, 川浦香奈子, 遠藤真咲, 大西一光, 林武司, 佐藤和広

    農研機構次世代作物開発研究センター成果情報(Web)   2019   2019

  • Functional analysis of a QTL gene controlling Cd accumulation in barley

    雷貴傑, 久野裕, 山地直樹, 佐藤和広, 馬建鋒

    日本土壌肥料学会講演要旨集(Web)   65   2019

  • オオムギの各種いもち病菌群に対する抵抗性遺伝子座Rmo2の1アリル候補配列の機能解析

    庭本大輔, 足助聡一郎, TAGLE Analiza Grubanzo, 久野裕, 佐藤和広, 土佐幸雄

    日本植物病理学会報   85 ( 1 )   2019

  • 塩ストレス条件で稔実歩合を高く維持するオオムギの量的形質遺伝子座(QTL)の同定

    児玉明日香, 成田亮平, 山口真功, 久野裕, 安達俊輔, 高木宏樹, 大川泰一郎, 佐藤和広, 平沢正

    日本作物学会講演会要旨集   245th   2018

  • うどんこ病菌の付着器分泌タンパク質とエフェクターの機能解析

    八丈野孝, 香口智宏, 菅井維之, 和原未季, 野村有子, 佐藤繭子, 若崎眞由美, 久野裕, 片岡創, 久保田直道, 小林括平, 西口正通, 豊岡公徳, 中神弘史, 山岡直人

    日本植物病理学会植物感染生理談話会論文集   ( 50 )   2015

  • 4倍体マメ科牧草シロクローバの連鎖地図作成と他マメ科植物との比較

    磯部祥子, 久野裕, 久野裕, 奥村健治, 佐藤修正, 白澤健太, 平川英樹, 田畑哲之

    育種学研究   13   2011

  • Genotype Matrix Mapping(GMM):QTLおよびQTL間の相互作用を検出する新たな解析法の開発

    磯部祥子, 中谷明弘, 矢野昌裕, 奥村健治, 久野裕, 田畑哲之

    生化学   2007

  • ゲノム構造比較と育種に向けたマメ科植物共通DNAマーカーの整備

    久野裕, 笹本茂美, 磯部祥子, 佐藤修正, 原田久也, 中村保一, 田畑哲之

    日本分子生物学会年会講演要旨集   28th   2005

  • ペレニアルライグラス (Lolium perenne L) におけるフルクタンネオシリーズおよびイヌリンを合成する酵素遺伝子の単離と同定

    久野 裕, 金澤 章, 吉田 みどり, 中嶋 博, 喜多村 啓介, 山田 敏彦

    日本草地学会誌   50   274 - 275   2004.3

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  • ペレニアルライグラス (Lolium perenne L.) における低温馴化冠部組織由来のcDNAライブラリーを用いたEST解析

    久野 裕, 山田 敏彦

    日本草地学会誌   50   272 - 273   2004.3

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  • Mapping of cold-induced genes in Lolium perenne L.

    SHINOZUKA H., HISANO H., YONEYAMA S., KANAZAWA A., JONES E. S., FORSTER J. W., SHIMAMOTO Y., YAMADA T.

    4   247 - 247   2002.8

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  • Production of transgenic perennial ryegrass (Lolium perenne L. ) carrying wheat genes encoding enzymes involved in fructan synthesis

    HISANO H., KANAZAWA A., KAWAKAMI K., YOSHIDA M., YAMADA T., NAKASHIMA H., ABE J., SHIMAMOTO Y.

    4   214 - 214   2002.8

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  • QTL analysis for winter hardiness in perennial ryegrass

    YAMADA T., NOMURA T., HISANO H., JONES E. S., FORSTER J. W., SHIMAMOTO Y.

    4 ( 1 )   279 - 279   2002.3

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Presentations

  • 品種の壁を越える:オオムギの形質転換や再分化を可能にするゲノム領域の同定 Invited

    久野 裕

    日本植物学会第82回大会  2018.9 

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    Language:Japanese   Presentation type:Symposium, workshop panel (public)  

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  • オオムギの形質転換効率を決めるゲノム領域の同定 Invited

    久野 裕

    第16回NC-CARP産学連携コンソーシアム講演会  2018.6.6 

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  • 品種の壁を越える: オオムギの形質転換に必要なゲノム領域の探索と利用 Invited

    第35回植物細胞分子生物学会(さいたま)大会  2017.8 

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  • Identification of TFA genomic regions that confer amenability to Agrobacterium-mediated transformation in barley Invited International conference

    Plant and Animal Genome XXV  2017.1 

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  • 遺伝子改変能を付与した低澱粉オオムギ変異系統の開発

    久野裕, 松島良, 佐藤和広

    第16回 ムギ類研究会  2021.12.25 

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    Event date: 2021.12.25

    Language:Japanese   Presentation type:Poster presentation  

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  • オオムギ種子休眠遺伝子の標的変異導入による発芽制御

    久野 裕, Hoffie Robert, 安倍 史高, 宗森 広美, 松浦 恭和, 遠藤 真咲, 三上 雅史, 中村 信吾, Kumlehn Jochen, 佐藤 和広

    日本育種学会第140回講演会  2021.9.24 

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  • オオムギ2H染色体に座乗する形質転換感受性に関わる遺伝子座TFA3の特性

    久野 裕, 宗森 広美, 佐藤 和広

    日本育種学会第139回講演会  2021.3.21 

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    Presentation type:Oral presentation (general)  

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  • Identification and application of TFA genomic regions that confer transformation amenability in barley International conference

    Hiroshi Hisano, Hiromi Munemori, Kazuhiro Sato

    Second International Barley Mutants Workshop 2018  2018.6 

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    Language:English   Presentation type:Poster presentation  

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  • オオムギの形質転換成否を決めるゲノム領域の特定とその利用 Invited

    久野 裕

    ストレス科学シンポジウム  2018.3 

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    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

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  • 形質転換能遺伝子座に関わるオオムギ準同質遺伝子系統の開発

    久野 裕, 宗森 広美, 佐藤 和広

    日本育種学会第133回講演会  2018.3 

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  • Baby boom1ならびにWuschel2に制御されるオオムギ遺伝子の単離

    久野 裕, 宗森 広美, 西村 秀希, 佐藤 和広

    日本育種学会第132回講演会  2017.10 

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  • オオムギの形質転換効率に関わる遺伝子単離に向けたDNAマーカー開発

    野 裕, 宗森 広美, 元井 由加, 新海 典夫, 瀬々 潤, 豊田 敦, 佐藤 和広

    日本育種学会第131回講演会  2017.3 

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  • TFA genomic regions confer amenability to Agrobacterium-mediated transformation in barley International conference

    Plant and Animal Genome XXV  2017.1 

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  • 形質転換可能なオオムギ系統を作る方法

    久野 裕, 安東広美, 元井由加, Patrick M. Hayes, 佐藤和広

    日本育種学会第130回講演会  2016.9 

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  • オオムギの形質転換効率に関わるゲノム領域の特定

    久野 裕, 元井 由加, 安東 広美, 佐藤 和広

    第34回植物細胞分子生物学会(上田)大会  2016.9 

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  • Identification of the genomic region responding to amenability of Agrobacterium-mediated transformation in barley International conference

    H. Hisano, Y. Motoi, K. Sato

    12th International Barley Genetics Symposium  2016.6 

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Industrial property rights

  • 形質転換感受性のオオムギの作出方法

    佐藤 和広, 久野 裕

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    Application no:特願2016-124749  Date applied:2016.6.23

    Patent/Registration no:特許第6876334号  Date registered:2021.4.28  Date issued:2021.5.26

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  • イネ科植物及びその作製方法

    松島良, 久野裕, 佐藤和広

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    Application no:特願2020-169225 

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  • マイクロサテライトマーカーを用いるシバ属植物品種判別法

    明石 良, 渡邉 学, 加藤 正広, 大越 一雄, 久野 裕, 田畑 哲之

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    Application no:特願2008-093038 

    Announcement no:特開2009-240268 

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  • マイクロサテライトマーカーを用いるラッカセイ品種識別法

    内藤 嘉磯, 福田 節也, 國方 俊智, 渡邉 学, 鈴木 茂, 大越 一雄, 田畑 哲之, 久野 裕, 久保山 勉

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    Application no:特願2009-196861 

    Announcement no:特開2010-075176 

    Patent/Registration no:特許第5190596号 

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  • マイクロサテライトマーカーを用いるバーミューダグラス品種判別法

    明石 良, 久野 裕, 田畑 哲之

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    Application no:特願2008-093039 

    Announcement no:特開2009-240269 

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Awards

  • Best poster award

    2018.6   International barley mutant workshop  

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  • 日本育種学会 優秀発表賞

    2015  

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    Country:Japan

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  • 論文賞

    2019.3   日本育種学会   QTLs maintaining grain fertility under salt stress detected by exome QTL-seq and interval mapping in barley

    Asuka Kodama, Ryouhei Narita, Makoto Yamaguchi, Hiroshi Hisano, Shunsuke Adachi, Hiroki Takagi, Taiichiro Ookawa, Kazuhiro Sato, Tadashi Hirasawa

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  • MBFT(牧草・芝草の国際分子育種学会) ベストポスター賞

    2007  

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    Country:Japan

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Research Projects

  • Investigation of the regulatory mechanism of plastid in the tissue transdifferentiation of grass species

    Grant number:22H05077  2022.05 - 2025.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Transformative Research Areas (B)  Grant-in-Aid for Transformative Research Areas (B)

    久野 裕

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    Authorship:Principal investigator 

    Grant amount:\23920000 ( Direct expense: \18400000 、 Indirect expense:\5520000 )

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  • Identification of genetic factors for tissue culture suitability and development of efficient haploid inducers to accelerate the haploid breeding

    Grant number:22H02313  2022.04 - 2025.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    久野 裕

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    Authorship:Principal investigator 

    Grant amount:\17680000 ( Direct expense: \13600000 、 Indirect expense:\4080000 )

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  • 遺伝子改変技術を利用したオオムギ機能性成分の高蓄積系の開発

    2021.01 - 2023.01

    NEDO 国立研究開発法人新エネルギー産業技術総合開発機構  官民による若手研究者発掘支援事業 

    久野裕, 松島良

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  • 作物超個体における根圏RNAウイルス叢の実体解明とその生態学的役割

    Grant number:23H02214  2023.04 - 2027.03

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    近藤 秀樹, 久野 裕, 兵頭 究, 鈴木 信弘

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    Grant amount:\18590000 ( Direct expense: \14300000 、 Indirect expense:\4290000 )

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  • ムギ類種子休眠性の精密制御と分子育種法の確立

    Grant number:23H00333  2023.04 - 2027.03

    日本学術振興会  科学研究費助成事業  基盤研究(A)

    佐藤 和広, 中村 信吾, 安倍 史高, 久野 裕

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    Grant amount:\46410000 ( Direct expense: \35700000 、 Indirect expense:\10710000 )

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  • 絶対寄生および半活物寄生に対するCa2+流動の制御機構と生理的意義の解明

    Grant number:22H02349  2022.04 - 2025.03

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    八丈野 孝, 久野 裕, 吉田 健太郎

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    Grant amount:\17680000 ( Direct expense: \13600000 、 Indirect expense:\4080000 )

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  • 作物根圏におけるウイルス叢の多様性とその感染動態から紐解く生態的意義

    Grant number:20H02987  2020.04 - 2023.03

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    近藤 秀樹, 久野 裕, 兵頭 究, 鈴木 信弘

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    Grant amount:\17680000 ( Direct expense: \13600000 、 Indirect expense:\4080000 )

    本課題では、ムギ類生存圏(特に根系)におけるウイルス叢の多様性・普遍性や動態を紐解き、さらにそれらの未知なる生態学的役割の一端に迫ることを目指している。以下に本年度の研究実績を示す。
    柱①根圏に存在する主要な菌類・昆虫ウイルス群の宿主探索:初年度に引き続き、ムギ類(オオムギ・コムギ)の地上部・根系、さらにアブラムシ、うどんこ病菌などのRNAseqデータを取得した。得られたデータセットはウイルス配列の確認作業や分子系統解析を進めている。さらに、北海道のコムギサンプルより見出された新規フレキシウイルス(WVQ)の解析の結果,当該圃場に三系統が存在し、ライムギにも感染できること、さらに土壌伝染性ウイルスの可能性が高いことを見出している。根系ウイルス叢のリザーバー探索については、本年度はオオムギ根のRNAseqデータに含まれる微生物リードの抽出と配列相同性解析により、微生物叢の概要を調査した。
    柱②根圏ウイルスの生物界を跨ぐ水平伝搬ポテンシャルと宿主への影響:昨年度より継続中のルテオウイルスなど新規植物ウイルスのゲノム解析を進めており、RT-PCRおよびRLM-RACE解析により全長ゲノムRNA配列を確認・決定した。また、圃場のうどんこ病菌のウイルスについては、今年度はdsRNAを鋳型にした全長cDNAのPCR増幅法の検討を進め、そのアンプリコン配列解析により未同定のゲノム末端配列の解析を進めている。
    柱③根圏ウイルス叢の年次変動とムギ類ウイルスの病原性の(再)評価:核酸抽出を伴わない簡易ウイルス診断法(RT-PCRをベースとする)の条件を設定し、一部圃場サンプル(葉)でルテオウイルス等の検出を進めている。現在、圃場オオムギサンプル(保存している過年度のを含む)について網羅的に検定すすめている。また、アブラムシ媒介性新規植物ウイルスについては、チャンバー内試験にて病原性評価を進めている。

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  • ムギ類に特異的な澱粉粒が形成される機構とその変異の利用

    Grant number:20K05970  2020.04 - 2023.03

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    松島 良, 久野 裕

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    イネ科植物の胚乳における澱粉粒は、複粒型澱粉粒と単粒型澱粉粒に大別される。複粒型澱粉粒は複数の小さい澱粉粒子が集合して形成されるのに対して、単粒型澱粉粒は1つの澱粉粒子から形成される。重要作物であるオオムギやコムギの胚乳の澱粉粒は、大きさが大小の二極性を示す単粒型であることから、二極性単粒型澱粉粒と呼ばれている。本研究では、オオムギを研究材料に用いて、不明な点が多い二極性単粒型澱粉粒の形成機構について新しい知見を得る事と、新しい澱粉特性を持つオオムギ系統の作出という2つの目的で研究を進めている。昨年度までの研究では、澱粉粒簡便観察法を用いて遺伝学的なスクリーニングを行い、二極性単粒型澱粉粒の形や大きさに異常を示すオオムギ突然変異体の単離と解析を行なってきた。本年度は、得られた変異体の中で、複粒型澱粉粒を発達させる変異体3系統に注目して解析を進めた。この3系統では同じ遺伝子に塩基置換が起こっていることを明らかにし、原因遺伝子がコードするタンパク質に対する抗体を作成した。western blotの結果、上記の3系統の変異体では、このタンパク質が種子に蓄積していないことが分かった。さらに、この3系統とは異なる表現型を示す突然変異体と交配し、多重変異体を作出した。その結果、複粒型澱粉粒を発達させる変異を亢進する変異や抑圧する変異を見出すことができた。本年度には澱粉科学の分野ではもっとも権威がある国際学会で招待講演を行った。また、得られた系統の一部で、国際特許出願を行った。オオムギは、醸造用、飲料用、食用、飼料用に利用される多用途作物であり、すべての用途において澱粉特性、種子特性は重要形質である。作出したオオムギ系統の社会実装を目指して、オオムギ関連企業に対して説明会も行なった。

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  • ムギ類種子休眠性遺伝子の分子進化機構の解明と精密育種技術の開発

    Grant number:19H00943  2019.04 - 2023.03

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    佐藤 和広, 安倍 史高, 大西 一光, 久野 裕

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    Grant amount:\45370000 ( Direct expense: \34900000 、 Indirect expense:\10470000 )

    1)Qsd1の解析と制御:a)野生オオムギと栽培オオムギのQsd1配列では4つのアミノ酸置換が確認されている。このアレル別の過剰発現個体の形質転換体の種子休眠性に対する効果を確認するための温室内栽培を継続実施した。b)オオムギQsd1のゲノム編集によって得られた変異体の効果を解析し、その効果について論文出版した。c)コムギのゲノム編集技術を用いて、作出した3つのサブゲノムのQsd1をそれぞれ改変した個体の野外栽培を2021年11月から実施中である。d)Qsd1変異体におけるタンパク質と休眠性の関連については継続実験中である。e)コムギの主要品種「春よ恋」と「きたほなみ」の人為突然変異集団からQsd1配列の変異体をTILLING法で選抜し、サブゲノム変異体の効果の確認作業を進めている。
    2)他の遺伝子の解析と制御:a)ゲノム編集によるオオムギQsd2遺伝子の改変によって、自然変異以外の変異体の作成と種子休眠性に対する効果を確認した。さらに、1)b)で作成したQsd1のゲノム編集個体との集積効果確認し、その成果を論文出版した。b)イネを中心に、ムギ類以外の種子休眠性同祖遺伝子のコムギのゲノム編集を試みているが、明確な効果は検出できていない。
    3)遺伝子進化相互作用解析:a)新規にGenotype by Sequence法で作出する約7万のマーカーによってオオムギ約4千系統についてのゲノムワイドアソシエーション解析(GWAS)のマーカー解析を終了し、解析のためのパイプライン構築が終了した。b)休眠型のqsd1を持っているにもかかわらず休眠しないエチオピア在来品種と「はるな二条」(休眠短)の交雑、野生オオムギ( 休眠長)と「はるな二条」の交雑、野生オオムギとエチオピア品種の交雑に由来するQTL解析を反復実施し、マーカー解析が終了した。

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  • Study on the regulation mechanism of transformation amenability in barley

    Grant number:19H02930  2019.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    HISANO Hiroshi

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\17550000 ( Direct expense: \13500000 、 Indirect expense:\4050000 )

    In order to fine map of the three TFA loci involved in the efficiency of genetic transformation of barley, the genetic experiments were conducted using two populations derived from a cross between transformable and non-transformable cultivars. The results revealed the presence of genes contributing to callus induction and differentiation in a region called TFA3. In addition, RNA-seq analysis revealed genes with differential expression between transformable and non-transformable cultivars. One gene with significant expression was cloned and introduced into barley, but no improvement in callus induction efficiency was observed. On the other hand, a genome editing experiment using a TFA-selected barley line succeeded in obtaining individuals with mutations in the target gene.

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  • Imaging Amyloplasts in the Developing Endosperm of Barley and Rice

    Grant number:17K07602  2017.04 - 2020.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Matsushima Ryo

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    Grant amount:\4680000 ( Direct expense: \3600000 、 Indirect expense:\1080000 )

    We generated transgenic plants expressing amyloplast-accumulating GFP in barley and rice. In this transgenic plant, amyloplasts and starch grains can be visualized directly and indirectly, respectively. We could observe amyloplasts and starch grains in developing endosperm to capture the live images how amyloplasts and starch grains are developed. In the case of barley, the multiple simple starch grains were present in a single amyloplast, and the large and small starch grains were also present in the same amyloplast. In the case of rice, some amyloplasts contained multiple compound starch grains especially in the early stage of development. These observations are the novel discovery of new morphological features for amyloplasts and starch grains in these cereals.The transgenic plants produced in this study will be an important tool for the analysis of cereal starch-related mutants in the future.

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  • Identification and functional analysis of loci involved in transformation amenability in barley

    Grant number:16K18634  2016.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)  Grant-in-Aid for Young Scientists (B)

    Hisano Hiroshi, SATO Kazuhiro, MUNEMORI Hiromi, SHINKAI Norio, SESE Jun, TOYODA Atsushi, SEKI Masahide, SUZUKI Yutaka

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    In this study, we tried to narrow down the regions of TFA (Transformation amenability) loci involved in transformation efficiency in barley in order to enable genetic transformation without limitation of their genotypes. First, SNPs array analysis and exome-sequence analysis were performed to develop the selection markers for TFAs. These markers were used to generate the near-isogenic lines of TFAs derived from a cross between barley cultivar “Haruna Nijo” and “Golden Promise” (GP). Subsequently, the transformation experiment was carried out using the doubled-haploid lines derived from a cross between barley cultivar "Full Pint" and GP to clarify that TFAs were necessary for barley transformation.

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  • 穂発芽耐性麦類の開発

    2014.11 - 2019.03

    独立行政法人農業・食品産業技術総合研究機構  戦略的イノベーション創造プログラム(SIP) 

    佐藤和広

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  • Genetic assay and study of crop germplasm introduced/originated in East Asia (2nd)

    Grant number:26257409  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)  Grant-in-Aid for Scientific Research (A)

    Kenji Kato

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    Grant amount:\33540000 ( Direct expense: \25800000 、 Indirect expense:\7740000 )

    Plant genetic resources of important crops, including barley, wheat, soybean and melon, have been explored in Asian countries through which crops were introduced to East Asia. Based on the international network constructed by our research group, we successfully conducted a total of twenty-one times of field expedition. In addition, we performed joint experiments with our counterpart scientists in respective countries, when international transfer of genetic resources was difficult. Several important new findings related with the evolution, transmission, adaptation of these crops have been obtained by the analysis of diverse plant genetic resources, and published in scientific papers and books. We also evaluated the agronomically important traits of these materials introduced which can be utilized in breeding program of each crop, as well as for basic research to identify novel genes.

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  • TALEN-based mutagenesis for target genes in barley

    Grant number:26850004  2014.04 - 2016.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)  Grant-in-Aid for Young Scientists (B)

    Hisano Hiroshi

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\2600000 ( Direct expense: \2000000 、 Indirect expense:\600000 )

    Transcription activator-like effector nuclease (TALEN)-vectors targeting for the genes encoding chalcone synthase (CHS) and dihydroflavonol 4-reductase (DFR) which were both involved in anthocyanin biosynthesis were constructed and applied to barley transformation. Although transgenic barley plants were obtained, any mutation was detected in target genes of these plants.
    On the other hand, 4 promoters regulating callus-specific gene expression were isolated in barley. For expression analysis, these promoters were cloned into expression vector of the gene encoding enhanced green fluorescent protein (EGFP). The transient expression of EGFP in callus was observed by all the promoters. In addition, 2 of 4 promoters showed stable expression of EGFP in transgenic callus in barley.

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  • オオムギの形質転換成否を左右する遺伝因子の同定

    2012.09 - 2014.03

    日本学術振興会  科学研究費補助金 研究活動スタート支援 

    久野 裕

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  • Identification of genetic factors affect the success or failure of the transformation in barley

    Grant number:24880025  2012.08 - 2014.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Research Activity start-up  Grant-in-Aid for Research Activity start-up

    HISANO Hiroshi

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    Grant amount:\2990000 ( Direct expense: \2300000 、 Indirect expense:\690000 )

    We explored the genomic regions required for genetic transformation by using a mapping populations generated from barley cv. Golden Promise, which was a reliable cultivar for transformation. As a result of this, the partial region of chromosome No. 1H, No. 2 H, No. 3H, No. 6 H and No. 7 H were estimated as the positions of candidate genes. Then we isolated 4308 genes which expression patterns were specific in Golden Promise by a comprehensive gene expression analysis using microarray. In the analysis of phytohormone, we identified that auxin showed a positive effect in regeneration of barley callus, although abscisic acid and salicylic acid showed negative.

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  • Genetic assay and study of crop germplasm introduced/originated in East Asia

    Grant number:23255007  2011.04 - 2014.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)  Grant-in-Aid for Scientific Research (A)

    KATO Kenji, SATO Kazuhiro, TSUJIMOTO Hisashi, TSUYUZAKI Hiroshi, ENOMOTO Takashi, TANAKA Hiroyuki, NISHIDA Hidetaka, ABE Jun, SASANUMA Tsuneo, KASHIWAGI Jun-ichi, KISHII Masahiro, TANAKA Katsunori, HISANO Hiroshi, SAISHO Daisuke

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    Grant amount:\38090000 ( Direct expense: \29300000 、 Indirect expense:\8790000 )

    Plant genetic resources of important crops, including barley, wheat, soybean and melon, have been explored in Asian countries through which crops were introduced to East Asia, and successfully introduced through a total of twelve times of field expedition. Several important new findings related with the evolution, transmission, adaptation of these crops have been obtained by the analysis of diverse plant genetic resources, and published in scientific papers and books. We also identified several accessions which can be used for the improvement of agronomically important traits. Such novel resources can be utilized in breeding program of each crop, as well as for basic research to identify novel genes.

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  • Development of genomics assisted breeding system of salt tolerant barley

    Grant number:23380007  2011.04 - 2014.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    SATO Kazuhiro, HISANO Hiroshi

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    Grant amount:\19240000 ( Direct expense: \14800000 、 Indirect expense:\4440000 )

    cDNA sequences are generated by an efficient analysis system and aligned with full length cDNA sequences of Haruna Nijo to develop genome wide SNP detection system. The array system is used to genotype recombinant inbred lines and recombinant chromosome substitution lines for genetic map development and estimation of substitution segments. Salt tolerance of these lines are evaluated to map tolerance QTLs and their responsible substitution segments. On the other hand, a previously isolated tolerance gene by subtraction method is used to make transgenic plant to evaluate its salt tolerance. Markers are also developed to narrow down tolerance genes. The recombinant chromosome substitution liens carrying tolerance gene segments are developed to be used as isogenic lines for salt tolerance.

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Class subject in charge

  • Plant Genome Dynamics (2023academic year) Second semester  - 月5,月6

  • Plant Molecular Breeding (2023academic year) Late  - その他

  • Plant Molecular Breeding (2023academic year) Late  - その他

  • Seminar in Plant diversity analysis (2023academic year) Prophase  - その他

  • Seminar in Plant Diversity Analysis (2023academic year) Late  - その他

  • Seminar in Plant diversity analysis (2023academic year) Late  - その他

  • Seminar in Plant diversity analysis (2023academic year) Late  - その他

  • Seminar in Plant Diversity Analysis (2023academic year) Prophase  - その他

  • Seminar in Plant diversity analysis (2023academic year) Prophase  - その他

  • Seminar in Plant diversity analysis (2023academic year) Year-round  - その他

  • Plant Diversity Genetics (2023academic year) Prophase  - 木5~8

  • Plant Diversity Genetics (2023academic year) Prophase  - 木5~8

  • Advanced Study (2023academic year) Other  - その他

  • Specific Research of Bioresources Science (2023academic year) Year-round  - その他

  • Plant Genome Dynamics (2022academic year) Second semester  - 月5,月6

  • Plant Molecular Breeding (2022academic year) Late  - その他

  • Seminar in Plant diversity analysis (2022academic year) Prophase  - その他

  • Seminar in Plant Diversity Analysis (2022academic year) Late  - その他

  • Seminar in Plant diversity analysis (2022academic year) Late  - その他

  • Seminar in Plant Diversity Analysis (2022academic year) Prophase  - その他

  • Plant Diversity Genetics (2022academic year) Prophase  - 木5~8

  • Specific Research of Bioresources Science (2022academic year) Year-round  - その他

  • Plant Genome Dynamics (2021academic year) Second semester  - 月5,月6

  • Plant Molecular Breeding (2021academic year) Late  - その他

  • Seminar in Plant diversity analysis (2021academic year) Prophase  - その他

  • Seminar in Plant Diversity Analysis (2021academic year) Late  - その他

  • Seminar in Plant diversity analysis (2021academic year) Late  - その他

  • Seminar in Plant Diversity Analysis (2021academic year) Prophase  - その他

  • Plant Diversity Genetics (2021academic year) Prophase  - 木5~8

  • Specific Research of Bioresources Science (2021academic year) Year-round  - その他

  • Plant Genome Dynamics (2020academic year) Second semester  - 月5,月6

  • Plant Molecular Breeding (2020academic year) special  - その他

  • Seminar in Plant diversity analysis (2020academic year) Prophase  - その他

  • Seminar in Plant Diversity Analysis (2020academic year) Late  - その他

  • Seminar in Plant diversity analysis (2020academic year) Late  - その他

  • Seminar in Plant Diversity Analysis (2020academic year) Prophase  - その他

  • Plant Diversity Genetics (2020academic year) Prophase  - その他

  • Specific Research of Bioresources Science (2020academic year) Year-round  - その他

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Social Activities

  • 倉敷青陵高校 サイエンストーク

    Role(s):Lecturer

    2019.12

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  • 植物からのバイオ燃料生産

    Role(s):Lecturer

    倉敷市大学連携講座  2013.8

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    Type:Lecture

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  • 裸麦が創る食と農の未来フォーラム

    Role(s):Panelist, Lecturer, Planner

    愛媛大学大学院農学研究科、岡山大学資源植物科学研究所、農林水産省中国四国農政局  第3回オオムギ資源開発研究セミナー 愛媛大学大学院農学研究科附属ハダカムギ開発研究センター設立シンポジウム  2021.12.12

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    Type:Lecture

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  • おかやまバイオアクティブ研究会

    Role(s):Lecturer

    おかやまバイオアクティブ研究会  2021.12.9

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    Type:Seminar, workshop

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  • 『ゲノム編集』作物が開く未来の可能性

    Role(s):Lecturer

    岡山大学  第76回岡大SDGsサイエンスカフェ  2020.9.29

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Media Coverage

  • 技術の森 出演 TV or radio program

    RSK山陽放送  天神ワイド 朝  2023.5

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  • オオムギ発芽 強く働く遺伝子解明 Newspaper, magazine

    山陽新聞社  山陽新聞  2022.1.16

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  • Cheers! Scientists have developed gene-edited barley that could better your beer Newspaper, magazine

    EurekAlert!  News Release  2021.11.12

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  • 技術の森 出演 TV or radio program

    RSK山陽放送  朝耳らじお5.5  2021.11

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  • オオムギの種子休眠の長さ、ゲノム編集技術で調節 岡山大など、ビール醸造に活用 Newspaper, magazine

    日刊工業新聞社  日刊工業新聞  2021.10.7

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Academic Activities

  • Review for grant

    Role(s):Peer review

    Biotechnology and Biological Sciences Research Council  2020.9.23 - 2020.11.9

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    Type:Peer review 

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