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Yamada Tetsuya

Research Faculty of Agriculture　Fundamental AgriScience Research　Agrobiology and BioresourcesAssociate Professor

Researcher basic information

■ Degree
  • 博士（学術）
■ URL
researchmap URLホームページURL■ Various IDs
Researcher number
  • 70374618
J-Global ID■ Research Keywords and Fields
Research Keyword
  • potato
  • クロロフィル
  • ゲノム編集
  • amiRNA
  • miRNA
  • 貯蔵タンパク質
  • ダイズ
  • 形質転換
  • フラボノイド
  • 転写因子
  • 大豆
  • Myb転写因子
  • 種子
  • アグロバクテリウム
  • α-トコフェロール
  • ルテイン
  • リノレン酸
  • γ-TMT
  • 高機能性
  • 遺伝子
  • 遺伝分析
  • QTL解析
  • 作用機序
Research Field
  • Life Science, Genetics
  • Life Science, Plant molecular biology and physiology
  • Environmental Science/Agriculture Science, Science in plant genetics and breeding
■ Educational Organization

Career

■ Career
Career
  • Jul. 2025 - Present
    Hokkaido University, Graduate School of Agriculture Research Faculty of Agriculture, 准教授
  • 2012 - Present
    Hokkaido University, Graduate School of Agriculture Research Faculty of Agriculture, 講師
  • 2008 - 2011
    北海道大学 (連合)農学研究科(研究院), 助教

Research activity information

■ Papers
  • Preferential induction of Dt2 gene in the absence of E1 function enhances semi-determinate stem growth in soybean
    Ryoma Takeshima; Shohei Nosaki; Meilan Xu; Kenta Nakashima; Shin Kato; Maria Stefanie Dwiyanti; Tetsuya Yamada; Baohui Liu; Jun Abe
    Journal of Experimental Botany, Oxford University Press (OUP), 16 Mar. 2026
    Scientific journal, Abstract

    Suitable plant architecture is key to maximizing crop yield. Stem growth habit is a crucial characteristic of soybean plant architecture. Here, we investigated the combined effect of the Dt2 gene (producing semi-determinate growth habit) and a loss-of-function e1-nl allele of the floral repressor E1. Comparisons among near-isogenic lines with different maturity genotypes indicated that the loss or repression of E1 function enhanced the effect of Dt2 on main stem node numbers. Dt2 expression in stem tips was upregulated in early growing stages under long-day conditions (LD) by the Dt2 and e1-nl alleles in an additive manner. Additionally, an Arabidopsis APETALA1 ortholog was highly upregulated in a Dt2/e1-nl line. The electrophoretic mobility shift assay revealed that the E1 DNA-binding domain bound to several genomic sites harboring key polymorphisms differentiating Dt2 from dt2 alleles. One of these sites had different transcription activities between Dt2 and dt2 alleles, and these activities were repressed by E1. Together, our data suggest that the Dt2 allele confers semi-determinacy by being preferentially induced when released from the repression by E1. The allelic combination of Dt2 and e1-nl confers a distinct semi-determinate phenotype, which would facilitate the use of the Dt2 allele, particularly under LD at high latitudes.
  • Soy protein selectively accumulates formaldehyde
    Masanori Yamada; Momoka Uchida; Tetsuya Yamada
    Scientific Reports, 15, 1, Springer Science and Business Media LLC, 18 Mar. 2025, [Peer-reviewed]
    Scientific journal
  • Simultaneous site-directed mutagenesis for soybean β-amyrin synthase genes via DNA-free CRISPR/Cas9 system using a single gRNA.
    Asa, H.; Kuwabara, C.; Matsumoto K.; Shigeta, R.; Yamamoto, T.; Masuda, Y.; Yamada, T.
    Plant Cell Reports, 44, 1, 13, Jan. 2025, [Peer-reviewed], [Corresponding author]
  • A DNA-free and genotype-independent CRISPR/Cas9 system in soybean
    Chikako Kuwabara; Ryuji Miki; Nobuyuki Maruyama; Masanori Yasui; Haruyasu Hamada; Yozo Nagira; Yumiko Hirayama; Wataru Ackley; Feng Li; Ryozo Imai; Naoaki Taoka; Tetsuya Yamada
    Plant Physiology, 196, 4, 2320, 2329, Oxford University Press (OUP), 23 Sep. 2024, [Peer-reviewed], [Corresponding author]
    Scientific journal, Abstract

    Here, we report a smart genome editing system for soybean (Glycine max) using the in planta bombardment-ribonucleoprotein (iPB-RNP) method without introducing foreign DNA or requiring traditional tissue culture processes such as embryogenesis and organogenesis. Shoot apical meristem (SAM) of embryonic axes was used as the target tissue for genome editing because the SAM in soybean mature seeds has stem cells and specific cell layers that develop germ cells during the reproductive growth stage. In the iPB-RNP method, the RNP complex of the CRISPR/Cas9 system was directly delivered into SAM stem cells via particle bombardment, and genome-edited plants were generated from these SAMs. Soybean allergenic gene Gly m Bd 30K was targeted in this study. Many E0 (the first generation of genome-edited) plants in this experiment harbored mutant alleles at the targeted locus. Editing frequency of inducing mutations transmissible to the E1 generation was approximately 0.4% to 4.6% of all E0 plants utilized in various soybean varieties. Furthermore, simultaneous mutagenesis by iPB-RNP method was also successfully performed at other loci. Our results offer a practical approach for both plant regeneration and DNA-free genome editing achieved by delivering RNP into the SAM of dicotyledonous plants.
  • The Accumulation of Metal Ions by a Soy Protein–Inorganic Composite Material
    Masanori Yamada; Maika Ujihara; Tetsuya Yamada
    Journal of Composites Science, 7, 10, 419, 419, MDPI AG, 07 Oct. 2023, [Peer-reviewed]
    Scientific journal, Water-soluble soy protein (SP), which contains many acidic amino acids in its structure, was complexed by mixing with a silane coupling agent, 3-glycidoxypropyltrimethoxysilane (GPTMS). These SP−GPTMS composite materials showed stability in water. This property is due to the cross-linking between SP and GPTMS through the ring cleavage reaction of the epoxy group in the GPTMS molecule and an encapsulation of SP into the 3D siloxane network of GPTMS. When the SP−GPTMS composite material was immersed in an aqueous Cu(II) ion solution, the composite material changed from light brown to blue green by the coordination of Cu(II) ions into the SP. Hence, we evaluated the accumulation of heavy ions, rare-earth ions, and light metal ions. The accumulating affinity of metal ions was Cd(II) << Zn(II), Cu(II), Pb(II) < La(III) < Al(III) < Nd(III), In(III) << Mg(II) < Ca(II) ions. In addition, the sorption capacities of Ca(II), Mg(II), In(III), Nd(III), Al(III), La(III), Pb(II), Cu(II), Zn(II), and Cd(II) ions were 700 nmol/mg, 660 nmol/mg, 470 nmol/mg, 470 nmol/mg, 410 nmol/mg, 380 nmol/mg, 350 nmol/mg, 350 nmol/mg, 300 nmol/mg, and 200 nmol/mg, respectively. These properties suggest that the SP−GPTMS composite material has a divalent light metal ion selectivity. Additionally, the accumulative mechanism of the light metal ions was related to the carboxylate group and the hydroxyl group in the composite material.
  • Site-directed mutagenesis of soybean <i>PEAPOD</i> genes using the CRISPR/Cas9 system alters tissue developmental transition
    Jaechol Sim; Yuhei Kanazashi; Tetsuya Yamada
    Plant Biotechnology, 40, 3, 247, 254, Japanese Society for Plant Cell and Molecular Biology, 25 Sep. 2023, [Peer-reviewed], [Corresponding author]
    Scientific journal
  • Recent advances in the improvement of soybean seed traits by genome editing
    Jaechol Sim; Chikako Kuwabara; Shota Sugano; Kohei Adachi; Tetsuya Yamada
    Plant Biotechnology, 40, 3, 193, 200, Japanese Society for Plant Cell and Molecular Biology, 25 Sep. 2023, [Peer-reviewed], [Corresponding author]
    Scientific journal
  • Preparation of bioplastic consisting of salmon milt DNA
    Masanori Yamada; Midori Kawamura; Tetsuya Yamada
    Scientific Reports, 12, 1, 7423, Springer Science and Business Media LLC, Dec. 2022, [Peer-reviewed]
    Scientific journal, Abstract

    The microplastic that pollutes the ocean is a serious problem around the world. The bioplastic consisting of biopolymers which is degraded in nature, is one of the strategies to solve this problem. Although the bioplastics consisting of protein, polysaccharide, polylactic acid, etc., have been reported, which consist of DNA, one of the most important materials in the genetic process, have not been reported to the best of our knowledge. In addition, a large amount of DNA-containing materials, such as salmon milts, is discarded as industrial waste around the world. Therefore, we demonstrated the preparation of a bioplastic consisting of salmon milt DNA. The DNA plastic was prepared by the immersion of a DNA pellet in a formaldehyde (HCHO) solution and heating. As a result, the water-stable DNA plastics were obtained at the HCHO concentration of 20% or more. Particularly, the DNA plastic with a 25% HCHO treatment showed water-insoluble, thermally stable, and highly mechanical properties. These are due to the formation of a three-dimensional network via the crosslinking reaction between the DNA chains. In addition, since DNA in plastic possesses the double-stranded structure, these plastics effectively accumulated the DNA intercalator, such as ethidium bromide. Furthermore, the DNA plastics indicated a biodegradable property in a nuclease-containing aqueous solution and the biodegradable stability was able to be controlled by the HCHO concentration. Therefore, salmon milt DNA has shown the potential to be a biodegradable plastic.
  • Oxicam-type nonsteroidal anti-inflammatory drugs enhance Agrobacterium-mediated transient transformation in plants
    Seung-won Choi; Kie Kumaishi; Reiko Motohashi; Harumi Enoki; Wiluk Chacuttayapong; Tadashi Takamizo; Hiroaki Saika; Masaki Endo; Tetsuya Yamada; Aya Hirose; Nobuya Koizuka; Seisuke Kimura; Yaichi Kawakatsu; Hiroyuki Koga; Emi Ito; Ken Shirasu; Yasunori Ichihashi
    Plant Biotechnology, 39, 3, 323, 327, Japanese Society for Plant Cell and Molecular Biology, 25 Sep. 2022, [Peer-reviewed], [Domestic magazines]
    English, Scientific journal, Agrobacterium-mediated transformation is a key innovation for plant breeding, and routinely used in basic researches and applied biology. However, the transformation efficiency is often the limiting factor of this technique. In this study, we discovered that oxicam-type nonsteroidal anti-inflammatory drugs, including tenoxicam (TNX), increase the efficiency of Agrobacterium-mediated transient transformation. TNX treatment increased the transformation efficiency of Agrobacterium-mediated transformation of Arabidopsis thaliana mature leaves by agroinfiltration. The increase of efficiency by TNX treatment was not observed in dde2/ein2/pad4/sid2 quadruple mutant, indicating that TNX inhibits the immune system mediated by jasmonic acid, ethylene, and salicylic acid against to Agrobacterium. We also found that TNX-treatment is applicable for the transient expression and subcellular localization analysis of fluorescent-tagged proteins in Arabidopsis leaf cells. In addition, we found that TNX increases the efficiency of Agrobacterium-mediated transient transformation of Jatropha. Given that treatment with oxicam compounds is a simple and cost effective method, our findings will provide a new option to overcome limitations associated with Agrobacterium-mediated transformation of various plant species.
  • Genetic analysis of chlorophyll synthesis and degradation regulated by BALANCE of CHLOROPHYLL METABOLISM
    Hiroshi Yamatani; Takeshi Ito; Kenji Nishimura; Tetsuya Yamada; Wataru Sakamoto; Makoto Kusaba
    Plant Physiology, Oxford University Press (OUP), 28 Mar. 2022, [Peer-reviewed]
    Scientific journal, Abstract

    Chlorophyll serves a number of essential functions, capturing and converting light energy as a component of photosystem supercomplexes. Chlorophyll degradation during leaf senescence is also required for adequate degeneration of chloroplasts and salvaging of nutrients from senescent leaves. In this study, we performed genetic analysis to determine the functions of BALANCE of CHLOROPHYLL METABOLISM1 (BCM1) and BCM2, which control chlorophyll levels by regulating synthesis and degradation, and STAY-GREEN1 (SGR1, also known as NON-YELLOWING1 [NYE1]) and SGR2, which encode Mg-dechelatase and catalyze chlorophyll a degradation in Arabidopsis (Arabidopsis thaliana). Analysis of bcm1 bcm2 revealed that both BCM1 and BCM2 are involved in the regulation of chlorophyll levels in presenescent leaves and chlorophyll degradation in senescing leaves. Analysis of bcm1 bcm2 nye1 nye2 suggested that BCMs repress chlorophyll-degrading activity in both presenescent and senescing leaves by regulating SGR activity. Furthermore, transactivation analysis and ChIP assay revealed that GOLDEN2-LIKE1 (GLK1), a central transcription factor regulating the expression of genes encoding photosystem-related proteins, such as light-harvesting Chl a/b-binding proteins (LHCPs), directly regulates the transcription of BCM1. LHCPs are stabilized by chlorophyll binding, suggesting that GLKs control the amount of LHCP through transcriptional and post-translational regulation via BCM-mediated chlorophyll-level regulation. Meanwhile, we generated a mutant of the BCM ortholog in lettuce (Lactuca sativa) by genome editing and found that it showed an early yellowing phenotype, but only a slight reduction in chlorophyll in presenescent leaves. Thus, this study revealed a conserved but slightly diversified regulation of chlorophyll and LHCP levels via the GLK-BCM pathway in eudicots.
  • Functional Divergence of G and Its Homologous Genes for Green Pigmentation in Soybean Seeds
    Yusuke Tokumitsu; Takuto Kozu; Hiroshi Yamatani; Takeshi Ito; Haruna Nakano; Ayaka Hase; Hiroki Sasada; Yoshitake Takada; Akito Kaga; Masao Ishimoto; Makoto Kusaba; Taiken Nakashima; Jun Abe; Tetsuya Yamada
    Frontiers in Plant Science, 12, Frontiers Media SA, 05 Jan. 2022, [Peer-reviewed], [Corresponding author]
    Scientific journal, The degradation of chlorophyll in mature soybean seeds is closely related to the development of their yellow color. In this study, we examined G, its homologue G-like (GL), and their mutant alleles and investigated the relationship between these genes and chlorophyll accumulation in the seed coats of mature seeds. Transient expression of G and GL proteins fused with green fluorescent protein revealed that both were localized in plastids. Overexpression of G resulted in the accumulation of chlorophyll in the seed coats and cotyledons of mature seeds, indicating that high expression levels of G result in chlorophyll accumulation that exceeds its metabolism in the seeds of yellow soybean. Analysis of near isogenic lines at the G locus demonstrated a significant difference in the chlorophyll content of the seed coats and cotyledons of mature seeds when G and mutant g alleles were expressed in the d1d2 stay-green genetic background, indicating that the G protein might repress the SGR-independent degradation of chlorophyll. We examined the distribution of mutant alleles at the G and GL loci among cultivated and wild soybean germplasm. The g allele was widely distributed in cultivated soybean germplasm, except for green seed coat soybean lines, all of which contained the G allele. The gl alleles were much fewer in number than the g alleles and were mainly distributed in the genetic resources of cultivated soybean from Japan. None of the landraces and breeding lines investigated in this study were observed to contain both the g and gl alleles. Therefore, in conclusion, the mutation of the G locus alone is essential for establishing yellow soybeans, which are major current soybean breeding lines.
  • Identification and Characterization of an Early Leaf Senescence Gene ELS1 in Soybean
    Hiroshi Yamatani; Titnarong Heng; Tetsuya Yamada; Makoto Kusaba; Akito Kaga
    Frontiers in Plant Science, 12, Frontiers Media SA, 16 Dec. 2021, [Peer-reviewed]
    Scientific journal, Early leaf senescence phenotype in soybean could be helpful to shorten the maturation period and prevent green stem disorder. From a high-density mutation library, we identified two early leaf senescence soybean mutant lines, els1-1 (early leaf senescence 1) and els1-2. The chlorophyll contents of both els1-1 and els1-2 were low in pre-senescent leaves. They degraded rapidly in senescent leaves, revealing that ELS1 is involved in chlorophyll biosynthesis during leaf development and chlorophyll degradation during leaf senescence. The causal mutations in els1 were identified by next-generation sequencing-based bulked segregant analysis. ELS1 encodes the ortholog of the Arabidopsis CaaX-like protease BCM1, which is localized in chloroplasts. Soybean ELS1 was highly expressed in green tissue, especially in mature leaves. The accumulation of photosystem I core proteins and light-harvesting proteins in els1 was low even in pre-senescent leaves, and their degradation was accelerated during leaf senescence. These results suggest that soybean ELS1 is involved in both chlorophyll synthesis and degradation, consistent with the findings in Arabidopsis BCM1. The gene els1, characterized by early leaf senescence and subsequent early maturation, does not affect the flowering time. Hence, the early leaf senescence trait regulated by els1 helps shorten the harvesting period because of early maturation characteristics. The els1-1 allele with weakly impaired function of ELS1 has only a small effect on agricultural traits and could contribute to practical breeding.
  • Anhydrous proton-conducting material consisting of basic protein protamine
    Masanori Yamada; Takaaki Sugihara; Tetsuya Yamada
    Journal of Electroanalytical Chemistry, 897, 115586, 115586, Elsevier BV, Sep. 2021, [Peer-reviewed]
    Scientific journal
  • Wheat α-gliadin and high-molecular-weight glutenin subunit accumulate in different storage compartments of transgenic soybean seed
    Yuki Matsuoka; Tetsuya Yamada; Nobuyuki Maruyama
    Transgenic Research, 31, 1, 43, 58, Springer Science and Business Media LLC, 24 Aug. 2021, [Peer-reviewed]
    Scientific journal
  • Site-directed mutagenesis by biolistic transformation efficiently generates inheritable mutations in a targeted locus in soybean somatic embryos and transgene-free descendants in the T1 generation
    Kohei Adachi; Aya Hirose; Yuhei Kanazashi; Miki Hibara; Toshiyuki Hirata; Masafumi Mikami; Masaki Endo; Sakiko Hirose; Nobuyuki Maruyama; Masao Ishimoto; Jun Abe; Tetsuya Yamada
    Transgenic Research, 30, 1, 77, 89, Springer Science and Business Media LLC, 01 Jan. 2021, [Peer-reviewed], [Corresponding author], [International Magazine]
    English, Scientific journal, The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated endonuclease 9 (Cas9) system is being rapidly developed for mutagenesis in higher plants. Ideally, foreign DNA introduced by this system is removed in the breeding of edible crops and vegetables. Here, we report an efficient generation of Cas9-free mutants lacking an allergenic gene, Gly m Bd 30K, using biolistic transformation and the CRISPR/Cas9 system. Five transgenic embryo lines were selected on the basis of hygromycin resistance. Cleaved amplified polymorphic sequence analysis detected only two different mutations in e all of the lines. These results indicate that mutations were induced in the target gene immediately after the delivery of the exogenous gene into the embryo cells. Soybean plantlets (T0 plants) were regenerated from two of the transgenic embryo lines. The segregation pattern of the Cas9 gene in the T1 generation, which included Cas9-free plants, revealed that a single copy number of transgene was integrated in both lines. Immunoblot analysis demonstrated that no Gly m Bd 30K protein accumulated in the Cas9-free plants. Gene expression analysis indicated that nonsense mRNA decay might have occurred in mature mutant seeds. Due to the efficient induction of inheritable mutations and the low integrated transgene copy number in the T0 plants, we could remove foreign DNA easily by genetic segregation in the T1 generation. Our results demonstrate that biolistic transformation of soybean embryos is useful for CRISPR/Cas9-mediated site-directed mutagenesis of soybean for human consumption.
  • Anhydrous Proton Conduction of Soy Protein
    Yamada M; Y Nagano; T Yamada
    International Journal of Electrochemical Science, 16, 1, 151046, 151046, Jan. 2021, [Peer-reviewed]
    English, Scientific journal
  • Artificially Edited Alleles of the Eukaryotic Translation Initiation Factor 4E1 Gene Differentially Reduce Susceptibility to Cucumber Mosaic Virus and Potato Virus Y in Tomato
    Hiroki Atarashi; Wikum Harshana Jayasinghe; Joon Kwon; Hangil Kim; Yosuke Taninaka; Manabu Igarashi; Kotaro Ito; Tetsuya Yamada; Chikara Masuta; Kenji S. Nakahara
    Frontiers in Microbiology, 11, 564310, 564310, Frontiers Media SA, 10 Dec. 2020, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Eukaryotic translation initiation factors, including eIF4E, are susceptibility factors for viral infection in host plants. Mutation and double-stranded RNA-mediated silencing of tomato eIF4E genes can confer resistance to viruses, particularly members of the Potyvirus genus. Here, we artificially mutated the eIF4E1 gene on chromosome 3 of a commercial cultivar of tomato (Solanum lycopersicum L.) by using CRISPR/Cas9. We obtained three alleles, comprising two deletions of three and nine nucleotides (3DEL and 9DEL) and a single nucleotide insertion (1INS), near regions that encode amino acid residues important for binding to the mRNA 5' cap structure and to eIF4G. Plants homozygous for these alleles were termed 3DEL, 9DEL, and 1INS plants, respectively. In accordance with previous studies, inoculation tests with potato virus Y (PVY; type member of the genus Potyvirus) yielded a significant reduction in susceptibility to the N strain (PVYN), but not to the ordinary strain (PVYO), in 1INS plants. 9DEL among three artificial alleles had a deleterious effect on infection by cucumber mosaic virus (CMV, type member of the genus Cucumovirus). When CMV was mechanically inoculated into tomato plants and viral coat accumulation was measured in the non-inoculated upper leaves, the level of viral coat protein was significantly lower in the 9DEL plants than in the parental cultivar. Tissue blotting of microperforated inoculated leaves of the 9DEL plants revealed significantly fewer infection foci compared with those of the parental cultivar, suggesting that 9DEL negatively affects the initial steps of infection with CMV in a mechanically inoculated leaf. In laboratory tests, viral aphid transmission from an infected susceptible plant to 9DEL plants was reduced compared with the parental control. Although many pathogen resistance genes have been discovered in tomato and its wild relatives, no CMV resistance genes have been used in practice. RNA silencing of eIF4E expression has previously been reported to not affect susceptibility to CMV in tomato. Our findings suggest that artificial gene editing can introduce additional resistance to that achieved with mutagenesis breeding, and that edited eIF4E alleles confer an alternative way to manage CMV in tomato fields.
  • Simultaneous induction of mutant alleles of two allergenic genes in soybean by using site-directed mutagenesis
    Shota Sugano; Aya Hirose; Yuhei Kanazashi; Kohei Adachi; Miki Hibara; Takeshi Itoh; Masafumi Mikami; Masaki Endo; Sakiko Hirose; Nobuyuki Maruyama; Jun Abe; Tetsuya Yamada
    BMC Plant Biology, 20, 1, 513, 513, Springer Science and Business Media LLC, Dec. 2020, [Peer-reviewed], [Corresponding author], [International Magazine]
    English, Scientific journal, Abstract

    Background
    Soybean (Glycine max) is a major protein crop, because soybean protein has an amino acid score comparable to that of beef and egg white. However, many allergens have been identified among soybean proteins. A decrease in allergenic protein levels would be useful for expanding the market for soybean proteins and processed foods. Recently, the CRISPR/Cas9 system has been adopted as a powerful tool for the site-directed mutagenesis in higher plants. This system is expected to generate hypoallergenic soybean varieties.




    Results
    We used two guide RNAs (gRNAs) and Agrobacterium-mediated transformation for simultaneous site-directed mutagenesis of two genes encoding the major allergens Gly m Bd 28 K and Gly m Bd 30 K in two Japanese soybean varieties, Enrei and Kariyutaka. We obtained two independent T0 Enrei plants and nine T0 Kariyutaka plants. Cleaved amplified polymorphic sequence (CAPS) analysis revealed that mutations were induced in both targeted loci of both soybean varieties. Sequencing analysis showed that deletions were the predominant mutation type in the targeted loci. The Cas9-free plants carrying the mutant alleles of the targeted loci with the transgenes excluded by genetic segregation were obtained in the T2 and T3 generations. Variable mutational spectra were observed in the targeted loci even in T2 and T3 progenies of the same T0 plant. Induction of multiple mutant alleles resulted in six haplotypes in the Cas9-free mutants derived from one T0 plant. Immunoblot analysis revealed that no Gly m Bd 28 K or Gly m Bd 30 K protein accumulated in the seeds of the Cas9-free plants. Whole-genome sequencing confirmed that a Cas9-free mutant had also no the other foreign DNA from the binary vector. Our results demonstrate the applicability of the CRISPR/Cas9 system for the production of hypoallergenic soybean plants.




    Conclusions
    Simultaneous site-directed mutagenesis by the CRISPR/Cas9 system removed two major allergenic proteins from mature soybean seeds. This system enables rapid and efficient modification of seed components in soybean varieties.


  • Characterization of chlorophyll-deficient soybean [Glycine max (L.) Merr.] mutants obtained by ion-beam irradiation reveals concomitant reduction in isoflavone levels
    Yui Shiroshita; Mashiro Yuhazu; Yoshihiro Hase; Tetsuya Yamada; Jun Abe; Akira Kanazawa
    Genetic Resources and Crop Evolution, 68, 3, 1213, 1223, 17 Nov. 2020, [Peer-reviewed]
    English, Scientific journal
  • Preparation of bioplastic using soy protein
    Masanori Yamada; Sakura Morimitsu; Eiji Hosono; Tetsuya Yamada
    International Journal of Biological Macromolecules, 149, 1077, 1083, Apr. 2020, [Peer-reviewed]
    English, Scientific journal
  • A space-saving visual screening method, Glycine max FAST, for generating transgenic soybean
    Kosei Iwabuchi; Takashi L. Shimada; Tetsuya Yamada; Ikuko Hara-Nishimura
    Plant Signaling & Behavior, 15, 2, 1722911, 1722911, Informa UK Limited, 01 Feb. 2020, [Peer-reviewed]
    Scientific journal
  • A Soybean Deletion Mutant That Moderates the Repression of Flowering by Cool Temperatures.
    Jingyu Zhang; Meilan Xu; Maria Stefanie Dwiyanti; Satoshi Watanabe; Tetsuya Yamada; Yoshihiro Hase; Akira Kanazawa; Takashi Sayama; Masao Ishimoto; Baohui Liu; Jun Abe
    Frontiers in plant science, 11, 429, 429, 2020, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Ambient growing temperature and photoperiod are major environmental stimuli that summer annual crops use to adjust their reproductive phenology so as to maximize yield. Variation in flowering time among soybean (Glycine max) cultivars results mainly from allelic diversity at loci that control photoperiod sensitivity and FLOWERING LOCUS T (FT) orthologs. However, variation in the thermal regulation of flowering and its underlying mechanisms are poorly understood. In this study, we identified a novel mutant (ef1) that confers altered thermal regulation of flowering in response to cool ambient temperatures. Mapping analysis with simple sequence repeat (SSR) markers located the mutation in the upper part of chromosome 19, where no QTL for flowering has been previously reported. Fine-mapping and re-sequencing revealed that the mutation was caused by deletion of a 214 kbp genomic region that contains 11 annotated genes, including CONSTANS-LIKE 2b (COL2b), a soybean ortholog of Arabidopsis CONSTANS. Comparison of flowering times under different photo-thermal conditions revealed that early flowering in the mutant lines was most distinct under cool ambient temperatures. The expression of two FT orthologs, FT2a and FT5a, was dramatically downregulated by cool temperature, but the magnitude of the downregulation was lower in the mutant lines. Cool temperatures upregulated COL2b expression or delayed peak expression, particularly at the fourth trifoliate-leaf stage. Intriguingly, they also upregulated E1, a soybean-specific repressor of FT orthologs. Our results suggest that the ef1 mutation is involved in thermal regulation of flowering in response to cool ambient temperature, and the lack of COL2b in the mutant likely alleviates the repression of flowering by cool temperature. The ef1 mutant can be used as a novel gene resource in breeding soybean cultivars adapted to cool climate and in research to improve our understanding of thermal regulation of flowering in soybean.
  • Identification of quantitative trait loci for increased α-tocopherol biosynthesis in wild soybean using a high-density genetic map.
    Park C; Dwiyanti MS; Nagano AJ; Liu B; Yamada T; Abe J
    BMC plant biology, 19, 1, 510, Nov. 2019, [Peer-reviewed]
    English, Scientific journal
  • Soybean (Glycine max L.) triacylglycerol lipase GmSDP1 regulates the quality and quantity of seed oil
    Kanai, Masatake; Yamada, Tetsuya; Hayashi, Makoto; Mano, Shoji; Nishimura, Mikio
    SCIENTIFIC REPORTS, 9, Jun. 2019, [Peer-reviewed]
    English, Scientific journal
  • Functional divergence between soybean FLOWERING LOCUS T orthologues, FT2a and FT5a, in post-flowering stem growth
    Ryoma Takeshima; Haiyang Nan; Kohei Harigai; Lidong Dong; Jianghui Zhu; Sijia Lu; Meilan Xu; Noriko Yamagishi; Nobuyuki Yoshikawa; Baohui Liu; Tetsuya Yamada; Fanjiang Kong; Jun Abe
    Journal of Experimental Botany, 70, 15, 3941, 3953, Oxford University Press ({OUP}), Apr. 2019, [Peer-reviewed]
  • Recessive resistance governed by a major quantitative trait locus restricts clover yellow vein virus in mechanically but not graft-inoculated cultivated soybeans.
    Abe J; Wang Y; Yamada T; Sato M; Ono T; Atsumi G; Abe J; Hajimorad MR; Nakahara KS
    Molecular plant-microbe interactions : MPMI, 32, 8, 1026, 1037, American Phytopathological Society(APS), Mar. 2019, [Peer-reviewed]
    English
  • Changes in storage components and organelles structure during filling of soybean seeds
    Yoda N; Osada A; Suzuki T; Hirata T; Ito T; Abe J; Yamada T
    Report of the Hokkaido Branch, the Japanese Society of Breeding and Hokkaido Branch, the Crop Science Society of Japan, 60, 4, 5, Hokkaido Branch, the Japanese Society of Breeding and Hokkaido Branch, the Crop Science Society of Japan, 2019
    Japanese
  • CRISPR/Cas9-Mediated Editing of Genes Encoding rgs-CaM-like Proteins in Transgenic Potato Plants
    Zhila Osmani; Shinnosuke Jin; Masafumi Mikami; Masaki Endo; Hiroki Atarashi; Kaien Fujino; Tetsuya Yamada; Kenji S. Nakahara
    Methods in Molecular Biology, 2028, 153, 165, Springer New York, 2019, [Peer-reviewed], [International Magazine]
    English, In book, A tobacco calmodulin-like protein, rgs-CaM, has been shown to interact with viruses in a variety of ways; it contributes to geminivirus infections but is also involved in primed immunity to the cucumber mosaic virus. Sequence similarity searches revealed several calmodulin-like proteins similar to rgs-CaM (rCML) in Arabidopsis and other Solanaceae plants, including potato (Solanum tuberosum). To analyze the functions of each rCML, mutations were introduced into potato rCMLs using the CRISPR/Cas9 system. Here, we describe our protocol of the CRISPR/Cas9-mediated targeted mutagenesis in stably transformed potato plants.
  • Structural features of the aleurone layer of the seed coat associated with imbibition injury in soybean
    Kei Sato; Yutaka Jitsuyama; Tetsuya Yamada; Baohui Liu; Jun Abe
    Breeding Science, 69, 2, 364, 370, Japanese Society of Breeding, 2019, [Peer-reviewed]
    Scientific journal
  • Characterization and quantitative trait locus mapping of late-flowering from a Thai soybean cultivar introduced into a photoperiod-insensitive genetic background.
    Sun F; Xu M; Park C; Dwiyanti MS; Nagano AJ; Zhu J; Watanabe S; Kong F; Liu B; Yamada T; Abe J
    PloS one, 14, 12, e0226116, 2019, [Peer-reviewed]
    English, Scientific journal
  • Loss of Function of the E1-Like-b Gene Associates With Early Flowering Under Long-Day Conditions in Soybean
    Jianghui Zhu; Ryoma Takeshima; Kohei Harigai; Meilan Xu; Fanjiang Kong; Baohui Liu; Akira Kanazawa; Tetsuya Yamada; Jun Abe
    Frontiers in Plant Science, 9, 1867, Frontiers Media {SA}, Jan. 2019, [Peer-reviewed]
  • A vacuolar sorting receptor-independent sorting mechanism for storage vacuoles in soybean seeds
    Nobuyuki Maruyama; Yuki Matsuoka; Kazunori Yokoyama; Kyoko Takagi; Tetsuya Yamada; Hisakazu Hasegawa; Teruhiko Terakawa; Masao Ishimoto
    Scientific Reports, 8, 1, 1108, Nature Publishing Group, 01 Dec. 2018, [Peer-reviewed]
    English, Scientific journal
  • Simultaneous site-directed mutagenesis of duplicated loci in soybean using a single guide RNA
    Yuhei Kanazashi; Aya Hirose; Ippei Takahashi; Masafumi Mikami; Masaki Endo; Sakiko Hirose; Seiichi Toki; Akito Kaga; Ken Naito; Masao Ishimoto; Jun Abe; Tetsuya Yamada
    Plant Cell Reports, 37, 3, 553, 563, Springer Verlag, 01 Mar. 2018, [Peer-reviewed]
    English, Scientific journal
  • Preparation of water-insoluble and biochemically-stable RNA hybrid material.
    Yamada M; C Tohyama; T Yamada
    Polymer for Advanced Technologies, 29, 12, 2890, 2898, 2018, [Peer-reviewed]
    English, Scientific journal
  • Development of a high-density linkage map and chromosome segment substitution lines for Japanese soybean cultivar Enrei.
    Watanabe S; Shimizu T; Machita K; Tsubokura Y; Xia Z; Yamada T; Hajika M; Ishimoto M; Katayose Y; Harada K; Kaga A
    DNA research : an international journal for rapid publication of reports on genes and genomes, 25, 2, 123, 136, Nov. 2017, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Using progeny of a cross between Japanese soybean Enrei and Chinese soybean Peking, we developed a high-density linkage map and chromosomal segment substitution lines (CSSLs). The map consists of 2,177 markers with polymorphism information for 32 accessions and provides a detailed genetic framework for these markers. The marker order on the linkage map revealed close agreement with that on the chromosome-scale assembly, Wm82.a2.v1. The differences, especially on Chr. 5 and Chr. 11, in the present map provides information to identify regions in the genome assembly where additional information is required to resolve marker order and assign remaining scaffolds. To cover the entire soybean genome, we used 999 BC3F2 backcross plants and selected 103 CSSLs carrying chromosomal segments from Peking in the genetic background of Enrei. Using these low-genetic-complexity resources, we dissected variation in traits related to flowering, maturity and yield into approximately 50 reproducible quantitative trait loci (QTLs) and evaluated QTLs with small genetic effects as single genetic factors in a uniform genetic background. CSSLs developed in this study may be good starting material for removing the unfavourable characteristics of Peking during pre-breeding and for isolation of genes conferring disease and stress resistance that have not yet been characterized.
  • Identification of quantitative trait loci for flowering time by a combination of restriction site-associated DNA sequencing and bulked segregant analysis in soybean
    Satoshi Watanabe; Chikaharu Tsukamoto; Tatsuki Oshita; Tetsuya Yamada; Toyoaki Anai; Akito Kaga
    BREEDING SCIENCE, 67, 3, 277, 285, Jun. 2017, [Peer-reviewed]
    English, Scientific journal
  • RNA silencing in the life cycle of soybean: multiple restriction systems and spatiotemporal variation associated with plant architecture
    Ayumi Mori; Hiroshi Sato; Megumi Kasai; Tetsuya Yamada; Akira Kanazawa
    TRANSGENIC RESEARCH, 26, 3, 349, 362, Jun. 2017, [Peer-reviewed]
    English, Scientific journal
  • The Non-Mendelian Green Cotyledon Gene in Soybean Encodes a Small Subunit of Photosystem II
    Kaori Kohzuma; Yutaka Sato; Hisashi Ito; Ayako Okuzaki; Mai Watanabe; Hideki Kobayashi; Michiharu Nakano; Hiroshi Yamatani; Yu Masuda; Yumi Nagashima; Hiroyuki Fukuoka; Tetsuya Yamada; Akira Kanazawa; Keisuke Kitamura; Yutaka Tabei; Masahiko Ikeuchi; Wataru Sakamoto; Ayumi Tanaka; Makoto Kusaba
    PLANT PHYSIOLOGY, 173, 4, 2138, 2147, Apr. 2017, [Peer-reviewed]
    English, Scientific journal
  • Isolation and molecular characterization of a Lotus japonicus R2R3-MYB subgroup 7 transcription factor gene
    Satoru Kunihiro; Daigo Tanabe; Yuiko Niwa; Keisuke Kitamura; Jun Abe; Tetsuya Yamada
    PLANT BIOTECHNOLOGY, 34, 1, 45, 49, Mar. 2017, [Peer-reviewed]
    English, Scientific journal
  • Frequent generation of mutants with coincidental changes in multiple traits via ion-beam irradiation in soybean
    Shun Mikuriya; Megumi Kasai; Kenta Nakashima; Natasia; Yoshihiro Hase; Tetsuya Yamada; Jun Abe; Akira Kanazawa
    Genes and Genetic Systems, 92, 3, 153, 161, Genetics Society of Japan, 2017, [Peer-reviewed]
    English, Scientific journal
  • A soybean quantitative trait locus that promotes flowering under long days is identified as FT5a, a FLOWERING LOCUS T ortholog
    Ryoma Takeshima; Takafumi Hayashi; Jianghui Zhu; Chen Zhao; Meilan Xu; Naoya Yamaguchi; Takashi Sayama; Masao Ishimoto; Lingping Kong; Xinyi Shi; Baohui Liu; Zhixi Tian; Tetsuya Yamada; Fanjiang Kong; Jun Abe
    JOURNAL OF EXPERIMENTAL BOTANY, 67, 17, 5247, 5258, Sep. 2016, [Peer-reviewed]
    English, Scientific journal
  • Natural diversity of seed alpha-tocopherol ratio in wild soybean (Glycine sofa) germplasm collection
    Maria Stefanie Dwiyanti; Shouhei Maruyama; Mari Hirono; Masako Sato; Euiho Park; Sei Hyung Yoon; Tetsuya Yamada; Jun Abe
    BREEDING SCIENCE, 66, 4, 653, 657, Sep. 2016, [Peer-reviewed]
    English, Scientific journal
  • Ncl Synchronously Regulates Na+, K+, and Cl- in Soybean and Greatly Increases the Grain Yield in Saline Field Conditions
    Tuyen Duc Do; Huatao Chen; Hien Thi Thu Vu; Aladdin Hamwieh; Tetsuya Yamada; Tadashi Sato; Yongliang Yan; Hua Cong; Mariko Shono; Kazuhiro Suenaga; Donghe Xu
    SCIENTIFIC REPORTS, 6, 19147, Jan. 2016, [Peer-reviewed]
    English, Scientific journal
  • A recessive allele for delayed flowering at the soybean maturity locus E9 is a leaky allele of FT2a, a FLOWERING LOCUS T ortholog
    Chen Zhao; Ryoma Takeshima; Jianghui Zhu; Meilan Xu; Masako Sato; Satoshi Watanabe; Akira Kanazawa; Baohui Liu; Fanjiang Kong; Tetsuya Yamada; Jun Abe
    BMC PLANT BIOLOGY, 16, 20, Jan. 2016, [Peer-reviewed]
    English, Scientific journal
  • Biochemical and molecular characterization of orange- and tangerine-colored rice calli
    Atsushi Ishihara; Kazuhiko Ohishi; Tetsuya Yamada; Mari Shibata-Hatta; Yuko Arai-Kichise; Satoru Watanabe; Hirofumi Yoshikawa; Kyo Wakasa
    PLANT BIOTECHNOLOGY, 32, 3, 193, +, Sep. 2015, [Peer-reviewed]
    English, Scientific journal
  • The Soybean-Specific Maturity Gene E1 Family of Floral Repressors Controls Night-Break Responses through Down-Regulation of FLOWERING LOCUS T Orthologs
    Meilan Xu; Noriko Yamagishi; Chen Zhao; Ryoma Takeshima; Megumi Kasai; Satoshi Watanabe; Akira Kanazawa; Nobuyuki Yoshikawa; Baohui Liu; Tetsuya Yamada; Jun Abe
    PLANT PHYSIOLOGY, 168, 4, 1735, U1080, Aug. 2015, [Peer-reviewed]
    English, Scientific journal
  • A Single-Nucleotide Polymorphism in an Endo-1,4-beta-Glucanase Gene Controls Seed Coat Permeability in Soybean
    Seong-Jin Jang; Masako Sato; Kei Sato; Yutaka Jitsuyama; Kaien Fujino; Haruhide Mori; Ryoji Takahashi; Eduardo R. Benitez; Baohui Liu; Tetsuya Yamada; Jun Abe
    PLOS ONE, 10, 6, e0128527, Jun. 2015, [Peer-reviewed]
    English, Scientific journal
  • Knockdown of the 7S globulin subunits shifts distribution of nitrogen sources to the residual protein fraction in transgenic soybean seeds
    Tetsuya Yamada; Yoshihiro Mori; Kazuho Yasue; Nobuyuki Maruyama; Keisuke Kitamura; Jun Abe
    PLANT CELL REPORTS, 33, 12, 1963, 1976, Dec. 2014, [Peer-reviewed]
    English, Scientific journal
  • Double-Stranded DNA Stereoselectively Promotes Aggregation of Amyloid-Like Fibrils and Generates Peptide/DNA Matrices
    Masanori Yamada; Sachiko Hara; Tetsuya Yamada; Fumihiko Katagiri; Kentaro Hozumi; Motoyoshi Nomizu
    BIOPOLYMERS, 102, 6, 465, 472, Nov. 2014, [Peer-reviewed]
    English, Scientific journal
  • A Green-Cotyledon/Stay-Green Mutant Exemplifies the Ancient Whole-Genome Duplications in Soybean
    Michiharu Nakano; Tetsuya Yamada; Yu Masuda; Yutaka Sato; Hideki Kobayashi; Hiroaki Ueda; Ryouhei Morita; Minoru Nishimura; Keisuke Kitamura; Makoto Kusaba
    PLANT AND CELL PHYSIOLOGY, 55, 10, 1763, 1771, Oct. 2014, [Peer-reviewed]
    English, Scientific journal
  • Immobilization of double-stranded DNA onto glass beads by psolaren
    Masanori Yamada; Yuka Kanamori; Tetsuya Yamada
    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 60, 39, 44, Sep. 2013, [Peer-reviewed]
    English, Scientific journal
  • Genetic variation in four maturity genes affects photoperiod insensitivity and PHYA-regulated post-flowering responses of soybean
    Meilan Xu; Zeheng Xu; Baohui Liu; Fanjiang Kong; Yasutaka Tsubokura; Satoshi Watanabe; Zhengjun Xia; Kyuya Harada; Akira Kanazawa; Testuya Yamada; Jun Abe
    BMC PLANT BIOLOGY, 13, 91, Jun. 2013, [Peer-reviewed]
    English, Scientific journal
  • Preparation of DNA-polyintercalator conjugate and its functional property
    Masanori Yamada; Ayaka Okumoto; Tetsuya Yamada
    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 51, 3, 215, 220, Oct. 2012, [Peer-reviewed]
    English, Scientific journal
  • Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1 that regulates photoperiodic flowering
    Zhengjun Xia; Satoshi Watanabe; Tetsuya Yamada; Yasutaka Tsubokura; Hiroko Nakashima; Hong Zhai; Toyoaki Anai; Shusei Sato; Toshimasa Yamazaki; Shixiang Lu; Hongyan Wu; Satoshi Tabata; Kyuya Harada
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 109, 32, E2155, E2164, Aug. 2012, [Peer-reviewed]
    English, Scientific journal
  • Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1, which regulates photoperiodic flowering
    Zhengjun Xia; Satoshi Watanabe; Tetsuya Yamada; Yasutaka Tsubokura; Hiroko Nakashima; Hong Zhai; Toyoaki Anai; Shusei Sato; Toshimasa Yamazaki; Shixiang Lu; Hongyan Wu; Satoshi Tabata; Kyuya Harada
    Proc. Natl. Acad. Sci. USA, 109, 32, E2155, E2164, Aug. 2012, [Peer-reviewed]
    English
  • Synthesis of DNA intercalator-immobilized cyclodextrin and interaction with double-stranded DNA: Utilization of DNA-cyclodextrin conjugated material as an environmental remediation material
    Masanori Yamada; Masamitsu Inoue; Tetsuya Yamada
    POLYMER CHEMISTRY, 3, 5, 1291, 1299, 2012, [Peer-reviewed]
    English, Scientific journal
  • Recent advances in soybean transformation and their application to molecular breeding and genomic analysis.
    Yamada T; Takagi K; Ishimoto M
    Breeding science, 61, 5, 480, 494, 5, Jan. 2012, [Peer-reviewed]
    English
  • Genetic variation of gamma-tocopherol methyltransferase gene contributes to elevated alpha-tocopherol content in soybean seeds
    Maria S. Dwiyanti; Tetsuya Yamada; Masako Sato; Jun Abe; Keisuke Kitamura
    BMC PLANT BIOLOGY, 11, 152, Nov. 2011, [Peer-reviewed]
    English, Scientific journal
  • Establishment of an efficient in vitro culture and particle bombardment-mediated transformation systems in Miscanthus sinensis Anderss., a potential bioenergy crop
    Xun Wang; Tetsuya Yamada; Fan-Jiang Kong; Yuki Abe; Yoichiro Hoshino; Hiroko Sato; Tadashi Takamizo; Akira Kanazawa; Toshihiko Yamada
    GLOBAL CHANGE BIOLOGY BIOENERGY, 3, 4, 322, 332, Aug. 2011, [Peer-reviewed]
    English, Scientific journal
  • Optimization of ion-beam irradiation for mutagenesis in soybean: effects on plant growth and production of visibly altered mutants
    Sachiko Arase; Yoshihiro Hase; Jun Abe; Megumi Kasai; Tetsuya Yamada; Keisuke Kitamura; Issay Narumi; Atsushi Tanaka; Akira Kanazawa
    PLANT BIOTECHNOLOGY, 28, 3, 323, 329, 2011, [Peer-reviewed]
    English, Scientific journal
  • Two Coordinately Regulated Homologs of FLOWERING LOCUS T Are Involved in the Control of Photoperiodic Flowering in Soybean
    Fanjiang Kong; Baohui Liu; Zhengjun Xia; Shusei Sato; Bo Min Kim; Satoshi Watanabe; Tetsuya Yamada; Satoshi Tabata; Akira Kanazawa; Kyuya Harada; Jun Abe
    PLANT PHYSIOLOGY, 154, 3, 1220, 1231, Nov. 2010, [Peer-reviewed]
    English, Scientific journal
  • Establishment of new method for analysis of starch contents and varietal differences in soybean seeds
    Woo-Hyeun Jeong; Kyuya Harada; Tetsuya Yamada; Jun Abe; Keisuke Kitamura
    BREEDING SCIENCE, 60, 2, 160, 163, Jun. 2010, [Peer-reviewed]
    English
  • The Soybean Stem Growth Habit Gene Dt1 Is an Ortholog of Arabidopsis TERMINAL FLOWER1
    Baohui Liu; Satoshi Watanabe; Tomoo Uchiyama; Fanjiang Kong; Akira Kanazawa; Zhengjun Xia; Atsushi Nagamatsu; Maiko Arai; Tetsuya Yamada; Keisuke Kitamura; Chikara Masuta; Kyuya Harada; Jun Abe
    PLANT PHYSIOLOGY, 153, 1, 198, 210, May 2010, [Peer-reviewed]
    English, Scientific journal
  • Cotyledonary node pre-wounding with a micro-brush increased frequency of Agrobacterium-mediated transformation in soybean
    Tetsuya Yamada; Satoshi Watanabe; Maiko Arai; Kyuya Harada; Keisuke Kitamura
    PLANT BIOTECHNOLOGY, 27, 2, 217, 220, 2010, [Peer-reviewed]
    English, Scientific journal
  • Spatial and temporal control of transcription of the soybean beta-conglycinin alpha subunit gene is conferred by its proximal promoter region and accounts for the unequal distribution of the protein during embryogenesis
    Yusuke Imoto; Tetsuya Yamada; Keisuke Kitamura; Akira Kanazawa
    GENES & GENETIC SYSTEMS, 83, 6, 469, 476, Dec. 2008, [Peer-reviewed]
    English, Scientific journal
  • Genetic analysis and biochemical characterization of the high lutein trait of wild soybean (Glycine soja Sieb. et Zucc.)
    Kyohei Kanamaru; Shaodong Wang; Tetsuya Yamada; Jun Abe; Keisuke Kitamura
    BREEDING SCIENCE, 58, 4, 393, 400, Dec. 2008, [Peer-reviewed]
    English, Scientific journal
  • Genetic relationship between lipid content and linolenic acid concentration in soybean seeds
    Masayuki Shibata; Kiyohiko Takayama; Aya Ujiie; Tetsuya Yamada; Jun Abe; Keisuke Kitamura
    BREEDING SCIENCE, 58, 4, 361, 366, Dec. 2008, [Peer-reviewed]
    English, Scientific journal
  • Mutation of a rice gene encoding a phenylalanine biosynthetic enzyme results in accumulation of phenylalanine and tryptophan
    Tetsuya Yamada; Fumio Matsuda; Koji Kasai; Shuichi Fukuoka; Keisuke Kitamura; Yuzuru Tozawa; Hisashi Miyagawa; Kyo Wakasa
    PLANT CELL, 20, 5, 1316, 1329, May 2008, [Peer-reviewed]
    English, Scientific journal
  • Simultaneous accumulation of high contents of alpha-tocopherol and lutein is possible in seeds of soybean (Glycine max (L.) merr.)
    Shaodong Wang; Kyohei Kanamaru; Wenbin Li; Jun Abe; Tetsuya Yamada; Keisuke Kitamura
    BREEDING SCIENCE, 57, 4, 297, 304, Dec. 2007, [Peer-reviewed]
    English, Scientific journal
  • Production of transgenic plants and their early seed set in Japanese soybean variety, Kariyutaka
    Hiroshi Sato; Tetsuya Yamada; Yohichi Kita; Masao Ishimoto; Keisuke Kitamura
    PLANT BIOTECHNOLOGY, 24, 5, 533, 536, Dec. 2007, [Peer-reviewed]
    English, Scientific journal
  • Genetic Analysis of High .ALPHA.-tocopherol Content in Soybean Seeds
    DWIYANTI Maria Stefanie; UJIIE Aya; THUY Le Thi Bich; YAMADA Tetsuya; KITAMURA Keisuke
    Breed Sci, 57, 1, 23-28 (J-STAGE), 28, Japanese Society of Breeding, 2007
    English, Tocopherols, known as members of vitamin E, are the major lipophilic antioxidants in soybean. Since soybean seeds contain a low concentration of α-tocopherol (α-Toc), the tocopherol isoform with the highest vitamin E activity, some breeding efforts have been implemented to increase the α-tocopherol content in the seeds. In the present study, we used F2 seed and F2 plant (F3 seeds) populations derived from a cross between a variety with a high α-tocopherol content, Keszthelyi A.S. (α-Toc 20–30%) and a Japanese cultivar, Ichihime (α-Toc < 10%) for genetic analysis of the high α-tocopherol concentration. There was no significant correlation between the α-tocopherol and total tocopherol contents in the F2 plant (F3 seeds) population, suggesting that the α-tocopherol and total tocopherol contents were regulated independently in the seeds. Therefore, it may be possible to increase the α-tocopherol concentration without affecting the total tocopherol content. Broad-sense heritability value for the α-tocopherol concentration trait was estimated to be 0.645 in the F2 seeds, suggesting that the α-tocopherol concentration trait was highly heritable. Chi-square goodness-of-fit test for evaluating the relationship between genotype of certain SSR markers and the α-tocopherol concentration showed that the Sat_243 and Sat_167 markers (linkage group K) were significantly associated with the α-tocopherol concentration (P < 0.05). These markers could be used for marker-assisted selection to breed soybean cultivars with high α-tocopherol concentrations. Analysis of the F2 seeds and the F2 plants (F3 seeds) for tocopherol composition showed that there was a significantly negative correlation between the α-tocopherol and γ-tocopherol concentrations, suggesting that the activity of γ-tocopherol methyltransferase, the enzyme that converts γ-tocopherol to α-tocopherol, is high in individuals with a high α-tocopherol concentration.
  • High-level tryptophan accumulation in seeds of transgenic rice and its limited effects on agronomic traits and seed metabolite profile
    Kyo Wakasa; Hisakazu Hasegawa; Hiroshi Nemoto; Fumio Matsuda; Haruna Miyazawa; Yuzuru Tozawa; Keiko Morino; Akira Komatsu; Tetsuya Yamada; Teruhiko Terakawa; Hisashi Miyagawa
    JOURNAL OF EXPERIMENTAL BOTANY, 57, 12, 3069, 3078, Sep. 2006, [Peer-reviewed]
    English, Scientific journal
  • Identification and characterization of wild soybean (Glycine soja Sieb. et Zecc.) strains with high lutein content
    Kyohei Kanamaru; Shaodong Wang; Jun Abe; Tetsuya Yamada; Keisuke Kitamura
    BREEDING SCIENCE, 56, 3, 231, 234, Sep. 2006, [Peer-reviewed]
    English, Scientific journal
  • Characterization of tryptophan-overproducing potato transgenic for a mutant rice anthranilate synthase alpha-subunit gene (OASA1D)
    F Matsuda; T Yamada; H Miyazawa; H Miyagawa; K Wakasa
    PLANTA, 222, 3, 535, 545, Oct. 2005, [Peer-reviewed]
    English, Scientific journal
  • Identification of soybean varieties with high α-tocopherol content
    Aya Ujiie; Tetsuya Yamada; Kenshiro Fujimoto; Yasushi Endo; Keisuke Kitamura
    Breeding Science, 55, 2, 123, 125, Jun. 2005, [Peer-reviewed]
    English, Scientific journal
  • Use of a feedback-insensitive α subunit of anthranilate synthase as a selectable marker for transformation of rice and potato
    Tetsuya Yamada; Yuzuru Tozawa; Hisakazu Hasegawa; Teruhiko Terakawa; Yasunobu Ohkawa; Kyo Wakasa
    Molecular Breeding, 14, 4, 363, 373, Springer Netherlands, 2005, [Peer-reviewed]
    English, Scientific journal
  • A selectable marker using cytochrome P450 monooxygenases for Arabidopsis transformation
    Hideyuki Inui; Ryu Yamada; Tetsuya Yamada; Yasunobu Ohkawa; Hideo Ohkawa
    Plant Biotechnology, 22, 4, 281, 286, Japanese Society for Plant Cell and Molecular Biology, 2005, [Peer-reviewed]
    English, Scientific journal
  • Enhancement of metabolizing herbicides in young tubers of transgenic potato plants with the rat CYP1A1 gene
    T Yamada; T Ishige; N Shiota; H Inui; H Ohkawa; Y Ohkawa
    THEORETICAL AND APPLIED GENETICS, 105, 4, 515, 520, Sep. 2002, [Peer-reviewed]
    English, Scientific journal
  • Inducible cross-tolerance to herbicides in transgenic potato plants with the rat CYP1A1 gene
    T. Yamada; Y. Ohashi; M. Ohshima; H. Inui; N. Shiota; H. Ohkawa; Y. Ohkawa
    Theoretical and Applied Genetics, 104, 2-3, 308, 314, 2002, [Peer-reviewed]
    English, Scientific journal
  • Ultra-simple DNA extraction method for marker-assisted selection using microsatellite markers in rice
    Nobuyuki Ikeda; Nonnatus S. Bautista; Tetsuya Yamada; Osamu Kamijima; Takashige Ishii
    Plant Molecular Biology Reporter, 19, 1, 27, 32, Springer Science and Business Media LLC, Mar. 2001, [Peer-reviewed]
    Scientific journal
  • Inheritance of Morphological Characters and Glycoalkaloids in Potatoes of Somatic Hybrids between DihaploidSolanum acauleand TetraploidSolanumtuberosum
    Nobuyuki Kozukue; Shuji Misoo; Tetsuya Yamada; Osamu Kamijima; Mendel Friedman
    Journal of Agricultural and Food Chemistry, 47, 10, 4478, 4483, American Chemical Society (ACS), Oct. 1999, [Peer-reviewed]
    Scientific journal
  • Cytological and molecular characterization of BC1 progeny from two somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum
    T Yamada; K Hosaka; N Kaide; K Nakagawa; S Misoo; O Kamijima
    Genome, 41, 6, 743, 750, Canadian Science Publishing, 01 Dec. 1998, [Peer-reviewed]
    Scientific journal, Two hexaploid somatic hybrids, DA12-3 (2n = 68) and DA28 (2n = 74), derived from cell fusion between dihaploid Solanum acaule and tetraploid Solanum tuberosum, were crossed with S. tuberosum. Two BC1 families, DA12T and DA28T, were generated, each containing 25 plants. Although the average of somatic chromosome numbers fitted hypothetical numbers assuming equal separation, considerable variation was found: 2n = 54-62 in DA12T and 2n = 58-65 in DA28T. Fifty-two of 80 RFLP single-copy probes generated 108 marker bands specific for S. acaule, by which BC1 progeny were analyzed. The segregation of marker bands from at least 44 probes was explained by two alleles per locus, which had to be heterozygous in dihaploid S. acaule and maintained in homozygous condition in homoeologous chromosomes of tetraploid S. acaule. On average, 73.7% of the chromosomes derived from S. acaule were detected as recombinants, suggesting that frequent homoeologous recombination occurred between S. acaule and S. tuberosum chromosomes. Therefore, S. acaule germplasm can be effectively transferred to the cultivated potato gene pool through backcrossing hexaploid somatic hybrids with S. tuberosum. In both BC1 families, S. acaule chromosomes 3, 4, 6, and 11 were detected as recombinants with higher frequencies (84-98%) than those of chromosomes 9 and 10 (60 and 58%, respectively), suggesting that chromosomal differentiation exists between S. acaule genomes.Key words: hexaploid somatic hybrids, Solanum acaule, homoeologous recombination, RFLP markers.
  • Nuclear genome constitution and other characteristics of somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum.
    Tetsuya Yamada; Kazuyoshi Hosaka; Kumi Nakagawa; Noriyo Kaide; Shuji Misoo; Osamu Kamijima
    Euphytica, 102, 2, 239, 246, Springer Science and Business Media LLC, 1998, [Peer-reviewed]
    Scientific journal
  • Characterization of Somatic Hybrids between Tetraploid Solanum tuberosum L. and Dihaploid S. acaule
    Tetsuya Yamada; Shuji Misoo; Takashige Ishii; Yasuhiro Ito; Keigo Takaoka; Osamu Kamijima
    Japanese Journal of Breeding, 47, 3, 229, 236, Japanese Society of Breeding, 01 Sep. 1997, [Peer-reviewed]
    Scientific journal
■ Other Activities and Achievements
■ Books and other publications
■ Syllabus
  • 農業生物学特論, 2024年, 修士課程, 農学院
  • 農業生物学特論演習, 2024年, 修士課程, 農学院
  • 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), 2024年, 学士課程, 全学教育
  • 生物資源科学実験Ⅰ, 2024年, 学士課程, 農学部
  • 工芸作物学, 2024年, 学士課程, 農学部
■ Affiliated academic society
  • JAPANESE SOCIETY FOR PLANT CELL AND MOLECULAR BIOLOGY
  • JAPANESE SOCIETY OF BREEDING
■ Research Themes
  • Soybean breeding learning from Azuki bean
    Grants-in-Aid for Scientific Research
    01 Apr. 2024 - 31 Mar. 2027
    高橋 有; 坂井 寛章; 山田 哲也
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), National Agriculture and Food Research Organization, 24K01736
  • Development of stress tolerant legume crops by Neo-domestication
    Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Jul. 2019 - Mar. 2022
    Takahashi Yu
    The aim of this study is to reverse the conventional idea of conferring stress tolerance to crops and develop new stress tolerant crops by conferring domestication traits to wild species. To speed up the process, this study identified three candidate genes responsible for three domestication traits in legume: loss of seed dormancy, loss of pod shattering, and organ gigantism.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), National Agriculture and Food Research Organization, 19KT0016
  • Creation of new biomass materials using defatted soy protein
    Grants-in-Aid for Scientific Research Challenging Research (Exploratory)
    Jun. 2019 - Mar. 2021
    Yamada Tetsuya
    In order to create a new biomass material from defatted soybean protein, a prototype bioplastic was fabricated using defatted soybean via the formaldehyde cross-linking. In addition, it was confirmed that the structure of bioplastic was significantly impaired by enzymatic treatment.
    To create a soybean material deviating from the conventional protein composition in soybean, we attempted to knock down 7S and 11S globulin subunit genes, which are major protein genes in mature soybean seeds, using RNA interference ( RNAi) technology. Protein analysis of the transgenic soybean seeds revealed that the accumulation of 7S and 11S globulin proteins was greatly reduced in the transgenic soybean plants compared to the conventional soybean plants.
    Japan Society for the Promotion of Science, Challenging Research (Exploratory), Hokkaido University, 19K22298
  • Physiological analysis of pericarp dehiscence mechanism involved in yield
    Grants-in-Aid for Scientific Research
    01 Apr. 2017 - 31 Mar. 2020
    Fujino Kaien
    Tartary buckwheat cultivars that allow easy dehulling are called rice-type cultivars. The rice and non-rice hull types are determined by a single gene, but this gene is unclear. Here, I show that cells underneath the epidermis in the proximity of ovary midribs undergo periclinal cell divisions in non-rice-type cultivars but do not in a rice-type cultivar. The cells that arose from the periclinal cell divisions later underwent lignification, which should increase mechanical strength of hulls. In RNA sequencing, a partial mRNA of an AGAMOUS ortholog in Tartary buckwheat (FtAG) was found to be absent in the rice-type cultivar. By RNA sequencing and linkage analysis, we also show that FtAG is a candidate for the gene that promotes differentiation of the cells undergoing the periclinal cell divisions and that thereby determines the RT and NT phenotypes in Tartary buckwheat.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 17H03754
  • Novel aspects of plant priming and its use for crop breeding
    Grants-in-Aid for Scientific Research
    01 Apr. 2016 - 31 Mar. 2020
    Nakahara Kenji
    We found that 6 calmodulin-like proteins (CML) and endogenous factors that binding to the CMLs interact with virus RNA silencing suppressors and are involved in systemic acquired resistance. Based on the results obtained in this study, we hypothize the molecular mechanism involving these CMLs and endogenous factors unldelying systemic acquired resistance. Moreover, genome editing the gene that were selected according to the results of this study confer antiviral resistance on tomato.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 16H04879
  • Molecular genetic and biochemical characterization of chlorophyll accumulation mutants in soybean
    Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Apr. 2016 - Mar. 2019
    Yamada Tetsuya; Tokumitsu Yusuke
    We isolated the responsible gene for the genetic factor Gsc1, which causes the stay green trait of chlorophyll accumulation in soybean seed coats. The sequencing analysis revealed that there was a substitution of one-base at the gene encoding the CAAX proteinase between green and colorless seed coat varieties. Transgenic soybean lines overexpressing this gene remarkably suppressed the decrease of chlorophylls in the seed coat organs during seed filling. When the distribution of the mutation in this gene was examined in soybean germplasm including green or colorless seed coat lines, this analysis revealed that all the seed coat green lines have a functional allele for this gene. We revealed also that almost all wild soybeans have functional allele for this gene. These results support the conclusion that the green seed coat traits was a trait derived from wild soybean.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 16K07550
  • 二瓶 直登
    科学研究費補助金(基盤研究(C))
    2014 - 2016
    二瓶 直登
    文部科学省, Competitive research funding
  • ダイズ遺伝資源に認められる耐塩性遺伝子の単離とその機能解析に関する研究
    科学研究費補助金(基盤研究(C))
    2014 - 2016
    許 東河
    文部科学省, Competitive research funding
  • Study on a network of genes related to flowering and growth habit and its responses to environments in soybean
    Grants-in-Aid for Scientific Research
    01 Apr. 2011 - 31 Mar. 2014
    ABE Jun; YAMADA Testuya
    Soybean is cultivated across a wide range of latitudes. Its wide adaptability is supported by a genetic diversity in the control of flowering. The purposes of this project were to reveal the genetic diversity at four flowering loci, E1 to E4 and to determine the genetic network of flowering genes and the responses to environments. There were three mechanisms in the control of photoperiod-insensitivity; dysfunction of phytochrome A genes, E3 and E4, dysfunction of E1, a possible transcriptional factor for FLOWERING LOCUS T (FT) orthologs, and an involvement of a novel gene(s). Two homologs of E1 inhibited flowering via down-regulation of FT, as in E1. Further, a light signal in a specific period of day induced the gene expression of E1 under the control of E3 and E4 to inhibit the induction of FT expressions, which consequently inhibited the floral induction. A pathway independent of FT may also be involved in the induction of flowering in soybean.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23380001
  • Establishment of highly gene transcriptional regulation mediated by artificial microRNA in soybean
    Grants-in-Aid for Scientific Research
    2011 - 2011
    YAMADA Tetsuya
    Artificial mircoRNA (amiRNA) was designed using the precursor sequence of the endogenous soybean miRNA to suppress the expression of target gene efficiently in soybean plants. The constructed amiRNA precursor contains a complementary sequence to the transcript of target gene. Transgenic soybean plants expressing the amiRNA were generated to suppress the gene expression of seed storage protein. The expression of these genes and accumulation of storage protein were significantly decreased in seeds of transgenic soybean. In addition, the function of glycosyltransferase gene for DDMP saponin, which is known to be one of functional chemical compounds in soybean seeds, was characterized by the amiRNA expression system. I demonstrated that the knock-down system established in this study was useful for the functional analysis of genes in soybean plant.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, Principal investigator, Competitive research funding, 23580002
  • Establishment and utilization of simple and rapid soybean transformation system in a Japanese variety, Kariyutaka
    Grants-in-Aid for Scientific Research(基盤研究(C))
    2008 - 2010
    Tetsuya YAMADA; Keisuke KITAMURA
    An efficient and stable transformation technology in higher plants is used to identify the gene function or to develop new varieties. Soybean transformation has continued to be laborious and the frequency of recovered transgenic plants is still low. We found that a Japanese soybean variety, Kariyutaka, possessed advantageous characters for transformation. We developed a simple and rapid transformation system in Kariyutaka. The transformation frequency in the method presented in this study was 4 times as high as in that obtained with our previous method.
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 20580003
  • Development of functional soybeans containing α-tocopherol, lutein and so on.
    Grants-in-Aid for Scientific Research(基盤研究(B))
    2007 - 2009
    Keisuke KITAMURA; Jyun ABE; Tetsuya YAMADA
    Soybean seeds contain low levels of α-tocopherol (Toc). Since α-Toc acts as vitamin E efficiently, increasing α-Toc content will improve nutritional value of soybean. Lutein shown to reduce risks for ocular diseases is found in low in the seeds. We have identified varieties with high α-Toc and wild soybean strains with high levels of lutein content. Genetic analysis of the high α-Toc and lutein traits, respectively, revealed that the high α-Toc and lutein contents are highly heritable. Two SSR markers, Sat_243 and Sat_167 were shown to be associated with high α-Toc content. We obtained some...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 北海道大学, Coinvestigator not use grants, Competitive research funding, 19380001
  • ダイズの高α-トコフェロール化遺伝子の同定と作用機序の解明
    科学研究費補助金(萌芽研究)
    2005 - 2006
    喜多村 啓介; 山田 哲也; 石本 政男
    高α-トコフェロール(Toc)品種「Keszthelyi A.S.」と普通品種「いちひめ」の交配後代の分離集団を用いた解析の結果、Sat_243とSat_167(連鎖群K)の遺伝子型とα-Toc含有率との問に密接な関係を認めた。また、α-Toc含有率とγ-Toc含有率の間に高い負の相関を認め,高α-Toc形質にはγ-Tocをα-Tocに変換するγ-Tocopherol methyltransferase(γ-TMT)が関与することが示唆された。次に,トコフェロール生合成関連酵素、Homogentisate phytyltransferase(HPT),2-Methyl-6-phytyl-1,4-benzoquinol methyltransferase(MPBQMT),Tocopherol cyclase(TC),及びγ-TMTの4種の酵素遺伝子について,ダイズTACライブラリーからPCRにより上記4酵素遺伝子を含むと推定されるクローンを単離した。さらに、上記の中でも特に高α-Toc化に強く影響を及ぼすと考えられるγ-TMTについて解析を進め,cDNAを用いた3'-RACEの結果から、γ-TMT遺伝子には少なくとも3コピー(γ-TMT1,γ-TMT2,γ-TMT3)が存在することを明らかにした。また、これらの3つの遺伝子の発現解析を行うためにリアルタイムRT-PCRを試みた。...
    文部科学省, 萌芽研究, 北海道大学, Coinvestigator not use grants, Competitive research funding, 17658001