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Ng H.M., Gondo T., Tanaka H., Akashi R.
Plant Cell Reports 43 ( 2 ) 2024年2月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Plant Cell Reports
Key Message: Genome editing by CRISPR/Cas9 can be applied to Z. matrella ‘Wakaba’, and knockout mutants of ZmNYC1 gene exhibited stay-green phenotype and reduced tillering. Abstract: Zoysia matrella is a widely used C4 warm-season turfgrass for landscaping, golf courses, and sports fields. Here, we used the CRISPR/Cas9 system to target the Non-Yellow Coloring1 (ZmNYC1) gene in the highly heterozygous allotetraploid Z. matrella ‘Wakaba’, aiming to generate a novel stay-green variety. Of 441 Agrobacterium-infected calli, 22 (5.0%) were transformed, and 14 of these (63.6%) showed targeted mutations through cleaved amplified polymorphic sequences analysis. Sequencing analysis revealed mutations mostly consisting of 1 or 2 bp indels, occurring 2 to 4 bp upstream of the PAM sequence. Regenerated plants exhibited five ZmNYC1 target locus genotypes, including homozygous mutants with a complete knockout of all four alleles in the T0 generation. Under dark treatment, ZmNYC1-mutated plants displayed suppressed chlorophyll b (Chl b) degradation, leading to higher chlorophyll content and Chl b, with a lower chlorophyll a/chlorophyll b ratio compared to the wild type (WT). However, the ZmNYC1 mutation also inhibited plant growth in homozygous mutant genotypes, exhibiting reduced tillering compared to WT. Additionally, during winter simulation, mutant with a complete knockout retained greenness longer than the WT. This is the first successful use of CRISPR/Cas9 genome editing in zoysiagrass. The mutants of the ZmNYC1 gene would serve as valuable breeding material for developing improved zoysiagrass varieties that can maintain their green color for longer periods, even during winter dormancy.
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Ng H.M., Gondo T., Yamada T., Tanaka H., Akashi R.
Grassland Science 2024年
掲載種別：研究論文（学術雑誌） 出版者・発行元：Grassland Science
Bahiagrass (Paspalum notatum Flügge) is a warm-season grass where high lignin content limits its forage quality, negatively affecting animal performance. To create a new breeding method by genome editing, isolation and characterization of lignin biosynthesis genes and identification of their molecular relationships are essential. The caffeic acid O-methyltransferase (COMT) and cinnamyl alcohol dehydrogenase (CAD) cDNA clones were isolated from bahiagrass, and their protein sequences showed high similarity to other C4 monocot species through phylogenetic analysis. Gene expression analysis of phenylalanine ammonia-lyase (PAL), COMT and CAD, involved in crucial stages of lignin biosynthesis and lignin content, was conducted at different plant developmental stages. The highest gene expression levels of these genes were observed in the vegetative (V) and early reproductive stages, while the lignin content increased until the middle reproductive stage and remained constant thereafter. Additionally, to further understand the molecular relationships in lignin biosynthesis, the effect of CAD downregulation was analyzed in transgenic bahiagrass lines introduced with sorghum CAD antisense and RNAi vector obtained from a previous report. This led to a reduced lignin content and affected the expression of PAL and COMT working upstream of CAD. In the V stage, PAL expression was lower in transgenic lines compared to wild type (WT), while COMT expression showed no significant differences. However, PAL and COMT expression of transgenic lines in the middle reproductive stage (R2) was significantly higher than in the WT. These findings suggest that the downregulation of CAD gene expression affects PAL and COMT expression and induces a feedback system in the R2 stage. Lignin content influenced the phenotype of the transgenic plants with significantly reduced lignin, exhibiting a dwarf phenotype with shorter plant heights. The findings of this study can be applied to genome editing for the development of practical new breeding materials with improved digestibility in bahiagrass.
DOI： 10.1111/grs.12419
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Development of a digital phenotyping system using 3D model reconstruction for zoysiagrass 査読あり
Pongpiyapaiboon S., Tanaka H., Hashiguchi M., Hashiguchi T., Hayashi A., Tanabata T., Isobe S., Akashi R.
Plant Phenome Journal 6 ( 1 ) 2023年6月
担当区分：責任著者 記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Plant Phenome Journal
Digital phenotyping, particularly the use of plant 3D models, is a promising method for high-throughput plant evaluation. Although many recent studies on the topic have been published, further research is needed to apply it to breeding research and other related fields. In this study, using a 3D model phenotyping system we developed, we reconstructed and analyzed 20 accessions of zoysiagrass (Zoysia spp.), including three species and their hybrid, over a period of 1 year. Artificial neural network with three hidden layers was able to effectively remove nonplant parts while retaining plant parts that were incorrectly removed using the cropping method, offering a robust and flexible approach for post-processing of 3D models. The system also demonstrated its ability to accurately evaluate a range of traits, including height, area, and color using red green blue (RGB)-based vegetation indices. The results showed a high correlation between the estimated volume obtained from voxel 3D model and dry weight, enabling its use as a non-destructive method for measuring plant volume. In addition, we found that the green red normalized difference index from RGB-based indices was similar to the commonly used normalized difference vegetation index in controlled illumination conditions. These results demonstrate the potential for three-dimensional model phenotyping to facilitate plant breeding, particularly in the field of turfgrass and feed crops.
DOI： 10.1002/ppj2.20076
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Jiamtae P., Hashiguchi T., Pongpiyapaiboon S., Lelapiyamit I., Tanaka H., Hashiguchi M., Akashi R.
Journal of Crop Science and Biotechnology 26 ( 4 ) 467 - 488 2023年
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Crop Science and Biotechnology
Soybean is a major crop that is rich in proteins, lipids, and isoflavones. Although the seeds are widely used for food and industrial purposes, soybean is recognized as a crop that does not store well, as storage conditions greatly affect seed quality. In this study, we selected 10 recombinant inbred lines (RILs) of soybean that are either sensitive or tolerant to storage, and examined the biochemical characteristics of the lines and their parents under storage-related stresses. The germination rates of all of the lines in the tolerant group were higher than those of the sensitive group for all storage conditions. Malondialdehyde (MDA) content was higher in the sensitive group than in the tolerant group, with the MDA content typically increasing under long-term storage conditions. Superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities were also more affected by long-term storage, but not by higher temperatures. Isoflavones were also assayed and it was found that the aglycones (except glycitein) increased with an increase of storage temperature. Malonyl isoflavone glucosides tended to decrease under long-term storage and high temperature conditions, suggesting that malonyl glucosides are converted to their aglycones. These results illustrate the mechanisms underlying storage-related stresses and the importance of screening for lines that can tolerate environmental stresses.
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Jiamtae P., Hashiguchi M., Lelapiyamit I., Harada K., Gondo T., Tanaka H., Akashi R.
Euphytica 218 ( 12 ) 2022年12月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Euphytica
Soybean (Glycine max L. Merr.) seeds are highly sensitive to the environmental conditions experienced in STORAGE. The objectives of this study were to identify QTLs for soybean seed storability by evaluating seed viability and seed vigor and investigating candidate genes for these traits in detected QTL regions. These objectives will aid in developing soybean cultivars with a high capacity for storability. The seeds of 109 and 90 recombinant inbred lines (RILs) derived from a cross between Misuzudaizu and Moshidou Gong 503 in 2010 and 2019, respectively, were used to evaluate seed viability by a germination test and seed vigor by an accelerated aging test, after storage under two temperature conditions (25 °C and 35 °C) for six months. Seed viability and seed vigor of Moshidou Gong 503 were higher than those of Misuzudaizu in both temperature conditions. The average seed viability and seed vigor of RILs decreased when stored at the higher temperature. A total of five QTLs were found for the two traits, seed viability and seed vigor, located in chromosomes 2, 6 and 8. The Misuzudaizu alleles decreased seed viability and seed vigor at all detected QTLs. Most QTLs in this study were found near loci controlling seed viability, maturity, germination, seed hardness, and seed surface micromorphology, indicating that seed storability is related to these traits. In addition, two new QTLs were found that are associated with seed storability.
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形質転換プロトコール《植物編》
田部井 豊 編（ 担当： 共著 , 範囲: セイヨウミヤコグサ）
化学同人 2012年9月
記述言語：日本語 著書種別：学術書
MISC 【 表示 ／ 非表示 】
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田中秀典
大学時報 ( 405 ) 78 - 83 2022年7月
担当区分：筆頭著者 記述言語：日本語 掲載種別：記事・総説・解説・論説等（商業誌、新聞、ウェブメディア）
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NBRPミヤコグサ・ダイズにおけるバイオリソース整備 招待あり
橋口 正嗣，田中 秀典，橋口 拓勇，明石 良
農業および園芸 96 ( 2 ) 153 - 162 2021年2月
記述言語：日本語 掲載種別：記事・総説・解説・論説等（学術雑誌）
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マメ科植物研究のためのミヤコグサバイオリソース 招待あり 査読あり
橋口正嗣，佐藤修正，橋口拓勇，田中秀典，明石 良
植物科学の最前線（BSJ-Review） 10 179 - 189 2019年7月
記述言語：日本語 掲載種別：記事・総説・解説・論説等（学術雑誌） 出版者・発行元：日本植物学会
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ダイズ成長過程における形質・環境計測値の相互関係のネットワーク解析
七夕高也, 磯部祥子, 林篤司, 田中秀典, 橋口正嗣, 明石良, 長谷川舞衣, 菊地正隆, 中谷明弘, 佐藤修正, 七夕小百合
育種学研究 20 97 2018年3月
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植物の成長環境の変化に関連する遺伝子機能を解明するための成長計測技術の開発
七夕高也, 林篤司, 高地伸夫, 磯部祥子, 田中秀典, 橋口正嗣, 明石良, 中谷明弘, 菊地正隆, 長谷川舞衣, 佐藤修正, 七夕小百合
電気学会全国大会講演論文集(CD-ROM) 2018 ROMBUNNO.S26‐4 2018年3月
講演・口頭発表等 【 表示 ／ 非表示 】
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イネのデジタル3Dモデルによる計測法の開発
Sorawich Pongpiyapaiboon, 田中 秀典, 平野 翔也, 貴島 祐治, 明石 良
日本育種学会第144回講演会 2023年9月17日
開催年月日： 2023年9月16日 - 2023年9月18日
会議種別：口頭発表（一般）
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デジタル3Dモデルを用いた同一イネ品種の2倍体と4倍体成長パターンの比較
平野 翔也, Sorawich Pongpiyapaiboon, 笹川 静香, 高牟禮 逸朗, 田中 秀典, 明石 良, 貴島 祐治
日本育種学会第144回講演会 2023年9月17日
開催年月日： 2023年9月16日 - 2023年9月18日
会議種別：口頭発表（一般）
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シバ属における3Dモデルから抽出した葉色情報とNDVIの相関
Pongpiyapaiboon Sorawich, 田中 秀典, 橋口 拓勇, 橋口 正嗣, 明石 良
日本育種学会第142回講演会 2022年9月23日
開催年月日： 2022年9月23日 - 2022年9月24日
記述言語：英語 会議種別：口頭発表（一般）
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シバ属における新たな形態計測法の検討 –全周囲画像による3Dモデルの構築と活用–
Pongpiyapaiboon Sorawich, 田中 秀典, 橋口 拓勇, 橋口 正嗣, 林 篤司, 七夕 高也, 兒玉 晋洋, 磯部 祥子, 明石 良
日本育種学会第141回講演会 2022年3月20日
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蛍光灯および LED 照明下におけるミヤコグサ実験系統の生育特性
橋口正嗣、橋口拓勇、田中秀典、明石 良
植物微生物研究会第30回研究交流会
科研費（文科省・学振・厚労省）獲得実績 【 表示 ／ 非表示 】
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ゲノム編集食品が遺伝子組み換え食品と同じ轍を踏まないために
研究課題/領域番号：23K05404 2023年04月 - 2027年03月
独立行政法人日本学術振興会 科学研究費補助金 基盤研究(C)
担当区分：研究分担者
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学生の学業継続および退学予測モデルの構築に関する実証的研究
研究課題/領域番号：22H01025 2022年04月 - 2025年03月
独立行政法人日本学術振興会 科学研究費補助金 基盤研究(B)
担当区分：研究分担者
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大学の数量的な「共通知」から分析マインドを涵養する人材育成プラットフォームの開発
研究課題/領域番号：18K02706 2018年04月 - 2022年03月
科学研究費補助金 基盤研究(C)
担当区分：研究分担者
社会貢献活動 【 表示 ／ 非表示 】
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山形大学・履修証明プログラム（IR担当者向け実践プログラム）担当講師
役割：講師
山形大学 2023年5月 - 2023年10月
対象： 教育関係者, 研究者, 社会人・一般, 行政機関
種別：資格認定講習
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山形大学・履修証明プログラム（IR担当者向け実践プログラム）担当講師
役割：講師
山形大学 2022年5月 - 2022年9月
対象： 教育関係者, 研究者, 社会人・一般, 行政機関
種別：資格認定講習
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山形大学・履修証明プログラム（IR担当者向け実践プログラム）担当講師
役割：講師
山形大学 2021年5月 - 2021年9月
対象： 教育関係者, 研究者, 社会人・一般, 行政機関
種別：資格認定講習
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山形大学・履修証明プログラム（IR担当者向け実践プログラム）ゲスト講師
役割：講師
山形大学 2020年11月28日
対象： 教育関係者, 研究者, 社会人・一般, 行政機関
種別：資格認定講習
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宮崎学園短期大学FD研修会
役割：講師
宮崎学園短期大学 2020年9月25日
対象： 教育関係者, 研究者
種別：セミナー・ワークショップ