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From School 【 display / non-display 】
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Tokyo University of Agriculture Graduated
2000 - 2005
Country：Japan
From Graduate School 【 display / non-display 】
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The Graduate University for Advanced Studies Graduate School, Division of Life Science Doctoral program (second term) Completed
2007 - 2010
Country：Japan
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Tokyo University of Agriculture Graduate School, Division of Agriculture Master Course Completed
2005 - 2007
Country：Japan
Studying abroad experiences 【 display / non-display 】
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2010.07 - 2010.09 リーズ大学 研究員
Degree 【 display / non-display 】
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博士（理学） ( 2010.03 総合研究大学院大学 )
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修士（農芸化学） ( 2007.03 東京農業大学 )
Employment Record in Research 【 display / non-display 】
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Tokyo University of Agriculture Faculty of Life Sciences Department of Chemistry for Life Sciences and Agriculture Associate Professor
2024.04
Professional Memberships 【 display / non-display 】
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日本植物バイオテクノロジー学会
2023
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日本分子生物学会
2012
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日本植物生理学会
2005
Research Areas 【 display / non-display 】
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Life Science / Plant molecular biology and physiology
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Life Science / Molecular biology
Papers 【 display / non-display 】
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昆虫病原糸状菌を利用した害虫防除の概要とホウ酸耐性を持つ新たな株の利用可能性 Invited
金井雅武、星理絵、重信秀治、野崎友成、小林裕樹、石井大輔、真野昌二
バイオインダストリー 41 ( 11 ) 25 - 29 2024.11
Authorship：Lead author, Corresponding author Language：Japanese Publishing type：Research paper (scientific journal) Publisher：シーエムシー出版
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Fusing the 3’UTR of seed storage protein genes leads to massive recombinant protein accumulation in seeds International journal
Masatake Kanai, Masaya Sugiyama, Maki Kondo, Kenji Yamada, Mikio Nishimura, Shoji Mano
Scientific Reports 13 ( 1 ) 12217 - 12217 2023.12
Language：English Publishing type：Research paper (scientific journal)
The demand for recombinant proteins is rising dramatically, and effective production systems are currently being developed. The production of recombinant proteins in plants is a promising approach due to its low cost and low risk of contamination of the proteins with endotoxins or infectious agents from the culture serum. Plant seeds primarily accumulate seed storage proteins (SSPs), which are transcribed and translated from a few genes; therefore, the mechanism underlying SSP accumulation has been studied to help devise ways to increase recombinant protein production. We found that the 3'UTR of SSP genes are essential for SSP accumulation and can be used in the production of recombinant proteins in Arabidopsis. Fusion of the 3'UTR of SSP genes to the 3' ends of DNA sequences encoding recombinant proteins enables massive accumulation of recombinant proteins with enzymatic activity in Arabidopsis seeds. This method is also applicable to the production of human Interferon Lambda-3 (IFN-lambda 3), a candidate biopharmaceutical compound against hepatitis C infection. Considering the low cost and ease of protein production in Arabidopsis, as well as the rapid growth of this plant, our method is useful for large-scale preparation of recombinant proteins for both academic research and biopharmaceutical production.
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Cloning and Functional Verification of Endogenous U6 Promoters for the Establishment of Efficient CRISPR/Cas9-Based Genome Editing in Castor (Ricinus communis)
Masatake Kanai, Kazumi Hikino, Shoji Mano
Genes 14 ( 7 ) 2023.07
Publishing type：Research paper (scientific journal)
Castor (Ricinus communis) seeds are rich in a type of hydroxy fatty acid called ricinoleic acid, which is in high demand for the production of plant-based plastics, lubricants, and hydraulic oils. However, the high content of ricin, a toxic protein, in these seeds has restricted further expansion in the area of castor cultivation. Therefore, the development of ricin-free castor is needed. Genome editing technology, although successfully applied in several plant species, is still in the developing stages in castor and awaits the identification of an endogenous U6 promoter with robust function. Here, we searched for U6 small nuclear RNA (snRNA) genes in the castor genome. This led to the identification of six U6 snRNA genes. The promoters of these U6 snRNA genes were cloned, and their function was examined in castor cells using the particle delivery method. The results showed that a U6 promoter length of approximately 300 bp from the transcription start site was sufficient to activate gene expression. This study provides insights into the endogenous castor U6 promoter sequences and outlines a method for verifying the function of U6 promoters in plants using the particle delivery system.
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Ubiquitin-conjugating activity by PEX4 is required for efficient protein transport to peroxisomes in Arabidopsis thaliana
Shoji Mano, Yasuko Hayashi, Kazumi Hikino, Masayoshi Otomo, Masatake Kanai, Mikio Nishimura
Journal of Biological Chemistry 298 ( 6 ) 102038 - 102038 2022.06
Publishing type：Research paper (scientific journal) Publisher：Elsevier BV
Protein transport to peroxisomes requires various proteins, such as receptors in the cytosol and components of the transport machinery on peroxisomal membranes. The Arabidopsis apem (aberrant peroxisome morphology) mutant apem7 shows decreased efficiency of peroxisome targeting signal 1–dependent protein transport to peroxisomes. In apem7 mutants, peroxisome targeting signal 2–dependent protein transport is also disturbed, and plant growth is repressed. The APEM7 gene encodes a protein homologous to peroxin 4 (PEX4), which belongs to the ubiquitin-conjugating (UBC) protein family; however, the UBC activity of Arabidopsis PEX4 remains to be investigated. Here, we show using electron microscopy and immunoblot analysis using specific PEX4 antibodies and in vitro transcription/translation assay that PEX4 localizes to peroxisomal membranes and possesses UBC activity. We found that the substitution of proline with leucine by apem7 mutation alters ubiquitination of PEX4. Furthermore, substitution of the active-site cysteine residue at position 90 in PEX4, which was predicted to be a ubiquitin-conjugation site, with alanine did not restore the apem7 phenotype. Taken together, these findings indicate that abnormal ubiquitination in the apem7 mutant alters ubiquitin signaling during the process of protein transport, suggesting that the UBC activity of PEX4 is indispensable for efficient protein transport to peroxisomes.
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Image-Based Analysis Revealing the Molecular Mechanism of Peroxisome Dynamics in Plants
Shino Goto-Yamada, Kazusato Oikawa, Katsuyuki T. Yamato, Masatake Kanai, Kazumi Hikino, Mikio Nishimura, Shoji Mano
Frontiers in Cell and Developmental Biology 10 2022.05
Publisher：Frontiers Media SA
Peroxisomes are present in eukaryotic cells and have essential roles in various biological processes. Plant peroxisomes proliferate by de novo biosynthesis or division of pre-existing peroxisomes, degrade, or replace metabolic enzymes, in response to developmental stages, environmental changes, or external stimuli. Defects of peroxisome functions and biogenesis alter a variety of biological processes and cause aberrant plant growth. Traditionally, peroxisomal function-based screening has been employed to isolate Arabidopsis thaliana mutants that are defective in peroxisomal metabolism, such as lipid degradation and photorespiration. These analyses have revealed that the number, subcellular localization, and activity of peroxisomes are closely related to their efficient function, and the molecular mechanisms underlying peroxisome dynamics including organelle biogenesis, protein transport, and organelle interactions must be understood. Various approaches have been adopted to identify factors involved in peroxisome dynamics. With the development of imaging techniques and fluorescent proteins, peroxisome research has been accelerated. Image-based analyses provide intriguing results concerning the movement, morphology, and number of peroxisomes that were hard to obtain by other approaches. This review addresses image-based analysis of peroxisome dynamics in plants, especially A. thaliana and Marchantia polymorpha.
Misc 【 display / non-display 】
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Does Guard Cell-Specific Loss of Cuticular Layer Contribute CO2 Absorption and Tolerance to Desiccation?
金井雅武, 星理絵, 真野昌二, 真野昌二
日本分子生物学会年会プログラム・要旨集(Web) 45th 2022
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Analysis of the mechanism and regulation of protein transport to peroxisomes using Arabidopsis apem mutants
真野昌二, 真野昌二, 林八寿子, 林八寿子, 曳野和美, 大友政義, 金井雅武, 西村幹夫
日本植物生理学会年会(Web) 63rd 2022
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Efficient protein transport to peroxisomes requires ubiquitin-conjugating activity by PEX4 in Arabidopsis thaliana
真野昌二, 真野昌二, 林八寿子, 林八寿子, 曳野和美, 大友政義, 金井雅武, 西村幹夫
日本分子生物学会年会プログラム・要旨集(Web) 45th 2022
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Screening for Arabidopsis mutants showing low CO2 resistance
金井雅武, 真野昌二, 真野昌二
日本分子生物学会年会プログラム・要旨集(Web) 44th 2021
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プラスチド局在性葉酸はシロイヌナズナの非光合成型プラスチドへ流入する糖によって引き起こされるデンプン合成を阻害する
林誠, 田中美名, 山本沙季, 中川太郎, 金井雅武, 姉川彩, 大西美輪, 三村徹郎, 西村幹夫
日本植物生理学会年会(Web) 60th 2019
Honours, Awards and Prizes 【 display / non-display 】
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日本植物生理学会 PCP 論文賞
2019.03 日本植物生理学会
Award type：Honored in official journal of a scientific society, scientific journal
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日本農芸化学会2013年度大会 トピックス賞
2012.03 日本農芸化学会
Award type：Award from Japanese society, conference, symposium, etc. Country：Japan
Scientific Research Funds Acquisition Results 【 display / non-display 】
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高温・乾燥地域での生育を可能にする植物の新規な蒸散制御システムの解明
Grant number：19K22438 2019.06 - 2024.03
日本学術振興会 科学研究費助成事業 挑戦的研究(萌芽)
金井 雅武
Grant amount：\6240000 （ Direct Cost: \4800000 、 Indirect Cost：\1440000 ）
ヒマは蒸散を迅速かつ厳密にコントロールすることで高温・乾燥地域においても高い生産性を示すと考えられる。ヒマ葉の気孔直下の海綿状組織には、他の植物には見られない特殊な細胞（バルブ細胞）が観察されており、このバルブ細胞の機能解析を進める。
これまでに確立したヒマ葉の細胞分画法を用いてバルブ細胞濃縮画分の大量調整を検討した。基礎生物学研究所内の人工気象器で4週間栽培したヒマの新葉から調整したところ、高い純度でバルブ細胞と考えられるサイズの大きい柔組織細胞が調整できた。この画分から全RNAおよび総タンパク質を抽出し、濃度を測定したところ今後の解析に必要な量に達しなかった。サンプル量を増やすために、新葉以外の葉からの分画を検討したところ、純度が低かった。これより、高い純度のサイズの大きい柔組織細胞画分を調整するためには4週間栽培したヒマの新葉が適していることが明らかとなった。十分量のヒマ新葉を用意する目的で、所内の複数の人工気象器、ガラス温室、圃場を利用して栽培し、それぞれから収穫した新葉を用いて画分を調整した。栽培場所によって生育が異なったものの、いずれも調整可能であったため、以降は気候に応じて最適な場所を選択可能であることを明らかとした。これより、精度の高いオミクスデータを取得するためのバルブ細胞画分の大量調整方法と条件を確立した。 -
Development of Fertilizer Management Technology for Tea Oil Production Using Abandoned Land
Grant number：18KT0095 2018.07 - 2022.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
Mano Shoji
Grant amount：\4420000 （ Direct Cost: \3400000 、 Indirect Cost：\1020000 ）
Component analysis of lipids and RNA-seq analysis using samples of tea seeds grown under different fertilization conditions in abandoned fields over time revealed that the type of fertilization affects the oleic acid content of seeds and that suppression of nitrogen fertilization in autumn and winter is necessary for the characteristic high oleation of tea oil. The expression levels of CsLipase1 and CsLipase2 are thought to reflect the amount of nitrogen fertilizer applied in the autumn and winter. The expression levels of CsLipase1 and CsLipase2 were considered to reflect the amount of nitrogen fertilizer applied in autumn and winter, and were considered to be candidate markers for determining the amount of nitrogen fertilizer.
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Analysis on molecular mechanism of protein transport to peroxisomes
Grant number：26440157 2014.04 - 2018.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
Mano Shoji, NISHIMURA MIKIO, KANAI MASATAKE, OIKAWA KAZUSATO, KAMIGAKI AKANE, KONDO MAKI, GOTO SHINO, HIKINO KAZUMI, NAGATA KYOKO, YAMAGUCHI CHINAMI, NAKAGAWA TSUYOSHI, KOUCHI TAKAYUKI, NISHIHAMA RYUICHI, YAMATO KATSUYUKI
Grant amount：\5070000 （ Direct Cost: \3900000 、 Indirect Cost：\1170000 ）
From imaging analysis using transgenic Marchantia polymorpha expressing PTS1 (Peroxisome targeting signal 1)- or PTS2-Citrine and immunoelectron microscopic analysis revealed the presence of PTS1- and PTS2-dependent protein transport in M. polymorpha. Bioinformatics analysis identified the orthologous genes in M. polymporpha genome. Using this information, we succeeded in generation of some mutants whose peroxisomal genes were disrupted by the CRISPR/Cas9 method. In addition, we generated the Gateway technology-compatible binary vectors, and these vectors functioned in M. polymorpha cells.
From the analysis of one of Arabidopsis apem (aberrant peroxisome morphology), the ubiquitin signaling pathway is involved in protein transport to peroxisomes, and that the defective of ubiquitin-dependent protein transport causes normal peroxisome functions, such as lipid metabolism and photorespiration. -
Molecular mechanisms on inhibition of flower initiation in shoot apical meristems controlled by developing seeds
Grant number：25650102 2013.04 - 2016.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
KANAI Masatake
Grant amount：\4160000 （ Direct Cost: \3200000 、 Indirect Cost：\960000 ）
Plants produce flowers for seed formation. Increase of flower number improves seed yields of crop plants. This study focuses on the molecular mechanisms on inhibitory mechanisms of flower initiation in shoot apical meristems controlled by developing seeds. We screened three mutants increased flower number of lateral branches. These mutants were deficient in chloroplast degradation and showed a stay green phenotype. These results indicate that flower number is closely related to leaf senescence.
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Elucidation of seed-specific stress tolerance
Grant number：23870034 2011 - 2012
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Research Activity Start-up
KANAI Masatake
Grant amount：\3250000 （ Direct Cost: \2500000 、 Indirect Cost：\750000 ）
Plants produce seeds for the next generation. Developing seeds are more sensitive to environmental stresses than plant (e. g., roots, leaves, stem). It is essential for developing seeds to be protected from environmental stresses, the protection system is largely unknown. The aim of this study was to identify genes that regulated seed-specific stress tolerance and DPS genes were identified. It was also shown that DPS1, one of the DPS genes, involved in seed-specific drought stress. This study leads to an improvement of crop yield in arid area