研究者詳細 - 入江　俊一

論文 - 入江　俊一

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High productivity of cellulase and xylanase enzymes in the mycelial-dispersed Pleurotus ostreatus Δpkac2 strain 査読

Yuitsu Otsuka, Moriyuki Kawauchi, Vladimir Elisashvili, Saori Endo, Kenya Tsuji, Akira Yoshimi, Chihiro Tanaka, Takehito Nakazawa, Toshikazu Irie, Yoichi Honda

Journal of Bioscience and Bioengineering 2025年8月

Homeobox transcription factor BmHox7 is required for sexual reproduction, but not for appressorial development and pathogenicity in Bipolaris maydis 査読

Seika Koya, Sae Shigeyoshi, Takuya Sumita, Toshikazu Irie, Kazumi Suzuki, Kosuke Izumitsu

Mycoscience 66 ( 4 ) 232 - 239 2025年7月

Dual regulation of appressorium development in the maize pathogen Bipolaris maydis: BmOPY2-mediated hydrophobic surface recognition and pectin recognition 査読

Hiroki Yoshida, Sae Shigeyoshi, Syunsuke Goto, Masashi Sassa, Kenya Tsuji, Akira Yoshimi, Chihiro Tanaka, Takuya Sumita, Toshikazu Irie, Kazumi Suzuki, Kosuke Izumitsu

Mycoscience 66 ( 4 ) 240 - 248 2025年7月

Disruption of the pkac2 gene in Pleurotus ostreatus alters cell wall structures and enables mycelial dispersion in liquid culture. 査読国際誌

Yuitsu Otsuka, Moriyuki Kawauchi, Kai Yoshizawa, Saori Endo, Kim Schiphof, Kenya Tsuji, Akira Yoshimi, Chihiro Tanaka, Shigekazu Yano, Vladimir Elisashvili, Takehito Nakazawa, Toshikazu Irie, Yoichi Honda

FEMS microbiology letters 2024年11月

記述言語：英語掲載種別：研究論文（学術雑誌）

In this study, we developed a mycelial dispersion strain by disrupting the pkac2 gene in the white-rot fungus Pleurotus ostreatus. pkac2 is a catalytic subunit gene of protein kinase A, which regulates several transcription factors related to cell wall synthesis. Liquid cultures of the Δpkac2 strains showed very high mycelial dispersibility and were visibly different from the wild-type strain (WT). Although growth on agar medium was slower than that of WT, Δpkac2 strains grew faster in liquid culture and had approximately twice the mycelial dry weight of WT after 5 d of culture. Microscopic observations showed that the cell walls of the Δpkac2 strains were thinner compared to WT. The β-glucan content in the cell walls decreased in the pkac2 disruptants, although the transcription levels of β-glucan synthase genes increased. Furthermore, the Δpkac2 strains showed decreased hydrophobicity and stress tolerance compared to WT. These results indicate that disruption of the pkac2 gene in P. ostreatus alters the structure of the cell wall surface layer, resulting in high-density cultures due to mycelial dispersion.

DOI： 10.1093/femsle/fnae101

PubMed

Marker recycling in Lentinula edodes via 5-fluoroorotic acid counter-selection. 査読国際誌

Takuma Narutaki, Ayane Kamiya, Yuma Tsujimoto, Masataka Naruse, Shota Nishida, Moriyuki Kawauchi, Yoichi Honda, Hisatoshi Kamitsuji, Toshitsugu Sato, Takuya Sumita, Kosuke Izumitsu, Toshikazu Irie

FEMS microbiology letters 371 2024年1月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Shiitake (Lentinula edodes) contains various beneficial compounds and possesses several notable properties. However, there are few reports on its molecular breeding due to delay in development of its gene-modifying technology. Therefore, here, strain UV30 (pyrG -) was bred from the UV-irradiated protoplasts of strain M2. Strain UV30 was uracil-auxotrophic, and the phenylalanine residue in the active centre of orotidine-5-phosphate decarboxylase encoded by pyrG in the strain was substituted with a serine residue. Next, a recycling marker consisting of the upstream sequence of ku80, a repeat sequence (a portion of the downstream sequence of ku80), pyrG, and the downstream sequence of ku80 was introduced into the strain UV30. Consequently, the prototrophic strain ckp2-1, in which ku80 was replaced with the recycling marker, was obtained. After cultivation in complete medium, mycelia from the edges of ckp2-1 colonies were inoculated into a complete medium containing 5-Fluoroorotic acid (5-FOA). A 5-FOA-resistant strain KaM2, in which pyrG sequence was spliced from the recycling marker sequence via homologous recombination, was obtained. In this study, we developed the first marker recycling system for multigene targeting in L. edodes. Moreover, the resulting ∆ku80 strain may serve as a non-homologous end-joining deficient strain for further genetic manipulations.

DOI： 10.1093/femsle/fnae073

PubMed

Development of a gene-targeting system using CRISPR/Cas9 and utilization of pyrG as a novel selectable marker in Lentinula edodes. 国際誌

Ayane Kamiya, Hiroki Ueshima, Shota Nishida, Yoichi Honda, Hisatoshi Kamitsuji, Toshitsugu Sato, Haruto Miyamoto, Takuya Sumita, Kosuke Izumitsu, Toshikazu Irie

FEMS microbiology letters 370 1 - 6 2023年3月

Marker-free genome editing in the edible mushroom, Pleurotus ostreatus, using transient expression of genes required for CRISPR/Cas9 and for selection. 査読

Koshi D., Ueshima H., Kawauchi M., Nakazawa T., Sakamoto M., Hirata M., Izumitsu K., Sumita T., Irie T., Honda Y.

Journal of Wood Science 68 ( 27 ) 2022年5月

Stable and transient transformation, and a promoter assay in the selective lignin-degrading fungus, Ceriporiopsis subvermispora 査読

Honda Y., Tanigawa E., Tsukihara T., Nguyen D., Kawabe H., Sakatoku N., Watari J., Sato H., Yano S., Tachiki T., Irie T., Watanabe T., Watanabe T.

AMB Express 9 ( 1 ) 2019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：AMB Express

© 2019, The Author(s). A genetic transformation system was developed for the selective white rot basidiomycete Ceriporiopsis subvermispora using a modified protocol with polyethylene glycol and CaCl2 treatment of the protoplasts and plasmids harboring recombinant hygromycin phosphotransferase (hph) driven by a homologous promoter. During repeated transfer on fresh potato dextrose agar plates containing 100 µg/ml hygromycin B, most transformants lost drug resistance, while the remaining isolates showed stable resistance over five transfers. No drug-resistant colonies appeared in control experiments without DNA or using a promoter-less derivative of the plasmid, indicating that a transient expression of the recombinant hph was driven by the promoter sequence in these unstable drug-resistant transformants. Southern blot analysis of the stable transformants revealed random integration of the plasmid DNA fragment in the chromosome at different copy numbers. This transformation system yielding mostly transient transformants was successfully used for promoter assay experiments, and only a 141-bp fragment was found to be essential for the basic promoter function of glyceraldehyde dehydrogenase gene (gpd) in this fungus. Subsequent mutational analyses suggested that a TATAA sequence is important for the basic promoter function of gpd gene. The promoter assay system will enable the functional analysis of gene expression control sequences quickly and easily, mostly in the absence of undesirable effects from differences in copy number and chromosomal position of an integrated reporter gene among stable transformants.

DOI： 10.1186/s13568-019-0818-1

Scopus

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85067881586&origin=inward

ウリ類炭疽病菌においてホメオボックス転写制御因子CoHox2は分生胞子形成および剛毛形成を制御する

小幡善和也, 横山綾, 泉津弘佑, 入江俊一, 鈴木一実

日本植物病理学会報 2019年4月

The homeobox transcription factor CoHox1 is required for the morphogenesis of infection hyphae in host plants and pathogenicity in Colletotrichum orbiculare. 査読

Yokoyama, A., Izumitsu, K., Irie, T., Suzuki, K.

Mycoscience 2019年2月

Homeobox transcription factor CoHox3 is essential for appressorium formation in the plant pathogenic fungus Colletotrichum orbiculare 査読

AyaYokoyama, Kosuke Izumitsu, Takuya Sumita, Chihiro Tanaka, Toshikazu Irie, Kazumi Suzuki

Mycoscience 59 ( 5 ) 353 - 362 2018年9月

Heterologous expression of the Pleurotus ostreatus MnP3 gene by the laccase gene promoter in Lentinula edodes 査読国際誌

Sato, T., Irie, T., Yoshino, F.

Bioscience, Biotechnology and Biochemistry 81 ( 8 ) 1553 - 1556 2017年8月

A development and an improvement of selectable markers in Pleurotus ostreatus transformation. 査読国際誌

Matsunaga, Y., Ando, M., Izumitsu, K., Suzuki, K., Honda, Y., Irie, T.

Journal of Microbiol Methods 134 27 - 29 2017年3月

Marker recycling via 5-fluoroorotic acid and 5-fluorocytosine counter-selection in the white-rot agaricomycete Pleurotus ostreatus. 査読国際誌

Nakazawa, T., Tsuzuki, M., Irie, T., Sakamoto, M., Honda, Y.

Fungal Biology 120 ( 9 ) 1146 - 1155 2016年9月

Effects of overexpression of PKAc genes on expressions of lignin modifying enzymes by Pleurotus ostreatus. 招待査読国際誌

Toyokawa C, Shobu M, Tsukamoto R, Okamura S, Honda Y, Kamitsuji H, Izumitsu K, Suzuki K, Irie T

Bioscience, Biotechnology and Biochemistry 80 ( 9 ) 1759 - 1767 2016年9月

Effects of calmodulin on expression of lignin-modifying enzymes in Pleurotus ostreatus 査読

Suetomi T, Sakamoto T, Tokunaga Y, Kameyama T, Honda Y, Kamitsuji H, Kameshita I, Izumitsu K, Suzuki K, Irie T

Curr Genet 61 ( 2 ) 127 - 140 2015年

The White-Rot Fungus Pleurotus ostreatus Transformant Overproduced Intracellular cAMP and Laccase 査読

Yuki Yao, Takaiku Sakamoto, Yoichi Honda, Yasuyuki Kagotani, Kousuke Izumitsu, Kazumi Suzuki, and Toshikazu Irie

Bioscience, Biotechnology and Biochemistry 77 ( 11 ) 2309 - 2311 2013年11月

Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene 査読

Takaiku Sakamoto, Yoichi Honda, Isamu Kameshita, Kazumi Suzuki, Toshikazu Irie

Mycoscience 2013年6月

A calmodulin inhibitor, W-7 influences the effect of cyclic adenosine 3', 5'-monophosphate signaling on ligninolytic enzyme gene expression in Phanerochaete chrysosporium. 査読

Takaiku Sakamoto, Yuki Yao1, Yoshifumi Hida, Yoichi Honda, Takashi Watanabe, Wataru Hashigaya, Kazumi Suzuki, Toshikazu Irie

AMB express 2 ( 1 ) 2012年7月

Transcriptional effect of a calmodulin inhibitor, W-7, on the ligninolytic enzyme genes in Phanerochaete chrysosporium 査読

Sakamoto T., H. Kitaura, M. Minami, Y. Honda, T. Watanabe, A. Ueda, K. Suzuki, T. Irie

Current Genetics 56 ( 5 ) 401 - 410 2010年5月

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