Faculty Profiles - IRIE Toshikazu

Papers - IRIE Toshikazu

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Disruption of the pkac2 gene in Pleurotus ostreatus alters cell wall structures and enables mycelial dispersion in liquid culture. Reviewed International journal

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 2024.11

Language：English Publishing type：Research paper (scientific journal)

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

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

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 2023.3

Language：English Publishing type：Research paper (scientific journal)

First, we attempted to recombine the Shiitake (Lentinula edodes) pyrG (ura3) gene homologously by introducing a donor vector containing a carboxin resistance gene (lecbxR) flanked by homologous sequences of pyrG into protoplasts of the fungus. However, all the carboxin-resistant transformants only contained ectopic insertions of the exogenous gene and no homologous insertions. Agaricomycetes are generally known for their low efficiency of homologous recombination, and a similar result was shown for L. edodes. We then co-introduced a Cas9 plasmid vector containing a CRISPR/Cas9 expression cassette targeting pyrG and donor plasmid vector. As a result, ∆pyrG strains containing the expected homologous recombination were obtained. However, only two of the seven ∆pyrG strains had the Cas9 sequence; the others did not. Our results suggest that genome editing occurred via the transient expression of the CRISPR/Cas9 cassette in the Cas9 plasmid vector introduced into the fungal cell. Transforming pyrG into a ∆pyrG strain (strain I8) resulted in prototrophic strains with an efficiency of 6.5 strains/experiment.

DOI： 10.1093/femsle/fnad042

PubMed

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

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 ) 2022.5

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

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 ) 2019.12

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

Other Link： 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 2019.4

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

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

Mycoscience 2019.2

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

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

Mycoscience 59 ( 5 ) 353 - 362 2018.9

Heterologous expression of the Pleurotus ostreatus MnP3 gene by the laccase gene promoter in Lentinula edodes Reviewed International journal

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

Bioscience, Biotechnology and Biochemistry 81 ( 8 ) 1553 - 1556 2017.8

A development and an improvement of selectable markers in Pleurotus ostreatus transformation. Reviewed International journal

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

Journal of Microbiol Methods 134 27 - 29 2017.3

Marker recycling via 5-fluoroorotic acid and 5-fluorocytosine counter-selection in the white-rot agaricomycete Pleurotus ostreatus. Reviewed International journal

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

Fungal Biology 120 ( 9 ) 1146 - 1155 2016.9

Effects of overexpression of PKAc genes on expressions of lignin modifying enzymes by Pleurotus ostreatus. Invited Reviewed International journal

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 2016.9

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

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

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

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

Bioscience, Biotechnology and Biochemistry 77 ( 11 ) 2309 - 2311 2013.11

Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene Reviewed

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

Mycoscience 2013.6

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

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

AMB express 2 ( 1 ) 2012.7

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

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

Current Genetics 56 ( 5 ) 401 - 410 2010.5

Changes in the gene expression of the white rot fungus Phanerochaete chrysosporium by the addition of atropine. Reviewed

Masahiko Minami, Kazumi Suzuki, Akifumi Shimizu, Tomohiro Hongo, Takaiku Sakamoto, Naoki Ohyama, Hironori Kitaura, Akiho Kusaka, Kenji Iwama, and Toshikazu Irie

Bioscience, Biotechnology and Biochemistry 73 ( 8 ) 1722 - 1731 2009.8

Changes in the Gene Expression of the White Rot Fungus Phanerochaete chrysosporium Due to the Addition of Atropine Reviewed

Masahiko Minami, Kazumi Suzuki, Akifumi Shimizu, Tomohiro Hongo, Takaiku Sakamoto, Naoki Ohyama, Hironori Kitaura, Akiho Kusaka, Kenji Iwama, Toshikazu Irie

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 73 ( 8 ) 1722 - 1731 2009.8

Language：English Publishing type：Research paper (scientific journal) Publisher：TAYLOR & FRANCIS LTD

We constructed a LongSAGE (Long Serial Analysis of Gene Expression) library from a 3-d culture of Phanerochaete chrysosporium supplemented with atropine, which inhibits the production of lignin-degrading enzymes. The library (the atropine library) contains 13,108 LongSAGE tags and 6,783 unique tags. The gene expression profile represented by the tags was compared with those of two previously constructed libraries, one of which was constructed using 2-d cultures in which the fungus had not yet produced ligninolytic enzymes (the 2-d library) and the other was constructed using 3-d cultures in which the fungus had just started to produce the enzymes (the 3-d library). We found a total of 595 genes that were at least twice more highly or at least twice less highly expressed in the 3-d library than in the 2-d library or the atropine library, and the fluctuations were statistically significant. The relationships among these 595 genes were considered using cluster analysis. Of the 595 genes, 164 showed expression patterns similar to those of four ligninolytic enzyme genes, which were more expressed on day 3 than under any other conditions. Many of these 164 genes comprised genes possibly involved in lignin degradation, lipid metabolism, xenobiotic degradation, stress response, or signal transduction pathways.

DOI： 10.1271/bbb.80870

Web of Science

Heterologous expression of lcc1 from Lentinula edodes in tobacco BY-2 cells results in the production an active, secreted form of fungal laccase Reviewed

Y. Sakamoto, K. Nakade, A. Yano, Y. Nakagawa, T. Hirano, T. Irie

Applied Microbiology and Biotechnology 96 ( 6 ) 971 - 980 2008.4

Long serial analysis of gene expression for transcriptome profiling during the initiation of ligninolytic enzymes production in Phanerochaete chrysosporium. Reviewed

Minami, M., O. Kureha, M. Mori, H. Kamitsuji, K. Suzuki and T. Irie.

Applied Microbiology and Biotechnology 75 ( 3 ) 609 - 618 2007.4

New choke diseases and their molecular phylogenetic analysis in Agropyron ciliare var. minus and Agropyron tsukushiense var. transiens. Reviewed

Yanagida, N., T. Irie, E. Tanaka, C. Teramoto, K. Kuwabara and A. Tajimi.

Mycologia 97 ( 6 ) 1313 - 1317 2005.4

Isolation and characterization of a fruiting body-specific exo-beta-1,3-glucanase-encoding gene, exg1, from Lentinula edodes. Reviewed

Sakamoto, Y., T. Irie, and T. Sato.

Current Genetics 47 ( 4 ) 244 - 252 2005.4

リアルタイムRT-PCRを用いたイネの翻訳開始遺伝子（eIF5A）の解析 Reviewed

藤山佳代、奥ゆか里、小西良司、入江俊一、長谷川博

近畿作物・育種研究会－ ( 50 ) 63 - 65 2005.4

SAGE (Serial Analysis of Gene Expression) of Magnaporthe grisea Reviewed

Irie, T., H. matsumura, R. Terauchi and H. Saitoh

Rice Blast: Interaction with Rice and Control 1 ( 1 ) 23 - 28 2004.4

Construction of a Homologous Selectable Marker Gene for Lentinula edodes Transformation Reviewed

Irie, T., T. Sato, K. Saito, Y. Honda, T. Watanabe, M. Kuwahara and H. Enei

Bioscience, Biotechnology and Biochemistry 67 ( 9 ) 2006 - 2009 2003.4

Serial Analysis of Gene Expression (SAGE) of Magnaporthe grisea: genes involved in appressorium formation Reviewed

Irie, T., H. matsumura, R. Terauchi and H. Saitoh

Molecular Genetics and Genomics 270 ( 2 ) 181 - 189 2003.4

イネのセシウム抵抗性突然変異株の翻訳開始因子（eIF-5A）遺伝子 Reviewed

小西良司、入江俊一、長谷川博

近畿作物・育種研究会 0 ( 48 ) 61 - 63 2003.4

Development of Transformation and Recombinant Gene Expression Systems in Pleurotus ostreatus Reviewed

Irie, T

Wood Research 1 ( 88 ) 1 - 18 2001.4

Efficient transformation of filamentous fungus Pleurotus ostreatus using single-strand carrier DNA Reviewed

Irie, T., Y. Honda, T. Watanabe and M. Kuwahara

Applied Microbiology and Biotechnology 55 ( 5 ) 563 - 565 2001.4

Homologous Expression of Recombinant Manganese Peroxidase Genes in ligninolytic fungus Pleurotus ostreatus Reviewed

Irie, T., Y. Honda, T. Watanabe and M. Kuwahara

Applied Microbiology and Biotechnology 55 ( 5 ) 566 - 570 2001.4

Stable transformation of Pleurotus ostreatus to hygromycin B resistance using Lentinus edodes GPD expression signals Reviewed

Irie, T., Y. Honda, T. Hirano, T. Sato, H. Enei, T. Watanabe and M. Kuwahara

Applied Microbiology and Biotechnology 56 ( 41400 ) 707 - 709 2001.4

Isolation of cDNA and genomic fragments encoding the major manganese peroxidase isozyme from the white rot basidiomycete Pleurotus ostreatus Reviewed

Irie, T., Y. Honda, H.-C. Ha, T. Watanabe and M. Kuwahara

Journal of Wood Science 41 ( 3 ) 230 - 233 2000.4

Carboxin resistance transformation of the homobasidiomycete fungus Pleurotus ostreatus Reviewed

Honda, Y., T. Matsuyama, T. Irie, T. Watanabe and M. Kuwahara

Current Genetics 37 ( 3 ) 209 - 212 2000.4

Isolation and sequence analysis of the promoter and allelic sequence of the iron-sulfur protein subunit gene from the white-rot fungus Pleurotus ostreatus Reviewed

Honda, Y., T. Matsuyama, T. Irie, T. Watanabe and M. Kuwahara

Journal of Wood Science 44 ( 6 ) 491 - 494 1998.4

Cloning and characterization of the gene encoding the iron-sulfur protein of succinate dehydrogenase from Pleurotus ostreatus Reviewed

Honda, Y., T. Matsuyama, T. Irie, T. Watanabe and M. Kuwahara

Biochemica et Biophysica Acta 1396 ( 1 ) 27 - 31 1998.4

Isolation and sequence analysis of the promoter and an allelic sequence of the iron-sulfur protein subunit gene from the white-rot fungus Pleurotus ostreatus Reviewed

T Irie, Y Honda, T Matsuyama, T Watanabe, M Kuwahara

JOURNAL OF WOOD SCIENCE 44 ( 6 ) 491 - 494 1998

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG TOKYO

We have isolated a structural gene of sdi1, which encodes the iron-sulfur protein (Ip) subunit of succinate dehydrogenase (EC 1.3.99.1), from a white-rot basidiomycete, Pleurotus ostreatus. Here we report isolation of the promoter region of sdi1 and an allelic sequence encoding the second-type cDNA fragment isolated in the former experiments. The nucleotide sequence analysis of the promoter region revealed the existence of putative CAAT and TATA boxes, which permits us to develop an expression system in this species. The Southern blot analysis and the restriction fragment length polymorphism assay using monokaryotic strains demonstrated that no family genes to sdi1 exist in the haploid genome of P. ostreatus. Moreover, a genetic analysis to detect a linkage between the sdi1 genotypes and flutolanil resistance in the mutant P. ostreatus strains was also developed.

Web of Science

Isolation and characterization of Pleurotus ostreatus mutant strains resistant to a carboxin-derived fungicide, flutolanil Reviewed

Honda, Y., T. Irie, M. Atsuji, T. Watanabe and M. Kuwahara

Mycoscience 37 ( 4 ) 459 - 461 1996.4

Structures of genomic and complementary DNAs coding for Pleurotus ostreatus manganese (II) peroxidase Reviewed

Asada, Y., A. Watanabe, T. Irie, T. Nakayama and M. Kuwahara

Biochemica et Biophysica Acta 1251 ( 2 ) 205 - 209 1995.4

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Papers - IRIE Toshikazu