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吉原 亮平（ヨシハラ リョウヘイ）
| 理工学研究科 生命科学部門 | 助教 |
| 理学部 生体制御学科 |
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■ 論文- A mutation in DNA polymerase γ harbors a shortened lifespan and high sensitivity to mutagens in the filamentous fungus Neurospora crassa.
Ryouhei Yoshihara; Yuzuki Shimakura; Satoshi Kitamura; Katsuya Satoh; Manami Sato; Taketo Aono; Yu Akiyama; Shin Hatakeyama; Shuuitsu Tanaka
Genetics, 巻：229, 号：2, 2025年02月, ［査読有り］, ［筆頭著者, 責任著者］, ［国際誌］
Hyphal elongation is the vegetative growth of filamentous fungi, and many species continuously elongate their hyphal tips over long periods. The details of the mechanisms for maintaining continuous growth are not yet clear. A novel short lifespan mutant of N. crassa that ceases hyphal elongation early was screened and analyzed to better understand the mechanisms for maintaining hyphal elongation in filamentous fungi. The mutant strain also exhibited high sensitivity to mutagens such as hydroxyurea and ultraviolet radiation. Based on these observations, we named the novel mutant "mutagen sensitive and short lifespan 1 (ms1)". The mutation responsible for the short lifespan and mutagen sensitivity in the ms1 strain was identified in DNA polymerase γ (mip-1:NCU00276). This mutation changed the amino acid at position 814 in the polymerase domain from leucine to arginine (MIP-1 L814R). A dosage analysis by next generation sequencing (NGS) reads suggested that mitochondrial DNA (mtDNA) sequences are decreased non-uniformly throughout the genome of the ms1 strain. This observation was confirmed by quantitative PCR for three representative loci and restriction fragment length polymorphisms in purified mtDNA. Direct repeat-mediated deletions, which had been reported previously, were not detected in the mitochondrial genome by our whole-genome sequencing analysis. These results imply the presence of novel mechanisms to induce the non-uniform decrease in the mitochondrial genome by DNA polymerase γ mutation. Some potential reasons for the non-uniform distribution of the mitochondrial genome are discussed in relation to the molecular functions of DNA polymerase γ.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1093/genetics/iyae201
DOI ID：10.1093/genetics/iyae201, PubMed ID：39611774 - Differential contributions of double-strand break repair pathways to DNA rearrangements following the irradiation of Arabidopsis seeds and seedlings with ion beams.
Satoshi Kitamura; Katsuya Satoh; Yoshihiro Hase; Ryouhei Yoshihara; Yutaka Oono; Naoya Shikazono
The Plant journal : for cell and molecular biology, 2024年09月, ［査読有り］, ［国際誌］
DNA rearrangements, including inversions, translocations, and large insertions/deletions (indels), are crucial for crop evolution, domestication, and improvement. The rearrangements are frequently induced by ion beams via the mis-repair of DNA double-strand breaks (DSBs). Unfortunately, how ion beam-induced DSBs are repaired has not been comprehensively analyzed and the mechanisms underlying DNA rearrangements remain unclear. In this study, clonal sectors originating from single mutated cells in carbon ion-irradiated plants were used for whole-genome sequencing analyses after Arabidopsis seeds and seedlings were irradiated. Comparative analyses of the induced mutations (e.g., size and frequency of indels and microhomology at the junctions of the rearrangements) in the irradiated materials suggested that the broken/rejoined DSB ends were more extensively processed in seedlings than in seeds. A mutation to canonical non-homologous end-joining (c-NHEJ), which is a DSB repair pathway with minimal processing of DSB ends, increased the sensitivity to ion beams more in the seeds than in the seedlings, which was consistent with the junction analysis results, indicative of the minor contribution of c-NHEJ to the carbon ion-induced DSB repair in seedlings. Considering the characteristics of the large templated insertions in irradiated seedlings, ion-beam-induced DSBs in seedlings are likely repaired primarily by a polymerase theta-mediated pathway. Polymerase theta-deficient seedlings were more sensitive to ion beams than the c-NHEJ-deficient seedlings, consistent with this hypothesis. This study revealed the key characteristics of ion beam-induced DSBs and the associated repair mechanisms related to the physiological status of the irradiated materials, with implications for elucidating the occurrence and induction of rearrangements.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1111/tpj.16955
DOI ID：10.1111/tpj.16955, PubMed ID：39312204 - Two high-mobility group domains of MHG1 are necessary to maintain mtDNA in Neurospora crassa
Hayami Seike; Keisuke Ishimori; Asagi Watanabe; Mao Kiryu; Shin Hatakeyama; Shuuitsu Tanaka
Fungal Biology, 巻：126, 号：11-12, 開始ページ：826, 終了ページ：833, 2022年11月, ［査読有り］, ［責任著者］
Elsevier {BV}, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.funbio.2022.11.001
DOI ID：10.1016/j.funbio.2022.11.001, ISSN：1878-6146, ORCID：124588814 - MLP-PG1, a major latex-like protein identified in Cucurbita pepo, confers resistance through the induction of pathogenesis-related genes
Fujita, K.; Asuke, S.; Isono, E.; Yoshihara, R.; Uno, Y.; Inui, H.
Planta, 巻：255, 号：1, 2022年11月, ［査読有り］
Springer Science and Business Media LLC, 研究論文（学術雑誌）
DOI：https://doi.org/10.1007/s00425-021-03795-x
DOI ID：10.1007/s00425-021-03795-x, ISSN：1432-2048, eISSN：1432-2048, ORCID：110618669, SCOPUS ID：85120576942 - A20/AN1 zinc-finger proteins positively regulate major latex-like proteins, transporting factors toward dioxin-like compounds in Cucurbita pepo
Kentaro Fujita; Ryouhei Yoshihara; Matashi Hirota; Junya Goto; Chihiro Sonoda; Hideyuki Inui
Chemosphere, 開始ページ：135536, 終了ページ：135536, 2022年06月, ［査読有り］
Elsevier BV, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.chemosphere.2022.135536
DOI ID：10.1016/j.chemosphere.2022.135536, ISSN：0045-6535 - A partial photoreactivation defect phenotype is not due to unrepaired ultraviolet-induced pyrimidine dimers in ultraviolet-sensitive mutants of Neurospora crassa.
Kotaro Tsukada; Ryouhei Yoshihara; Shin Hatakeyama; Akihiko Ichiishi; Shuuitsu Tanaka
Genes & genetic systems, 巻：95, 号：6, 開始ページ：281, 終了ページ：289, 2021年02月, ［査読有り］, ［国内誌］
Photoreactivation is a mechanism in which photolyase directly repairs either cyclobutane pyrimidine dimers (CPDs) or (6-4) photoproducts [(6-4) PPs] caused by ultraviolet (UV) light. In the filamentous fungus Neurospora crassa, some UV-sensitive mutants such as mus-44 have been reported to exhibit a partial photoreactivation defect (PPD) phenotype, but its mechanism has not been elucidated for a long time. In this study, the N. crassa CPD photolyase PHR was overexpressed in the mus-44 strain, but photoreactivation ability was not increased. Furthermore, Escherichia coli CPD photolyase or Arabidopsis thaliana (6-4) PP photolyase was also introduced into mus-44; however, the PPD phenotype was not complemented. These results suggested that the PPD phenotype in N. crassa is not caused by residual unrepaired pyrimidine dimers, which are the main type of DNA damage caused by UV irradiation. Finally, we revealed that mus-44, but not the mus-43 strain, which does not show the PPD phenotype, displayed higher sensitivity with increasing dose rate of UV. Moreover, mus-44 was also sensitive to an interstrand crosslinking agent. This indicates that the high dose of UV in our experimental condition induces DNA damage other than pyrimidine dimers, and that such damage is a likely cause of the PPD phenotype.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1266/ggs.20-00022
DOI ID：10.1266/ggs.20-00022, PubMed ID：33551431 - The msh1 gene is responsible for short life span mutant natural death and functions to maintain mitochondrial DNA integrity.
Mitsuyoshi Endo; Takato Yokoi; Suguru Hatazawa; Yuna Kojima; Shiena Takahama; Ryouhei Yoshihara; Shuuitsu Tanaka; Shin Hatakeyama
Fungal genetics and biology : FG & B, 巻：144, 開始ページ：103465, 終了ページ：103465, 2020年11月, ［査読有り］, ［国際誌］
Wild-type filamentous fungus Neurospora crassa continues to grow its hyphae for a very lengthy period of time (>2 years), whereas mutations at the natural death (nd) locus shorten life span (approximately 20 days). By positional cloning based on heat augmented mutagen sensitivity of the nd strain, we identified a nonsense mutation in the msh1 gene, an eukaryotic homolog of bacterial MutS, and this mutation resulted in encoding non-functional polypeptide. By tagging with GFP, subcellular localization of the MSH1 protein in the mitochondria was observed, and knock out of the msh1 gene caused severe growth deficiency accompanying mitochondrial DNA (mtDNA) aberrations such as large-scale mtDNA deletions and rearrangements as seen in the nd strain. These results suggested that MSH1 may maintain mtDNA integrity. Thus, loss of function compromises mtDNA, leading to the acceleration of cellular aging.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.fgb.2020.103465
DOI ID：10.1016/j.fgb.2020.103465, PubMed ID：32949723 - Suppression of the genes responsible for transporting hydrophobic pollutants leads to the production of safer crops.
Kentaro Fujita; Yuki Haga; Ryouhei Yoshihara; Chisato Matsumura; Hideyuki Inui
The Science of the total environment, 巻：741, 開始ページ：140439, 終了ページ：140439, 2020年11月, ［査読有り］, ［国際誌］
Hydrophobic pollutants have become widely distributed across the world. From an agricultural perspective, their accumulation in crops from contaminated soil threatens food security and quality, leading to many diseases in humans. The Cucurbitaceae family can accumulate high concentrations of hydrophobic pollutants in their aerial parts. The Cucurbitaceae family contains major latex-like proteins (MLPs) as transporting factors for hydrophobic pollutants. MLP genes are expressed in the roots in which the MLPs bind hydrophobic pollutants. MLPs transport these hydrophobic pollutants to the aerial parts of the plant through the xylem vessels. As a result, hydrophobic pollutant contamination occurs in the Cucurbitaceae family. In this study, we suppressed the expression of MLP genes in the roots and reduced the amounts of MLPs with pesticide treatments. First, the fungicides Benlate and Daconil that deceased the hydrophobic pollutant, perylene, concentration in the xylem sap of zucchini plants were selected. Daconil suppressed the transcription activity of MLP in the roots. In the Daconil treatment, the amount of MLPs in the roots and xylem sap of zucchini plants was decreased, and the concentrations of the hydrophobic pollutants, pyrene and dieldrin, were significantly decreased. Our research contributes to the production of safer crops.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.scitotenv.2020.140439
DOI ID：10.1016/j.scitotenv.2020.140439, PubMed ID：32887003 - Effects of Arabidopsis Ku80 deletion on the integration of the left border of T-DNA into plant chromosomal DNA via Agrobacterium tumefaciens.
Ryouhei Yoshihara; Yuka Mitomi; Maki Okada; Hanako Shibata; Mai Tanokami; Yurie Nakajima; Hideyuki Inui; Yutaka Oono; Hiroyuki Furudate; Shuuitsu Tanaka
Genes & genetic systems, 巻：95, 号：4, 開始ページ：173, 終了ページ：182, 2020年08月, ［査読有り］, ［筆頭著者, 責任著者］, ［国内誌］
T-DNA integration into plant chromosomal DNA via Agrobacterium tumefaciens can be achieved by exploiting the double-strand break repair system of the host's DNA. However, the detailed mechanism of T-DNA integration remains unclear. Here, a sequence analysis of the junction sequences of T-DNA and chromosomal DNA was performed to assess the mechanism of T-DNA integration. T-DNA was introduced into Arabidopsis wild-type and NHEJ-deficient ku80 mutant plants using the floral dip method; the junctions of the left border (LB) of T-DNA were subsequently analyzed by adapter PCR. The most frequent junction of the LB of T-DNA with chromosomal DNA was of the filler DNA type in both lines. The lengths of direct or inverted repeat sequences within or around the filler DNA sequence were greater in the ku80 mutant. In addition, the frequency of T-DNA integration near a transcription start site was significantly higher in the ku80 mutant. Our observations suggest that the presence of the Ku80 protein affects the location of the integration of T-DNA and the pattern of formation of repeat sequences within or around the filler DNA during LB integration into chromosomal DNA.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1266/ggs.19-00064
DOI ID：10.1266/ggs.19-00064, PubMed ID：32848122 - Analysis of localization of cell-cycle regulators in Neurospora crassa.
Kazuki Kuwabara; Ryouhei Yoshihara; Shin Hatakeyama; Shuuitsu Tanaka
Fungal biology, 巻：124, 号：7, 開始ページ：613, 終了ページ：618, 2020年07月, ［査読有り］, ［国際誌］
Most fungi are multinucleated organisms. In some fungi, they have asynchronous nuclei in the same cytoplasm. We analyzed a cell-cycle regulation mechanism using a model fungus Neurospora crassa, which can make heterokaryon cells. G1/S cyclin CLN-1 and cyclin-dependent kinase CDC-2 were tagged with different fluorescence in different strains and expressed. By forming a heterokaryon strain of these, two different fluorescence-tagged proteins were expressed in the same cytoplasm. CDC-2 was localized in all nuclei, whereas CLN-1 was not detected in most of the nuclei and was dispersed in the cytoplasm with small granular clusters. This indicates that in multinucleated fungi, cell-cycle regulators, similar to other proteins, are shared around the nuclei regardless of different cell-cycle stages. Moreover, each nucleus can select and use a special cell-cycle regulator only when it is necessary. Fungal nuclei may have a novel pickup mechanism of necessary proteins from their cytoplasm at the point of use.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.funbio.2020.02.017
DOI ID：10.1016/j.funbio.2020.02.017, PubMed ID：32540184 - Uptake mechanisms of polychlorinated biphenyls in Cucurbita pepo via xylem sap containing major latex-like proteins
Goto J; Iwabuchi A; Yoshihara R; Kodama N; Matsui T; Hirota M; Eun H; Inui H
Environmental and Experimental Botany, 巻：162, 開始ページ：399, 終了ページ：405, 2019年06月, ［査読有り］
Elsevier BV, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.envexpbot.2019.03.019
DOI ID：10.1016/j.envexpbot.2019.03.019, ISSN：0098-8472 - Phenotypic analysis of newly isolated short-lifespan Neurospora crassa mutant deficient in a high mobility group box protein
Ryouhei Yoshihara; ZhengHao Li; Keisuke Ishimori; Kazuki Kuwabara; Shin Hatakeyama; Shuuitsu Tanaka
FUNGAL GENETICS AND BIOLOGY, 巻：105, 開始ページ：28, 終了ページ：36, 2017年08月, ［査読有り］, ［筆頭著者, 責任著者］
To elucidate genetic mechanisms affecting the lifespan of the filamentous fungus Neurospora crassa, we attempted to identify a gene of which a defect causes a short-lifespan. By screening a Neurospora knockout library, provided by the Fungal Genetics Stock Center at Kansas State University, several KO strains with a short-lifespan were isolated. FGSC#11693 is one of these, which shows similar phenotypes to known Neurospora short-lifespan mutants as follows: 1) hyphal growth ceases after about 2 weeks of cultivation, despite that of the wild-type continuing for over 2 years, 2) viability of conidia is lower than that of the wild-type, and 3) high sensitivity to mutagens such as methyl methanesulfonate, ultraviolet radiation, and hydroxyl urea is exhibited. The NCU number of the knocked-out gene in the KO strain is NCU02695, and recovery from the short-lifespan and mutagen sensitivity was achieved by the introduction of this gene from the wild-type. The putative amino acid sequence of the knocked-out gene contains two high mobility group box domains and a mitochondrial localization signal is found at the N-terminal of this sequence. Upon analyzing the subcellular localization of the gene product fused with GFP, GFP signals were detected in mitochondria. From these observations, the gene and KO strain were named mitochondrial high mobility group box protein 1 (MHG1) and mhgl(KO) strain, respectively. The amount of mtDNA relative to the nuclear amount was lower in the mhgl(KO) strain than in the wild-type. mtDNA aberration was also observed in the mhgl(KO) strain. These results suggest that the MHG1 protein plays an important role in the maintenance of mitochondrial DNA, and mitochondrial abnormality caused by mtDNA aberration is responsible for the short-lifespan of the mhgl(KO) strain.
ACADEMIC PRESS INC ELSEVIER SCIENCE, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.fgb.2017.06.001
DOI ID：10.1016/j.fgb.2017.06.001, ISSN：1087-1845, eISSN：1096-0937, Web of Science ID：WOS:000406827300004 - Zinc finger protein genes from Cucurbita pepo are promising tools for conferring non-Cucurbitaceae plants with ability to accumulate persistent organic pollutants
Hideyuki Inui; Matashi Hirota; Junya Goto; Ryouhei Yoshihara; Noriko Kodama; Tomomi Matsui; Kiyoshi Yamazaki; Heesoo Eun
CHEMOSPHERE, 巻：123, 開始ページ：48, 終了ページ：54, 2015年03月, ［査読有り］
Some cultivars of cucumbers, melons, pumpkins, and zucchini, which are members of the Cucurbitaceae family, are uniquely subject to contamination by hydrophobic pollutants such as the organohalogen insecticides DDT. However, the molecular mechanisms for the accumulation of these pollutants in cucurbits have not been determined. Here, cDNA subtraction analysis of Cucurbita pepo cultivars that are low and high accumulators of hydrophobic contaminants revealed that a gene for zinc finger proteins (ZFPs) are preferentially expressed in high accumulators. The cloned CpZFP genes were classified into 2 types: (1) the PEG type, which were expressed in C. pepo cultivars Patty Green, Black Beauty, and Gold Rush, and (2) the BG type, which were expressed in Black Beauty and Gold Rush. Expression of these CpZFP genes in transgenic tobacco plants carrying an aryl hydrocarbon receptor-based inducible gene expression system significantly induced E-glucuronidase activity when the plants were treated with a polychlorinated biphenyl (PCB) compound, indicating that highly hydrophobic PCBs accumulated in the plants. In transgenic tobacco plants carrying CpZFPs, accumulation of dioxins and dioxin-like compounds increased in their aerial parts when they were cultivated in the dioxin-contaminated soil. In summary, we propose that addition of CpZFP genes is a promising tool for conferring noncucurbits with the ability to accumulate hydrophobic contaminants. (C) 2014 Elsevier Ltd. All rights reserved.
PERGAMON-ELSEVIER SCIENCE LTD, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.chemosphere.2014.11.068
DOI ID：10.1016/j.chemosphere.2014.11.068, ISSN：0045-6535, eISSN：1879-1298, Web of Science ID：WOS:000349271800007 - Effect of amending soil with organic acids on perylene uptake into Cucurbita pepo
Ryouhei Yoshihara; Shinryo Matsuo; Toru Iwata; Hideyuki Inui
JOURNAL OF PESTICIDE SCIENCE, 巻：39, 号：3-4, 開始ページ：162, 終了ページ：164, 2014年08月, ［査読有り］, ［筆頭著者］
The solubilization of hydrophobic organic pollutants from soil is important for the phytoremediation of contaminated soil. In this study, we used perylene as a model pollutant to evaluate the abilities of 11 organic acids to solubilize hydrophobic compounds from soil. Oxalic acid and citric acid showed high solubilization abilities, and when added to soil, they enhanced perylene uptake into Cucurbita pepo, likely owing to the chelating ability of these acids. (C) Pesticide Science Society of Japan
PESTICIDE SCI SOC JAPAN, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1584/jpestics.D14-054
DOI ID：10.1584/jpestics.D14-054, ISSN：1348-589X, eISSN：1349-0923, Web of Science ID：WOS:000346892300008 - Mutational effects of γ-rays and carbon ion beams in Arabidopsis seedlings
Yoshihara R; Nozawa S; Hase Y; Narumi I; Hidema J; Sakamoto AN
Journal of Radiation Research, 巻：54, 号：6, 開始ページ：1050, 終了ページ：1056, 2013年11月, ［査読有り］, ［筆頭著者］
To assess the mutational effects of radiation on vigorously proliferating plant tissue, the mutation spectrum was analyzed with Arabidopsis seedlings using the plasmid-rescue method. Transgenic plants containing the Escherichia coli rpsL gene were irradiated with gamma-rays and carbon ion beams (320-MeV C-12(6+)), and mutations in the rpsL gene were analyzed. Mutant frequency increased significantly following irradiation by gamma-rays, but not by 320-MeV C-12(6+). Mutation spectra showed that both radiations increased the frequency of frameshifts and other mutations, including deletions and insertions, but only gamma-rays increased the frequency of total base substitutions. These results suggest that the type of DNA lesions which cause base substitutions were less often induced by 320-MeV C-12(6+) than by gamma-rays in Arabidopsis seedlings. Furthermore, gamma-rays never increased the frequencies of G:C to T:A or A:T to C:G transversions, which are caused by oxidized guanine; 320-MeV C-12(6+), however, produced a slight increase in both transversions. Instead, gamma-rays produced a significant increase in the frequency of G:C to A:T transitions. These results suggest that 8-oxoguanine has little effect on mutagenesis in Arabidopsis cells.
OXFORD UNIV PRESS, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1093/jrr/rrt074
DOI ID：10.1093/jrr/rrt074, ISSN：0449-3060, eISSN：1349-9157, Web of Science ID：WOS:000327458900010 - Arabidopsis sos1 mutant in a salt-tolerant accession revealed an importance of salt acclimation ability in plant salt tolerance.
Hirotaka Ariga; Taku Katori; Ryouhei Yoshihara; Yoshihiro Hase; Shigeki Nozawa; Issay Narumi; Satoshi Iuchi; Masatomo Kobayashi; Kenji Tezuka; Yoichi Sakata; Takahisa Hayashi; Teruaki Taji
Plant signaling & behavior, 巻：8, 号：7, 開始ページ：e24779, 2013年07月, ［査読有り］, ［国際誌］
An analysis of the salinity tolerance of 354 Arabidopsis thaliana accessions showed that some accessions were more tolerant to salt shock than the reference accession, Col-0, when transferred from 0 to 225 mM NaCl. In addition, several accessions, including Zu-0, showed marked acquired salt tolerance after exposure to moderate salt stress. It is likely therefore that Arabidopsis plants have at least two types of tolerance, salt shock tolerance and acquired salt tolerance. To evaluate a role of well-known salt shock tolerant gene SOS1 in acquired salt tolerance, we isolated a sos1 mutant from ion-beam-mutagenized Zu-0 seedlings. The mutant showed severe growth inhibition under salt shock stress owing to a single base deletion in the SOS1 gene and was even more salt sensitive than Col-0. Nevertheless, it was able to survive after acclimation on 100 mM NaCl for 7 d followed by 750 mM sorbitol for 20 d, whereas Col-0 became chlorotic under the same conditions. We propose that genes for salt acclimation ability are different from genes for salt shock tolerance and play an important role in the acquisition of salt or osmotic tolerance.
英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.4161/psb.24779
DOI ID：10.4161/psb.24779, PubMed ID：23656872, PubMed Central ID：PMC3908940 - Mutagenic effects of carbon ions near the range end in plants
Yoshihiro Hase; Ryouhei Yoshihara; Shigeki Nozawa; Issay Narumi
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 巻：731, 号：1-2, 開始ページ：41, 終了ページ：47, 2012年03月, ［査読有り］
To gain insight into the mutagenic effects of accelerated heavy ions in plants, the mutagenic effects of carbon ions near the range end (mean linear energy transfer (LET): 425 keV/mu m) were compared with the effects of carbon ions penetrating the seeds (mean LET: 113 keV/mu m). Mutational analysis by plasmid rescue of Escherichia coli rpsL from irradiated Arabidopsis plants showed a 2.7-fold increase in mutant frequency for 113 keV/mu m carbon ions, whereas no enhancement of mutant frequency was observed for carbon ions near the range end. This suggested that carbon ions near the range end induced mutations that were not recovered by plasmid rescue. An Arabidopsis DNA ligase IV mutant, deficient in non-homologous end-joining repair, showed hyper-sensitivity to both types of carbon-ion irradiation. The difference in radiation sensitivity between the wild type and the repair-deficient mutant was greatly diminished for carbon ions near the range end, suggesting that these ions induce irreparable DNA damage. Mutational analysis of the Arabidopsis GL1 locus showed that while the frequency of generation of glabrous mutant sectors was not different between the two types of carbon-ion irradiation, large deletions (>similar to 30 kb) were six times more frequently induced by carbon ions near the range end. When 352 keV/mu m neon ions were used, these showed a 6.4 times increase in the frequency of induced large deletions compared with the 113 keV/mu m carbon ions. We suggest that the proportion of large deletions increases with LET in plants, as has been reported for mammalian cells. The nature of mutations induced in plants by carbon ions near the range end is discussed in relation to mutation detection by plasmid rescue and transmissibility to progeny. (C) 2011 Elsevier B.V. All rights reserved.
ELSEVIER SCIENCE BV, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1016/j.mrfmmm.2011.10.004
DOI ID：10.1016/j.mrfmmm.2011.10.004, ISSN：0027-5107, eISSN：1873-135X, PubMed ID：22027091, Web of Science ID：WOS:000300918800005 - Mutational effects of different LET radiations in rpsL transgenic Arabidopsis
Ryouhei Yoshihara; Yoshihiro Hase; Ryohei Sato; Koichi Takimoto; Issay Narumi
INTERNATIONAL JOURNAL OF RADIATION BIOLOGY, 巻：86, 号：2, 開始ページ：125, 終了ページ：131, 2010年02月, ［査読有り］, ［筆頭著者, 責任著者］
Purpose: In an effort to assess the characteristics of mutation induced by different linear energy transfer (LET) radiation in higher plants, the mutational effects of carbon-ion beams and gamma-rays were investigated in Arabidopsis.
Materials and methods: The rpsL (Escherichia coli ribosomal protein small subunit S 12) transgenic Arabidopsis (Arabidopsis/rpsL) mutation detection system was adopted. Dry seeds of Arabidopsis/rpsL were irradiated with gamma-rays and 208-MeV carbon ions (208-MeV (12)C(5+)), and the mutation frequency and mutation spectrum were examined.
Results: The frequency of mutant clones increased following irradiation with 208-MeV (12)C(5+) and gamma-rays. Mutation spectrum analysis showed that G:C to A:T transitions and > 2 bp deletions/insertions were significantly induced by both 208-MeV (12)C(5+) and gamma-rays. -1 and -2 frameshift mutations were characteristic in the gamma-ray irradiated group.
Conclusions: 208-Mev (12)C(5+) and gamma-rays induced different intragenic mutations in respect to the size of deletions, reflecting differences in the nature of the DNA damage induced. Our results also suggested that base substitutions derived from the generation of 8-oxoguanine were low in dry seeds. The mutation spectrum obtained in this study might have reflected the characteristic conditions of plant dry seeds such as low water content and cell proliferation activity.
TAYLOR & FRANCIS LTD, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.3109/09553000903336826
DOI ID：10.3109/09553000903336826, ISSN：0955-3002, CiNii Articles ID：10026470848, PubMed ID：20148698, Web of Science ID：WOS:000274629400005 - Silencing of CPD Photolyase Makes Arabidopsis Hypersensitive and Hypermutable in Response to UV-B Radiation.
Yoshihara, R; Nakane, C; Sato, R; Yasuda, A; Takimoto, K
Genes and Environment, 巻：30, 号：2, 開始ページ：53, 終了ページ：61, 2008年06月, ［査読有り］, ［筆頭著者, 責任著者］
Plants are exposed to solar ultraviolet radiation (UV), which has deleterious effects on plant growth, development and physiology. Cyclobutane pyrimidine dimers (CPDs) are a major form of UV-induced DNA damage. It is conceivable that the reversal of CPDs is important for the reduction of lethal and mutagenic effects. Photoreactivation catalyzed by CPD photolyase is an efficient CPD repair system with a mechanism dependent on UV-A/visible light, which is contained in solar radiation. Photoreactivation presumably functions to protect plants from solar UV. We generated a CPD photolyase knock-down in Arabidopsis thaliana by RNA interference (RNAi) to investigate the role of CPD photorepair for protection of plants from solar UV. These knock-down lines exhibited hypersensitivity to UV-B and an increased occurrence of mutation induced by UV-B radiation compared with Arabidopsis proficient in CPD photolyase. Mutations induced by UV-B were determined by an rpsL mutation assay system. G:C to A:T transitions were frequently observed in CPD photolyase knock-down lines at the site of dipyrimidine sequences. A high incidence of frameshifts was observed in irradiated knock-down lines. These results indicate that CPD photoreactivation plays an important role for UV resistance of Arabidopsis and suppression of UV-induced mutagenesis.
日本環境変異原学会, 英語
DOI：https://doi.org/10.3123/jemsge.30.53
DOI ID：10.3123/jemsge.30.53, ISSN：1880-7046, CiNii Articles ID：110006781782, CiNii Books ID：AA1212552X - A new system for detecting mutations in Arabidopsis thaliana and the mutational spectra resulting from ethylmethanesulfonate treatment
Ryouhei Yoshihara; Chiyoko Nakane; Koichi Takimoto
JOURNAL OF RADIATION RESEARCH, 巻：47, 号：3-4, 開始ページ：223, 終了ページ：228, 2006年11月, ［査読有り］, ［筆頭著者］
A system was developed for the detection and analysis of mutations occurring on chromosomal DNA in plants. The plasmid pML4, carrying the Escherichia coli rpsL gene, a target gene for mutagenesis, was inserted into a shuttle vector, pCGN5138, to construct a plasmid which could be used for the transformation of plants. pML4 sequences were introduced into Arabidopsis thaliana mediated by Agrobacterium. The pML4 DNA was rescued from transgenic Arabidopsis plants exposed to mutagens, and the plasmids were introduced into Escherichia coli DH10B to isolate mutant clones. In this system, any form of inactivation mutation in the rpsL gene can be positively selected since it makes the E. coli cells resistant to streptomycin. Here we report that the system could detect the mutagenic effect of ethylmethanesulfonate (EMS). Further characterization of the mutants revealed that G:C to A:T transitions predominated among the EMS-induced mutations. This assay system is useful for the detection and analysis of mutations arising on chromosomal DNA in plants, and should be useful for evaluating analysis of the effects of environmental mutagens.
JAPAN RADIATION RESEARCH SOC, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1269/jrr.0623
DOI ID：10.1269/jrr.0623, ISSN：0449-3060, CiNii Articles ID：110004850768, Web of Science ID：WOS:000243613000001 - CPD photolyase gene from Spinacia oleracea: repair of UV-damaged DNA and expression in plant organs
R Yoshihara; T Imaki; M Hori; C Watanabe; K Yamamoto; K Takimoto
JOURNAL OF RADIATION RESEARCH, 巻：46, 号：2, 開始ページ：157, 終了ページ：164, 2005年06月, ［査読有り］, ［筆頭著者］
The UV-B radiation contained in solar radiation has deleterious effects on plant growth, development and physiology. Specific damage to DNA caused by UV radiation involves the cyclobutyl pyrimidine dimers (CPD) and the pyrimidine (6-4) pyrimidone photoproducts. CPDs are repaired by CPD photolyase via a UV-A/blue light-dependent mechanism. The gene for the class II CPD photolyase has been cloned from higher plants such as Arabidopsis, cucumbers and rice. We isolated and characterized the cDNA and a genomic clone encoding the spinach class II CPD photolyase. The gene consisted of 3777 bases and 9 exons. The sequence of amino acids predicted from the nucleotide sequence of the cDNA of the gene was highly homologous to that of the higher plants listed above. When a photolyase-deficient Escherichia coli strain was transformed with the cDNA, photoreactivation activity was partially restored, by the illumination with photoreactivating light, resulting in an increased survival and decreased content of CPDs in the Escherichia coli genome. In both the male and female plants, the gene was highly expressed in leaves and flowers under the condition of 14-h light and 10-h dark cycle. The expression in the roots was quite low compared with the other organs.
JAPAN RADIATION RESEARCH SOC, 英語, 研究論文（学術雑誌）
DOI：https://doi.org/10.1269/jrr.46.157
DOI ID：10.1269/jrr.46.157, ISSN：0449-3060, CiNii Articles ID：110004041111, PubMed ID：15988133, Web of Science ID：WOS:000231293000004
- Generating new ornamental plant varieties using ion beams
Zaiton Ahmand; Affrida Abu Hassan; Shakinah Salleh; Nurul Hidayah Mahmud; Yutaka Oono; Yoshihiro Hase; Shigeki Nozawa; Ryouhei Yoshihara; Issay Narumi; Atsushi Tanaka
JAEA-Review 2015-037, 開始ページ：27, 終了ページ：120, 2016年03月
英語 - Gamma-ray and ion beam irradiated plants showed unique mutation spectra
Yoshihara R; Nozawa S; Hase Y; Narumi I; Hidema J; Sakamoto AN
ANNUAL REPORT QuBS 2014, 開始ページ：28, 2015年
英語 - Mutational effects of carbon ions near the range end and development of an efficient mutagenesis technique using ion beams
Hase Y; Yoshihara R; Nozawa S; Narumi I
JAEA Takasaki Annual Report 2011, 開始ページ：98, 2013年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - Effect of carbon ion beam irradiation on callus growth in arabidopsis and rice
Saika H; Yoshihara R; Sakamoto AN; Toki S
JAEA Takasaki Annual Report 2011, 開始ページ：99, 2013年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - Selective uptake of polychlorinated biphenyls by Cucurbita pepo
Inui, H; Yamazaki, K; Matsuo, S; Yoshihara, R; Eun, H
Organohalogen Compounds, 巻：74, 開始ページ：1213, 終了ページ：1216, 2012年
英語 - Mutational Effects of carbon ions near range end in Arabidopsis
Hase Y; Yoshihara R; Nozawa S; Narumi I
JAEA Takasaki Annual Report 2010, 開始ページ：103, 2012年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - イオンビーム照射を行った‘弘前在来’トウガラシの照射次代における変異の発現
佐藤悠輔; 前田智雄; 本多和茂; 嵯峨紘一; 石川隆二; 秋田祐介; 吉原亮平; 野澤樹; 鳴海一成
園芸学研究 別冊, 巻：10, 号：2, 開始ページ：421, 2011年09月24日
日本語
DOI：https://doi.org/10.2503/hrj.10.421
DOI ID：10.2503/hrj.10.421, ISSN：1881-8307, J-Global ID：201102294601607135 - Effect of different LET radiations on Root Growth of Arabidopsis thaliana
Yoshihara R; Nozawa S; Saika H; Teranishi M; Toki S; Hidema J; Sakamoto AN
JAEA Takasaki Annual Report 2009, 開始ページ：74, 2011年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - Mutational effect of gamma-rays and carbon ion beams on Arabidopsis thaliana seedlings
Yoshihara R; Hase Y; Nozawa S; Sakamoto AN; Narumi I
JAEA Takasaki Annual Report 2009, 開始ページ：59, 2011年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - シロイヌナズナにおいて異なるLETの炭素イオンによって誘発される突然変異の特徴
長谷純宏; 吉原亮平; 野澤樹; 玉置雅紀; 鳴海一成
日本放射線影響学会大会講演要旨集, 巻：53rd, 2010年
ISSN：1347-8680, J-Global ID：201002289881003131 - Mutation spectrum induced by γ-rays and carbon ion beams in plant
Yoshihara R; Hase Y; Nozawa S; Narumi I
JAEA Takasaki Annual Report 2008, 開始ページ：67, 2009年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - Analysis on optimum irradiation condition for the ion beam breeding.
Yoshihara R; Hase Y; Narumi I
JAEA Takasaki Annual Report 2007, 開始ページ：59, 2008年
英語, 速報，短報，研究ノート等（大学，研究機関紀要） - Specificity of UV-induced mutation in Arabidopsis thaliana
Ryouhei Yoshihara; Chiyoko Nakane; Ryohei Sato; Ai Yasuda; Koichi Takimoto
巻：48, 開始ページ：S234, 終了ページ：S234, 2007年
英語, 研究発表ペーパー・要旨（国際会議）
ISSN：0032-0781, Web of Science ID：WOS:000245922701414 - <総説>高等植物における突然変異スペクトル解析システムの現状
吉原亮平; 滝本晃一
放射線生物研究, 巻：41, 号：1, 開始ページ：61, 終了ページ：76, 2006年
放射線生物研究会, 日本語
ISSN：0441-747X, CiNii Articles ID：40007245056, CiNii Books ID：AN00225482 - Development of the assay system for the detection of mutation occurring on DNA in higher plants
R Yoshihara; M Ezawa; K Takimoto
巻：47, 開始ページ：S219, 終了ページ：S219, 2006年
英語, 研究発表ペーパー・要旨（国際会議）
ISSN：0032-0781, Web of Science ID：WOS:000236401401362
- ミトコンドリアDNA安定維持機構の破綻が⽷状菌の菌⽷⽣⻑に及ぼす影響
吉原亮平; 石森啓右; 野剛人; 清家快海; 渡部あさぎ; 島倉柚貴; 北村智; 畠山晋; 田中秀逸
第45回 日本分子生物学会年会, 2022年11月
ポスター発表 - 高等植物におけるDSB修復に対するPOLQ遺伝子の寄与
戸田美波子; 北村智; 柴田晴菜子; 大圖美世; 島倉柚貴; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第94回大会, 2022年09月
ポスター発表 - 早期に菌糸生長を停止させるアカパンカビ変異株ms-1の遺伝学的解析
島倉柚貴; 北村智; 大圖美世; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第94回大会, 2022年09月
ポスター発表 - シロイヌナズナにおける NHEJ 因子 Ku70 の局在解析
大圖美世; 北村智; 柴田晴菜子; 戸田美波子; 島倉柚貴; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第94回大会, 2022年09月
ポスター発表 - アグロバクテリウムによる植物染色体DNAへのT-DNA挿入に対するNHEJ経路の関与
戸田美波子; 廣石歩; 中嶋友里恵; 柴田晴菜子; 大野豊; 北村智; 乾秀之; 畠山晋; 田中秀逸; 吉原亮平
日本分子生物学会年会プログラム・要旨集(Web), 2021年
2021年 - 2021年 - MHG1タンパク質の推定機能ドメインはNeurospora crassaのmtDNA安定性に重要な役割を果たす
吉原亮平; 清家快海; 石森啓右; 桐生真緒; 渡部あさぎ; 廣石歩; 畠山晋; 田中秀逸
日本分子生物学会年会プログラム・要旨集(Web), 2020年
2020年 - 2020年 - シロイヌナズナにおけるNHEJ主要因子Kuタンパク質の局在解析
柴田晴菜子; 小池学; 田中秀逸; 吉原亮平
日本分子生物学会年会プログラム・要旨集(Web), 2020年
2020年 - 2020年 - 複製ストレス応答におけるヒストンH3K4メチル化の重要性
柳澤健斗; 吉原亮平; 畠山晋; 田中秀逸
日本遺伝学会大会プログラム・予稿集, 2020年
2020年 - 2020年 - DNA鎖間架橋修復に関与するヌクレアーゼの遺伝学的解析
塚田耕太郎; 吉原亮平; 畠山晋; 田中秀逸
日本遺伝学会大会プログラム・予稿集, 2020年
2020年 - 2020年 - カビや植物で見られるDNA修復が関わる生命現象の解析
吉原亮平
変異機構研究会・第32回夏の学校, 2019年09月, ［招待有り］, ［国内会議］
日本語, 口頭発表（招待・特別） - アカパンカビにおける寿命関連遺伝子mhg1の機能ドメイン解析
清家快海; 石森啓右; 桐生真緒; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第91回大会, 2019年09月, ［国内会議］
日本語, 口頭発表（一般） - アカパンカビ新規短寿命変異原感受性株の責任遺伝子の解析
秋山優; 青野剛人; 清家快海; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第91回大会, 2019年09月, ［国内会議］
日本語, ポスター発表 - アカパンカビにおける光回復の部分的な欠損はDNA鎖間架橋修復機構の機能不全によって生じる
塚田耕太郎; 吉原亮平; 畠山晋; 田中秀逸
日本遺伝学会第91回大会, 2019年09月, ［国内会議］
日本語, 口頭発表（一般） - DNA二本鎖切断修復機構がRhizobium radiobacterによる植物染色体へのDNA挿入に与える影響
岡田真希; 三富優香; 田上舞; 乾秀之; 田中秀逸; 吉原亮平
日本遺伝学会第90回大会, 2018年09月, ［国内会議］
日本語, ポスター発表 - アカパンカビにおける寿命関連遺伝子mhg1の機能解析
清家快海; 石森啓右; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第90回大会, 2018年09月, ［国内会議］
日本語, ポスター発表 - アカパンカビにおけるミトコンドリア維持に関わるmhg1の機能解析
石森啓右; 清家快海; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第90回大会, 2018年09月, ［国内会議］
日本語, 口頭発表（一般） - アカパンカビ新規短寿命変異原感受性株の表現型解析および原因遺伝子の特定
青野剛人; 秋山優; 清家快海; 畠山晋; 田中秀逸; 吉原亮平
日本遺伝学会第90回大会, 2018年09月, ［国内会議］
日本語, ポスター発表 - アカパンカビにおける寿命関連遺伝子mhg1の機能解析
石森啓右; 吉原亮平; 清家快海; 畠山晋; 田中秀逸
ConBio2017, 2017年12月, ［国内会議］
日本語, ポスター発表 - アカパンカビ新規短寿命株の表現型解析および短寿命原因遺伝子の特定
青野剛人; 吉原亮平; 清家快海; 畠山晋; 田中秀逸
ConBio2017, 2017年12月, ［国内会議］
日本語, ポスター発表 - Analysis of newly isolated gene affecting to life span of Neurospora crassa
吉原 亮平; 李 正皓; 桑原 一樹; 石森 啓右; 畠山 晋; 田中 秀逸
第39回日本分子生物学会, 2016年11月, ［国内会議］
ポスター発表 - アカパンカビ早期老化変異株の解析
吉原 亮平; 李 正皓; 畠山 晋; 田中 秀逸
第38回日本分子生物学会, 2015年12月, ［国内会議］
日本語, ポスター発表 - Screening and analysis of short life span strains of Neurospora crassa
Yoshihara R; Li ZH; Hatakeyama S; Tanaka S
15th International Congress of Radiation Research, 2015年05月, ［国際会議］
英語, ポスター発表 - ウリ科植物の根における脂溶性物質結合タンパク質の局在性
吉原 亮平; 後藤 純弥; 岩渕 彩; 池田 健一; 井上 加奈子; 乾 秀之
日本農芸化学会2014年度大会, 2014年03月, ［国内会議］
日本語, 口頭発表（一般） - High accumulation and congener specific uptake of dioxins and dioxin-like compounds in Cucurbita pepo
Yoshihara R; Yamazaki K; Eun H; Inui H
SETAC Asia/Pacific 2012, 2012年09月, ［国際会議］
英語, 口頭発表（一般） - Mutation Spectrum Analysis in Higher Plants
Yoshihara R; Hase Y; Nozawa S; Narumi I; Takimoto K; Sakamoto AN
EMBO Workshop, GENETIC STABILITY & CHANGE: Genome maintenance mechanisms in plants, 2012年05月, ［国際会議］
英語, ポスター発表 - 放射線誘発ＤＮＡ酸化損傷がシロイヌナズナの突然変異に与える影響
吉原 亮平; 長谷 純宏; 野澤 樹; 鳴海 一成; 滝本 晃一; 日出間 純; 坂本 綾子
日本放射線影響学会 第54回大会, 2011年11月, ［国内会議］
日本語, ポスター発表 - 変異誘発効果から見る紫外線および電離放射線の高等植物に対する影響 –シロイヌナズナを用いた突然変異スペクトル解析–
吉原 亮平; 滝本 晃一; 長谷 純宏; 野澤 樹; 坂本 綾子; 鳴海 一成
日本放射線影響学会 第53回大会, 2010年10月, ［国内会議］
日本語, 口頭発表（一般） - シロイヌナズナのM1個体における放射線誘発DNA損傷の推定 -Arabidopsis/rpsLを用いた遺伝子内変異の解析-
吉原亮平; 長谷純宏; 坂本綾子; 野澤樹; 滝本晃一; 鳴海一成
第51回 日本植物生理学会, 2010年03月, ［国内会議］
日本語, ポスター発表 - 高等植物における放射線誘発変異の特徴解析
吉原亮平; 長谷純宏; 野澤樹; 滝本晃一; 鳴海一成
日本放射線影響学会 第52回大会, 2009年11月, ［国内会議］
日本語, ポスター発表 - シロイヌナズナ乾燥種子および幼植物体における放射線誘発変異の解析
吉原亮平; 長谷純宏; 滝本晃一; 鳴海一成
日本放射線影響学会 第51回大会, 2008年11月, ［国内会議］
日本語, ポスター発表 - 放射線により高等植物内で誘発される変異の特徴
吉原亮平; 長谷純宏; 滝本晃一; 鳴海一成
変異機構研究会・第21回夏の学校, 2008年06月, ［国内会議］
日本語, 口頭発表（一般） - 高等植物におけるイオンビームおよびガンマ線誘発突然変異スペクトル解析
吉原亮平; 長谷純宏; 滝本晃一; 鳴海一成
第5回イオンビーム育種研究会大会, 2008年05月, ［国内会議］
日本語, 口頭発表（一般） - シロイヌナズナにおけるイオンビームおよびガンマ線誘発突然変異の解析
吉原亮平; 長谷純宏; 滝本晃一; 鳴海一成
第49回 日本植物生理学会, 2008年03月, ［国内会議］
日本語, ポスター発表 - rpsL遺伝子を用いた植物における突然変異解析
吉原亮平; 安田愛; 佐藤良平; 長谷純宏; 鳴海一成; 滝本晃一
変異機構研究会・第20回夏の学校, 2007年07月, ［国内会議］
日本語, 口頭発表（一般） - シロイヌナズナにおける紫外線誘発遺伝子突然変異の特異性
吉原亮平; 中根千陽子; 佐藤良平; 安田愛; 滝本晃一
第48回 日本植物生理学会, 2007年03月, ［国内会議］
日本語, ポスター発表 - シロイヌナズナにおけるEMS誘発突然変異
吉原亮平; 滝本晃一
日本放射線影響学会 第49回大会, 2006年09月, ［国内会議］
日本語, ポスター発表 - 高等植物における遺伝子突然変異の検出システムの開発
吉原亮平; 江澤美智; 滝本晃一
第47回 日本植物生理学会, 2006年03月, ［国内会議］
日本語, ポスター発表 - 高等植物のもつ光回復機能とその紫外線耐性への寄与の検討
吉原亮平; 渡辺千尋; 中根千陽子; 森 俊雄; 滝本晃一
日本農芸化学会 中四国支部 第12回例会, 2005年05月, ［国内会議］
日本語, 口頭発表（一般） - 植物の染色体DNAに生じた突然変異解析システムの開発
吉原亮平; 渡辺千尋; 滝本晃一
日本放射線影響学会 第47回大会, 2004年11月, ［国内会議］
日本語, ポスター発表 - ホウレンソウ CPD 光回復遺伝子(II) 機能と特性
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