研究者総覧

佐久間 洋 (サクマ ヨウ)

  • 大学院理工学研究科 環境機能科学専攻 准教授
Last Updated :2020/11/10

研究者情報

J-Global ID

研究分野

  • ライフサイエンス / 植物分子、生理科学

研究活動情報

論文

  • Modifications of water status, growth rate and antioxidant system in two wheat cultivars as affected by salinity stress and salicylic acid
    Naglaa Loutfy, Yoh Sakuma, Dharmendra K Gupta, Masahiro Inouhe
    Journal of Plant Research 2020年04月 [査読有り]
     研究論文(学術雑誌)
  • Sakae Horisawa, Hiromasa Ando, Osamu Ariga, Yoh Sakuma
    BIORESOURCE TECHNOLOGY 197 37 - 41 2015年12月 [査読有り]
     研究論文(学術雑誌) 
    In the present study, ethanol production from polysaccharides or wood chips was conducted in a single reactor under anaerobic conditions using the white rot fungus Schizophyllum commune NBRC 4928, which produces enzymes that degrade lignin, cellulose and hemicellulose. The ethanol yields produced from glucose and xylose were 80.5%, and 52.5%, respectively. The absolute yields of ethanol per microcrystalline cellulose (MCC), xylan and arabinogalactan were 0.26 g/g(-MCC), 0.0419 g/g(-xylan) and 0.0508 g/g(-arabinogalactan), respectively. By comparing the actual ethanol yields from polysaccharides with monosaccharide fermentation, it was shown that the rate of saccharification was slower than that in fermentation. S. commune NBRC 4928 is concluded to be suitable for CBP because it can produce ethanol from various types of sugar. From the autoclaved cedar chip, only little ethanol was produced by S. commune NBRC 4928 alone but ethanol production was enhanced by combined use of ethanol fermenting and lignin degrading fungi. (C) 2015 Elsevier Ltd. All rights reserved.
  • Sakae Horisawa, Yoh Sakuma, Shuichi Doi
    JOURNAL OF WOOD SCIENCE 59 5 432 - 441 2013年10月 [査読有り]
     研究論文(学術雑誌) 
    A method for identification and typing of wood rotting fungi using the melting temperature [T(m)] of DNA fragments coding rRNA (rDNA) was examined. The T(m)s of four DNA fragments, inter transcribed spacer (ITS) I, ITS II, and two partial fragments of 28S rDNA from each of 20 species of wood rotting fungi, were measured by melting curve analysis. The T(m) variation was large enough between species to enable identification based on the T(m) values. A pair of T(m)s of the ITS I region (between the primers ITS1 and ITS2) and the ITS II region (between the primers ITS3 and ITS4) had the highest resolution for identifying wood rotting fungi. To assess about the diversity of the T(m), intraspecific diversity of these DNA fragment sequences was evaluated using test strain sequences and data from the GenBank/EMBL/DDBJ biological database. The intraspecific diversity of T(m) was considered to be small because the nucleotide diversity of each fragment was small within the species.
  • Coordinative roles of organic and inorganic solutes in plant responses to drought and salinity stresses
    Inouhe M, Ichi T, Matsumoto A, Sakuma Y, Loutfy N
    Abstract of VIPCA II Conference for Plant Abiotic Stress Tolerance II, Vienna, Austria 2012年02月 [査読有り][招待有り]
     研究論文(学術雑誌)
  • Naglaa Loutfy, Mohamed A. El-Tayeb, Ahmed M. Hassanen, Mahmoud F. M. Moustafa, Yoh Sakuma, Masahiro Inouhe
    JOURNAL OF PLANT RESEARCH 125 1 173 - 184 2012年01月 [査読有り]
     研究論文(学術雑誌) 
    Salicylic acid (SA) controls growth and stress responses in plants. It also induces drought tolerance in plants. In this paper, four wheat (Triticum aestivum L.) cultivars with different drought responses were treated with SA in three levels of drain (90, 60, 30% of maximum field capacity) to examine its interactive effects on drought responses and contents of osmotic solutes that may be involved in growth and osmotic adjustment. Under drought condition, the cultivars Geza 164 and Sakha 69 had the plant biomass and leaf relative water content (LRWC) greater than the cultivars Gemaza 1 and Gemaza 3. In all cultivars, drought stress decreased the biomass, LRWC, and the contents of inorganic solutes (Ca, K, Mg) and largely increased the contents of organic solutes (soluble sugars and proline). By contrast, SA increased the biomass, LRWC and the inorganic and organic solute contents, except proline. Correlation analysis revealed that the LRWC correlated positively with the inorganic solute contents but negatively with proline in all cultivars. SA caused maximum accumulations of soluble sugars in roots under drought. These results indicated that SA-enhanced tolerance might involve solute accumulations but independently of proline biosynthesis. Drought-sensitive cultivars had a trait lowering Ca and K levels especially in shoots. Possible functions of the ions and different traits of cultivars were discussed.
  • Takumi Yoshida, Naohiko Ohama, Jun Nakajima, Satoshi Kidokoro, Junya Mizoi, Kazuo Nakashima, Kyonoshin Maruyama, Jong-Myong Kim, Motoaki Seki, Daisuke Todaka, Yuriko Osakabe, Yoh Sakuma, Friedrich Schoeffl, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    MOLECULAR GENETICS AND GENOMICS 286 5-6 321 - 332 2011年12月 [査読有り]
     研究論文(学術雑誌) 
    Arabidopsis DREB2A is a key transcription factor of heat- and drought-responsive gene expression, and DREB2A expression is induced by these stresses. We analyzed the DREB2A promoter and found a heat shock element that functions as a cis-acting element in the heat shock (HS)-responsive expression of DREB2A. Among the 21 Arabidopsis heat shock factors, we chose 4 HsfA1-type proteins as candidate transcriptional activators (HsfA1a, HsfA1b, HsfA1d, and HsfA1e) based on transactivation activity and expression patterns. We generated multiple mutants and found that the HS-responsive expression of DREB2A disappeared in hsfa1a/b/d triple and hsfa1a/b/d/e quadruple mutants. Moreover, HS-responsive gene expression, including that of molecular chaperones and transcription factors, was globally and drastically impaired in the hsfa1a/b/d triple mutant, which exhibited greatly reduced tolerance to HS stress. HsfA1 protein accumulation in the nucleus was negatively regulated by their interactions with HSP90, and other factors potentially strongly activate the HsfA1 proteins under HS stress. The hsfa1a/b/d/e quadruple mutant showed severe growth retardation, and many genes were downregulated in this mutant even under non-stress conditions. Our study indicates that HsfA1a, HsfA1b, and HsfA1d function as main positive regulators in HS-responsive gene expression and four HsfA1-type proteins are important in gene expression for normal plant growth.
  • Kyonoshin Maruyama, Migiwa Takeda, Satoshi Kidokoro, Kohji Yamada, Yoh Sakuma, Kaoru Urano, Miki Fujita, Kyouko Yoshiwara, Satoko Matsukura, Yoshihiko Morishita, Ryosuke Sasaki, Hideyuki Suzuki, Kazuki Saito, Daisuke Shibata, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT PHYSIOLOGY 150 4 1972 - 1980 2009年08月 [査読有り]
     研究論文(学術雑誌) 
    DREB1A/CBF3 and DREB2A are transcription factors that specifically interact with a cis-acting dehydration-responsive element (DRE), which is involved in cold-and dehydration-responsive gene expression in Arabidopsis (Arabidopsis thaliana). Overexpression of DREB1A improves stress tolerance to both freezing and dehydration in transgenic plants. In contrast, overexpression of an active form of DREB2A results in significant stress tolerance to dehydration but only slight tolerance to freezing in transgenic plants. The downstream gene products for DREB1A and DREB2A are reported to have similar putative functions, but downstream genes encoding enzymes for carbohydrate metabolism are very different between DREB1A and DREB2A. We demonstrate that under cold and dehydration conditions, the expression of many genes encoding starch-degrading enzymes, sucrose metabolism enzymes, and sugar alcohol synthases changes dynamically; consequently, many kinds of monosaccharides, disaccharides, trisaccharides, and sugar alcohols accumulate in Arabidopsis. We also show that DREB1A overexpression can cause almost the same changes in these metabolic processes and that these changes seem to improve freezing and dehydration stress tolerance in transgenic plants. In contrast, DREB2A overexpression did not increase the level of any of these metabolites in transgenic plants. Strong freezing stress tolerance of the transgenic plants overexpressing DREB1A may depend on accumulation of these metabolites.
  • Sakae Horisawa, Yoh Sakuma, Shuichi Doi
    JOURNAL OF WOOD SCIENCE 55 2 133 - 138 2009年04月 [査読有り]
     研究論文(学術雑誌) 
    Species-specific oligonucleotide primers for detecting wood rot fungi, Gloeophyllum trabeum, Trametes versicolor, Coniophora puteana, and Serpula lacrymans, and the primer detecting a group of related fungi to G. sepiarium were developed. These primer sequences were picked up from the internal transcribed spacer region between small-subunit rDNA and large-subunit rDNA. The species selectivities of the developed primers were checked. Real-time polymerase chain reaction (PCR) was carried out using these highly specific primers to quantitatively detect at least of 0.01 ng genome DNA of the target species. This quantitative PCR was also used to differentiate the target species DNA from mixed species DNA. A PCR-based technique using the species-specific primers would be applicable to multiple-sample assay in diagnosis of wood decay and to investigation of environmental fungal populations.
  • Feng Qin, Yoh Sakuma, Lam-Son Phan Tran, Kyonoshin Maruyama, Satoshi Kidokoro, Yasunari Fujita, Miki Fujita, Taishi Umezawa, Yoriko Sawano, Ken-ichi Miyazono, Masaru Tanokura, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT CELL 20 6 1693 - 1707 2008年06月 [査読有り]
     研究論文(学術雑誌) 
    The DEHYDRATION-RESPONSIVE ELEMENT BINDING PROTEIN2A (DREB2A) transcription factor controls water deficit inducible gene expression and requires posttranslational modification for its activation. The activation mechanism is not well understood; however, the stability of this protein in the nucleus was recently found to be important for its activation. Here, we report the isolation of Arabidopsis thaliana DREB2A-INTERACTING PROTEIN1 (DRIP1) and DRIP2, C3HC4 RING domain-containing proteins that interact with the DREB2A protein in the nucleus. An in vitro ubiquitination assay showed that they function as E3 ubiquitin ligases and are capable of mediating DREB2A ubiquitination. Overexpression of DRIP1 in Arabidopsis delayed the expression of DREB2A-regulated drought-responsive genes. Drought-inducible gene expression was slightly enhanced in the single T-DNA mutants of drip1-1 and drip2-1. By contrast, significantly enhanced gene expression was revealed in the drip1 drip2 double mutant under dehydration stress. Collectively, these data imply that DRIP1 and DRIP2 function negatively in the response of plants to drought stress. Moreover, overexpression of full-length DREB2A protein was more stable in drip1-1 than in the wild-type background. These results suggest that DRIP1 and DRIP2 act as novel negative regulators in drought-responsive gene expression by targeting DREB2A to 26S proteasome proteolysis.
  • Sakae Horisawa, Yoh Sakuma, Yasunorl Nakamura, Shuichi Doi
    BIORESOURCE TECHNOLOGY 99 8 3084 - 3093 2008年05月 [査読有り]
     研究論文(学術雑誌) 
    In order to determine the conditions for the maximum performance of a fed-batch composting (FBC) reactor, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the microbial communities established under the confined conditions of moisture content and environmental temperature. To evaluate the effects of microbial community structures on the performance of FBC reactors, degradation experiments using small-scale reactors and model waste were conducted under confined environmental conditions. A high degradation rate was observed under a wide range of MC conditions (30-60%) and at higher than usual temperatures (30-50 degrees C). The microbial communities that formed in the experimental FBC reactors were analyzed by DGGE of PCR-amplified 16S rRNA genes. The DGGE banding patterns at the same level as the degradation rates were similar even if the environmental conditions were different. Sequence analysis of the DGGE bands revealed the primary microbes which act in the reactor. (c) 2007 Elsevier Ltd. All rights reserved.
  • Takumi Yoshida, Yoh Sakuma, Daisuke Todaka, Kyonoshin Maruyama, Feng Qin, Junya Mizoi, Satoshi Kidokoro, Yasunari Fujita, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 368 3 515 - 521 2008年04月 [査読有り]
     研究論文(学術雑誌) 
    A transcription factor DREB2A functions as a key regulator not only in drought stress responses but also in heat stress (HS) responses, and activates expression of many abiotic stress-responsive-genes involved in drought and HS tolerance. HsfA3 is one of the most up-regulated heat-inducible genes in transgenic plants overexpressing DREB2A. In this study, the analyses of HsfA3 expression profile and the transactivation analysis of HsfA3 showed that the expression of HsfA3 was directly regulated by DREB2A under HS. Microarray analysis using transgenic plants overexpressing HsfA3 also showed that overexpression of HsfA3 induces many heat-inducible genes. Furthermore, we showed that thermotolerance of the HsfA3 overexpressors was increased, and that of the hsfA3 T-DNA tagged mutants was decreased. These results indicate that HsfA3 regulates expression of many heat-inducible genes in the transcriptional cascade downstream of the DREB2A stress-regulatory system and functions in acquisition of thermotolerance under the control of the DREB2A cascade. (c) 2008 Elsevier Inc. All rights reserved.
  • Feng Qin, Masayuki Kakimoto, Yoh Sakuma, Kyonoshin Maruyama, Yuriko Osakabe, Lam-Son Phan Tran, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT JOURNAL 50 1 54 - 69 2007年04月 [査読有り]
     研究論文(学術雑誌) 
    DREB1/CBFs and DREB2s are transcription factors that specifically interact with a cis-acting element, DRE/CRT, which is involved in the expression of genes responsive to cold and drought stress in Arabidopsis thaliana. The function of DREB1/CBFs has been precisely analyzed and it has been found to activate the expression of many genes responsive to cold stress containing a DRE/CRT sequence in their promoters. However, the regulation and function of DREB2-type transcription factors remained to be elucidated. In this research, we report the cloning of a DREB2 homolog from maize, ZmDREB2A, whose transcripts were accumulated by cold, dehydration, salt and heat stresses in maize seedlings. Unlike Arabidopsis DREB2A, ZmDREB2A produced two forms of transcripts, and quantitative real-time PCR analyses demonstrated that only the functional transcription form of ZmDREB2A was significantly induced by stresses. Moreover, the ZmDREB2A protein exhibited considerably high transactivation activity compared with DREB2A in Arabidopsis protoplasts, suggesting that protein modification is not necessary for ZmDREB2A to be active. Constitutive or stress-inducible expression of ZmDREB2A resulted in an improved drought stress tolerance in plants. Microarray analyses of transgenic plants overexpressing ZmDREB2A revealed that in addition to genes encoding late embryogenesis abundant (LEA) proteins, some genes related to heat shock and detoxification were also upregulated. Furthermore, overexpression of ZmDREB2A also enhanced thermotolerance in transgenic plants, implying that ZmDREB2A may play a dual functional role in mediating the expression of genes responsive to both water stress and heat stress.
  • Satoshi Kidokoro, Kyonoshin Maruyama, Kazuo Nakashima, Yoh Sakuma, Yoshiyuki Imura, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT AND CELL PHYSIOLOGY 48 S238 - S238 2007年 [査読有り]
  • Lam-Son Phan Tran, Kazuo Nakashima, Yoh Sakuma, Yuriko Osakabe, Feng Qin, Sean D. Simpson, Kyonoshin Maruyama, Yasunari Fujita, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT JOURNAL 49 1 46 - 63 2007年01月 [査読有り]
     研究論文(学術雑誌) 
    The ZFHD recognition sequence (ZFHDRS) and NAC recognition sequence (NACRS) play an important role in the dehydration-inducible expression of the Arabidopsis thaliana EARLY RESPONSIVE TO DEHYDRATION STRESS 1 (ERD1) gene. Using the yeast one-hybrid system, we isolated a cDNA encoding the ZFHD1 transcriptional activator that specifically binds to the 62 bp promoter region of ERD1, which contains the ZFHDRS. Both in vitro and in vivo analyses confirmed specific binding of the ZFHD1 to ZFHDRS, and the expression of ZFHD1 was induced by drought, high salinity and abscisic acid. The DNA-binding and activation domains of ZFHD1 were localized on the C-terminal homeodomain and N-terminal zinc finger domain, respectively. Microarray analysis of transgenic plants over-expressing ZFHD1 revealed that several stress-induciblew genes were upregulated in the transgenic plants. Transgenic plants exhibited a smaller morphological phenotype and had a significant improvement of drought stress tolerance. Using the yeast two-hybrid system, we detected an interaction between ZFHD1 and NACRS- binding NAC proteins. Moreover, co-overexpression of the ZFHD1 and NAC genes restored the morphological phenotype of the transgenic plants to a near wild-type state and enhanced expression of ERD1 in both Arabidopsis T87 protoplasts and transgenic Arabidopsis plants.
  • Feng Qin, Yoh Sakuma, Lam-Son Phan Tran, Yuriko Osakabe, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PLANT AND CELL PHYSIOLOGY 48 S241 - S241 2007年 [査読有り]
  • Yoh Sakuma, Kyonoshin Maruyama, Feng Qin, Yuriko Osakabe, Kazuo Shinozaki, Kazuko Yamaguchi-Shinozaki
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 103 49 18822 - 18827 2006年12月 [査読有り]
     研究論文(学術雑誌) 
    Transcription factor DRIEB2A interacts with a cis-acting dehydration-responsive element (DRE) sequence and activates expression of downstream genes involved in drought- and salt-stress response in Arabidopsis thaliana. Intact DREB2A expression does not activate downstream genes under normal growth conditions. A negative regulatory domain exists in the central region of DREB2A, and deletion of this region transforms DREB2A to a constitutive active form (DREB2A CA). We carried out microarray analysis of transgenic Arabidopsis-overexpressing DREB2A CA and found that the overexpression of DREB2A CA induces not only drought- and salt-responsive genes but also heat-shock (HS)-related genes. Moreover, we found that transient induction of the DREB2A occurs rapidly by HS stress, and that the sGFP-DREB2A protein accumulates in nuclei of HS-stressed cells. DREB2A up-regulated genes were classified into three groups based on their expression patterns: genes induced by HS, genes induced by drought stress, and genes induced by both HS and drought stress. DREB2A up-regulated genes were down-regulated in DREB2A knockout mutants under stress conditions. Thermotolerance was significantly increased in plants overexpressing DREB2A CA and decreased in DREB2A knockout plants. Collectively, these results indicate that DREB2A functions in both water and HS-stress responses.
  • Y Sakuma, K Maruyama, Y Osakabe, F Qin, M Seki, K Shinozaki, K Yamaguchi-Shinozaki
    PLANT CELL 18 5 1292 - 1309 2006年05月 [査読有り]
     研究論文(学術雑誌) 
    Transcription factors DREB1A/CBF3 and DREB2A specifically interact with cis-acting dehydration-responsive element/C-repeat (DRE/CRT) involved in cold and drought stress-responsive gene expression in Arabidopsis thaliana. Intact DREB2A expression does not activate downstream genes under normal growth conditions, suggesting that DREB2A requires posttranslational modification for activation, but the activation mechanism has not been clarified. DREB2A domain analysis using Arabidopsis protoplasts identified a transcriptional activation domain between residues 254 and 335, and deletion of a region between residues 136 and 165 transforms DREB2A to a constitutive active form. Overexpression of constitutive active DREB2A resulted in significant drought stress tolerance but only slight freezing tolerance in transgenic Arabidopsis plants. Microarray and RNA gel blot analyses revealed that DREB2A regulates expression of many water stress inducible genes. However, some genes downstream of DREB2A are not downstream of DREB1A, which also recognizes DRE/CRT but functions in cold stress-responsive gene expression. Synthetic green fluorescent protein gave a strong signal in the nucleus under unstressed control conditions when fused to constitutive active DREB2A but only a weak signal when fused to full-length DREB2A. The region between DREB2A residues 136 and 165 plays a role in the stability of this protein in the nucleus, which is important for protein activation.
  • LSP Tran, K Nakashima, Y Sakuma, SD Simpson, Y Fujita, K Maruyama, M Fujita, M Seki, K Shinozaki, K Yamaguchi-Shinozaki
    PLANT CELL 16 9 2481 - 2498 2004年09月 [査読有り]
     研究論文(学術雑誌) 
    The MYC-like sequence CATGTG plays an important role in the dehydration-inducible expression of the Arabidopsis thaliana EARLY RESPONSIVE TO DEHYDRATION STRESS 1 (ERD1) gene, which encodes a ClpA (ATP binding subunit of the caseinolytic ATP-dependent protease) homologous protein. Using the yeast one-hybrid system, we isolated three cDNA clones encoding proteins that bind to the 63-bp promoter region of erd1, which contains the CATGTG motif. These three cDNA clones encode proteins named ANAC019, ANAC055, and ANAC072, which belong to the NAC transcription factor family. The NAC proteins bound specifically to the CATGTG motif both in vitro and in vivo and activated the transcription of a beta-glucurdnidise (GUS) 'reporter gene driven by the 63-bp region containing the CATGTG motif in Arabidopsis T87 protoplasts. The expression of ANAC019, ANAC055, and ANAC072 was induced by drought, high salinity, and abscisic acid. A histochemical assay using P(NAC)-GUS fusion constructs showed that expression of the GUS reporter gene was localized mainly to the leaves of transgenic Arabidopsis plants. Using the yeast'one-hybrid system, we determined the complete NAC recognition sequence, containing CATGT and harboring CACG as the core DNA binding site. Microarray analysis of transgenic plants overexpressing eitherANAC019, ANAC055, orANAC072 revealed that several stress-inducible genes were upregulated in the transgenic plants, and the plants showed significantly increased drought tolerance. However, erdl was not upregulated in the transgenic plants. Other interacting factors may be necessary for the induction of erdl in Arabidopsis under stress conditions.
  • H Sakamoto, K Maruyama, Y Sakuma, T Meshi, M Iwabuchi, K Shinozaki, K Yamaguchi-Shinozaki
    PLANT PHYSIOLOGY 136 1 2734 - 2746 2004年09月 [査読有り]
     研究論文(学術雑誌) 
    ZPT2-related proteins that have two canonical Cys-2/His-2-type zinc-finger motifs in their molecules are members of a family of plant transcription factors. To characterize the role of this type of protein, we analyzed the function of Arabidopsis L, Heynh. genes encoding four different ZPT2-related proteins (AZF1, AZF2, AZF3, and STZ). Gel-shift analysis showed that the AZFs and STZ bind to A(G/C)T repeats within an EP2 sequence, known as a target sequence of some petunia (Petunia hybrida) ZPT2 proteins. Transient expression analysis using synthetic green fluorescent protein fusion genes indicated that the AZFs and STZ are preferentially localized to the nucleus. These four ZPT2-related proteins were shown to act as transcriptional repressors that down-regulate the transactivation activity of other transcription factors. RNA gel-blot analysis showed that expression of AZF2 and STZ was strongly induced by dehydration, high-salt and cold stresses, and abscisic acid treatment. Histochemical analysis of beta-glucuronidase activities driven by the AZF2 or STZ promoters revealed that both genes are induced in leaves rather than roots of rosette plants by the stresses. Transgenic Arabidopsis overexpressing STZ showed growth retardation and tolerance to drought stress. These results suggest that AZF2 and STZ function as transcriptional repressors to increase stress tolerance following growth retardation.
  • F Qin, Y Sakuma, J Li, Q Liu, YQ Li, K Shinozaki, KY Yamagushi-Shinozaki
    PLANT AND CELL PHYSIOLOGY 45 8 1042 - 1052 2004年08月 [査読有り]
     研究論文(学術雑誌) 
    The transcription factors DREB1s/CBFs specifically interact with the DRE/CRT cis-acting element (core motif: G/ACCGAC) and control the expression of many stress-inducible genes in Arabidopsis. We isolated a cDNA for a DREB1/CBF homolog, ZmDREB1A in maize using a yeast one-hybrid system. The ZmDREB1A proteins specifically bound to DRE and the highly conserved valine at the 14th residue in the ERF/AP2 DNA binding domain was a key to determining the specific interaction between this protein and the DRE sequence. Expression of ZmDREB1A was induced by cold stress and slightly increased by high-salinity stress. This gene was also transiently expressed by mechanical attack. ZmDREB1A activated the transcription of the GUS reporter gene driven by DRE in rice protoplasts. Overexpression of ZmDREB1A in transgenic Arabidopsis induced overexpression of target stress-inducible genes of Arabidopsis DREB1A resulting in plants with higher tolerance to drought and freezing stresses. This indicated that ZmDREB1A has functional similarity to DREB1s/CBFs in Arabidopsis. The structure of the ERF/AP2 domain of ZmDREB1A in maize is closely related to DREB1-type ERF/AP2 domains in the monocots as compared with that in the dicots. ZmDREB1A is suggested to be potentially useful for producing transgenic plants that is tolerant to drought, high-salinity and/or cold stresses.
  • K Maruyama, Y Sakuma, M Kasuga, Y Ito, M Seki, H Goda, Y Shimada, S Yoshida, K Shinozaki, K Yamaguchi-Shinozaki
    PLANT JOURNAL 38 6 982 - 993 2004年06月 [査読有り]
     研究論文(学術雑誌) 
    The transcriptional factor DREB/CBF (dehydration-responsive element/C-repeat-binding) specifically interacts with the dehydration-responsive element (DRE)/C-repeat (CRT) cis-acting element (A/GCCGAC) and controls the expression of many stress-inducible genes in Arabidopsis. Transgenic plants overexpressing DREB1A showed activated expression of many stress-inducible genes and improved tolerance to not only drought, salinity, and freezing but also growth retardation. We searched for downstream genes in transgenic plants overexpressing DREB1A using the full-length cDNA microarray and Affymetrix GeneChip array. We confirmed candidate genes selected by array analyses using RNA gel blot and identified 38 genes as the DREB1A downstream genes, including 20 unreported new downstream genes. Many of the products of these genes were proteins known to function against stress and were probably responsible for the stress tolerance of the transgenic plants. The downstream genes also included genes for protein factors involved in further regulation of signal transduction and gene expression in response to stress. The identified genes were classified into direct downstream genes of DREB1A and the others based on their expression patterns in response to cold stress. We also searched for conserved sequences in the promoter regions of the direct downstream genes and found A/GCCGACNT in their promoter regions from -51 to -450 as a consensus DRE. The recombinant DREB1A protein bound to A/GCCGACNT more efficiently than to A/GCCGACNA/G/C.
  • S Horisawa, Y Sakuma, K Takata, S Doi
    JOURNAL OF WOOD SCIENCE 50 5 427 - 432 2004年 [査読有り]
     研究論文(学術雑誌) 
    We investigated a genotype-based assay to discriminate the dry rot fungi Serpula lacrymans. DNAs were extracted from 74 isolates from the northern half of Japan. and internal transcribed spacers (ITS) were amplified by polymerase chain reaction. Genotypes of isolates were checked by restriction fragment length polymorphism (RFLP) using two enzymes. Tag I and Hha I. Among the 74 isolates identified as S. lacrymans in terms of morphologic features. 5 isolates were shown to have been misidentified. Random amplified polymorphic DNA (RAPD) analysis was conducted in order to detect the intraspecific diversity of S. lacrymans isolated in Japan. Because no relation between geographical origin and genetic distances was observed. the intraspecific diversity of S. lacrymans is suggested to be small.
  • Narusaka Y, Nakashima K, Shinwari ZK, Sakuma Y, Furihata T, Abe H, Narusaka M, Shinozaki K, Yamaguchi-Shinozaki K
    The Plant journal : for cell and molecular biology 34 2 137 - 148 2003年04月 [査読有り]
  • Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K
    The Plant journal : for cell and molecular biology 33 4 751 - 763 2003年02月 [査読有り]
  • S Horisawa, Y Sakuma, KL Chen, S Doi
    JOURNAL OF WOOD SCIENCE 48 3 232 - 236 2002年 研究論文(学術雑誌) 
    Simulated organic waste was biodegraded in a laboratory-scale machine using matrices prepared from four wood species to investigate the effects of wood species on the degradation rate and the bacterial community. The degradation rate, estimated by measuring weight loss and CO, evolution, was found to be equal among the four wood species. Changes in viable cell counts and microbial communities over time were examined. Viable cell counts were also similar among the wood species, but initial bacterial communities differed owing to differences in wood species, although these communities became similar with time. The sensitivity of isolates to wood extractives was examined using paper discs. The extractive-insensitive bacteria species were dominant at the initial stage of biodegradation. However, occupancy of sensitive bacteria increased with time. It was thought that antibacterial extractives were degraded or inactivated after some time.
  • Y Sakuma, Q Liu, JG Dubouzet, H Abe, K Shinozaki, K Yamaguchi-Shinozaki
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 290 3 998 - 1009 2002年01月 [査読有り]
     研究論文(学術雑誌) 
    DRE/CRT is a cis-acting element that is involved in gene expression responsive to drought and low-temperature stress in higher plants. DREB1A/CBF3 and DREB2A are transcription factors that specifically bind to DRE/CRT in Arabidopsis. We precisely analyzed the DNA-binding specificity of DREBs. Both DREBs specifically bound to six nucleotides (A/GCCGAC) of DRE. However, these proteins had different binding specificities to the second or third nucleotides of DRE. Gel mobility shift assay using mutant DREB proteins showed that the two amino acids, valine and glutamic acid conserved in the ERF/AP2 domains, especially valine, have important roles in DNA-binding specificity. In the Arabidopsis genome, 145 DREB/ERF-related proteins are encoded. These proteins were classified into five groups-AP-2 subfamily, RAV subfamily, DREB subfamily, ERF subfamily, and others. The DREB subfamily included three novel DREB1A- and six DREB2A-related proteins. We analyzed expression of novel genes for these proteins and discuss their roles in stress-responsive gene expression. (C) 2002 Elsevier Science (USA).
  • Yoshiaki Kato, Yoh Sakuma, Tomonori Azuma, Akira Ando, Kiyoshi Miura, Keiji Takabe, Yutaka Tamai
    Journal of Wood Science 47 3 165 - 170 2001年 [査読有り]
     研究論文(学術雑誌) 
    A cDNA clone from Populus nigra L. var. italica Koehne, denoted PCY2-6, coding for an anionic peroxidase has been isolated, cloned, and characterized. PCY2-6 is 1160 bp long and its deduced product, PnC26, contains 343 amino acid residues. The mature protein has a calculated isoelectric point of 4.09. The protein contains two motifs typical of peroxidase and 10 potential W-glycosylation sites. PnC26 is therefore classified as an anionic peroxidase. The mRNA of the PCY2-6 gene family was detected in immature and mature leaves and in two parts of current-year stems: the shoot tip and the older stem. The mRNA of PCY2-6 gene family was found to localize in the phloem and cortex of the current-year stems. We therefore conclude that expression of the PCY2-6 gene family is related to bark development.
  • Sakae Horisawa, Yoh Sakuma, Yutaka Tamai, Shuichi Doi, Minoru Terazawa
    Journal of Wood Science 47 2 154 - 158 2001年 [査読有り]
     研究論文(学術雑誌) 
    The optimum environmental temperature for a biodegrading machine using wood particles as a matrix was investigated using a small-scale degradation reactor and model waste. The biodegradation rate was evaluated by weight loss of waste and CO2 evolution. The degradation reaction was restricted only by adjusting the environmental temperature while sufficient oxygen and substrates were supplied. Results suggested that the optimum temperature for degradation was 30°-40°C for exploiting biological activity effectively with the lowest use of energy. Bacteria from the environment propagated in the reactor with no inoculum added. The microbial flora changed during the operation time but had no effect on the biodegradation rate.
  • K Nakashima, ZK Shinwari, Y Sakuma, M Seki, S Miura, K Shinozaki, K Yamaguchi-Shinozaki
    PLANT MOLECULAR BIOLOGY 42 4 657 - 665 2000年03月 [査読有り]
     研究論文(学術雑誌) 
    In plants, a cis-acting element, DRE/CRT, is involved in ABA-independent gene expression in response to dehydration and low-temperature stress. To understand signal transduction pathways from perception of the dehydration stress signal to gene expression, we characterized a gene family for DRE/CRT-binding proteins DREB2A and DREB2B in Arabidopsis thaliana. Northern analysis showed that both genes are induced by dehydration and high-salt stress. Organ-specific northern analysis with gene-specific probes showed that these genes are strongly induced in roots by high-salt stress and in stems and roots by dehydration stress. The DREB2A gene is located on chromosome 5, and DREB2B on chromosome 3. We screened an Arabidopsis genomic DNA library with cDNA fragments of DREB2A and DREB2B as probes, and isolated DNA fragments that contained 5'-flanking regions of these genes. Sequence analysis showed that both genes are interrupted by a single intron at identical positions in their leader sequence. Several conserved sequences were found in the promoter regions of both genes. The beta-glucuronidase (GUS) reporter gene driven by the DREB2 promoters was induced by dehydration and high-salt stress in transgenic Arabidopsis plants.
  • Takeshi Yokono, Yutaka Tamai, Tomonori Azuma, Yoh Sakuma, Kiyoshi Miura, Yasuo Kojima, Masahide Sunagawa, Masatake Ohmasa
    Journal of Wood Science 46 6 480 - 484 2000年 [査読有り]
     研究論文(学術雑誌) 
    The purpose of this research was to analyze the karyotype of the interspecific fusants of two Pleurotus species. Auxotrophic mutants derived from the cultivated strain of P. ostreatus and P. cornucopiae were used. Protoplasts were fused electrically, and the fusants were selected under auxotrophic complementation. Esterase isozyme analysis showed that several fusants had isozyme bands originating from both parental strains, and others had unilateral isozyme bands. The fusant that had expressed isozyme bands of both parental strains showed chromosomal DNA bands of both of the parental strains in pulsed-field gel electrophoresis analysis. Despite the above results, the chromosomal composition of the fusants obtained by the pulsed-field gel electrophoresis did not exhibit all of the bands of both fusion parents.
  • Sakae Horisawa, Yutaka Tamai, Yoh Sakuma, Shuichi Doi, Minoru Terazawa
    Journal of Wood Science 46 4 317 - 321 2000年 [査読有り]
     研究論文(学術雑誌) 
    The optimum working moisture content of a wood matrix for the garbage automatic decomposer-extinguisher (GADE) machine was investigated using a small-scale degradation reactor. A formula feed for rabbits was used as the model waste. The degradation experiment was conducted under controlled conditions such as moisture content, environmental temperature, and airflow rate. The degradation rate was estimated precisely from weight loss and the CO2 evolution rate. The degradation rate were nearly constant at a moisture content of 30%-80% on a wet-weight basis. Microorganisms from the environment propagated in the reactor with no inoculums added. The number of microorganisms showed a trend similar to that of the degradation rate. The microorganism community changed according to the moisture content of the matrix and were considered to attain a constant degradation rate at a wide range of moisture content of a matrix.
  • Doolyi Kim, Yutaka Tamai, Tomonori Azuma, Akira Harada, Akira Ando, Yoh Sakuma, Kiyoshi Miura
    Journal of Wood Science 46 6 466 - 469 2000年 [査読有り]
     研究論文(学術雑誌) 
    The karyotype of Flammulina velutipes (Curt.: Fr.) Sing, was analyzed electrophoretically using contour-clamped homogeneous electric field gel electrophoresis and hybridization with DNA probes. The chromosomal DNA from the monokaryon (Fv-4K) and the dikaryon (Fv-4) were resolved into six and eight bands, respectively. The sizes of the chromosomes ranged from 1.9 to 6.0 megabase (Mb) pairs. Each of the separated bands of chromosomal DNA was identified by use of five cloned probes. The number of these chromosomes was estimated to be 6 and 12, respectively and the size of the entire genome was estimated to be about 20.1 and 38.6Mb, respectively. From a comparison of the hybridization patterns, the existence of allelic chromosomes of different sizes was deduced in the Fv-4 strain.
  • Molecular responses to drought stress in plants: regulation of gene expression and signal transduction.
    Shinozaki, K, Yamaguchi-Shinozaki, K, Liu, Q, Kasuga, M, Ichimura, K, Mizoguchi, T, Urao, T, Miyata, S, Nakashima, K, Shinwari, Z.K, Abe, H, Sakuma, Y, Ito, T, Seki, M
    Plant Responses to Environmental Stress. Edited by Smallwood, M.F., Calvert, C.M. and Bowles, D.J. BIOS Scientific Publishers, Oxford. 133 - 143 1999年 [査読有り]
  • Q Liu, M Kasuga, Y Sakuma, H Abe, S Miura, K Yamaguchi-Shinozaki, K Shinozaki
    PLANT CELL 10 8 1391 - 1406 1998年08月 [査読有り]
     研究論文(学術雑誌) 
    Plant growth is greatly affected by drought and low temperature. Expression of a number of genes is induced by both drought and low temperature, although these stresses are quite different. Previous experiments have established that a cia-acting element named DRE (for dehydration-responsive element) plays an important role in both dehydration- and low-temperature-induced gene expression in Arabidopsis. Two cDNA clones that encode DRE binding proteins, DREB1A and DREB2A, were isolated by using the yeast one-hybrid screening technique. The two cDNA libraries were prepared from dehydrated and cold-treated rosette plants, respectively. The deduced amino acid sequences of DREB1A and DREB2A showed no significant sequence similarity, except in the conserved DNA binding domains found in the EREBP and APETALA2 proteins that function in ethylene-responsive expression and floral morphogenesis, respectively. Both the DREB1A and DREB2A proteins specifically bound to the DRE sequence in vitro and activated the transcription of the beta-glucuronidase reporter gene driven by the DRE sequence in Arabidopsis leaf protoplasts. Expression of the DREB1A gene and its two homologs was induced by low-temperature stress, whereas expression of the DREB2A gene and its single homolog was induced by dehydration. Overexpression of the DREB1A cDNA in transgenic Arabidopsis plants not only induced strong expression of the target genes under unstressed conditions but also caused dwarfed phenotypes in the transgenic plants. These transgenic plants also revealed freezing and dehydration tolerance. In contrast, overexpression of the DREB2A cDNA induced weak expression of the target genes under unstressed conditions and caused growth retardation of the transgenic plants. These results indicate that two independent families of DREB proteins, DREB1 and DREB2, function as trans-acting factors in two separate signal transduction pathways under low-temperature and dehydration conditions, respectively.

講演・口頭発表等

  • イネの種子発芽と芽生え成長に対するサリチル酸と重金属イオン(Fe2+,Cu2+,Zn2+) の影響
    大銅優太郎, 堀怜美, 佐久間洋, 井上雅裕
    第61回日本植物生理学会年会 2020年03月 ポスター発表
  • 半陰性蘚類コツボゴケ (Plagiomnium acutum) の可溶性および細胞壁糖組成に対する水・温度ストレス の影響  [通常講演]
    中西亮介, 佐久間洋, 井上雅裕
    第61回日本植物生理学会年会 2020年03月
  • 分裂酵母の細胞増殖と細胞壁形成に対するガジュマルラテックスと各種キチナーゼの影響  [通常講演]
    寺尾桃子, 仁木宏典, 佐久間洋, 井上雅裕
    第61回日本植物生理学会年会 2020年03月 ポスター発表
  • 四国における Google Map を用いたダンチクの分布調査 ―Azure Custom Vision Service を用いた評価―  [通常講演]
    永瀬 真, 吉川 朝彦, 森脇 直人, 堀澤 栄, 井上 雅裕, 佐久間 洋
    日本植物学会第83回大会 2019年09月 ポスター発表
  • 水面から水底生活に移行して増殖するウキクサ葉状体の生理学的・形態学的特徴  [通常講演]
    小野 浩之, 荒井 希望, 佐久間 洋, 井上 雅裕
    中四国植物学会 第76回大会 2019年05月 ポスター発表
  • 分裂酵母と他種菌類の細胞増殖と細胞壁多糖類に及ぼすガジュマルラテックスの影響  [通常講演]
    寺尾 桃子, 佐久間 洋, 井上 雅裕, 徳重 遥也
    中四国植物学会 第76回大会 2019年05月 ポスター発表
  • ダンチク懸濁培養細胞の一過的質転換条件の検討  [通常講演]
    佐久間 洋, 阿部 友紀子, 今塩屋 賢, 堀澤 栄, 井上 雅裕
    中四国植物学会 第76回大会 2019年05月 ポスター発表
  • 四国における Google Map を用いた大型イネ科植物ダンチクの分布調査  [通常講演]
    吉川 朝彦, 永瀬 真, 森脇 直人, 堀澤 栄, 井上 雅裕, 佐久間 洋
    中四国植物学会 第76回大会 2019年05月 ポスター発表
  • 藻類・細菌・繊毛虫からなる合成生態系を用いた進化の解析:大腸菌の進化と種間関係  [通常講演]
    中島敏幸, 安部雄一, 坪井睦枝, 松浦正幸, 松本沙千, 堀澤栄, 佐久間洋
    日本ゲノム微生物学会 2019年03月
  • 合成生態系における藻類・細菌間の種間関係の進化と大腸菌の全ゲノム解析  [通常講演]
    安部雄一, 坪井睦枝, 松浦正幸, 藤川佳之, 松本沙千, 堀澤栄, 佐久間洋, 中島敏幸
    日本ゲノム微生物学会 2019年03月 ポスター発表
  • 藻類・細菌・繊毛虫からなるモデル微生物生態系における細菌の進化と種間関係  [通常講演]
    安部雄, 坪井睦枝, 松浦正幸, 松本沙千, 堀澤栄, 佐久間洋, 中島敏幸
    第9回 愛媛微生物学ネットワーク・フォーラム 2018年11月
  • 長期培養モデル生態系における藻類・細菌間共生の進化  [通常講演]
    安部雄一, 藤井陽介, 松浦正幸, 坪井睦枝, 松本沙千, 堀澤栄, 佐久間洋, 中島敏幸
    日本原生生物学会 2018年10月
  • 高知県中東部沿岸および愛媛県西部における Google マップを用いたダンチクの分布調査 ―ダンチクは津波避難の目印に使えるのか?―  [通常講演]
    佐久間 洋, 森脇 直人, 堀澤 栄, 井上 雅裕
    日本植物学会 第82回大会 2018年09月 ポスター発表
  • スベリヒユの多肉化と茎肥大に及ぼす環境ストレスと無機栄養塩類の影響  [通常講演]
    佐久間 洋
    日本植物学会 第82回大会 2018年09月 ポスター発表
  • 分裂酵母(S. japonicus)の細胞形態と増殖曲線に対するガジュマルラテックス成分の影響  [通常講演]
    寺尾 桃子, 仁木 宏典, 佐久間 洋, 井上 雅裕
    日本植物学会 第82回大会 2018年09月 ポスター発表
  • スベリヒユの多肉化と茎肥大に及ぼす環境ストレスと植物ホルモンの影響  [通常講演]
    梅崎 昇太, 佐久間 洋, 井上 雅裕
    中四国植物学会 第75回大会 2018年05月 ポスター発表
  • 分裂酵母と出芽酵母の細胞増殖と形態形成に及ぼすガジュマルラテックス可用性成分の影響  [通常講演]
    寺尾 桃子, 佐久間 洋, 井上 雅裕
    中四国植物学会 第75回大会 2018年05月 ポスター発表
  • 大型海浜植物ダンチクのGoogleマップを用いた分布調査  [通常講演]
    佐久間 洋, 森脇 直人, 堀澤 栄, 井上 雅裕
    中四国植物学会 第75回大会 2018年05月 ポスター発表
  • コカナダモの食害応答におけるジャスモン酸伝達とフラボノイド生成の影響  [通常講演]
    門倉 祐太, 佐久間 洋, 宮本 健助, 井上 雅裕
    日本植物学会・第81回大会 2017年09月 ポスター発表
  • 大型海浜植物ダンチクのGoogleマップを用いた分布調査 —分布に影響を与える要因—  [通常講演]
    佐久間 洋, 高木方隆, 堀澤 栄, 井上雅裕
    日本植物学会・第81回大会 2017年09月 ポスター発表
  • Community structure analysis of wood-inhabiting fungi by DNA based methods  [通常講演]
    堀沢 栄, 濱口航大, 佐久間洋
    International Union of Microbiological Societies 2017年07月
  • 水中食害に応答するコカナダモの成長パターンとジャスモン酸濃度の変化  [通常講演]
    門倉 祐太, 宮本 健助, 佐久間 洋, 井上 雅裕
    中四国植物学会第74回大会 2017年05月 ポスター発表
  • 植物の障害ストレスによって分泌されるラテックス成分の特徴と役割について  [通常講演]
    寺尾桃子, 門倉祐太, 佐久間 洋, 井上雅裕
    中四国植物学会第74回大会 2017年05月 ポスター発表
  • 大型海浜植物ダンチクのGoogleマップを用いた分布調査  [通常講演]
    佐久間 洋, 高木方隆, 堀澤 栄, 井上雅裕
    中四国植物学会第74回大会 2017年05月 ポスター発表
  • トマト植物の成長と土壌微生物相に対するヨーグルト乳酸菌と培地・乳成分の影響  [通常講演]
    渡邊由喜, 金平満美, 多田真実子, 佐久間洋, 仁木宏典, 伊藤みさご, 徳本勇人, 井上雅裕
    大阪府立大学学内報告会 2017年03月
  • トマトの成長と土壌微生物層に及ぼすヨーグルトの影響  [通常講演]
    井上雅裕, 渡邊由喜, 金平満美, 多田真実子, 佐久間洋, 仁木宏典
    第58回植物生理学会年会 2017年03月 ポスター発表
  • 乳酸菌ヨーグルト処理によるトマト植物の成長促進と土壌微生物相の変  [通常講演]
    井上雅裕, 佐久間洋, 多田真実子, 金平満実, 渡邊由喜, 仁木宏典
    日本植物学会第80回大会 2016年09月
  • トチカガミ科沈水植物のマキガイによる食害応答に関する研究  [通常講演]
    門倉祐太, 佐久間 洋, 井上雅裕
    中国四国植物学会第73回大会 2016年05月 ポスター発表
  • イネの成長と金属結合物質生成に及ぼす重金属イオン(Cd2+,Zn2+,Cu2+)の影響  [通常講演]
    堀 怜美, 佐久間 洋, 井上雅裕
    中国四国植物学会第73回大会 2016年05月 ポスター発表
  • イネ科大型海浜植物ダンチクの塩耐性に関する遺伝子の単離と解析  [通常講演]
    坂井佑衣, 亀井奈穗, 井上雅裕, 佐久間 洋
    中国四国植物学会第73回大会 2016年05月 ポスター発表

MISC

産業財産権

受賞

  • 2016年 Clarivate Analytics Highly Cited Researcher
     
    受賞者: 佐久間 洋
  • 2015年 Thomson Reuter Highly Cited Researcher
     
    受賞者: 佐久間 洋

共同研究・競争的資金等の研究課題

  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2010年 -2012年 
    代表者 : 井上 雅裕, 佐久間 洋
     
    複合的環境汚染を想定し有用植物トマトの細胞と植物体における各種重金属イオンの集積・輸送能と各種リガンド間相互作用を検証し、以下の知見を得た: (1)トマト細胞の Cd 集積・耐性には液胞でのフィトケラチンリガンドによる Cd-PC 複合体が関係し、その複合体形成を Cu 等の重金属や無機アニオンが阻害する。これらは Cd-PC 複合体には殆ど取込まれない。 (2)地上部への Cd・As 輸送には PC 以外に有機酸と無機リガンド(ホウ酸)も関る。重金属混合実験では Zn と Cd は Cu や As より地上部に早く輸送され、その輸送量は有機酸と PC 濃度に比例する。As・Cu では地上部に PC は検出されず、主に無機リガンドが輸送に関る。 (3)他金属(Ca, Mg, Na, Cs, K)の吸収・輸送実験から、上記と異なる輸送(リガンド)と再分配の機構の存在が示唆される。以上の知見と研究成果は新たな発展課題も提示する。

愛媛大学教員活動実績

教育活動(B)

担当授業科目(B01)

  • 2019, 前期, 学部, 分子遺伝学
  • 2019, 前期, 学部, 生物学ゼミナールⅠ
  • 2019, 前期, 学部, 生物学特別演習Ⅰ
  • 2019, 前期, 学部, 卒業研究Ⅰ
  • 2019, 前期, 学部, 基礎生物学実験
  • 2019, 前期, 学部, 基礎生物学実験
  • 2019, 前期, 学部, 生物学実験IV
  • 2019, 前期, 学部, 基礎生物学実験
  • 2019, 前期, 学部, 分子遺伝学


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