2017年12月, 「第1回茨城テックグランプリ」 リバネス賞, 葉緑体のチカラで創る未来の社会, 株式会社リバネス
中平洋一(クロロ・ドリームズ)
その他の賞

〔主要な業績〕Novel bovine viral diarrhea virus (BVDV) virus-like particle vaccine candidates presenting the E2 protein using the SpyTag/SpyCatcher system induce a robust neutralizing antibody response in mice
Miki Katsura; Masaki Fukushima; Ken-Ichiro Kameyama; Takehiko Kokuho; Yoichi Nakahira; Kaoru Takeuchi
Archives of Virology, 2023年01月07日, ［査読有り］

〔主要な業績〕Lack of plastid‐encoded Ycf10, a homolog of the nuclear‐encoded DLDG1 and the cyanobacterial PxcA, enhances the induction of non‐photochemical quenching in tobacco
Mai Duy Luu Trinh; Akira Hashimoto; Masaru Kono; Shinichi Takaichi; Yoichi Nakahira; Shinji Masuda, Wiley
Plant Direct, 2021年12月14日, ［査読有り］

〔主要な業績〕Mass Production of Virus-Like Particles Using Chloroplast Genetic Engineering for Highly Immunogenic Oral Vaccine Against Fish Disease.
Yoichi Nakahira; Kaori Mizuno; Hirofumi Yamashita; Minami Tsuchikura; Kaoru Takeuchi; Takashi Shiina; and Hidemasa Kawakami, 筆頭著者, Nervous necrosis virus (NNV) is the causative agent of viral nervous necrosis (VNN), which is one of the most serious fish diseases leading to mass mortality in a wide range of fish species worldwide. Although a few injectable inactivated vaccines are commercially available, there is a need for more labor-saving, cost-effective, and fish-friendly immunization methods. The use of transgenic plants expressing pathogen-derived recombinant antigens as edible vaccines is an ideal way to meet these requirements. In this study, chloroplast genetic engineering was successfully utilized to overexpress the red-spotted grouper NNV capsid protein (RGNNV-CP). The RGNNV-CP accumulated at high levels in all young, mature, and old senescent leaves of transplastomic tobacco plants (averaging approximately 3 mg/g leaf fresh weight). The RGNNV-CP efficiently self-assembled into virus-like particles (RGNNV-VLPs) in the chloroplast stroma of the transgenic lines, which could be readily observed by in situ transmission electron microscopy. Furthermore, intraperitoneal injection and oral administration of the crudely purified protein extract containing chloroplast-derived RGNNV-VLPs provided the sevenband grouper fish with sufficient protection against RGNNV challenge, and its immunogenicity was comparable to that of a commercial injectable vaccine. These findings indicate that chloroplast-derived VLP vaccines may play a promising role in the prevention of various diseases, not only in fish but also in other animals, including humans., frontiers
frontiers in Plant Science, 2021年08月23日, ［査読有り］

〔主要な業績〕Selective activation of chloroplast psbD light-responsive promoter and psaA/B promoter in transplastomic tobacco plants overexpressing Arabidopsis sigma factor AtSIG5.
Nozoe M; Tsunoyama Y; Ishizaki Y; Nakahira Y; Shiina T, BACKGROUND: Plastid-encoded eubacterial-type RNA polymerase (PEP) plays a critical role in the transcription of photosynthesis genes in chloroplasts. Notably, some of the reaction center genes, including psaA, psaB, psbA, and psbD genes, are differentially transcribed by PEP in mature chloroplasts. However, the molecular mechanism of promoter selection in the reaction center gene transcription by PEP is not well understood. OBJECTIVE: Sigma factor proteins direct promoter selection by a core PEP in chloroplasts as well as bacteria. AtSIG5 is a unique chloroplast sigma factor essential for psbD light-responsive promoter (psbD LRP) activity. To analyze the role of AtSIG5 in chloroplast transcription in more detail, we assessed the effect of AtSIG5 hyper-expression on the transcription of plastid-encoded genes in chloroplast transgenic plants. RESULTS: The chloroplast transgenic tobacco (CpOX-AtSIG5) accumulates AtSIG5 protein at extremely high levels in chloroplasts. Due to the extremely high-level expression of recombinant AtSIG5, most PEP holoenzymes are most likely to include the recombinant AtSIG5 in the CpOXAtSIG5 chloroplasts. Thus, we can assess the promoter preference of AtSIG5 in vivo. The overexpression of AtSIG5 significantly increased the expression of psbD LRP transcripts encoding PSII reaction center D2 protein and psaA/B operon transcripts encoding PSI core proteins. Furthermore, run-on transcription analyses revealed that AtSIG5 preferentially recognizes the psaA/B promoter, as well as the psbD LRP. Moreover, we found that psbD LRP is constitutively active in CpOX-AtSIG5 plants irrespective of light and dark. CONCLUSION: AtSIG5 probably plays a significant role in differential transcription of reaction center genes in mature chloroplasts.
Protein Pept Lett., 2019年10月14日, ［査読有り］

〔主要な業績〕A Ycf2-FtsHi heteromeric AAA-ATPase complex is required for chloroplast protein import.
Kikuchi S; Asakura Y; Imai M; Nakahira Y; Kotani Y; Hashiguchi Y; Nakai Y; Takafuji K; Bédard J; Hirabayashi-Ishioka Y; Mori H; Shiina T; Nakai M, Chloroplasts import thousands of nucleus-encoded preproteins synthesized in the cytosol through the TOC and TIC translocons on the outer and inner envelope membranes, respectively. Preprotein translocation across the inner membrane requires ATP; however, the import motor has remained unclear. Here, we report that a 2-MD heteromeric AAA-ATPase complex associates with the TIC complex and functions as the import motor, directly interacting with various translocating preproteins. This 2-MD complex consists of a protein encoded by the previously enigmatic chloroplast gene ycf2 and five related nuclear-encoded FtsH-like proteins, namely, FtsHi1, FtsHi2, FtsHi4, FtsHi5, and FtsH12. These components are each essential for plant viability and retain the AAA-type ATPase domain, but only FtsH12 contains the zinc binding active site generally conserved among FtsH-type metalloproteases. Furthermore, even the FtsH12 zinc binding site is dispensable for its essential function. Phylogenetic analyses suggest that all AAA-type members of the Ycf2/FtsHi complex including Ycf2 evolved from the chloroplast-encoded membrane-bound AAA-protease FtsH of the ancestral endosymbiont. The Ycf2/FtsHi complex also contains an NAD-malate dehydrogenase, a proposed key enzyme for ATP production in chloroplasts in darkness or in nonphotosynthetic plastids. These findings advance our understanding of this ATP-driven protein translocation system that is unique to the green lineage of photosynthetic eukaryotes.
Plant Cell, 2018年10月11日, ［査読有り］

〔主要な業績〕Comparative Analysis of Chloroplast psbD Promoters in Terrestrial Plants
Shuichi Shimmura; Mikio Nozoe; Shota Kitora; Satoko Kin; Shigeru Matsutani; Yoko Ishizaki; Yoichi Nakahira; Takashi Shiina, The transcription of photosynthesis genes encoded by the plastid genome is mainly mediated by a prokaryotic-type RNA polymerase called plastid-encoded plastid RNA polymerase (PEP). Standard PEP-dependent promoters resemble bacterial sigma-70-type promoters containing the so-called 10 and 35 elements. On the other hand, an unusual light-and stress-responsive promoter (psbD LRP) that is regulated by a 19-bp AAG-box immediately upstream of the -35 element has been mapped upstream of the psbD-psbC operon in some angiosperms. However, the occurrence of the AAG-box containing psbD LRP in plant evolution remains elusive. We have mapped the psbD promoters in eleven embryophytes at different evolutionary stages from liverworts to angiosperms. The psbD promoters were mostly mapped around 500-900 bp upstream of the psbD translational start sites, indicating that the psbD mRNAs have unusually long 5'-UTR extensions in common. The -10 elements of the psbD promoter are well-conserved in all embryophytes, but not the -35 elements. We found that the AAG-box sequences are highly conserved in angiosperms and gymnosperms except for gnetaceae plants. Furthermore, partial AAG-box-like sequences have been identified in the psbD promoters of some basal embryophytes such as moss, hornwort, and lycophyte, whereas liverwort has the standard PEP promoter without the AAG-box. These results suggest that the AAG-box sequences of the psbD LRP may have evolved from a primitive type of AAG-box of basal embryophytes. On the other hand, monilophytes (ferns) use another type of psbD promoter composed of a distinct cis-element upstream of the potential -35 element. Furthermore, we found that psbD expression is not regulated by light in gymnosperms or basal angiosperms, although they have the well-conserved AAG-box sequences. Thus, it is unlikely that acquisition of the AAG-box containing psbD promoter is directly associated with light-induced transcription of the psbD-psbC operon. Light-and stress-induced transcription may have evolved independently and multiple times during terrestrial plant evolution., FRONTIERS MEDIA SA
FRONTIERS IN PLANT SCIENCE, 2017年07月, ［査読有り］

〔主要な業績〕Genetic engineering and metabolite profiling for overproduction of polyhydroxybutyrate in cyanobacteria
Sayaka Hondo; Masatoshi Takahashi; Takashi Osanai; Mami Matsuda; Tomohisa Hasunuma; Akio Tazuke; Yoichi Nakahira; Shigeru Chohnan; Morifumi Hasegawa; Munehiko Asayama, Genetic engineering and metabolite profiling for the overproduction of polyhydroxybutyrate (PHB), which is a carbon material in biodegradable plastics, were examined in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Transconjugants harboring cyanobacterial expression vectors that carried the pha genes for PHB biosynthesis were constructed. The overproduction of PHB by the engineering cells was confirmed through microscopic observations using Nile red, transmission electron microscopy (TEM), or nuclear magnetic resonance (NMR). We successfully recovered PHB from transconjugants prepared from nitrogen-depleted medium without sugar supplementation in which PHB reached approximately 7% (w/w) of the dry cell weight, showing a value of 12-fold higher productivity in the transconjugant than that in the control strain. We also measured the intracellular levels of acetyl-CoA, acetoacetyl-CoA, and 3-hydroxybutyryl-CoA (3HB-CoA), which are intermediate products for PHB. The results obtained indicated that these products were absent or at markedly low levels when cells were subjected to the steady-state growth phase of cultivation under nitrogen depletion for the overproduction of bioplastics. Based on these results, efficient factors were discussed for the overproduction of PHB in recombinant cyanobacteria. (C) 2015, The Society for Biotechnology, Japan. All rights reserved., SOC BIOSCIENCE BIOENGINEERING JAPAN
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 2015年11月, ［査読有り］

Diversity in Guanosine 3 ',5 '-Bisdiphosphate (ppGpp) Sensitivity among Guanylate Kinases of Bacteria and Plants
Yuhta Nomura; Atsushi Izumi; Yoshinori Fukunaga; Kensuke Kusumi; Koh Iba; Seiya Watanabe; Yoichi Nakahira; Andreas P. M. Weber; Akira Nozawa; Yuzuru Tozawa, The guanosine 3,5-bisdiphosphate (ppGpp) signaling system is shared by bacteria and plant chloroplasts, but its role in plants has remained unclear. Here we show that guanylate kinase (GK), a key enzyme in guanine nucleotide biosynthesis that catalyzes the conversion of GMP to GDP, is a target of regulation by ppGpp in chloroplasts of rice, pea, and Arabidopsis. Plants have two distinct types of GK that are localized to organelles (GKpm) or to the cytosol (GKc), with both enzymes being essential for growth and development. We found that the activity of rice GKpm in vitro was inhibited by ppGpp with a K-i of 2.8 m relative to the substrate GMP, whereas the K-m of this enzyme for GMP was 73 m. The IC50 of ppGpp for GKpm was approximate to 10 m. In contrast, the activity of rice GKc was insensitive to ppGpp, as was that of GK from bakers' yeast, which is also a cytosolic enzyme. These observations suggest that ppGpp plays a pivotal role in the regulation of GTP biosynthesis in chloroplasts through specific inhibition of GKpm activity, with the regulation of GTP biosynthesis in chloroplasts thus being independent of that in the cytosol. We also found that GKs of Escherichia coli and Synechococcus elongatus PCC 7942 are insensitive to ppGpp, in contrast to the ppGpp sensitivity of the Bacillus subtilis enzyme. Our biochemical characterization of GK enzymes has thus revealed a novel target of ppGpp in chloroplasts and has uncovered diversity among bacterial GKs with regard to regulation by ppGpp., AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
JOURNAL OF BIOLOGICAL CHEMISTRY, 2014年05月, ［査読有り］

〔主要な業績〕Theophylline-dependent riboswitch as a novel genetic tool for strict regulation of protein expression in cyanobacterium synechococcus elongatus PCC 7942
Yoichi Nakahira; Atsushi Ogawa; Hiroyuki Asano; Tokitaka Oyama; Yuzuru Tozawa, 筆頭著者, The cyanobacterium Synechococcus elongatus PCC 7942 is a major model species for studies of photosynthesis. It is are also a potential cell factory for the production of renewable biofuels and valuable chemicals. We employed engineered riboswitches to control translational initiation of target genes in this cyanobacterium. A firefly luciferase reporter assay revealed that three theophylline riboswitches performed as expected in the cyanobacterium. Riboswitch-E* exhibited very low leaky expression of luciferase and superior and dose-dependent on/off regulation of protein expression by theophylline. The maximum magnitude of the induction vs. basal level was ∼190-fold. Furthermore, the induction level was responsive to a wide range of theophylline concentrations in the medium, from 0 to 2 mM, facilitating the fine-tuning of luciferase expression. We adapted this riboswitch to another gene regulation system, in which expression of the circadian clock kaiC gene product is controlled by the theophylline concentration in the culture medium. The results demonstrated that the adequately adjusted expression level of KaiC restored complete circadian rhythm in the kaiC-deficient arrhythmic mutant. This theophylline-dependent riboswitch system has potential for various applications as a useful genetic tool in cyanobacteria. © 2013 The Author.
Plant and Cell Physiology, 2013年10月, ［査読有り］

Vascular plant one-zinc-finger protein 1/2 transcription factors regulate abiotic and biotic stress responses in Arabidopsis
Yusuke Nakai; Yoichi Nakahira; Hiroki Sumida; Kosuke Takebayashi; Yumiko Nagasawa; Kanako Yamasaki; Masako Akiyama; Masaru Ohme-Takagi; Sumire Fujiwara; Takashi Shiina; Nobutaka Mitsuda; Eiichiro Fukusaki; Yasuyuki Kubo; Masa H. Sato, Plants adapt to abiotic and biotic stresses by activating abscisic acid-mediated (ABA) abiotic stress-responsive and salicylic acid-(SA) or jasmonic acid-mediated (JA) biotic stress-responsive pathways, respectively. Although the abiotic stress-responsive pathway interacts antagonistically with the biotic stress-responsive pathways, the mechanisms that regulate these pathways remain largely unknown. In this study, we provide insight into the function of vascular plant one-zinc-finger proteins (VOZs) that modulate various stress responses in Arabidopsis. The expression of many stress-responsive genes was changed in the voz1voz2 double mutant under normal growth conditions. Consistent with altered stress-responsive gene expression, freezing- and drought-stress tolerances were increased in the voz1voz2 double mutant. In contrast, resistance to a fungal pathogen, Colletotrichum higginsianum, and to a bacterial pathogen, Pseudomonas syringae, was severely impaired. Thus, impairing VOZ function simultaneously conferred increased abiotic tolerance and biotic stress susceptibility. In a chilling stress condition, both the VOZ1 and VOZ2 mRNA expression levels and the VOZ2 protein level gradually decreased. VOZ2 degradation during cold exposure was completely inhibited by the addition of the 26S proteasome inhibitor, MG132, a finding that suggested that VOZ2 degradation is dependent on the ubiquitin/26S proteasome system. In voz1voz2, ABA-inducible transcription factor CBF4 expression was enhanced significantly even under normal growth conditions, despite an unchanged endogenous ABA content. A finding that suggested that VOZs negatively affect CBF4 expression in an ABA-independent manner. These results suggest that VOZs function as both negative and positive regulators of the abiotic and biotic stress-responsive pathways, and control Arabidopsis adaptation to various stress conditions. © 2012 The Authors The Plant Journal © 2012 Blackwell Publishing Ltd.
Plant Journal, 2013年03月, ［査読有り］

Erratum: Chloroplast-mediated activation of plant immune signalling in Arabidopsis (Nature Communications (2012) 3 (926) DOI: 10.1038/ncomms1926)
Hironari Nomura; Teiko Komori; Shuhei Uemura; Yui Kanda; Koji Shimotani; Kana Nakai; Takuya Furuichi; Kohsuke Takebayashi; Takanori Sugimoto; Satoshi Sano; I. Nengah Suwastika; Eiichiro Fukusaki; Hirofumi Yoshioka; Yoichi Nakahira; Takashi Shiina
Nature Communications, 2013年, ［査読有り］

〔主要な業績〕Overproduction of hyperthermostable β-1,4-endoglucanase from the archaeon Pyrococcus horikoshii by tobacco chloroplast engineering.
Nakahira Y; Ishikawa K; Tanaka K; Tozawa Y; Shiina T, 筆頭著者, One of the most cost-effective methods of producing industrial enzymes is by the use of transgenic plants. We demonstrated successful high-level expression of a hyperthermostable archaeal beta-1,4-endoglucanase in mature tobacco leaves by transformation of chloroplasts by homologous recombination. The active recombinant enzyme was readily recovered not only from fresh but also from dried leaves., TAYLOR & FRANCIS LTD
Biosci. Biotechnol. Biochem., 2013年, ［査読有り］

Chloroplast-mediated activation of plant immune signalling in Arabidopsis
Hironari Nomura; Teiko Komori; Shuhei Uemura; Yui Kanda; Koji Shimotani; Kana Nakai; Takuya Furuichi; Kohsuke Takebayashi; Takanori Sugimoto; Satoshi Sano; I. Nengah Suwastika; Eiichiro Fukusaki; Hirofumi Yoshioka; Yoichi Nakahira; Takashi Shiina, Chloroplasts have a critical role in plant immunity as a site for the production for salicylic acid and jasmonic acid, important mediators of plant immunity. However, the molecular link between chloroplasts and the cytoplasmic-nuclear immune system remains largely unknown. Here we show that pathogen-associated molecular pattern (PAMP) signals are quickly relayed to chloroplasts and evoke specific Ca2+ signatures in the stroma. We further demonstrate that a chloroplast-localized protein, named calcium-sensing receptor (CAS), is involved in stromal Ca2+ transients and responsible for both PAMP-induced basal resistance and R gene-mediated hypersensitive cell death. CAS acts upstream of salicylic acid accumulation. Transcriptome analysis demonstrates that CAS is involved in PAMP-induced expression of defence genes and suppression of chloroplast gene expression possibly through O-1(2)-mediated retrograde signalling, allowing chloroplast-mediated transcriptional reprogramming during plant immune responses. The present study reveals a previously unknown chloroplast-mediated signalling pathway linking chloroplasts to cytoplasmic-nuclear immune responses., NATURE PUBLISHING GROUP
NATURE COMMUNICATIONS, 2012年06月, ［査読有り］

Eukaryotic-type plastid nucleoid protein pTAC3 is essential for transcription by the bacterial-type plastid RNA polymerase
Yusuke Yagi; Yoko Ishizaki; Yoichi Nakahira; Yuzuru Tozawa; Takashi Shiina, Plastid transcription is mediated by two distinct types of RNA polymerases (RNAPs), bacterial-type RNAP (PEP) and phage-type RNAP (NEP). Recent genomic and proteomic studies revealed that higher plants have lost most prokaryotic transcription regulators and have acquired eukaryotic-type proteins during plant evolution. However, in vivo dynamics of chloroplast RNA polymerases and eukaryotic-type plastid nucleoid proteins have not been directly characterized experimentally. Here, we examine the association of the a-subunit of PEP and eukaryotic-type protein, plastid transcriptionally active chromosome 3 (pTAC3) with transcribed regions in vivo by using chloroplast chromatin immunoprecipitation (cpChIP) assays. PEP a-subunit preferentially associates with PEP promoters of photosynthesis and rRNA genes, but not with NEP promoter regions, suggesting selective and accurate recognition of PEP promoters by PEP. The cpChIP assays further demonstrate that the peak of PEP association occurs at the promoterproximal region and declines gradually along the transcribed region. pTAC3 is a putative DNA-binding protein that is localized to chloroplast nucleoids and is essential for PEP-dependent transcription. Density gradient and immunoprecipitation analyses of PEP revealed that pTAC3 is associated with the PEP complex. Interestingly, pTAC3 associates with the PEP complex not only during transcription initiation, but also during elongation and termination. These results suggest that pTAC3 is an essential component of the chloroplast PEP complex. In addition, we demonstrate that light-dependent chloroplast transcription is mediated by light-induced association of the PEP-pTAC3 complex with promoters. This study illustrates unique dynamics of PEP and its associated protein pTAC3 during light-dependent transcription in chloroplasts., NATL ACAD SCIENCES
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2012年05月, ［査読有り］

Function and evolution of plastid sigma factors.
Shiina T; Ishizaki Y; Yagi Y; and Nakahira Y, ラスト(シニア)オーサー, Chloroplasts are plant-specific organelles that perform photosynthesis and responsible for the world's primary productively. Using light energy, chloroplasts produce many important products, including starch, amino acids, lipids, pigments and various secondary products. Therefore, chloroplasts are essential to the lives of all plants and animals alike. Chloroplast transformation is a unique technology to produce huge amount of valuable materials in chloroplasts using photosynthetic energy. In order to control chloroplasts at will, we need more information on molecular basis of chloroplast gene expression and communication between the chloroplast and nucleus. Chloroplast sigma factors are key regulators of the chloroplast gene expression and chloroplast differentiation. This review summarized recent findings on roles of chloroplast sigma factors in the chloroplast differentiation and environmental responses., JAPANESE SOC PLANT CELL & MOLECULAR BIOL
Plant Biotechnol., 2009年, ［査読有り］

Evidence for chloroplast control of external Ca2+-induced cytosolic Ca2+ transients and stomatal closure.
Nomura H; Komori T; Kobori M; Nakahira Y; and Shiina T, The role of guard cell chloroplasts in stomatal function is controversial. It is usually assumed that stomatal closure is preceded by a transient increase in cytosolic free Ca2+ concentration ([Ca2+](cyt)) in the guard cells. Here, we provide the evidence that chloroplasts play a critical role in the generation of extracellular Ca2+ ([Ca2+](ext))-induced [Ca2+](cyt) transients and stomatal closure in Arabidopsis. CAS (Ca2+ sensing receptor) is a plant-specific putative Ca2+-binding protein that was originally proposed to be a plasma membrane-localized external Ca2+ sensor. In the present study, we characterized the intracellular localization of CAS in Arabidopsis with a combination of techniques, including (i) in vivo localization of green fluorescent protein (GFP) fused gene expression, (ii) subcellular fractionation and fractional analysis of CAS with Western blots, and (iii) database analysis of thylakoid membrane proteomes. Each technique produced consistent results. CAS was localized mainly to chloroplasts. It is an integral thylakoid membrane protein, and the N-terminus acidic Ca2+-binding region is likely exposed to the stromal side of the membrane. The phenotype of T-DNA insertion CAS knockout mutants and cDNA mutant-complemented plants revealed that CAS is essential for stomatal closure induced by external Ca2+. In contrast, overexpression of CAS promoted stomatal closure in the absence of externally applied Ca2+. Furthermore, using the transgenic aequorin system, we showed that [Ca2+](ext)-induced [Ca2+](cyt) transients were significantly reduced in CAS knockout mutants. Our results suggest that thylakoid membrane-localized CAS is essential for [Ca2+](ext)-induced [Ca2+](cyt) transients and stomatal closure., WILEY-BLACKWELL
Plant J., 2008年, ［査読有り］

Regulation of external calcium-induced stomatal closure by chloroplast localized CAS
Nomura Hironari; Komori Teiko; Nakahira Yoichi; Shiina Takashi
PLANT AND CELL PHYSIOLOGY, 2007年, ［査読有り］

Coordinated functions of plastid sigma factors in chloroplast development and gene expression
Shiina T; Ishizaki Y; Ozono K; Takenaka C; Hanaoka M; Kanamaru K; Tanaka K; Nakahira Y
Photosynthesis Research, 2007年, ［査読有り］

[Plastid transformation in higher plants: application for chloroplast factory].
Nakahira Y; Shiina T
Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme, 2005年11月, ［査読有り］

Characterization of a plastid sigma factor AtSIG5 responsible for psbDlight-responsive transcription.　　　　　　　　　　　　　　　
Nozoe M; Ishizaki Y; Tsunoyama Y; Tsubokura Y; Kato K; Shinnmyo A; Nakahira Y; Shiina T
"Photosynthesis: Fundamental Aspects to Global Perspectives ”A. van der Est, D. Bruce, eds., Kluwer Academic Publishers, Dordrecht., 2005年, ［査読有り］

A nuclear-encoded sigma factor, Arabidopsis SIG6, recognizes sigma-70 type chloroplast promoters and regulates early chloroplast development in cotyledons.
Ishizaki Y; Tsunoyama Y; Hatano K; Ando K; Kato K; Shinmyo A; Kobori M; Takeba G; Nakahira Y; and Shiina T, Eubacterial-type multi-subunit plastid RNA polymerase (PEP) is responsible for the principal transcription activity in chloroplasts. PEP is composed of plastid-encoded core subunits and one of multiple nuclear-encoded sigma factors that confer promoter specificity on PEP. Thus, the replacement of sigma factors associated with PEP has been assumed to be a major mechanism for the switching of transcription patterns during chloroplast development. The null mutant (sig6-1) of plastid sigma factor gene AtSIG6 exhibited a cotyledon-specific pale green phenotype. Light-dependent chloroplast development was significantly delayed in the sig6-1 mutant. Genetic complementation of the mutant phenotype by the AtSIG6 cDNA demonstrated that AtSIG6 plays a key role in light-dependent chloroplast development. Northern and array-based global analyses for plastid transcripts revealed that the transcript levels of most PEP-dependent genes were greatly reduced in the sig6-1 mutant, but that the accumulation of nuclear-encoded RNA polymerase (NEP)-dependent transcripts generally increased. As the PEP alpha subunit and PEP-dependent trnV accumulated at normal levels in the sig6-1 mutant, the AtSIG6 knockout mutant probably retained functional PEP, and the transcriptional defects are likely to have been directly caused by AtSIG6 deficiency. Most of the AtSIG6-dependent genes are preceded by sigma(70)-type promoters comprised of conserved -35/-10 elements. Thus, AtSIG6 may act as a major general sigma factor in chloroplasts during early plant development. On the other hand, the mutant phenotype was restored in older seedlings. Arabidopsis probably contains another late general sigma factor, the promoter specificity of which widely overlaps with that of AtSIG6., BLACKWELL PUBLISHING LTD
Plant J., 2005年, ［査読有り］

Transcriptional regulation of the circadian clock operon kaiBC by upstream regions in cyanobacteria.
Kutsuna S; Nakahira Y; Katayama M; Ishiura M; Kondo T, In the cyanobacterium, Synechococcus elongatus PCC 7942, the kaiBC operon is upregulated by the KaiA protein and downregulated by the KaiC protein to generate circadian oscillation. We investigated the regulation of kaiBC transcription. A primer extension and deletion analyses of the upstream region mapped the sufficient promoter region (SPR) to base pairs -55 to +1 (the transcription start site, TSS) and identified a constitutive negative regulatory region upstream of the SPR (base pairs -897 to -56) that extended into the coding sequence of kaiA. Base-pair substitution within the SPR identified a sequence from -52 to -28 that was the essential element for transcription. Most of the examined sequences drove rhythmic expression of a luxAB reporter that was similar to the expression driven by the kaiBC promoter (PkaiBC) and responded to the overexpression of kaiA or kaiC, even in a promoter activity range of 1-8000%. These results indicate that circadian feedback regulation by KaiA and KaiC is addressed to a global step preceding transcription driven by PkaiBC. However, increasing or decreasing the intrinsic activity of PkaiBC greatly affected the rhythm, suggesting that constitutive adjustment of PkaiBC activity by the sequences identified here is essential for the oscillator.
Mol. Microbiol., 2005年, ［査読有り］

〔主要な業績〕Global gene repression by KaiC as a master process of prokaryotic circadian system.
Nakahira Y; Katayama M; Miyashita H; Kutsuna S; Iwasaki H; Oyama T; and Kondo T, 筆頭著者, A kaiABC clock gene cluster was previously identified from cyanobacterium Synechococcus elongatus PCC 7942, and the feedback regulation of kai genes was proposed as the core mechanism generating circadian oscillation. In this study, we confirmed that the Kai-based oscillator is the dominant circadian oscillator functioning in cyanobacteria. We probed the nature of this regulation and found that excess KaiC represses not only kaiBC but also the rhythmic components of all genes in the genome. This result strongly suggests that the KaiC protein primarily coordinates genomewide gene expression, including its own expression. We also found that a promoter derived from E. coli is feedback controlled by KaiC and restores the complete circadian rhythm in kaiBC-inactivated arrhythmic mutants, provided it can express kaiB and kaiC genes at an appropriate level. Unlike eukaryotic models, specific regulation of the kaiBC promoter is not essential for cyanobacterial circadian oscillations., NATL ACAD SCIENCES
Proc. Natl. Acad. Sci. USA, 2004年, ［査読有り］

An in vivo dual-reporter system of cyanobacteria using two railroad-worm luciferases with different color emissions.
Kitayama Y; Kondo T; Nakahira Y; Nishimura H; Ohmiya Y; and Oyama T, In vivo genetic reporter systems using luciferase enzymes enable the real-time monitoring of gene expression in living cells. We have challenged concurrent monitoring of two independent promoter activities within the same cells to precisely compare their characteristics in vivo. In this report, we describe a simple dual-reporter system capable of simultaneously monitoring two promoter activities in living cyanobacterial cells. Two railroad-worm luciferases catalyzing the bioluminescent emissions of different colors served as the dual reporters; each emission was successfully separated by interference filters to estimate the individual bioluminescence signals using photomultiplier tubes. Using this system, we clearly demonstrated the difference in the expression profiles between promoters in the same cells., OXFORD UNIV PRESS
Plant Cell Physiol., 2004年, ［査読有り］

Blue light-induced transcription of plastid-encoded psbD gene is mediated by a nuclear-encoded transcription initiation factor, AtSig5.
Tsunoyama Y; Ishizaki Y; Morikawa K; Kobori M; Nakahira Y; Takeba G; Toyoshima Y; and Shiina T, Light is one of the most important environmental factors regulating expression of photosynthesis genes. The plastid psbD gene encoding the photosystem II reaction center protein D2 is under the control of a unique blue light responsive promoter (BLRP) that is transcribed by a bacterial-type plastid RNA polymerase (PEP). Promoter recognition of PEP is mediated by one of the six nuclear-encoded sigma factors in Arabidopsis. The replacement of the plastid sigma factor associated with PEP may be the major mechanism for switching of plastid transcription pattern in response to environmental and developmental signals. This study demonstrates that AtSig5 is a unique or factor that is essential for psbD BLRP activity. A T-DNA insertional mutant with reduced AtSIG5 expression resulted in loss of primary transcripts from the psbD BLRP. Furthermore, transient overexpression of AtSig5 in dark-adapted protoplasts specifically elevated psbD and psbA transcription activities. On the other hand, overproduction of AtSig2 enhanced the transcription of psbA gene and trnE operon, but not psbD transcription. The AtSIG5 gene is phylogenetically distinct from other plastid or factors, and its expression is induced exclusively by blue light. We propose that AtSig5 acts as a mediator of blue light signaling that specifically activates the psbD BLRP in response to blue light in Arabidopsis., NATL ACAD SCIENCES
Proc. Natl. Acad. Sci. USA, 2004年, ［査読有り］

〔主要な業績〕Mutations in KaiA, a clock protein, extend the period of circadian rhythm in the cyanobacterium Synechococcus elongatus PCC 7942.
Nishimura H; Nakahira Y; Imai K; Tsuruhara A; Kondo H; Hayashi H; Hirai M; Saito H; and Kondo T, 筆頭著者, KaiA KaiB and KaiC are essential circadian clock proteins in the unicellular cyanobacterium Synechococcus elongatus PCC 7942. KaiA protein activates transcription of the kaiBC operon, which is believed to be a crucial step in the oscillating feedback loop of cyanobacteria. In this study, similar to400 mutations were introduced into kaiA by PCR-based mutagenesis, and rhythmic phenotypes of these mutants were studied by a bioluminescence reporter. In contrast to mutations in KaiB or KaiC, the vast majority of KaiA mutations extended the period and only rarely shortened it. The period could be extended to 35 h without lowering the mean or peak levels of kaiBC expression. However, several mutations resulted in low-amplitude oscillations or arrhythmia, which were accompanied by lowered kaiBC transcription. These results imply that the KaiA protein can change the period length of the circadian rhythm directly (through an unknown biochemical mechanism) or indirectly (by lowering kaiBC expression). Specific mutations of KaiA were identified in 34 mutants. While mutations mapped to various locations of the KaiA sequence, two clusters of period-altering mutations were found. This suggested that these regions are important domains of the KaiA protein for defining the period length. On the other hand, different sequences within KaiA to which arrhythmic mutations were mapped are important to enhance kaiBC expression., SOC GENERAL MICROBIOLOGY
Microbiology, 2002年, ［査読有り］

シアノバクテリアにおけるkai時計遺伝子作用モデルの再検討
岩崎 秀雄; 中平 洋一; 片山 光徳; 近藤 孝男
日本時間生物学会会誌: Journal of Chronobiology, 2001年10月01日

Circadian-regulated expression of a nuclear-encoded plastid sigma factor gene (sigA) in wheat seedlings.
Morikawa K; Ito S; Tsunoyama Y; Nakahira Y; Shiina T; and Toyoshima Y, The activity of a light-responsive psbD promoter in plastids is known to be regulated by a circadian clock, However, the mechanism of the circadian regulation of the psbD light-responsive promotor, which is recognized by an Escherichia coli-type RNA polymerase, is not yet known. We examined the time course of mRNA accumulation of two E. coli-type RNA polymerase subunit genes, sigA and rpoA, under a continuous light condition after 12 h light/12 h dark entrainment. Accumulation of the sigA mRNA was found to be regulated by a circadian clock, while rpoA mRNA did not show any significant oscillation throughout the experiment. (C) 1999 Federation of European Biochemical Societies., ELSEVIER SCIENCE BV
FEBS Lett., 1999年, ［査読有り］

Developmental stage-specific multi-subunit plastid RNA polymerases (PEP) in wheat.
Satoh J; Baba K; Nakahira Y; Tsunoyama Y; Shiina T; and Toyoshima Y, Most photosystem I and II plastid genes are transcribed by a plastid encoded Escherichia coli-like RNA polymerase (PEP). In this study, we show that both promoter selectivity and light-dependency of PEP change dramatically during development in wheat leaves. In the leaf tip, psbA and psbD promoter activities are light induced, whilst psbC, psbE and 16S rRNA promoters do not function efficiently irrespective of light conditions. In contrast to the leaf tip, in the basal portion all PEP promoters studied function in the dark as well as the light, except for psbD. Using in vitro transcription, we found that PEP in the illuminated leaf tip can initiate transcription from the -35 destructed psbA promoter, but the -35 element is essential for transcription in the basal portion. There is an extended -10 element in the psbA promoter, recognized by the PEP in the illuminated leaf tip or purified sigma(70)-type Escherichia coli RNA polymerase but not by the PEP in the leaf base. These results suggest that during wheat leaf development, PEP in the leaf base that is functional for most PEP promoters even in the dark is replaced by the light-dependent PEP selectively transcribing the psbA and psbD promoters., WILEY-BLACKWELL
Plant J., 1999年, ［査読有り］

〔主要な業績〕Circadian-regulated transcription of the psbD light-responsive promoter in wheat chloroplasts seedlings.
Nakahira Y; Baba K; Yoneda A; Shiina T; and Toyoshima Y, 筆頭著者, The level of mRNAs derived from the plastid-encoded psbD light-responsive promoter (LRP) is controlled by a circadian clock(s) in wheat ( Triticum aestivum). The circadian oscillations in the psbD LRP mRNA level persisted for at least three cycles in continuous light and for one cycle in continuous dark, with maxima in subjective morning and minima in subjective early night. In vitro transcription in chloroplast extracts revealed that the circadian cycles in the psbD LRP mRNA level were dominantly attributed to the circadian-regulated transcription of the psbD LRP. The effects of various mutations introduced into the promoter region on the psbD LRP activity in vitro suggest the existence of two positive elements located between -54 and -36, which generally enhance the transcription activity, and an anomalous core promoter structure lacking the functional "-35" element, which plays a crucial role in the circadian fluctuation and light dependency of psbD LRP transcription activity., AMER SOC PLANT PHYSIOLOGISTS
Plant Physiol., 1998年, ［査読有り］

〔主要な業績〕Circadian-regulated transcription of the psbD light-responsive promoter (psbD LRP) in wheat chloroplasts.
Nakahira Y; Morikawa K; Shiina T; and Toyoshima Y, 筆頭著者, SPRINGER
In Photosynthesis: Mechanisms and Effects (Garab, G eds.) Kluwer Academic publishers, 1998年

Characterization of dynamics of the psbD light-induced transcription in mature wheat chloroplasts.
Satoh J; Baba K; Nakahira Y; Shiina T; and Toyoshima Y, Dynamical aspects of three chloroplast promoters responding to change in light condition were examined in mature chloroplasts of wheat (Triticum aestivum) by in vitro transcription. The wheat psbD/C operon has four distinct promoters, two of which named as D/C-3 and D/C-4 promoters dominantly function in mature chloroplasts to produce the mRNAs encoding D2/CP43 and CP43 alone, respectively. Activity of the D/C-3 promoter in mature chloroplasts was reduced to less than 30% by 24 h dark adaptation and recovered by re-illumination to the original level within 30 to 60 min. The activation of the D/C-3 promoter which requires de novo cytoplasmic protein synthesis was induced by low fluence of light (e.g. 16 mu E m(-2) s(-1)), but the extent of activation increased with increasing light fluence. The accumulation of mRNAs from the D/C-3 promoter saturated at 2- to 3-fold higher level within 2 h when the dark-adapted seedlings were transferred to the light at 72 mu E m(-2) s(-1), concomitant with the increase in rate of D2 synthesis, suggesting that synthesis of D2 in mature chloroplasts is controlled via the D/C-3 promoter activity in a light-dependent way. Activity of the D/C-4 promoter slightly increased in the dark and decreased in the light. Effect of light on the psbA promoter activity was not observed at all in mature chloroplasts. In vitro transcriptional analysis of the D/C-3 promoter with 5' deletion mutations revealed that at least two cis elements which are located within the sequences of -78 to -47 and -46 to -29 of the transcription initiation site, respectively, act as enhancing elements in the D/C-3 promoter. The light-switching element of the transcription, however, was suggested to be located in the core promoter sequence downstream of the -35 element., KLUWER ACADEMIC PUBL
Plant Mol. Biol., 1997年, ［査読有り］

Cis and trans factor(s) of a light-responsive promoter of psbD/C gene cluster in wheat.
Tsunoyama Y; Nakahira Y; Shiina T; and Toyoshima Y, KLUWER ACADEMIC PUBL
"Photosynthesis: from Light to Biosphere" edited by P. Mathis Kluwer Academic Publishers, 1995年

Possible roles of light-responsive psb promoters in regulation of turnover of PSII proteins.
Baba K; Satoh J; Nakahira Y; Shiina T; and Toyoshima Y, KLUWER ACADEMIC PUBL
"Photosynthesis: from Light to Biosphere" edited by P. Mathis Kluwer Academic Publishers, 1995年

〔主要な業績〕葉緑体工学でつくる水産用ワクチン植物
中平 洋一
アグリバイオ, 2024年11月, ［招待有り］
筆頭著者

〔主要な業績〕葉緑体工学によるモノづくり,高等植物葉緑体ゲノム改変技術の進展とその応用
中平 洋一
化学と生物, 2024年03月01日, ［査読有り］, ［招待有り］
筆頭著者

植物の感染防御応答におけるサリチル酸誘導と葉緑体チラコイド膜タンパク質CAS
椎名 隆; 吉岡 博文; 福崎 英一郎; 植村 周平; 神田 ゆい; 佐野 智; 竹林 宏祐; 杉本 孝徳; 馬場 健史; 中平 洋一; 野村 裕也
日本植物生理学会年会およびシンポジウム 講演要旨集, 2011年

NAC-like transcription factor,VOZは生物学的ストレスと非生物学的ストレスに対する適応制御の切り換えに関与する。
中井勇介; 中平洋一; 隅田浩規; 安居佑季子; 河内孝之; 高木優; 光田展隆; 久保康之; 佐藤雅彦
日本植物学会大会研究発表記録, 2011年

シロイヌナズナNAC関連転写因子VOZは非生物学的ストレス応答の制御に関与している
中井勇介; 中平洋一; 安居佑季子; 河内孝之; 椎名隆; 高木優; 光田展隆; 佐藤雅彦
日本植物生理学会年会要旨集, 2011年

フィトクロムシグナル伝達において機能する花成促進因子VOZの局在解析
安居佑季子; 西谷亜依子; 上本充大; 硯亮太; 向川佳子; 中井勇介; 中平洋一; 佐藤雅彦; 河内孝之
日本植物生理学会年会要旨集, 2010年03月12日

植物特異的DNA結合タンパク質VOZは転写コアクチベーターMBF1bと相互作用する
中井勇介; 佐藤雅彦; 安居佑季子; 河内孝之; 山崎健一; 中平洋一
日本植物生理学会年会要旨集, 2010年

Involvement of a chloroplast-localized protein CAS in the external Ca2+-induced stomatal closure.
H Nomura; M Kobori; Y Tsunoyama; E Iwagishi; Y Nakahira; T Shiina
PLANT AND CELL PHYSIOLOGY, 2006年

葉緑体シグマ因子二重変異体（sig2sig6）はアルビノになる
石崎 陽子; 尾園 加奈子; 竹中 智佳子; 角山 雄一; 中平 洋一; 田中 寛; 金丸 研吾; 華岡 光正; 椎名 隆
日本植物生理学会年会およびシンポジウム 講演要旨集, 2006年

Characterization of a specific plastid sigma factor AtSIG5 using chloroplast transformation
M Nozoe; Y Ishizaki; Y Tsunoyama; Y Tsubokura; Y Nakahira; T Shiina
PLANT AND CELL PHYSIOLOGY, 2005年

〔主要な業績〕葉緑体の秘めたるパワー 〜高等植物の葉緑体形質転換法が持つ可能性〜
中平 洋一; 椎名 隆
蛋白質 核酸 酵素, 2005年
筆頭著者

〔主要な業績〕Plastid transcription in higher plants.
Toyoshima Y; Onda Y; Shiina T; and Nakahira Y
Critical Reviews in Plant Sciences, 2005年
ラスト(シニア)オーサー

〔主要な業績〕Plastid RNA polymerases, promoters and transcription regulators in higher plants.
Shiina T; Tsunoyama Y; Nakahira Y; and Khan MS
International Review of Cytology, 2005年

Arabidopsis Sig6 is a general sigma factor in chloroplasts, important for transcription of photosynthesis-related genes at an early stage of seedling growth
Y Ishizaki; Y Tsunoyama; K Hatano; M Kobori; G Takeba; Y Nakahira; T Shiina
PLANT AND CELL PHYSIOLOGY, 2004年

Monitoring of circadian gene expression by rail-road worm luciferases dual-reporter system
Y Kitayama; T Oyama; Y Nakahira; Viviani, V; Y Ohmiya; T Kondo
PLANT AND CELL PHYSIOLOGY, 2003年

〔主要な業績〕Transcriptional regulation of the kaiABC clock genes in cyanobacteria
Y Nakahira; M Katayama; H Iwasaki; T Kondo
PLANT AND CELL PHYSIOLOGY, 2002年

〔主要な業績〕植物の概日時計遺伝子
中平 洋一; 近藤 孝男
化学と生物, 2000年
筆頭著者

〔主要な業績〕Light-induced switching of promoter recognition specificity of chloroplast polymerase.
T Shiina; K Baba; Y Tsunoyama; Y Nakahira; A Yoneda; K Morikawa; Y Toyoshima
PLANT PHYSIOLOGY, 1997年07月

〔主要な業績〕葉緑体における転写制御機構
中平 洋一; 豊島 喜則
日本農芸化学会誌, 1997年
筆頭著者

〔主要な業績〕高活性アルカン生合成酵素を用いたシアノバクテリアにおける持続可能な航空燃料（SAF）の生産性向上　　　　　　　　　　　　　　　
石田梨紗子; 金子太樹; 中平洋一
第66回 日本植物生理学会年会（金沢）, 2025年03月15日

〔主要な業績〕ポジティブ・ネガティブ選抜マーカーを併用した新規「マーカーフリー葉緑体形質転換植物作出法」の開発　　　　　　　　　　　　　　　
小薗江彩唯; 石川奈緒美; 松村優斗; 桑田小夜子; 中平洋一
第66回 日本植物生理学会年会（金沢）, 2025年03月14日

〔主要な業績〕シアノバクテリア用「テオフィリン誘導型人工リボスイッチ」のシステマティックな改変　　　　　　　　　　　　　　　
藤原 未来; 大嶋 真由子; 中平 洋一
第41回 日本植物バイオテクノロジー学会（仙台）大会, 2024年09月01日

〔主要な業績〕耐熱性セルラーゼ類を大量発現する葉緑体形質転換タバコ　　　　　　　　　　　　　　　
坂本 晴那; 磯野 真秀; 中平 洋一
第41回 日本植物バイオテクノロジー学会（仙台）大会, 2024年08月31日

〔主要な業績〕シアノバクテリアでの有用物質生産に適したテオフィリン依存型人工リボスイッチの改変　　　　　　　　　　　　　　　
藤原 未来、 大嶋 真由子、 中平 洋一
第65回 日本植物生理学会年会（神戸）, 2024年03月17日

〔主要な業績〕魚病防除用ウイルス様粒子（VLP）ワクチンを大量発現する葉緑体形質転換レタス　　　　　　　　　　　　　　　
平山 美羽、原川 翔伍、川上 秀昌、中平 洋一
第40回 日本植物バイオテクノロジー 学会（千葉）大会, 2023年09月12日

マロニル-CoA生合成を強化した大腸菌での遊離脂肪酸生産　　　　　　　　　　　　　　　
濵田 美志、中平 洋一、西澤 智康、長南 茂
第74回日本生物工学会大会, 2022年10月20日

〔主要な業績〕葉緑体工学で創る「経口ワクチン植物」 - 水産用ワクチンを例に -　　　　　　　　　　　　　　　
中平 洋一
第38回日本植物バイオテクノロジー学会（つくば）大会,シンポジウム 「植物オルガネラゲノム工学の新展開」, 2021年09月10日, ［招待有り］

〔主要な業績〕茨城大など、タバコ葉で魚病ウイルスの経口ワクチン開発 大量生産でコストダウン、今後は食用植物で実用化目指す　　　　　　　　　　　　　　　
日経バイオテクONLINE, 2021年08月31日, ［招待有り］

〔主要な業績〕養殖魚に経口ワクチン 茨城大学など開発 疾病予防に効果　　　　　　　　　　　　　　　
日本経済新聞（北関東地域面）, 2021年08月26日, ［招待有り］

〔主要な業績〕養殖魚に経口ワクチン 愛媛県農水研など開発 疾病予防に　　　　　　　　　　　　　　　
日本経済新聞（四国版）, 2021年08月26日, ［招待有り］

〔主要な業績〕植物で魚病ワクチン作出 茨城大など研究チーム開発 経口投与で免疫効果確認　　　　　　　　　　　　　　　
日刊水産経済新聞, 2021年08月24日, ［招待有り］

〔主要な業績〕養魚に｢食べるワクチン植物｣ ウイルス疾病予防に有効 既存注射型と同等の高免疫原性　　　　　　　　　　　　　　　
みなと新聞, 2021年08月24日, ［招待有り］

〔主要な業績〕Development of a strictly controlled inducible gene expression system for plastids by using synthetic riboswitches　　　　　　　　　　　　　　　
RIKA YAMANE; ATSUSHI OGAWA; YUZURU TOZAWA; TAKASHI SHIINA; YOICHI NAKAHIRA
The 2019 Gordon Research Conference on Chloroplast Biotechnology, 2019年01月08日

〔主要な業績〕Overproduction of a virus-like-particle (VLP) vaccine against a fish disease by plastid engineering　　　　　　　　　　　　　　　
Yoichi Nakahira; Hirofumi Yamashita; Minami Tsuchikura; Kaoru Takeuchi; Takashi Shiina
The 2019 Gordon Research Conference on Chloroplast Biotechnology, 2019年01月07日

〔主要な業績〕人工リボスイッチを基盤とした葉緑体遺伝子発現誘導系の改良　　　　　　　　　　　　　　　
山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
第41回日本分子生物学会年会, 2018年11月28日

〔主要な業績〕葉緑体工学を用いた水産用VLPワクチンの大量生産　　　　　　　　　　　　　　　
中平 洋一、山下 浩史、土倉 みなみ、竹内 薫、椎名 隆
第36回日本植物細胞分子生物学会（金沢）大会, 2018年08月28日

〔主要な業績〕EVALUATION OF THEOPHYLLINE-DEPENDENT SYNTHETIC RIBOSWITCHES FOR STRICT CONTROL OF PLASTID GENE EXPRESSION　　　　　　　　　　　　　　　
RIKA YAMANE; ATSUSHI OGAWA; YUZURU TOZAWA; TAKASHI SHIINA; YOICHI NAKAHIRA
International Plant Molecular Biology 2018, 2018年08月07日

〔主要な業績〕HIGH-LEVEL PRODUCTION OF A VIRUS-LIKE PARTICLE VACCINE AGAINST A FISH NERVOUS NECROSIS VIRUS IN TRANSPLASTOMIC TOBACCO PLANTS　　　　　　　　　　　　　　　
YOICHI NAKAHIRA; HIROFUMI YAMASHITA; MINAMI TSUCHIKURA; KAORU TAKEUCHI; TAKASHI SHIINA
International Plant Molecular Biology 2018, 2018年08月06日

〔主要な業績〕テオフィリン依存型人工リボスイッチを用いたシアノバクテリアにおけるアルカン増産の試み　　　　　　　　　　　　　　　
金子 太樹、福田 寛史、武波 洋志、朝山 宗彦、中平 洋一
2017年度 生命科学系学会合同年次大会, 2017年12月07日

〔主要な業績〕人工リボスイッチを活用した葉緑体遺伝子発現誘導系の評価　　　　　　　　　　　　　　　
山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
2017年度 生命科学系学会合同年次大会, 2017年12月07日

〔主要な業績〕人工リボスイッチを活用した葉緑体遺伝子発現制御系の評価　　　　　　　　　　　　　　　
山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
第35回 日本植物細胞分子生物学会（さいたま）大会, 2017年08月29日

〔主要な業績〕Metabolic engineering for improved production of drop-in fuels in the cyanobacterium Synechococcus elongatus PCC 7942　　　　　　　　　　　　　　　
Hiroki Kaneko; Hirofumi Fukuda; MunehikoAsayama; and Yoichi Nakahira
第58回 日本植物生理学会年会, 2017年03月16日

〔主要な業績〕Utilization of a theophylline-dependent engineered riboswitch for enhanced alkane production in the cyanobacterium Synechocystis sp. PCC 6803　　　　　　　　　　　　　　　
Hirofumi Fukuda; Hiroki Kaneko; MunehikoAsayama; and Yoichi Nakahira
第58回 日本植物生理学会年会, 2017年03月16日

〔主要な業績〕シアノバクテリアにおけるドロップイン燃料生産性強化に向けた遺伝子改変技術の開発　　　　　　　　　　　　　　　
金子太樹、山根里佳、櫻井大貴、福田寛史、朝山宗彦、中平洋一
第39回 日本分子生物学会年会, 2016年12月02日

〔主要な業績〕シアノバクテリアのアルカンと脂肪生産に与える栄養源欠乏培地の影響　　　　　　　　　　　　　　　
福田寛史、中平洋一、長谷川守文、宮口右二、朝山宗彦
第39回 日本分子生物学会年会, 2016年12月02日

〔主要な業績〕人工リボスイッチを基盤とした葉緑体遺伝子発現誘導系　　　　　　　　　　　　　　　
中平洋一、山根里佳、小川敦司、戸澤譲、椎名隆
第39回 日本分子生物学会年会, 2016年12月01日

〔主要な業績〕人工リボスイッチを用いたタバコ葉緑体遺伝子発現誘導系　　　　　　　　　　　　　　　
中平洋一、小川敦司、戸澤譲、椎名隆
第34回 日本植物細胞分子生物学会（上田）大会, 2016年09月01日

〔主要な業績〕Genetic engineering for enhanced alkane production in the model cyanobacterium Synechococcus elongatus PCC 7942　　　　　　　　　　　　　　　
金子太樹、福田寛史、朝山宗彦、中平洋一
第57回日本植物生理学会年会, 2016年03月20日

〔主要な業績〕Utilization of the theophylline-dependent engineered riboswitches for strict control of plastid gene expression in tobacco.　　　　　　　　　　　　　　　
中平 洋一; 小川 敦司; 戸澤 譲; 椎名 隆
第57回日本植物生理学会年会, 2016年03月18日

Genetic Engineering and Overproduction of Polyhydroxybutyrate in Cyanobacteria　　　　　　　　　　　　　　　
Sayaka Hondo; Masatoshi Takahashi; Takashi Osanai; Mami Matsuda; Tomohisa Hasunuma; Akio Tazuke; Yoichi Nakahira; Shigeru Chonan; Morifumi Hasegawa; Munehiko Asayama
THE INTERNATIONAL CHEMICAL CONGRESS OF PACIFIC BASIN SOCIETIES 2015, 2015年12月

藻類バイオプラスチックPHB高生産株の創生と特徴付け　　　　　　　　　　　　　　　
本堂彩花、高橋正俊、小山内崇、松田真実、蓮沼誠久、長谷川守文、中平洋一、朝山宗彦
第３７回 日本分子生物学会年会, 2014年11月27日

〔主要な業績〕人工リボスイッチを基盤としたラン藻の新規遺伝子発現制御技術　　　　　　　　　　　　　　　
中平 洋一、小川 敦司、浅野 宏幸、小山 時隆、戸澤 譲
第55回 日本植物生理学会年会, 2014年03月20日

〔主要な業績〕人工リボスイッチを用いたラン藻のための新規遺伝子発現制御技術　　　　　　　　　　　　　　　
中平 洋一、小川 敦司、浅野 宏幸、小山 時隆、戸澤 譲
第36回 日本分子生物学会年会, 2013年12月

〔主要な業績〕Overproduction of Thermostable Biomass-Saccharizing Enzymes in Tobacco Chloroplasts: Towards Developing the Self-Saccharizing Crops　　　　　　　　　　　　　　　
Nakahira Y; Kashima Y; Tozawa Y; Shiina T
3rd International Symposium on Chloroplast Genomics and Genetic Engineering, 2013年05月

〔主要な業績〕ラン藻のための新規遺伝子発現制御系の開発　　　　　　　　　　　　　　　
中平 洋一、小川 敦司、浅野 宏幸、小山 時隆、戸澤 譲
第54回 日本植物生理学会年会, 2013年03月

〔主要な業績〕葉緑体工学による有用植物の開発　　　　　　　　　　　　　　　
中平 洋一、鹿島 康浩、椎名 隆、戸澤 譲
日本農芸化学会中四国支部大会（第34回講演会）, 2012年09月

〔主要な業績〕葉緑体工学による自己糖化型タバコの作出　　　　　　　　　　　　　　　
第29回日本植物細胞分子生物学会大会, 2011年09月

〔主要な業績〕High-Level Expression of a Set of Six Thermostable Cell Wall-Degrading Enzymes in Tobacco Chloroplasts: A First Step Towards Development of the Auto-Saccharification System of Bioenergy Crops.　　　　　　　　　　　　　　　
22nd International Conference on Arabidopsis Research, 2011年05月

〔主要な業績〕葉緑体工学による耐熱性糖化酵素群の大量発現　　　　　　　　　　　　　　　
中平 洋一、鹿島 康浩
日本農芸化学会2011年度大会, 2011年03月

〔主要な業績〕超耐熱性セルラーゼを大量発現する葉緑体形質転換植物　　　　　　　　　　　　　　　
中平 洋一、田中 國介、石川 一彦、椎名 隆
日本農芸化学会2009年度大会, 2009年03月

〔主要な業績〕葉緑体工学による超耐熱性セルラーゼの大量発現　　　　　　　　　　　　　　　
中平 洋一、田中 國介、石川 一彦、椎名 隆
第50回 日本植物生理学会年会, 2009年03月

〔主要な業績〕葉緑体工学を用いた超耐熱性セルラーゼの大量発現　　　　　　　　　　　　　　　
第31回 日本分子生物学会年会・第81回 日本生化学会大会 合同大会, 2008年12月

〔主要な業績〕プラスチド核様体に関連するSWIBドメイン蛋白質の解析　　　　　　　　　　　　　　　
中平 洋一、矢田 和正、椎名 隆
第47回 日本植物生理学会年会, 2006年03月

〔主要な業績〕Firefly luciferase as a vital reporter for gene expression in tobacco chloroplasts.　　　　　　　　　　　　　　　
Nakahira Y; Takeba G; and Shiina T
XIIIth International Congress on Photosynthesis, 2004年08月

〔主要な業績〕ルシフェラーゼを利用した葉緑体の遺伝子発現モニター系　　　　　　　　　　　　　　　
中平 洋一、野添 幹雄、竹場 剛、椎名 隆
第45回 日本植物生理学会年会, 2004年03月

〔主要な業績〕ルシフェラーゼ遺伝子を用いた葉緑体遺伝子発現モニタ系の開発　　　　　　　　　　　　　　　
中平 洋一、野添 幹雄、椎名 隆
第26回 日本分子生物学会年会, 2003年12月

植物生理学実験　　　　　　　　　　　　　　　
茨城大学

植物遺伝子工学実験　　　　　　　　　　　　　　　
茨城大学

植物生理学Ⅰ　　　　　　　　　　　　　　　
茨城大学

植物生理学Ⅱ　　　　　　　　　　　　　　　
茨城大学

植物分子遺伝学　　　　　　　　　　　　　　　
茨城大学

植物科学実験Ⅰ　　　　　　　　　　　　　　　
茨城大学

植物科学実験Ⅱ　　　　　　　　　　　　　　　
茨城大学

2008年05月, 日本植物バイオテクノロジー学会

2008年, 日本農芸化学会

2001年08月, 日本分子生物学会

1994年03月, 日本植物生理学会

〔主要な業績〕バイオジェット燃料を“一気通貫生産”するエネルギー作物の開発　　　　　　　　　　　　　　　
2023年04月 - 2024年03月

〔主要な業績〕さまざまな魚病の防除に適用可能な「水産用ワクチン植物」創出基盤技術の開発　　　　　　　　　　　　　　　
トライアウト
2022年10月 - 2024年03月

〔主要な業績〕タンパク質“自己結合”システムを用いた多機能型ウイルス様中空粒子ワクチンの開発
基盤研究(C)
筑波大学
2020年04月 - 2023年03月

〔主要な業績〕“低コスト”・“省力”・“高免疫原性”を兼ね備えた「水産用ワクチン植物」の開発　　　　　　　　　　　　　　　
研究成果最適展開支援プログラム(A-STEP) 機能検証フェーズ
2019年11月 - 2021年03月

〔主要な業績〕医食同源により万病予防を実現できる「経口ワクチン植物」創製基盤技術の開発　　　　　　　　　　　　　　　
挑戦的研究(萌芽)
2018年06月 - 2021年03月

〔主要な業績〕完全人工合成された葉緑体ゲノムを有する遺伝子組換え植物の創出　　　　　　　　　　　　　　　
萌芽研究
2015年04月 - 2018年03月

色素体―核コミュニケーションを介した植物機能統御の新機構
基盤研究(B)
京都府立大学
2013年04月01日 - 2017年03月31日

〔主要な業績〕葉緑体工学を用いた養殖魚のための疾病予防植物の創出　　　　　　　　　　　　　　　
2012年11月 - 2013年10月

〔主要な業績〕葉緑体工学を用いた自己糖化型エネルギー作物の開発　　　　　　　　　　　　　　　
2008年10月 - 2011年03月

植物の感染防御応答に関わる葉緑体カルシウムシグナル伝達ネットワーク
新学術領域研究(研究課題提案型)
京都府立大学
2008年 - 2010年

〔主要な業績〕概日時計による葉緑体遺伝子の転写制御
特別研究員奨励費
京都府立大学
2002年 - 2004年