Jun. 2024, iMOSS - international molecular moss science society Propelling Flagellated Plant Research, Golden Spore Award for 2024　　　　　　　　　　　　　　　
Tomomichi Fujita

2018, 日本植物学会, Journal of Plant Research Best paper論文賞　　　　　　　　　　　　　　　
藤田 知道

2014, 北海道大学, 北海道大学教育総長賞　　　　　　　　　　　　　　　
藤田 知道

2014, 日本植物学会, Journal of Plant Research Best paper論文賞　　　　　　　　　　　　　　　
藤田 知道

2014, 日本学術振興会, 特別研究員等審査会専門委員及び国際事業委員会書面審査委員表彰者　　　　　　　　　　　　　　　
藤田 知道

ABA signaling converts stem cell fate by substantiating a tradeoff between cell polarity, growth and cell cycle progression and abiotic stress responses in the moss Physcomitrium patens
Marcel Pascal Beier, Chiyo Jinno, Natsumi Noda, Kohei Nakamura, Sumio Sugano, Yutaka Suzuki, Tomomichi Fujita
Frontiers in Plant Science, 14:1303195, 01 Nov. 2023, [Peer-reviewed], [Corresponding author]

Surface-localized glycoproteins act through class C ARFs to fine-tune gametophore initiation in Physcomitrium patens
Ooi Kock Teh, Prerna Singh, Junling Ren, Lin Tzu Huang, Menaka Ariyarathne, Benjamin Prethiviraj Salamon, Yu Wang, Toshihisa Kotake, Tomomichi Fujita
Development, 149, 24, dev200370, The Company of Biologists, 15 Dec. 2022, [Peer-reviewed], [Corresponding author]
Scientific journal, ABSTRACT

Arabinogalactan proteins are functionally diverse cell wall structural glycoproteins that have been implicated in cell wall remodeling, although the mechanistic actions remain elusive. Here, we identify and characterize two AGP glycoproteins, SLEEPING BEAUTY (SB) and SB-like (SBL), that negatively regulate the gametophore bud initiation in Physcomitrium patens by dampening cell wall loosening/softening. Disruption of SB and SBL led to accelerated gametophore formation and altered cell wall compositions. The function of SB is glycosylation dependent and genetically connected with the class C auxin response factor (ARF) transcription factors PpARFC1B and PpARFC2. Transcriptomics profiling showed that SB upregulates PpARFC2, which in turn suppresses a range of cell wall-modifying genes that are required for cell wall loosening/softening. We further show that PpARFC2 binds directly to multiple AuxRE motifs on the cis-regulatory sequences of PECTIN METHYLESTERASE to suppress its expression. Hence, our results demonstrate a mechanism by which the SB modulates the strength of intracellular auxin signaling output, which is necessary to fine-tune the timing of gametophore initials formation.

Three-Dimensionally Visualized Rhizoid System of Moss, Physcomitrium Patens, by Refraction-Contrast X-ray Micro-Computed Tomography.
Ryohei Yamaura, Daisuke Tamaoki, Hiroyuki Kamachi, Daisuke Yamauchi, Yoshinobu Mineyuki, Kentaro Uesugi, Masato Hoshino, Tomomi Suzuki, Toru Shimazu, Haruo Kasahara, Motoshi Kamada, Yuko T Hanba, Atsushi Kume, Tomomichi Fujita, Ichirou Karahara
Microscopy (Oxford, England), 22 Aug. 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Land plants have two types of shoot-supporting systems, root system and rhizoid system, in vascular plants and bryophytes. However, since the evolutionary origin of the systems are different, how much they exploit common systems or distinct systems to architect their structures are largely unknown. To understand the regulatory mechanism how bryophytes architect rhizoid system responding to environmental factors, we have developed the methodology to visualize and quantitatively analyze the rhizoid system of the moss, Physcomitrium patens in 3D. The rhizoids having the diameter of 21.3 µm on the average were visualized by refraction-contrast X-ray micro-CT using coherent X-ray optics available at synchrotron radiation facility SPring-8. Three types of shape (ring-shape, line, black circle) observed in tomographic slices of specimens embedded in paraffin were confirmed to be the rhizoids by optical and electron microscopy. Comprehensive automatic segmentation of the rhizoids which appeared in different three form types in tomograms was tested by a method using Canny edge detector or machine learning. Accuracy of output images was evaluated by comparing with the manually-segmented ground truth images using measures such as F1 score and IoU, revealing that the automatic segmentation using the machine learning was more effective than that using Canny edge detector. Thus, machine learning-based skeletonized 3D model revealed quite dense distribution of rhizoids. We successfully visualized the moss rhizoid system in 3D for the first time. High resolution refraction-contrast X-ray micro-CT using coherent X-ray optics successfully visualized 3D architecture of rhizoid system of moss, Physcomitrium patens, which is composed of cellular filaments having the diameter of 21.3 µm on the average, for the first time by using machine learning for segmentation.

The cellular function of ROP GTPase prenylation is important for multicellularity in the moss Physcomitrium patens.
Liang Bao, Junling Ren, Mary Nguyen, Arkadiusz Slawomir Slusarczyk, Julie Thole, Susana Perez Martinez, Jinling Huang, Tomomichi Fujita, Mark Running
Development, 149, 12, dev200279, Jun. 2022, [Peer-reviewed]

Abscisic acid switches cell division modes of asymmetric cell division and symmetric cell division in stem cells of protonemal filaments in the moss Physcomitrium patens.
Akihiko Hiroguchi, Kohei Nakamura, Tomomichi Fujita
Plant Biotechnology, 39, 1, 13, 17, Mar. 2022, [Peer-reviewed], [Corresponding author]

A PSTAIRE-type cyclin-dependent kinase controls light responses in land plants.
Liang Bao, Natsumi Inoue, Masaki Ishikawa, Eiji Gotoh, Ooi-Kock Teh, Takeshi Higa, Tomoro Morimoto, Eggie Febrianto Ginanjar, Hirofumi Harashima, Natsumi Noda, Masaaki Watahiki, Yuji Hiwatashi, Masami Sekine, Mitsuyasu Hasebe, Masamitsu Wada, Tomomichi Fujita
Science Advances, 8, 4, eabk2116, American Association for the Advancement of Science (AAAS), 28 Jan. 2022, [Peer-reviewed], [Corresponding author]
Scientific journal, Light is a critical signal perceived by plants to adapt their growth rate and direction. Although many signaling components have been studied, how plants respond to constantly fluctuating light remains underexplored. Here, we showed that in the moss
Physcomitrium
(
Physcomitrella
)
patens
, the PSTAIRE-type cyclin-dependent kinase PpCDKA is dispensable for growth. Instead, PpCDKA and its homolog in
Arabidopsis thaliana
control light-induced tropisms and chloroplast movements by probably influencing the cytoskeleton organization independently of the cell cycle. In addition, lower PpCDKA kinase activity was required to elicit light responses relative to cell cycle regulation. Thus, our study suggests that plant CDKAs may have been co-opted to control multiple light responses, and owing to the bistable switch properties of PSTAIRE-type CDKs, the noncanonical functions are widely conserved for eukaryotic environmental adaptation.

Characterisation of Rapid Alkalinisation Factors (RAFLs) in Physcomitrium patens Reveals Functional Conservation in Tip Growth.
Eggie Febrianto Ginanjar, Ooi-Kock Teh, Tomomichi Fujita
The New phytologist, 233, 2442, 2457, 25 Dec. 2021, [Peer-reviewed], [Corresponding author], [International Magazine]
English, Scientific journal, Small signalling peptides are key molecules for cell-to-cell communications in plants. The cysteine-rich signalling peptide, rapid alkalinisation factors (RALFs) family are involved in diverse developmental and stress responses and have expanded considerably during land plant evolution, implying neofunctionalisations in the RALF family. However, the ancestral roles of RALFs when land plant first acquired them remain unknown. Here, we functionally characterised two of the three RALFs in bryophyte Physcomitrium patens using loss-of-function mutants, overexpressors as well as fluorescent proteins tagged reporter lines. We showed that PpRALF1 and PpRALF2 have overlapping functions in promoting protonema tip growth and elongation, showing a homologous function as the Arabidopsis RALF1 in promoting root hair tip growth. Although both PpRALFs are secreted to the plasma membrane on which PpRALF1 symmetrically localised, PpRALF2 showed a polarized localisation at the growing tip. Notably, proteolytic cleavage of PpRALF1 is necessary for its function. Our data reveals a possible evolutionary origin of the RALF functions and suggests that functional divergence of RALFs is essential to drive complex morphogenesis and to facilitate other novel processes in land plants.

Molecular biology of mosses
Tomomichi Fujita, Fabien Nogué, Stefan A. Rensing, Daisuke Takezawa, Luis Vidali
Plant Molecular Biology, 107, 4-5, 209, 211, Springer Science and Business Media LLC, Nov. 2021, [Invited], [Lead author, Corresponding author]
Scientific journal

Practical application of proximal sensing for monitoring the growth of Physcomitrium patens.
Hiroki Wakabayashi, Osamu Matsuda, Tomomichi Fujita, Atsushi Kume
Biological Sciences in Space, 35, 32, 40, Japanese Society for Biological Sciences in Space, Sep. 2021, [Peer-reviewed]
Scientific journal

The bryophytes Physcomitrium patens and Marchantia polymorpha as model systems for studying evolutionary cell and developmental biology in plants
Satoshi Naramoto, Yuki Hata, Tomomichi Fujita, Junko Kyozuka
The Plant Cell, 38, 1, 228, 246, Oxford University Press (OUP), 30 Aug. 2021, [Peer-reviewed]
Scientific journal,
Bryophytes are nonvascular spore-forming plants. Unlike in flowering plants, the gametophyte (haploid) generation of bryophytes dominates the sporophyte (diploid) generation. A comparison of bryophytes with flowering plants allows us to answer some fundamental questions raised in evolutionary cell and developmental biology. The moss Physcomitrium patens was the first bryophyte with a sequenced genome. Many cell and developmental studies have been conducted in this species using gene targeting by homologous recombination. The liverwort Marchantia polymorpha has recently emerged as an excellent model system with low genomic redundancy in most of its regulatory pathways. With the development of molecular genetic tools such as efficient genome editing, both P. patens and M. polymorpha have provided many valuable insights. Here, we review these advances with a special focus on polarity formation at the cell and tissue levels. We examine current knowledge regarding the cellular mechanisms of polarized cell elongation and cell division, including symmetric and asymmetric cell division. We also examine the role of polar auxin transport in mosses and liverworts. Finally, we discuss the future of evolutionary cell and developmental biological studies in plants.

Substitution of Deoxycholate with the Amphiphilic Polymer Amphipol A8-35 Improves the Stability of Large Protein Complexes during Native Electrophoresis
Shinsa Kameo, Michiki Aso, Ryo Furukawa, Renon Matsumae, Makio Yokono, Tomomichi Fujita, Ayumi Tanaka, Ryouichi Tanaka, Atsushi Takabayashi
Plant and Cell Physiology, 62, 2, 348, 355, Oxford University Press (OUP), 11 May 2021, [Peer-reviewed], [Domestic magazines]
English, Scientific journal,
Native polyacrylamide gel electrophoresis (PAGE) is a powerful technique for protein complex separation that retains both their activity and structure. In photosynthetic research, native-PAGE is particularly useful given that photosynthetic complexes are generally large in size, ranging from 200 kD to 1 MD or more. Recently, it has been reported that the addition of amphipol A8-35 to solubilized protein samples improved protein complex stability. In a previous study, we found that amphipol A8-35 could substitute sodium deoxycholate (DOC), a conventional electrophoretic carrier, in clear-native (CN)-PAGE. In this study, we present the optimization of amphipol-based CN-PAGE. We found that the ratio of amphipol A8-35 to α-dodecyl maltoside, a detergent commonly used to solubilize photosynthetic complexes, was critical for resolving photosynthetic machinery in CN-PAGE. In addition, LHCII dissociation from PSII–LHCII was effectively prevented by amphipol-based CN-PAGE compared with that of DOC-based CN-PAGE. Our data strongly suggest that majority of the PSII–LHCII in vivo forms C2S2M2 at least in Arabidopsis and Physcomitrella. The other forms might appear owing to the dissociation of LHCII from PSII during sample preparation and electrophoresis, which could be prevented by the addition of amphipol A8-35 after solubilization from thylakoid membranes. These results suggest that amphipol-based CN-PAGE may be a better alternative to DOC-based CN-PAGE for the study of labile protein complexes.

How plants grow under gravity conditions besides 1 g: perspectives from hypergravity and space experiments that employ bryophytes as a model organism
Atsushi Kume, Hiroyuki Kamachi, Yusuke Onoda, Yuko T. Hanba, Yuji Hiwatashi, Ichirou Karahara, Tomomichi Fujita
Plant Molecular Biology, 107, 4-5, 279, 291, Springer Science and Business Media LLC, 14 Apr. 2021, [Peer-reviewed], [Corresponding author]
English, Scientific journal, Plants have evolved and grown under the selection pressure of gravitational force at 1 g on Earth. In response to this selection pressure, plants have acquired gravitropism to sense gravity and change their growth direction. In addition, plants also adjust their morphogenesis in response to different gravitational forces in a phenomenon known as gravity resistance. However, the gravity resistance phenomenon in plants is poorly understood due to the prevalence of 1 g gravitational force on Earth: not only it is difficult to culture plants at gravity > 1 g(hypergravity) for a long period of time but it is also impossible to create a < 1 genvironment (mu g, micro g) on Earth without specialized facilities. Despite these technical challenges, it is important to understand how plants grow in different gravity conditions in order to understand land plant adaptation to the 1 g environment or for outer space exploration. To address this, we have developed a centrifugal device for a prolonged duration of plant culture in hypergravity conditions, and a project to grow plants under the mu g environment in the International Space Station is also underway. Our plant material of choice is Physcomitrium (Physcomitrella) patens, one of the pioneer plants on land and a model bryophyte often used in plant biology. In this review, we summarize our latest findings regarding P. patens growth response to hypergravity, with reference to our on-going "Space moss" project. In our ground-based hypergravity experiments, we analyzed the morphological and physiological changes and found unexpected increments of chloroplast size and photosynthesis rate, which might underlie the enhancement of growth and increase in the number of gametophores and rhizoids. We further discussed our approaches at the cellular level and compare the gravity resistance in mosses and that in angiosperms. Finally, we highlight the advantages and perspectives from the space experiments and conclude that research with bryophytes is beneficial to comprehensively and precisely understand gravitational responses in plants.

Metabolic Control of Gametophore Shoot Formation through Arginine in the Moss Physcomitrium patens
Kensuke Kawade, Gorou Horiguchi, Yuu Hirose, Akira Oikawa, Masami Yokota Hirai, Kazuki Saito, Tomomichi Fujita, Hirokazu Tsukaya
Cell Reports, 32, 10, 108127, 108127, Elsevier BV, Sep. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, Shoot formation is accompanied by active cell proliferation and expansion, requiring that metabolic state adapts to developmental control. Despite the importance of such metabolic reprogramming, it remains unclear how development and metabolism are integrated. Here, we show that disruption of ANGUSTIFOLIA3 orthologs (PpAN3s) compromises gametophore shoot formation in the moss Physcomitrium patens due to defective cell proliferation and expansion. Trans-omics analysis reveals that the downstream activity of PpAN3 is linked to arginine metabolism. Elevating arginine level by chemical treatment leads to stunted gametophores and causes Ppan3 mutant-like transcriptional changes in the wild-type plant. Furthermore, ectopic expression of AtAN3 from Arabidopsis thaliana ameliorates the defective arginine metabolism and promotes gametophore formation in Ppan3 mutants. Together, these findings indicate that arginine metabolism is a key pathway associated with gametophore formation and provide evolutionary insights into the establishment of the shoot system in land plants through the integration of developmental and metabolic processes.

AP2/ERF transcription factors regulate salt-induced chloroplast division in the moss Physcomitrella patens
Thi Huong Do, Prapaporn Pongthai, Menaka Ariyarathne, Ooi-Kock Teh, Tomomichi Fujita
Journal of Plant Research, 133, 4, 537, 548, Springer Science and Business Media LLC, Jul. 2020, [Peer-reviewed], [Corresponding author]
Scientific journal

Quantitative Imaging Reveals Distinct Contributions of SnRK2 and ABI3 in Plasmodesmatal Permeability in Physcomitrella patens
Takumi Tomoi, Kensuke Kawade, Munenori Kitagawa, Yoichi Sakata, Hirokazu Tsukaya, Tomomichi Fujita
Plant and Cell Physiology, 61, 5, 942, 956, Oxford University Press (OUP), 01 May 2020, [Peer-reviewed], [Corresponding author]
English, Scientific journal,
Cell-to-cell communication is tightly regulated in response to environmental stimuli in plants. We previously used a photoconvertible fluorescent protein Dendra2 as a model reporter to study this process. This experiment revealed that macromolecular trafficking between protonemal cells in Physcomitrella patens is suppressed in response to abscisic acid (ABA). However, it remains unknown which ABA signaling components contribute to this suppression and how. Here, we show that ABA signaling components SUCROSE NON-FERMENTING 1-RELATED PROTEIN KINASE 2 (PpSnRK2) and ABA INSENSITIVE 3 (PpABI3) play roles as an essential and promotive factor, respectively, in regulating ABA-induced suppression of Dendra2 diffusion between cells (ASD). Our quantitative imaging analysis revealed that disruption of PpSnRK2 resulted in defective ASD onset itself, whereas disruption of PpABI3 caused an 81-min delay in the initiation of ASD. Live-cell imaging of callose deposition using aniline blue staining showed that, despite this onset delay, callose deposition on cross walls remained constant in the PpABI3 disruptant, suggesting that PpABI3 facilitates ASD in a callose-independent manner. Given that ABA is an important phytohormone to cope with abiotic stresses, we further explored cellular physiological responses. We found that the acquisition of salt stress tolerance is promoted by PpABI3 in a quantitative manner similar to ASD. Our results suggest that PpABI3-mediated ABA signaling may effectively coordinate cell-to-cell communication during the acquisition of salt stress tolerance. This study will accelerate the quantitative study for ABA signaling mechanism and function in response to various abiotic stresses.

Formation of a PSI-PSII megacomplex containing LHCSR and PsbS in the moss Physcomitrella patens
Furukawa, R, Aso M, Fujita, T, Akimoto, S, Tanaka, R, Tanaka, A, Yokono, M, Takabayashi, A
Journal of Plant Research, 132, 6, 867, 880, Nov. 2019, [Peer-reviewed]
English, Scientific journal

The PSI-PSII megacomplex in green plants
Yokono, M, Takabayashi, A, Kishimoto, J, Fujita, T, Iwai, M, Murakami, A, Akimoto, S, Tanaka, A
Plant and Cell Physiology, 60, 5, 1098, 1108, May 2019, [Peer-reviewed]
English, Scientific journal

Abscisic Acid Acts as a Regulator of Molecular Trafficking through Plasmodesmata in the Moss Physcomitrella patens.
Munenori Kitagawa, Takumi Tomoi, Tomoki Fukushima, Yoichi Sakata, Mayuko Sato, Kiminori Toyooka, Tomomichi Fujita, Hitoshi Sakakibara
Plant and Cell Physiology, 60, 4, 738, 751, 01 Apr. 2019, [Peer-reviewed], [Corresponding author], [Domestic magazines]
English, Scientific journal, In multi-cellular organisms, cell-to-cell communication is crucial for adapting to changes in the surrounding environment. In plants, plasmodesmata (PD) provide a unique pathway for cell-to-cell communication. PD interconnect most cells and generate a cytoplasmic continuum, allowing the trafficking of various micro- and macromolecules between cells. This molecular trafficking through PD is dynamically regulated by altering PD permeability dependent on environmental changes, thereby leading to an appropriate response to various stresses; however, how PD permeability is dynamically regulated is still largely unknown. Moreover, studies on the regulation of PD permeability have been conducted primarily in a limited number of angiosperms. Here, we studied the regulation of PD permeability in the moss Physcomitrella patens and report that molecular trafficking through PD is rapidly and reversibly restricted by abscisic acid (ABA). Since ABA plays a key role in various stress responses in the moss, PD permeability can be controlled by ABA to adapt to surrounding environmental changes. This ABA-dependent restriction of PD trafficking correlates with a reduction in PD pore size. Furthermore, we also found that the rate of macromolecular trafficking is higher in an ABA-synthesis defective mutant, suggesting that the endogenous level of ABA is also important for PD-mediated macromolecular trafficking. Thus, our study provides compelling evidence that P. patens exploits ABA as one of the key regulators of PD function.

An experimental system for examining phototropic response of gametophytic shoots in the moss physcomitrella patens
Liang Bao, Kotaro T. Yamamoto, Tomomichi Fujita
Methods in Molecular Biology, 1924, 45, 51, Humana Press Inc., 2019
English, In book, Shoot phototropism benefits growth and metabolism in land plants by enabling them to position their photosynthetic organs in favorable light conditions. Nonvascular land plants, like the ancestors of modern mosses, are believed to have been among the first plants to occupy the land. To understand the evolutional history of shoot phototropism in land plants, we have established a system for experimentally studying phototropism in gametophores of the moss Physcomitrella patens. Here we will describe the key points in our system, including obtaining etiolated gametophores, the light sources used for inducing bending, and the methods for evaluation of phototropic responses.

Correction to: A hypergravity environment increases chloroplast size, photosynthesis, and plant growth in the moss Physcomitrella patens.
Takemura K, Kamachi H, Kume A, Fujita T, Karahara I, Hanba YT
Journal of Plant Research, 131, 5, 887, 887, SPRINGER JAPAN KK, Sep. 2018, [Peer-reviewed]
English, The original article can be found online.

The Physcomitrella patens gene atlas project: large-scale RNA-seq based expression data
Pierre-François Perroud, Fabian B. Haas, Manuel Hiss, Kristian K. Ullrich, Alessandro Alboresi, Mojgan Amirebrahimi, Kerrie Barry, Roberto Bassi, Sandrine Bonhomme, Haodong Chen, Juliet C. Coates, Tomomichi Fujita, Anouchka Guyon-Debast, Daniel Lang, Junyan Lin, Anna Lipzen, Fabien Nogué, Melvin J. Oliver, Inés Ponce de León, Ralph S. Quatrano, Catherine Rameau, Bernd Reiss, Ralf Reski, Mariana Ricca, Younousse Saidi, Ning Sun, Péter Szövényi, Avinash Sreedasyam, Jane Grimwood, Gary Stacey, Jeremy Schmutz, Stefan A. Rensing
Plant Journal, 95, 1, 168, 182, Blackwell Publishing Ltd, 01 Jul. 2018, [Peer-reviewed], ［Internationally co-authored］, [International Magazine]
English, Scientific journal, High-throughput RNA sequencing (RNA-seq) has recently become the method of choice to define and analyze transcriptomes. For the model moss Physcomitrella patens, although this method has been used to help analyze specific perturbations, no overall reference dataset has yet been established. In the framework of the Gene Atlas project, the Joint Genome Institute selected P. patens as a flagship genome, opening the way to generate the first comprehensive transcriptome dataset for this moss. The first round of sequencing described here is composed of 99 independent libraries spanning 34 different developmental stages and conditions. Upon dataset quality control and processing through read mapping, 28 509 of the 34 361 v3.3 gene models (83%) were detected to be expressed across the samples. Differentially expressed genes (DEGs) were calculated across the dataset to permit perturbation comparisons between conditions. The analysis of the three most distinct and abundant P. patens growth stages – protonema, gametophore and sporophyte – allowed us to define both general transcriptional patterns and stage-specific transcripts. As an example of variation of physico-chemical growth conditions, we detail here the impact of ammonium supplementation under standard growth conditions on the protonemal transcriptome. Finally, the cooperative nature of this project allowed us to analyze inter-laboratory variation, as 13 different laboratories around the world provided samples. We compare differences in the replication of experiments in a single laboratory and between different laboratories.

緑色植物のPSI-PSII複合体　　　　　　　　　　　　　　　
横野 牧生, 高林 厚史, 岸本 純子, 藤田 知道, 岩井 優和, MURAKAMI AKIO, AKIMOTO SEIJI, 田中 歩
光合成研究, 28, 1, 15, 19, Apr. 2018, [Peer-reviewed]
Japanese, Scientific journal

Hypergravity of 10g Changes Plant Growth, Anatomy, Chloroplast Size, and Photosynthesis in the Moss Physcomitrella patens
Kaori Takemura, Rina Watanabe, Ryuji Kameishi, Naoya Sakaguchi, Hiroyuki Kamachi, Atsushi Kume, Ichirou Karahara, Yuko T. Hanba, Tomomichi Fujita
MICROGRAVITY SCIENCE AND TECHNOLOGY, 29, 6, 467, 473, SPRINGER, Dec. 2017, [Peer-reviewed]
English, Scientific journal, The photosynthetic and anatomical responses of bryophytes to changes in gravity will provide crucial information for estimating how these plant traits evolved to adapt to changes in gravity in land plant history. We performed long-term hypergravity experiments at 10g for 4 and 8 weeks using the moss Physcomitrella patens with two centrifuges equipped with lighting systems that enable long-term plant growth under hypergravity with irradiance. The aims of this study are (1) to quantify changes in the anatomy and morphology of P. patens, and (2) to analyze the post-effects of hypergravity on photosynthesis by P. patens in relation to these changes. We measured photosynthesis by P. patens for a population of gametophores (e.g., canopy) in Petri dishes and plant culture boxes. Gametophore numbers increased by 9% for a canopy of P. patens, with 24-27% increases in chloroplast sizes (diameter and thickness) in leaf cells. In a canopy of P. patens, the area-based photosynthesis rate (A (canopy)) was increased by 57% at 10g. The increase observed in A (canopy) was associated with greater plant numbers and chloroplast sizes, both of which involved enhanced CO2 diffusion from the atmosphere to chloroplasts in the canopies of P. patens. These results suggest that changes in gravity are important environmental stimuli to induce changes in plant growth and photosynthesis by P. patens, in which an alteration in chloroplast size is one of the key traits. We are now planning an ISS experiment to investigate the responses of P. patens to microgravity.

Occurrence of brassinosteroids in non-flowering land plants, liverwort, moss, lycophyte and fern
Takao Yokota, Toshiyuki Ohnishi, Kyomi Shibata, Masashi Asahina, Takahito Nomura, Tomomichi Fujita, Kimitsune Ishizaki, Takayuki Kohchi
PHYTOCHEMISTRY, 136, 46, 55, PERGAMON-ELSEVIER SCIENCE LTD, Apr. 2017, [Peer-reviewed]
English, Scientific journal, Endogenous brassinosteroids (BRs) in non-flowering land plants were analyzed. BRs were found in a liverwort (Marchantia polymorpha), a moss (Physcomitrella patens), lycophytes (Selaginella moellendorffii and S. uncinata) and 13 fern species. A biologically active BR, castasterone (CS), was identified in most of these non-flowering plants but another biologically active BR, brassinolide, was not. It may be distinctive that levels of CS in non-flowering plants were orders of magnitude lower than those in flowering plants. 22-Hydroxycampesterol and its metabolites were identified in most of the non-flowering plants suggesting that the biosynthesis of BRs via 22-hydroxylation of campesterol occurs as in flowering plants. Phylogenetic analyses indicated that M. polymorpha, P. patens and S. moellendorffii have cytochrome P450s in the CYP85 clans which harbors BR biosynthesis enzymes, although the P450 profiles are simpler as compared with Arabidopsis and rice. Furthermore, these basal land plants were found to have multiple P450s in the CYP72 clan which harbors enzymes to catabolize BRs. These findings indicate that green plants were able to synthesize and inactivate BRs from the land-transition stage. (C) 2016 Elsevier Ltd. All rights reserved.

Genome of the pitcher plant Cephalotus reveals genetic changes associated with carnivory
Kenji Fukushima, Xiaodong Fang, David Alvarez-Ponce, Huimin Cai, Lorenzo Carretero-Paulet, Cui Chen, Tien-Hao Chang, Kimberly M. Farr, Tomomichi Fujita, Yuji Hiwatashi, Yoshikazu Hoshi, Takamasa Imai, Masahiro Kasahara, Pablo Librado, Likai Mao, Hitoshi Mori, Tomoaki Nishiyama, Masafumi Nozawa, Gergo Palfalvi, Stephen T. Pollard, Julio Rozas, Alejandro Sanchez-Gracia, David Sankoff, Tomoko F. Shibata, Shuji Shigenobu, Naomi Sumikawa, Taketoshi Uzawa, Meiying Xie, Chunfang Zheng, David D. Pollock, Victor A. Albert, Shuaicheng Li, Mitsuyasu Hasebe
NATURE ECOLOGY & EVOLUTION, 1, 3, 0059, NATURE PUBLISHING GROUP, Mar. 2017, [Peer-reviewed]
English, Scientific journal, Carnivorous plants exploit animals as a nutritional source and have inspired long-standing questions about the origin and evolution of carnivory-related traits. To investigate the molecular bases of carnivory, we sequenced the genome of the heterophyllous pitcher plant Cephalotus follicularis, in which we succeeded in regulating the developmental switch between carnivorous and non-carnivorous leaves. Transcriptome comparison of the two leaf types and gene repertoire analysis identified genetic changes associated with prey attraction, capture, digestion and nutrient absorption. Analysis of digestive fluid proteins from C. follicularis and three other carnivorous plants with independent carnivorous origins revealed repeated co-options of stress-responsive protein lineages coupled with convergent amino acid substitutions to acquire digestive physiology. These results imply constraints on the available routes to evolve plant carnivory.

A hypergravity environment increases chloroplast size, photosynthesis, and plant growth in the moss Physcomitrella patens
Kaori Takemura, Hiroyuki Kamachi, Atsushi Kume, Tomomichi Fujita, Ichirou Karahara, Yuko T. Hanba
JOURNAL OF PLANT RESEARCH, 130, 1, 181, 192, SPRINGER JAPAN KK, Jan. 2017, [Peer-reviewed]
English, Scientific journal, The physiological and anatomical responses of bryophytes to altered gravity conditions will provide crucial information for estimating how plant physiological traits have evolved to adapt to significant increases in the effects of gravity in land plant history. We quantified changes in plant growth and photosynthesis in the model plant of mosses, Physcomitrella patens, grown under a hypergravity environment for 25 days or 8 weeks using a custom-built centrifuge equipped with a lighting system. This is the first study to examine the response of bryophytes to hypergravity conditions. Canopy-based plant growth was significantly increased at 10xg, and was strongly affected by increases in plant numbers. Rhizoid lengths for individual gametophores were significantly increased at 10xg. Chloroplast diameters (major axis) and thicknesses (minor axis) in the leaves of P. patens were also increased at 10xg. The area-based photosynthesis rate of P. patens was also enhanced at 10xg. Increases in shoot numbers and chloroplast sizes may elevate the area-based photosynthesis rate under hypergravity conditions. We observed a decrease in leaf cell wall thickness under hypergravity conditions, which is in contrast to previous findings obtained using angiosperms. Since mosses including P. patens live in dense populations, an increase in canopy-based plant numbers may be effective to enhance the toughness of the population, and, thus, represents an effective adaptation strategy to a hypergravity environment for P. patens.

PCoM-DB Update: A Protein Co-Migration Database for Photosynthetic Organisms
Atsushi Takabayashi, Saeka Takabayashi, Kaori Takahashi, Mai Watanabe, Hiroko Uchida, Akio Murakami, Tomomichi Fujita, Masahiko Ikeuchi, Ayumi Tanaka
PLANT AND CELL PHYSIOLOGY, 58, 1, OXFORD UNIV PRESS, Jan. 2017, [Peer-reviewed]
English, Scientific journal, The identification of protein complexes is important for the understanding of protein structure and function and the regulation of cellular processes. We used blue-native PAGE and tandemmass spectrometry to identify protein complexes systematically, and built a web database, the protein co-migration database (PCoM-DB, http://pcomdb.lowtem.hokudai.ac.jp/proteins/top), to provide prediction tools for protein complexes. PCoM-DB provides migration profiles for any given protein of interest, and allows users to compare them with migration profiles of other proteins, showing the oligomeric states of proteins and thus identifying potential interaction partners. The initial version of PCoM-DB (launched in January 2013) included protein complex data for Synechocystis whole cells and Arabidopsis thaliana thylakoid membranes. Here we report PCoM-DB version 2.0, which includes new data sets and analytical tools. Additional data are included from whole cells of the pelagic marine picocya-nobacterium Prochlorococcus marinus, the thermophilic cyanobacterium Thermosynechococcus elongatus, the unicellular green alga Chlamydomonas reinhardtii and the bryophyte Physcomitrella patens. The Arabidopsis protein data now include data for intact mitochondria, intact chloroplasts, chloroplast stroma and chloroplast envelopes. The new tools comprise a multiple-protein search form and a heat map viewer for protein migration profiles. Users can compare migration profiles of a protein of interest among different organelles or compare migration profiles among different proteins within the same sample. For Arabidopsis proteins, users can compare migration profiles of a protein of interest with putative homologous proteins from non-Arabidopsis organisms. The updated PCoM-DB will help researchers find novel protein complexes and estimate their evolutionary changes in the green lineage.

Comparisons of the effects of vibration of two centrifugal systems on the growth and morphological parameters of the moss Physcomitrella patens
Mori, A, Kamachi, H, Karahara, I, Kume, A, Hanba, Y.-T, Takemura, K, Fujita, T
Biological Sciences in Space, 31, 9, 13, 2017, [Peer-reviewed]

Phototropism in gametophytic shoots of the moss Physcomitrella patens
Liang Bao, Kotaro T. Yamamoto, Tomomichi Fujita
Plant Signaling and Behavior, 10, 3, Taylor and Francis Inc., 07 Apr. 2015, [Peer-reviewed]
English, Scientific journal, Shoot phototropism enables plants to position their photosynthetic organs in favorable light conditions and thus benefits growth and metabolism in land plants. To understand the evolution of this response, we established an experimental system to study phototropism in gametophores of the moss Physcomitrella patens. The phototropic response of gametophores occurs slowly
a clear response takes place more than 24 hours after the onset of unilateral light irradiation, likely due to the slow growth rate of gametophores. We also found that red and far-red light can induce phototropism, with blue light being less effective. These results suggest that plants used a broad range of light wavelengths as phototropic signals during the early evolution of land plants.

A model system for analyzing intercellular communication through plasmodesmata using moss protonemata and leaves
Munenori Kitagawa, Tomomichi Fujita
JOURNAL OF PLANT RESEARCH, 128, 1, 63, 72, SPRINGER JAPAN KK, Jan. 2015, [Peer-reviewed]
English, Plant growth, development, and environmental responses require the proper regulation of intercellular movement of signals and nutrients. For this, plants have specialized cytoplasmic channels, the plasmodesmata (PD), which allow the symplasmic movement of micro- and macromolecules between neighboring cells. Internal and external signals spatio-temporally regulate the movement of molecules through the PD to control plant development and environmental responses. Although some aspects of targeted movement of molecules have been revealed, the mechanisms of non-targeted, diffusible flow of molecules through PD, and its regulation and function, remain poorly understood, particularly at the cellular level. Previously, we developed a system to quantitatively analyze non-targeted movement of a photoconvertible fluorescent protein, Dendra2, at the single-cell level in the filamentous protonemata tissue of the moss Physcomitrella patens. In protonemata, one-dimensional intercellular communication can be easily observed and quantitatively analyzed at the cellular level. In this review, we describe how protonemata and leaves of P. patens can be used to study symplasmic movement through PD, and discuss how this system can help improve our understanding of PD regulation and function in development and environmental responses in plants.

Plasmodesmata: function and diversity in plant intercellular communication
Tomomichi Fujita
JOURNAL OF PLANT RESEARCH, 128, 1, 3, 5, SPRINGER JAPAN KK, Jan. 2015, [Peer-reviewed]
English

ヒメツリガネゴケにおけるアブシジン酸処理時の光合成の変化
藤田拓矢, 高林厚史, 田中歩, 藤田知道
せん苔類研究, 11, 3, 92, Dec. 2014
Japanese

Conserved function of Rho-related Rop/RAC GTPase signaling in regulation of cell polarity in Physcomitrella patens
Kanako Ito, Junling Ren, Tomomichi Fujita
GENE, 544, 2, 241, 247, ELSEVIER SCIENCE BV, Jul. 2014, [Peer-reviewed]
English, Scientific journal, Cell polarity is fundamentally important to growth and development in higher plants, from pollen tubes to root hairs. Basal land plants (mosses and ferns) also have cell polarity, developing protonemal apical cells that show polar tip growth. Flowering plants have a distinct group of Rho GTPases that regulate polarity in polarized cell growth. Rop/RAC signaling module components have been identified in non-flowering plants, but their roles remain unclear. To understand the importance and evolution of Rop/RAC signaling in polarity regulation in land plants, we examined the functions of PpRop and PpRopGEF in protonemal apical cells of the moss Physcomitrella patens. Inducible overexpression of PpRop2 or PpRopGEF3 caused depolarized growth of tip-growing apical cells. PpRop2 overexpression also caused aberrant cross wall formation. Fluorescent protein-tagged PpRop2 localized to the plasma membrane, including the cross wall membrane, and fluorescent-tagged PpRopGEF3 showed polarized localization to the tip region in apical cells. Thus, our results suggest common functions of PpRop and PpRopGEF in the tip-growing apical cells and the importance of a conserved Rop/RAC signaling module in the control of cell polarity in land plants. (C) 2014 Elsevier B.V. All rights reserved.

Quantitative imaging of directional transport through plasmodesmata in moss protonemata via single-cell photoconversion of Dendra2
Munenori Kitagawa, Tomomichi Fujita
JOURNAL OF PLANT RESEARCH, 126, 4, 577, 585, SPRINGER JAPAN KK, Jul. 2013, [Peer-reviewed]
English, Scientific journal, Cell-to-cell transport of molecules in plants must be properly regulated for plant growth and development. One specialized mechanism that plants have evolved involves transport through plasmodesmata (PD), but when and how transport of molecules via PD is regulated among individual cells remains largely unknown, particularly at the single-cell level. Here, we developed a tool for quantitatively analyzing cell-to-cell transport via PD at a single-cell level using protonemata of Physcomitrella patens and a photoconvertible fluorescent protein, Dendra2. In the filamentous protonemal tissues, one-dimensional intercellular communication can be observed easily. Using this system, we found that Dendra2 was directionally transported toward the apex of the growing protonemata. However, this directional transport could be eliminated by incubation in the dark or treatment with a metabolic inhibitor. Thus, we propose that directional transport of macromolecules can occur via PD in moss protonemata, and may be affected by the photosynthetic and metabolic activity of cells.

The Selaginella Genome Identifies Genetic Changes Associated with the Evolution of Vascular Plants
Jo Ann Banks, Tomoaki Nishiyama, Mitsuyasu Hasebe, John L. Bowman, Michael Gribskov, Claude dePamphilis, Victor A. Albert, Naoki Aono, Tsuyoshi Aoyama, Barbara A. Ambrose, Neil W. Ashton, Michael J. Axtell, Elizabeth Barker, Michael S. Barker, Jeffrey L. Bennetzen, Nicholas D. Bonawitz, Clint Chapple, Chaoyang Cheng, Luiz Gustavo Guedes Correa, Michael Dacre, Jeremy DeBarry, Ingo Dreyer, Marek Elias, Eric M. Engstrom, Mark Estelle, Liang Feng, Cedric Finet, Sandra K. Floyd, Wolf B. Frommer, Tomomichi Fujita, Lydia Gramzow, Michael Gutensohn, Jesper Harholt, Mitsuru Hattori, Alexander Heyl, Tadayoshi Hirai, Yuji Hiwatashi, Masaki Ishikawa, Mineko Iwata, Kenneth G. Karol, Barbara Koehler, Uener Kolukisaoglu, Minoru Kubo, Tetsuya Kurata, Sylvie Lalonde, Kejie Li, Ying Li, Amy Litt, Eric Lyons, Gerard Manning, Takeshi Maruyama, Todd P. Michael, Koji Mikami, Saori Miyazaki, Shin-ichi Morinaga, Takashi Murata, Bernd Mueller-Roeber, David R. Nelson, Mari Obara, Yasuko Oguri, Richard G. Olmstead, Naoko Onodera, Bent Larsen Petersen, Birgit Pils, Michael Prigge, Stefan A. Rensing, Diego Mauricio Riano-Pachon, Alison W. Roberts, Yoshikatsu Sato, Henrik Vibe Scheller, Burkhard Schulz, Christian Schulz, Eugene V. Shakirov, Nakako Shibagaki, Naoki Shinohara, Dorothy E. Shippen, Iben Sorensen, Ryo Sotooka, Nagisa Sugimoto, Mamoru Sugita, Naomi Sumikawa, Milos Tanurdzic, Guenter Theissen, Peter Ulvskov, Sachiko Wakazuki, Jing-Ke Weng, William W. G. T. Willats, Daniel Wipf, Paul G. Wolf, Lixing Yang, Andreas D. Zimmer, Qihui Zhu, Therese Mitros, Uffe Hellsten, Dominique Loque, Robert Otillar, Asaf Salamov, Jeremy Schmutz, Harris Shapiro, Erika Lindquist, Susan Lucas, Daniel Rokhsar, Igor V. Grigoriev
SCIENCE, 332, 6032, 960, 963, AMER ASSOC ADVANCEMENT SCIENCE, May 2011, [Peer-reviewed]
English, Scientific journal, Vascular plants appeared similar to 410 million years ago, then diverged into several lineages of which only two survive: the euphyllophytes (ferns and seed plants) and the lycophytes. We report here the genome sequence of the lycophyte Selaginella moellendorffii (Selaginella), the first nonseed vascular plant genome reported. By comparing gene content in evolutionarily diverse taxa, we found that the transition from a gametophyte- to a sporophyte-dominated life cycle required far fewer new genes than the transition from a nonseed vascular to a flowering plant, whereas secondary metabolic genes expanded extensively and in parallel in the lycophyte and angiosperm lineages. Selaginella differs in posttranscriptional gene regulation, including small RNA regulation of repetitive elements, an absence of the trans-acting small interfering RNA pathway, and extensive RNA editing of organellar genes.

Microtubules Regulate Dynamic Organization of Vacuoles in Physcomitrella patens
Yoshihisa Oda, Aiko Hirata, Toshio Sano, Tomomichi Fujita, Yuji Hiwatashi, Yoshikatsu Sato, Akeo Kadota, Mitsuyasu Hasebe, Seiichiro Hasezawa
PLANT AND CELL PHYSIOLOGY, 50, 4, 855, 868, OXFORD UNIV PRESS, Apr. 2009, [Peer-reviewed]
English, Scientific journal, Eukaryotic cells have developed several essential membrane components. In flowering plants, appropriate structures and distributions of the major membrane components are predominantly regulated by actin microfilaments. In this study, we have focused on the regulatory mechanism of vacuolar structures in the moss, Physcomitrella patens. The high ability of P. patens to undergo homologous recombination enabled us stably to express green fluorescent protein (GFP) or red fluorescent protein (RFP) fusion proteins, and the simple body structure of P. patens enabled us to perform detailed visualization of the intracellular vacuolar and cytoskeletal structures. Three-dimensional analysis and high-speed time-lapse observations revealed surprisingly complex structures and dynamics of the vacuole, with inner sheets and tubular protrusions, and frequent rearrangements by separation and fusion of the membranes. Depolymerization of microtubules dramatically affected these structures and movements. Dual observation of microtubules and vacuolar membranes revealed that microtubules induced tubular protrusions and cytoplasmic strands of the vacuoles, indicative of interactions between microtubules and vacuolar membranes. These results demonstrate a novel function of microtubules in maintaining the distribution of the vacuole and suggest a functional divergence of cytoskeletal functions in land plant evolution.

Convergences and divergences in polar auxin transport and shoot development in land plant evolution　　　　　　　　　　　　　　　
Fujita, T, Hasebe, M
Plant Signaling & Behavior, 4, 313, 315, 2009, [Peer-reviewed]

Kinesins Are Indispensable for Interdigitation of Phragmoplast Microtubules in the Moss Physcomitrella patens
Yuji Hiwatashi, Mari Obara, Yoshikatsu Sato, Tomomichi Fujita, Takashi Murata, Mitsuyasu Hasebe
PLANT CELL, 20, 11, 3094, 3106, AMER SOC PLANT BIOLOGISTS, Nov. 2008, [Peer-reviewed]
English, Scientific journal, Microtubules form arrays with parallel and antiparallel bundles and function in various cellular processes, including subcellular transport and cell division. The antiparallel bundles in phragmoplasts, plant-unique microtubule arrays, are mostly unexplored and potentially offer new cellular insights. Here, we report that the Physcomitrella patens kinesins KINID1a and KINID1b (for kinesin for interdigitated microtubules 1a and 1b), which are specific to land plants and orthologous to Arabidopsis thaliana PAKRP2, are novel factors indispensable for the generation of interdigitated antiparallel microtubules in the phragmoplasts of the moss P. patens. KINID1a and KINID1b are predominantly localized to the putative interdigitated parts of antiparallel microtubules. This interdigitation disappeared in double-deletion mutants of both genes, indicating that both KINID1a and 1b are indispensable for interdigitation of the antiparallel microtubule array. Furthermore, cell plates formed by these phragmoplasts did not reach the plasma membrane in; 20% of the mutant cells examined. We observed that in the double-deletion mutant lines, chloroplasts remained between the plasma membrane and the expanding margins of the cell plate, while chloroplasts were absent from the margins of the cell plates in the wild type. This suggests that the kinesins, the antiparallel microtubule bundles with interdigitation, or both are necessary for proper progression of cell wall expansion.

Expression and complementation analyses of a chloroplast-localized homolog of bacterial RecA in the moss Physcomitrella patens
Takayuki Inouye, Masaki Odahara, Tomomichi Fujita, Mitsuyasu Hasebe, Yasuhiko Sekine
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 72, 5, 1340, 1347, TAYLOR & FRANCIS LTD, May 2008, [Peer-reviewed]
English, Scientific journal, RecA protein is widespread in bacteria, and it plays a crucial role in homologous recombination. We have identified two bacterial-type recA gene homologs (PprecA1, PprecA2) in the cDNA library of the moss Physcomitrella patens. N-terminal fusion of the putative organellar targeting sequence of PpRecA2 to the green fluorescent protein (GFP) caused a targeting of PpRecA2 to the chloroplasts. Mutational analysis showed that the first AUG codon acts as initiation codon. Fusion of the full-length PpRecA2 to GFP caused the formation of foci that were colocalized with chloroplast nucleoids. The amounts of PprecA2 mRNA and protein in the cells were increased by treatment, with DNA damaging agents. PprecA2 partially complemented the recA mutation in Escherichia coli. These results suggest the involvement of PpRecA2 in the repair of chloroplast DNA.

Convergent evolution of shoots in land plants: lack of auxin polar transport in moss shoots
Tomomichi Fujita, Hisako Sakaguchi, Yuji Hiwatashi, Steven J. Wagstaff, Motomi Ito, Hironori Deguchi, Toshiyuki Sato, Mitsuyasu Hasebe
EVOLUTION & DEVELOPMENT, 10, 2, 176, 186, BLACKWELL PUBLISHING, Mar. 2008, [Peer-reviewed]
English, Scientific journal, The shoot is a repeated structure made up of stems and leaves and is the basic body plan in land plants. Vascular plants form a shoot in the diploid generation, whereas nonvascular plants such as mosses form a shoot in the haploid generation. It is not clear whether all land plants use similar molecular mechanisms in shoot development or how the genetic networks for shoot development evolved. The control of auxin distribution, especially by polar auxin transport, is essential for shoot development in flowering plants. We did not detect polar auxin transport in the gametophytic shoots of several mosses, but did detect it in the sporophytes of mosses without shoot structure. Treatment with auxin transport inhibitors resulted in abnormal embryo development, as in flowering plants, but did not cause any morphological changes in the haploid shoots. We fused the soybean auxin-inducible promoter GH3 with a GUS reporter gene and used it to indirectly detect auxin distribution in the moss Physcomitrella patens. An auxin transport inhibitor NPA did not cause any changes in the putative distribution of auxin in the haploid shoot. These results indicate that polar auxin transport is not involved in haploid shoot development in mosses and that shoots in vascular plants and mosses are most likely regulated differently during development.

The Physcomitrella genome reveals evolutionary insights into the conquest of land by plants
Stefan A. Rensing, Daniel Lang, Andreas D. Zimmer, Astrid Terry, Asaf Salamov, Harris Shapiro, Tomoaki Nishiyama, Pierre-Francois Perroud, Erika A. Lindquist, Yasuko Kamisugi, Takako Tanahashi, Keiko Sakakibara, Tomomichi Fujita, Kazuko Oishi, Tadasu Shin-I, Yoko Kuroki, Atsushi Toyoda, Yutaka Suzuki, Shin-ichi Hashimoto, Kazuo Yamaguchi, Sumio Sugano, Yuji Kohara, Asao Fujiyama, Aldwin Anterola, Setsuyuki Aoki, Neil Ashton, W. Brad Barbazuk, Elizabeth Barker, Jeffrey L. Bennetzen, Robert Blankenship, Sung Hyun Cho, Susan K. Dutcher, Mark Estelle, Jeffrey A. Fawcett, Heidrun Gundlach, Kousuke Hanada, Alexander Heyl, Karen A. Hicks, Jon Hughes, Martin Lohr, Klaus Mayer, Alexander Melkozernov, Takashi Murata, David R. Nelson, Birgit Pils, Michael Prigge, Bernd Reiss, Tanya Renner, Stephane Rombauts, Paul J. Rushton, Anton Sanderfoot, Gabriele Schween, Shin-Han Shiu, Kurt Stueber, Frederica L. Theodoulou, Hank Tu, Yves Van de Peer, Paul J. Verrier, Elizabeth Waters, Andrew Wood, Lixing Yang, David Cove, Andrew C. Cuming, Mitsuyasu Hasebe, Susan Lucas, Brent D. Mishler, Ralf Reski, Igor V. Grigoriev, Ralph S. Quatrano, Jeffrey L. Boore
SCIENCE, 319, 5859, 64, 69, AMER ASSOC ADVANCEMENT SCIENCE, Jan. 2008, [Peer-reviewed]
English, Scientific journal, We report the draft genome sequence of the model moss Physcomitrella patens and compare its features with those of flowering plants, from which it is separated by more than 400 million years, and unicellular aquatic algae. This comparison reveals genomic changes concomitant with the evolutionary movement to land, including a general increase in gene family complexity; loss of genes associated with aquatic environments ( e. g., flagellar arms); acquisition of genes for tolerating terrestrial stresses ( e. g., variation in temperature and water availability); and the development of the auxin and abscisic acid signaling pathways for coordinating multicellular growth and dehydration response. The Physcomitrella genome provides a resource for phylogenetic inferences about gene function and for experimental analysis of plant processes through this plant's unique facility for reverse genetics.

Involvement of mitochondrial-targeted RecA in the repair of mitochondrial DNA in the moss, Physcomitrella patens
Masaki Odahara, Takayuki Inouye, Tomomichi Fujita, Mitsuyasu Hasebe, Yasuhiko Sekine
GENES & GENETIC SYSTEMS, 82, 1, 43, 51, GENETICS SOC JAPAN, Feb. 2007, [Peer-reviewed]
English, Scientific journal, Homologous recombination is a universal process that contributes to genetic diversity and genomic integrity. Bacterial-type RecA generally exists in all bacteria and plays a crucial role in homologous recombination. Although RecA homologues also exist in plant mitochondria, there have been few reports about the in vivo functions of these homologues. We identified a recA gene orthologue (named PprecA1) in a cDNA library of the moss, Physcomitrella patens. N-terminal fusion of the putative organellar targeting sequence of PpRecA1 to GFP caused a targeting of PpRecA1 to mitochondria. PprecA1 partially complemented the effects of a DNA damaging agent in an Escherichia coli recA deficient strain. Additionally, the expression of PprecA1 was induced by treating the plants with DNA damaging agents. Disruption of PprecA1 by targeted replacement resulted lower rate of the recovery of the mitochondrial DNA from methyl methan sulfonate damage. This is the first report about the characteristics of a null mutant of bacterial-type recA gene in plant. The data suggest that PprecA1 participates in the repair of mitochondrial DNA in P. patens.

Microtubule dependent regulation of vacuolar morphogenesis in the moss, Physcomitrella patens
Yoshihisa Oda, Toshio Sano, Tomomichi Fujita, Yuji Hiwatashi, Yoshikatsu Sato, Natsumaro Kutsuna, Aiko Hirata, Mitsuyasu Hasebe, Seiichiro Hasezawa
PLANT AND CELL PHYSIOLOGY, 48, S24, S24, OXFORD UNIV PRESS, 2007, [Peer-reviewed]
English

Identification of proteins which accumulated preferentially in stem cells of Physcomitrella patens
Tomomichi Fujita, Kaoru Hashimoto, Yuji Hiwatashi, Yoshikatsu Sato, Takashi Murata, Mitsuyasu Hasebe
PLANT AND CELL PHYSIOLOGY, 48, S50, S50, OXFORD UNIV PRESS, 2007, [Peer-reviewed]
English

Molecular mechanism of type II ubiquitin-like proteins, PUBL1 and PUBL2, which are involved in cytokinesis, in the moss Physcomitrella patens.
Yuji Hiwatashi, Tomomichi Fujita, Takashi Murata, Mitsuyasu Hasebe
PLANT AND CELL PHYSIOLOGY, 48, S88, S88, OXFORD UNIV PRESS, 2007, [Peer-reviewed]
English

Genome-wide comparison of developmental genes in land plants
Mitsuyasu Hasebe, Tomoaki Nishiyama, Takako Tanahashi, Naoki Aono, Tsuyoshi Aoyama, Chaoyang Cheng, Tomomichi Fujita, Kaoru Hashimoto, Tadayoshi Hirai, Yuji Hiwatashi, Masaki Ishikawa, Mineko Iwata, Minoru Kubo, Tetsuya Kurata, Koij Mikami, Saori Miyazaki, Shin-Ichi Morinaga, Takashi Murata, Mari Obara, Yasuko Oguri, Naoko Onodera, Yoshikatsu Sato, Naomi Sumikawa, Naoki Shinohara, Sachiko Wakaduki, Nagisa Sugimoto
PLANT AND CELL PHYSIOLOGY, 48, S50, S50, OXFORD UNIV PRESS, 2007, [Peer-reviewed]
English

Oncogene 6b from Agrobacterium tumefaciens induces abaxial cell division at late stages of leaf development and modifies vascular development in petioles.
Terakura S, Kitakura S, Ishikawa M, Ueno Y, Fujita T, Machida C, Wabiko H, Machida Y
Plant & cell physiology, 47, 5, 664, 672, May 2006, [Peer-reviewed], [Domestic magazines]
English, Scientific journal, The 6b gene in the T-DNA region of the Ti plasmids of Agrobacterium tumefaciens and A. vitis is able to generate shooty calli in phytohormone-free culture of leaf sections of tobacco transformed with 6b. In the present study, we report characteristic morphological abnormalities of the leaves of transgenic tobacco and Arabidopsis that express 6b from pTiAKE10 (AK-6b), and altered expression of genes related to cell division and meristem formation in the transgenic plants. Cotyledons and leaves of both transgenic tobacco and Arabidopsis exhibited various abnormalities including upward curling of leaf blades, and transgenic tobacco leaves produced leaf-like outgrowths from the abaxial side. Transcripts of some class 1 KNOX homeobox genes, which are thought to be related to meristem functions, and cell cycle regulating genes were ectopically accumulated in mature leaves. M phase-specific genes were also ectopically expressed at the abaxial sides of mature leaves. These results suggest that the AK-6b gene stimulates the cellular potential for division and meristematic functions preferentially in the abaxial side of leaves and that the leaf phenotypes generated by AK-6b are at least in part due to such biased cell division during polar development of leaves. The results of the present experiments with a fusion gene between the AK-6b gene and the glucocorticoid receptor gene showed that nuclear import of the AK-6b protein was essential for upward curling of leaves and hormone-free callus formation, suggesting a role for AK-6b in nuclear events.

Genes for the peptidoglycan synthesis pathway are essential for chloroplast division in moss.
Machida M, Takechi K, Sato H, Chung SJ, Kuroiwa H, Takio S, Seki M, Shinozaki K, Fujita T, Hasebe M, Takano H
Proceedings of the National Academy of Sciences of the United States of America, 103, 17, 6753, 6758, Apr. 2006, [Peer-reviewed], [International Magazine]
English, Scientific journal, The general consensus is that a cyanobacterium phagocytosed by a host cell evolved into the plastids of red and green algae, land plants, and glaucophytes. In contrast to the plastids of glaucophytes, which retain a cyanobacterial-type peptidoglycan layer, no wall-like structures have been detected in plastids from other sources. Although the genome of Arabidopsis thaliana contains five genes that are essential for peptidoglycan synthesis, MurE, MurG, two genes for D-Ala-D-Ala ligase (Ddl), and the gene for translocase I (MraY), their functions have not been determined. We report that the moss Physcomitrella patens has nine homologous genes related to peptidoglycan biosynthesis: MurA, B, C, D, E, and F, Ddl, genes for the penicillin-binding protein Pbp, and dd-carboxypeptidase (Dac). Corroborating a computer prediction, analysis of the GFP fusion proteins with the N terminus of PpMurE or of PpPbp suggests that these proteins are located in the chloroplasts. Gene disruption of the PpMurE gene in P. patens resulted in the appearance of macrochloroplasts both in protonema and in leaf cells. Moreover, gene knockout of the P. patens Pbp gene showed inhibition of chloroplast division in this moss; however, no Pbp gene was found in A. thaliana.

P. patens protoplast asymmetric cell division : from cell polarity to cell fate determination in plant stem cells
Fujita Tomomichi
Regulation of Plant Growth & Development, 41, 2, 156, 162, The Japanese Society for Chemical Regulation of Plants, 2006, [Corresponding author]
Japanese

Isolation of mutant lines with decreased numbers of chloroplasts per cell from a tagged mutant library of the moss Physcomitrella patens
A Hayashida, K Takechi, M Sugiyama, M Kubo, RD Itoh, S Takio, T Fujita, Y Hiwatashi, M Hasebe, H Takano
PLANT BIOLOGY, 7, 3, 300, 306, GEORG THIEME VERLAG KG, May 2005, [Peer-reviewed]
English, Scientific journal, Eleven mutant lines exhibiting decreased numbers of chloroplasts per cell were isolated from 8800 tagged mutant lines of Physcomitrella patens by microscopic observations. Chloronema subapical cells in wild-type plants had a mean of 48 chloroplasts, whereas chloroplast numbers in subapical cells in mutant lines 215 and 222 decreased to 75% of that in the wild type. Seven mutant lines - 473, 122, 221, 129, 492, 207, and 138 had about half as many chloroplasts as the wild type. Mutant line 11 had a few remarkably enlarged chloroplasts, and mutant line 347 had chloroplasts of various sizes. Whereas the cell volume was the same as in the wild type in mutant lines 222, 473, 221, 129, 492, and 207, the cell volume of the other mutants increased. The chloroplast number of leaf cells was the same as that of chloronema cells in each mutant line when gametophores could be formed. Treatment with ampicillin decreased the number of chloroplasts in all mutant lines. Southern hybridization using DNA in tags as probes showed that only one insertion occurred in mutant lines 473 and 221. To determine whether the tagged DNA inserted into the known genes for plastid division, we isolated the PpMinD1, PpMinD2, and PPMinE1 genes. Genomic polymerase chain reaction analysis showed that the PpFtsZ and PpMinD/E genes were not disrupted by the insertion of the tags in mutant lines 11 and 347, respectively.

Monoclonal antibodies against regions topologically surrounding the homodimeric β-barrel interface of Epstein-Barr virus nuclear antigen-1
Hiroyuki Eda, Yasuyuki Ishii, Maya Obayashi, Shizuko Harada, Sayuri Ito, Tomomichi Fujita, Masato Ikeda, Shuichi Kusano, Ryo Kitamura, Chieko Suzuki, Takahiko Hara, Motoo Watanabe, Hiroshi Satoh, Keisuke Sugihara, Kazuo Yanagi
Virus Research, 109, 1, 87, 94, Elsevier BV, Apr. 2005
Scientific journal

Gene tagging, gene- and enhancer-trap systems, and full-length cDNA overexpression in Physcomitrella patens.　　　　　　　　　　　　　　　
Fujita, T., Nishiyama, T., Hiwatashi, Y., and Hasebe, M.
In New Frontiers in Bryology:Physiology, Molecular Biology & Functional Genomics (eds. by Wood, AJ., Oliver, MJ. and Cove, DJ.), Kluwer Academ・・・, 2004
Gene tagging, gene- and enhancer-trap systems, and full-length cDNA overexpression in Physcomitrella patens. In New Frontiers in Bryology:Physiology, Molecular Biology & Functional Genomics (eds. by Wood, AJ., Oliver, MJ. and Cove, DJ.), Kluwer Academic Publishers, Netherlands, pp. 111-132.

Comparative genomics of Physcomitrella patens gametophytic transcriptome and Arabidopsis thaliana: Implication for land plant evolution
T Nishiyama, T Fujita, T Shin-I, M Seki, H Nishide, Uchiyama, I, A Kamiya, P Carninci, Y Hayashizaki, K Shinozaki, Y Kohara, M Hasebe
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 100, 13, 8007, 8012, NATL ACAD SCIENCES, Jun. 2003, [Peer-reviewed]
English, Scientific journal, The mosses and flowering plants diverged >400 million years ago. The mosses have haploid-dominant life cycles, whereas the flowering plants are diploid-dominant. The common ancestors of land plants have been inferred to be haploid-dominant, suggesting that genes used in the diploid body of flowering plants were recruited from the genes used in the haploid body of the ancestors during the evolution of land plants. To assess this evolutionary hypothesis, we constructed an EST library of the moss Physcomitrella patens, and compared the moss transcriptome to the genome of Arabidopsis thaliana. We constructed full-length enriched cDNA libraries from auxin-treated, cytokinin-treated, and untreated gametophytes of P. patens, and sequenced both ends of >40,000 clones. These data, together with the mRNA sequences in the public databases, were assembled into 15,883 putative transcripts. Sequence comparisons of A. thaliana and P. patens showed that at least 66% of the A. thaliana genes had homologues in A patens. Comparison of the A patens putative transcripts with all known proteins, revealed 9,907 putative transcripts with high levels of similarity to vascular plant genes, and 850 putative transcripts with high levels of similarity to other organisms. The haploid transcriptome of A patens appears to be quite similar to the A. thaliana genome, supporting the evolutionary hypothesis. Our study also revealed that a number of genes are moss specific and were lost in the flowering plant lineage.

Identification of Regions of the Tomato γ-Glutamyl Kinase That Are Involved in Allosteric Regulation by Proline
Tomomichi Fujita, Albino Maggio, Mario Garcı́a-Rı́os, Cynthia Stauffacher, Ray A. Bressan, Laszlo N. Csonka
Journal of Biological Chemistry, 278, 16, 14203, 14210, American Society for Biochemistry & Molecular Biology (ASBMB), 18 Apr. 2003
Scientific journal

Does proline accumulation play an active role in stress-induced growth reduction?
Maggio A, Miyazaki S, Veronese P, Fujita T, Ibeas JI, Damsz B, Narasimhan ML, Hasegawa PM, Joly RJ, Bressan RA
PLANT JOURNAL, 31, 6, 699, 712, Sep. 2002, [Peer-reviewed]

The protein encoded by oncogene 6b from Agrobacterium tumefaciens interacts with a nuclear protein of tobacco
S Kitakura, T Fujita, Y Ueno, S Terakura, H Wabiko, Y Machida
PLANT CELL, 14, 2, 451, 463, AMER SOC PLANT BIOLOGISTS, Feb. 2002
English, Scientific journal, The 6b gene in the T-DNA from Agrobacterium has oncogenic activity in plant cells, inducing tumor formation, the phytohormone-independent division of cells, and alterations in leaf morphology. The product of the 6b gene appears to promote some aspects of the proliferation of plant cells, but the molecullar mechanism of its action remains unknown. We report here that the 6b protein associates with a nuclear protein in tobacco that we have designated NtSI1 (for Nicotiana tabacum 6b-interacting protein 1). NtSIP1 appears to be a transcription factor because its predicted amino acid sequence includes two regions that resemble a nuclear localization signal and a putative DNA binding motif, which is similar in terms of amino acid sequence to the triple helix motif of rice transcription factor GT-2. Expression in tobacco cells of a fusion protein composed of the DNA binding domain of the yeast GAL4 protein and the 6b protein activated the transcription of a reporter gene that was under the control of a chimeric promoter that included the GAL4 upstream activating sequence and the 35S minimal promoter of Cauliflower mosaic virus. Furthermore, nuclear localization of green fluorescent protein-fused 6b protein was enhanced by NtSIP1. A cluster of acidic residues in the 6b protein appeared to be essential for nuclear localization and for transactivation as well as for the hormone-independent growth of tobacco cells. Thus, it seems possible that the 6b protein might function in the proliferation of plant cells, at least in part, through an association with NtSIP1.

Establishment of gene-trap and enhancer-trap systems in the moss Physcomitrella patens
Y Hiwatashi, T Nishiyama, T Fujita, M Hasebe
PLANT JOURNAL, 28, 1, 105, 116, BLACKWELL SCIENCE LTD, Oct. 2001, [Peer-reviewed]
English, Scientific journal, Because of its simple body plan and ease of gene knockout and allele replacement, the moss Physcomitrella patens is often used as a model system for studies in plant physiology and developmental biology. Gene-trap and enhancer-trap systems are useful techniques for cloning genes and enhancers that function in specific tissues or cells. Additionally, these systems are convenient for obtaining molecular markers specific for certain developmental processes. Elements for gene-trap and enhancer-trap systems were constructed using the uidA reporter gene with either a splice acceptor or a minimal promoter. Through a high rate of transformation conferred by a method utilizing homologous recombination, 235 gene-trap and 1073 enhancer-trap lines were obtained from 5637 and 3726 transgenic lines, respectively. The expression patterns of these trap lines in the moss gametophyte varied. The candidate gene trapped in a gene-trap line YH209, which shows rhizoid-specific expression, was obtained by 5' and 3' RACE. This gene was named PpGLU, and forms a clade with plant acidic alpha -glucosidase genes. Thus, these gene-trap and enhancer-trap systems should prove useful to identify tissue- and cell-specific genes in Physcomitrella.

第17回国際植物生長物質会議に参加して
岩井 美穂, 斎藤 茂樹, CHON Nguyen Minh, 桑原 明日香, 原田 英美子, 朝比奈 雅志, 小田 篤, DA SILVA Jaime A. T., 野村 崇人, 関 原明, 藤田 知道
植物の生長調節, 36, 2, 222, 228, 一般社団法人植物化学調節学会, 2001
Japanese

Amino Acid Substitution Analyses of the DNA Contact Region, Two Amphipathic α-Helices and a Recognition-Helix-Like Helix outside the Dimeric β-Barrel of Epstein-Barr Virus Nuclear Antigen 1
Tomomichi Fujita, Masato Ikeda, Shuichi Kusano, Makoto Yamazaki, Sayuri Ito, Maya Obayashi, Kazuo Yanagi
Intervirology, 44, 5, 271, 282, S. Karger AG, 2001
Scientific journal

An improved RNA isolation method for plant tissues containing high levels of phenolic compounds or carbohydrates
R.A. Salzman, T. Fujita, K. Zhu-Salzman, P.M. Hasegawa, R.A. Bressan
Plant Molecular Biology Reporter, 17, 1, 11, 17, Springer Science and Business Media LLC, 1999
Scientific journal

Comparative Analysis of the Regulation of Expression and Structures of Two Evolutionarily Divergent Genes for Δ1-Pyrroline-5-Carboxylate Synthetase from Tomato
Tomomichi Fujita, Albino Maggio, Mario Garcia-Rios, Ray A. Bressan, Laszlo N. Csonka
Plant Physiology, 118, 2, 661, 674, American Society of Plant Biologists (ASPB), 01 Oct. 1998
Scientific journal

Cloning of a polycistronic cDNA from tomato encoding  -glutamyl kinase and  -glutamyl phosphate reductase
M. Garcia-Rios, T. Fujita, P. C. LaRosa, R. D. Locy, J. M. Clithero, R. A. Bressan, L. N. Csonka
Proceedings of the National Academy of Sciences, 94, 15, 8249, 8254, Proceedings of the National Academy of Sciences, 22 Jul. 1997
Scientific journal

Cloning of tomPRO1 and tomPRO2 from Lycopersicon esculentum L.: coexistence of polycistronic and monocistronic genes which encode the enzymes catalyzing the first two steps of proline biosynthesis.　　　　　　　　　　　　　　　
A. Maggio, M. Garcia-Rios, T. Fujita, R.A. Bressan, R.J. Joly, P.M. Hasegawa, L.N. Csonka
112, 862, 862, 1996

Sequence of the cellular T-DNA in the untransformed genome of Nicotiana glauca that is homologous to ORFs 13 and 14 of the Ri plasmid and analysis of its expression in genetic tumors of N. glauca x N. langsdorffii
Seishiro Aoki, Akiyoshi Kawaoka, Masami Sekine, Takanari Ichikawa, Tomomichi Fujita, Atsuhiko Shinmyo, Kunihiko Syono
MGG Molecular & General Genetics, 243, 6, 706, 710, Springer-Verlag, Nov. 1994
English, Scientific journal, A region homologous to the TL-DNA of Agrobacterium rhizogenes was previously detected in the genome of untransformed Nicotiana glauca and designated cellular T-DNA (cT-DNA). Subsequently, part of this region was sequenced and two genes, which corresponded to rolB and rolC and were named Ng rolB and Ng rolC, were found. We have now sequenced a region of the cT-DNA other than the region that includes Ng rolB and C and we have found two other open reading frames (ORFs), NgORF13 and NgORF14. These ORFs correspond to ORFs 13 and 14 of the TL-DNA of A. rhizogenes and exhibit a high degree of homology to these ORFs, without having a nonsense codon. We have not found any sequence homologous to rolD (ORF15). The two genes, NgORF13 and 14, as well as the Ng rolB and C genes, are expressed in genetic tumors of hybrids between N. glauca and N. langsdorffii but not in leaf tissues of the hybrid. © 1994 Springer-Verlag.

Cloning of cDNAs for genes that are specifically or preferentially expressed during the development of tobacco genetic tumors
Tomomichi Fujita, Hiroshi Kouchi, Takanari Ichikawa, Kunihiko Syono
The Plant Journal, 5, 5, 645, 654, Wiley, May 1994, [Peer-reviewed], [Lead author]
Scientific journal

Screening of Genes Related to Tumor Formation in Tobacco Genetic Tumors.
FUJITA Tomomichi
Plant Biotechnology, 11, 3, 171, 177, Japanese Society for Plant Cell and Molecular Biology, 1994
Japanese

Isolation and Characterization of a cDNA That Encodes a Novel Proteinase Inhibitor I from a Tobacco Genetic Tumor
T. Fujita, H. Kouchi, T. Ichikawa, K. Syono
Plant and Cell Physiology, 34, 1, 137, 142, Oxford University Press (OUP), Jan. 1993, [Peer-reviewed], [Lead author]
English, Scientific journal, We have isolated a cDNA clone, designated GTI, by screening a tobacco genetic tumor cDNA library with a tumor-specific "subtracted" cDNA probe. The cDNA contained the entire coding sequence for a 94-amino-acid polypeptide that exhibited significant homology to members of the proteinase inhibitor I family from tomato and potato. The predicted protein has a pre-sequence of 22 amino acids but lacks a pro-sequence, unlike genes for proteinase inhibitor I isolated to date. Furthermore, the protein encoded by GTI cDNA has a novel reactive site, having glutamine as the P_1 reactive residue. These results suggest that the GTI protein is a novel member of the proteinase inhibitor I family. The mRNA for GTI accumulated at a high level but only transiently after the wounding of tobacco plants. Thus, it appears that the GTI protein has a function that is related to the protection of tissues against damage due to wounding.

Changes in Morphology, Levels of Endogenous IAA and Protein Composition in Relation to the Development of Tobacco Genetic Tumor Induced in the Dark
Tomomichi Fujita, Takanari Ichikawa, Kunihiko Syōno
Plant and Cell Physiology, 32, 2, 169, 177, Oxford University Press (OUP), Mar. 1991, [Peer-reviewed], [Lead author]
English, Scientific journal, When normal shoots which had been regenerated from tobacco genetic tumor cultured in the dark were cut, tumorous tissues were again induced on the cut segments. Morphological examination of the segments, 5 days and 15days after cutting, in a comparison with normally regenerated shoots, revealed active cell division in vascular-bundle tissue and formation of primordial teratoma-like structures in the induced tumorous tissues. Endogeneous IAA remained at a constant, low level throughout the tumorigenetic process. Proteins from each step of tumorigenesis were separated by two-dimensional electrophoresis on mini-gels and visualized by silver staining. Twenty-seven polypeptides showed qualitative or quantitative changes during the tumorigenetic process. These results are discussed in relation to the development of tobacco genetic tumor.

ヒメツリガネゴケが重力の大きさに応答し成長量を変化させることに関する研究
青木真太郎, 山下祐輝, 半場祐子, 蒲池浩之, 唐原一郎, 久米篤, 藤田知道, 日本植物学会大会研究発表記録(CD-ROM), 87th, 2023

スペース・モス宇宙実験で得たヒメツリガネゴケ仮根系のX線マイクロCTによる3D可視化
唐原一郎, 若林孝尚, 山浦遼平, 玉置大介, 蒲池浩之, 山内大輔, 峰雪芳宣, 星野真人, 上杉健太朗, 嶋津徹, 笠原春夫, 鎌田源司, 鈴木智美, 日渡祐二, 半場祐子, 久米篤, 藤田知道, 日本植物学会大会研究発表記録(CD-ROM), 87th, 2023

転写因子AP2はヒメツリガネゴケの重力変化に対する光合成・成長応答に関与する—Photosynthetic and Growth Responses of the Gametophore of Physcomitrium Patens to Changes in Gravity
半場, 祐子, 竹村, 香里, 北島, 佐紀人, 山下, 祐輝, 横井, 真希, 篠澤, 章久, 前田, 彩友子, 安井, 祐太郎, 坂田, 洋一, 蒲池, 浩之, 小野田, 雄介, 唐原, 一郎, 久米, 篤, 笠原, 春夫, 鎌田, 源司, 嶋津, 徹, 鈴木, 智美, 矢野, 幸子, 藤田, 知道, HANBA, Yuko T., TAKEMURA, Kaori, KITAJIMA, Sakihito, YAMASHITA, Yuki, YOKOI, Maki, SHINOZAWA, Akihisa, MAEDA, Ayuko, YASUI, Yutaro, SAKATA, Yoichi, KAMACHI, Hiroyuki, ONODA, Yusuke, KARAHARA, Ichiro, KUME, Atsushi, KASAHARA, Haruo, KAMADA, Motoshi, SHIMAZU, Toru, SUZUKI, Tomomi, YANO, Sachiko, FUJITA, Tomomichi, 宇宙環境利用シンポジウム 第37回： 令和四年度 = Space Utilization Research, Vol. 37 2022: Proceedings of The Thirty-seventh Space Utilization Symposium, Jan. 2023
第37回宇宙環境利用シンポジウム (2023年1月17日-18日. オンライン開催)
Space Utilization Research (January 17-18, 2023. Online Meeting)
著者人数: 19名
資料番号: SA6000180014
G-8, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

パラボリックフライトにおける重力に応じたヒメツリガネゴケ細胞内カルシウムイオン動態のライブイメージング解析—Live-Imaging of Cytoplasmic Ca2+ Concentration of the Physcomitrium Cells in Response to Gravity During Parabolic Flights
日渡, 祐二, 逵, ローレンス かおる, 蒲池, 浩之, 唐原, 一郎, 半場, 祐子, 久米, 篤, 藤田, 知道, 鈴木, 智美, 嶋津, 徹, HIWATARI, Yuji, TSUJI, Kaoru Lorence, KAMACHI, Hiroyuki, KARAHARA, Ichiro, HANBA, Yuko T., KUME, Atsushi, FUJITA, Tomomichi, SUZUKi, Tomomi, SHIMAZU, Toru, 宇宙環境利用シンポジウム 第37回： 令和四年度 = Space Utilization Research, Vol. 37 2022: Proceedings of The Thirty-seventh Space Utilization Symposium, Jan. 2023
第37回宇宙環境利用シンポジウム (2023年1月17日-18日. オンライン開催)
Space Utilization Research (January 17-18, 2023. Online Meeting)
資料番号: SA6000180013
G-7, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

パラボリックフライトにおける重力に応じたヒメツリガネゴケ細胞内カルシウムイオン動態のライブイメージング解析　　　　　　　　　　　　　　　
日渡 祐二, 逵 ローレンス かおる, 蒲池 浩之, 唐原 一郎, 半場 祐子, 久米 篤, 藤田 知道, 鈴木 智美, 嶋津 徹, 宇宙環境利用シンポジウム 第37回： 令和四年度/Space Utilization Research, Vol. 37 2022: Proceedings of The Thirty-seventh Space Utilization Symposium, (2023-01), Jan. 2023

「スペース・モス」の活動報告：ISS における宇宙微小重力実験から地上過重力実験まで—Report on Space Moss project: from microgravity experiments on the ISS to hyper-gravity experiments
藤田, 知道, 久米, 篤, 蒲池, 浩之, 半場, 祐子, 日渡, 祐二, 唐原, 一郎, 小野田, 雄介, 横井, 真希, ヴィアチェスラヴォヴァ, アリサ, 山下, 祐輝, バイヤー, マルセル, 安田, 柚里, 中澤, 誠, 新濱, 梨奈, 佐々木, 智哉, 逵, かおる, 平山, 桃菜, 笠原, 春夫, 鈴木, 智美, 嶋津, 徹, 鎌田, 源司, 矢野, 幸子, FUJITA, Tomomichi, KUME, Atsushi, KAMACHI, Hiroyuki, HANBA, T. Yuko, HIWATASHI, Yuji, KARAHARA, Ichiro, ONODA, Yusuke, YOKOI, Maki, ALISA, Vyacheslavova, YAMASHITA, Yuki, BEIER, Marcel, YASUDA, Yuri, NAKAZAWA, Makoto, SHINHAMA, Rina, SASAKI, Tomoya, TSUJI, Kaoru, HIRAYAMA, Momona, KASAHARA, Haruo, SUZUKI, Tomomi, SHIMAZU, Toru, KAMADA, Motoshi, YANO, Sachiko, 宇宙環境利用シンポジウム 第36回： 令和三年度 = Space Utilization Research, Vol. 36 2021: Proceedings of The Thirty-sixth Space Utilization Symposium, Jan. 2022
第36回宇宙環境利用シンポジウム (2022年1月18日-19日. オンライン開催)
Space Utilization Research (January 18-19, 2022. Online Meeting)
著者人数: 22名
資料番号: SA6000168005
G-04, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

国際宇宙ステーションで生育したヒメツリガネゴケ茎葉体の機械的特性—Mechanical Properties of the Moss Physcomitrium patens Gametophores Grown in the International Space Station
蒲池, 浩之, 小野田, 雄介, 新濱, 梨奈, 浅野, 加杜己, 森, 耀久, 佐々木, 智哉, 唐原, 一郎, 久米, 篤, 半場, 祐子, 笠原, 春夫, 鎌田, 源司, 嶋津, 徹, 鈴木, 智美, 矢野, 幸子, 藤田, 知道, KAMACHI, Hiroyuki, ONODA, Yusuke, SHINHAMA, Rina, ASANO, Kazuki, MORI, Akihisa, SASAKI, Tomoya, KARAHARA, Ichirou, KUME, Atsushi, HANBA, Yuko, KASAHARA, Haruo, KAMADA, Motoshi, SHIMAZU, Toru, SUZUKI, Tomomi, YANO, Sachiko, FUJITA, Tomomichi, 宇宙環境利用シンポジウム 第35回： 令和二年度 = Space Utilization Research, Vol. 35 2020: Proceedings of The Thirty-fifth Space Utilization Symposium, 35, Jan. 2021
第35回宇宙環境利用シンポジウム(2021年1月19日-20日. オンライン開催)
Space Utilization Research (January 19-20, 2021. Online Meeting)
著者人数: 15名
資料番号: SA6000156012
レポート番号: G-04, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

国際宇宙ステーション(ISS) で生育したヒメツリガネゴケ茎葉体の光合成・成長特性—Photosynthesis and Growth Traits of the Moss Physcomitrium patens Gametophores Grown in the International Space Station
半場, 祐子, 安田, 柚里, 中澤, 誠, 蒲池, 浩之, 小野田, 雄介, 唐原, 一郎, 久米, 篤, 笠原, 春夫, 鎌田, 源司, 嶋津, 徹, 鈴木, 智美, 矢野, 幸子, 藤田, 知道, HANBA, Yuko, YASUDA, Yuri, NAKAZAWA, Makoto, KAMACHI, Hiroyuki, ONODA, Yusuke, KARAHARA, Ichirou, KUME, Atsushi, KASAHARA, Haruo, KAMADA, Motoshi, SHIMIZU, Toru, SUZUKI, Tomomi, YANO, Sachiko, FUJITA, Tomomichi, 宇宙環境利用シンポジウム 第35回： 令和二年度 = Space Utilization Research, Vol. 35 2020: Proceedings of The Thirty-fifth Space Utilization Symposium, 35, Jan. 2021
第35回宇宙環境利用シンポジウム(2021年1月19日-20日. オンライン開催)
Space Utilization Research (January 19-20, 2021. Online Meeting)
著者人数: 13名
資料番号: SA6000156011
レポート番号: G-03, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

Life cycle of plants in space - Optimization of plant cultivation conditions under low gravity through evaluation of 3-
KASAHARA Ichirou, YAMAMURA Ryohei, KUROGANE Tomofumi, YAMAUCHI Daisuke, MINEYUKI Yoshinobu, KAMACHI Hiroyuki, HASHIMOTO Hirofumi, HOSHINO Makoto, UESUGI Kentaro, NAKAI Yusuke, NAKANO Akimasa, TANIHATA Koshiro, TAMAOKI Daisuke, NISHIUCHI Takumi, TAKAO Yasumasa, TAURA Futoshi, YANO Sachiko, TANIGAKI Fumiaki, SHIMAZU Toru, KASAHARA Haruo, KAMADA Motoshi, SUZUKI Tomomi, ONODA Yusuke, KUME Atsushi, HANBA Yuko, FUJITA Tomomichi, KAMISAKA Seiichiro, 宇宙環境利用シンポジウム 第35回： 令和二年度 = Space Utilization Research, Vol. 35 2020: Proceedings of The Thirty-fourth Space Utilization Symposium, 35, Jan. 2021
Space Utilization Research (January 19-20, 2021. Online Meeting), Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

ヒメツリガネゴケ茎葉体の発生とアルギニン代謝
川出健介, 川出健介, 川出健介, 川出健介, 堀口吾朗, 及川彰, 及川彰, 平井優美, 斉藤和季, 斉藤和季, 藤田知道, 塚谷裕一, Plant Morphology, 32, 1, 2020

茎葉体の発生と代謝をつなぐ転写調節ネットワーク
川出健介, 川出健介, 川出健介, 広瀬侑, 堀口吾朗, 堀口吾朗, 及川彰, 及川彰, 平井優美, 斉藤和季, 斉藤和季, 藤田知道, 塚谷裕一, 日本植物学会大会研究発表記録(CD-ROM), 84th, 2020

Response of Physcomitrella patens under environmental stress
中澤誠, 横井真希, 藤田知道, 新濱梨奈, 浅野加杜己, 蒲池浩之, 唐原一郎, 久米篤, 小野田雄介, 笠原春夫, 鈴木智美, 嶋津徹, 半場祐子, 日本生態学会大会講演要旨(Web), 67th, 2020

Hypergravity response in Physcomitrella patens mutant (cdka)
安田柚里, 久米篤, 森耀久, 蒲池浩之, 藤田知道, 半場祐子, 日本生態学会大会講演要旨(Web), 67th, 2020

共生と競争の狭間で：植物の声を聞いてみる　　　　　　　　　　　　　　　
藤田 知道, 特集「30年後の植物科学」植物科学の最前線（BSJ-Review）発行10周年記念（日本植物学会）, 2020, [Lead author, Corresponding author]

1gとは異なる重力環境で植物はどのように育つのだろうか －コケ植物を用いた宇宙実験(スペース・モス)から期待できること－
藤田知道, 久米篤, 蒲池浩之, 小野田雄介, 半場祐子, 日渡祐二, 唐原一郎, 植物科学の最前線 BSJ-Review, 「宇宙から識る植物科学」（日本植物学会 ）, 11,, 60, 74, 2020, [Peer-reviewed], [Invited], [Lead author, Corresponding author]

Space microgravity experiments in moss: The model land plant, Physcomitrella patens on International Space Station
Kume Asushi, Fujita Tomomichi, Kamachi Hiroyuki, Hanba Yuko T, Hiwatari Yuji, Karahara Ichiro, Onoda Yusuke, Yokoi Maki, Vyacheslavova Alisa, Yamashita Yuki, Yasuda Yuri, Nakazawa Makoto, Shinhama Rina, Asano Kazuki, Tsuji Kaoru, Hirayama Momona, Kasahara Haruo, Suzuki Tomomi, Shimazu Toru, Kamata Motoshi, 宇宙環境利用シンポジウム 第34回： 令和元年度 = Space Utilization Research, Vol. 34 2019: Proceedings of The Thirty-fourth Space Utilization Symposium, 34, Jan. 2020
第34回宇宙環境利用シンポジウム (2020年1月21日-22日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県著者人数: 20名資料番号: SA6000145014, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

Life cycle of plants in space - Effects of gravity on reproductive organs and root system of Arabidopsis, and morphology of leguminous plants-
Karahara Ichiro, Sawada Ryota, Tanihata Koshiro, Yamaura Ryohei, Kurogane Tomofumi, Tamaoki Daisuke, Yano Sachiko, Tanigaki Fumiaki, Shimazu Toru, Kasahara Haruo, Yamauchi Daisuke, Uesugi Kentaro, Hoshino Makoto, Mineyuki Yoshinobu, Takao Yasumasa, Taura Futoshi, Kurosaki Fumiya, Chin Piow WONG, Morita Hiroyuki, Kamachi Hiroyuki, Kume Atsushi, Nishiuchi Takumi, Soga Koichi, Yoshida Kumi, Hanba Yuko T, Fujita Tomomichi, Kamisaka Seiichiro, 宇宙環境利用シンポジウム 第34回： 令和元年度 = Space Utilization Research, Vol. 34 2019: Proceedings of The Thirty-fourth Space Utilization Symposium, 34, Jan. 2020
Space Utilization Research (January 21-22, 2020. Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS)), Sagamihara, Kanagawa JapanG-07, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

ヒメツリガネゴケ変異体(CDKA)の過重力応答
安田柚里, 久米篤, 森耀久, 蒲池浩之, 藤田知道, 半場祐子, 日本植物学会大会研究発表記録, 83rd, 2019

陸上植物におけるCDKAによる葉縁体運動制御
井上夏実, BAO Liang, 菅原駿人, 石川雅樹, 石川雅樹, 後藤栄治, 関根政実, TEH Ooikock, 長谷部光泰, 長谷部光泰, 和田正三, 藤田知道, 日本植物学会大会研究発表記録, 83rd, 2019

Mechanical Properties of the Physcomitrella patens Gametophores Grown at 10 G
Shinhama Rina, Mori Akihisa, Onoda Yusuke, Nagashima Hisae, Kume Atsushi, Karahara Ichiro, Hanba Yuko T, Fujita Tomomichi, Kamachi Hiroyuki, 宇宙環境利用シンポジウム 第33回： 平成30年度 = Space Utilization Research, Vol. 33 2018: Proceedings of The Thirty-third Space Utilization Symposium, 33, Jan. 2019
第33回宇宙環境利用シンポジウム (2019年1月24日-25日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県資料番号: SA6000132008, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

Practical application of proximal remote sensing for plant growth monitoring in space　　　　　　　　　　　　　　　
久米 篤, 松田 修, 藤田 知道, Proceedings of The Thirty-second Space Utilization Symposium, 32, 2019
Japanese, Summary national conference

Recommendation of [Tylewicz S et al., Science 2018 360(6385):212-215]
Tomomichi Fujita, F1000Prime, Jun. 2018

微小重力の宇宙実験下におけるヒメツリガネゴケの栽培環境の検討
安田柚里, 久米篤, 蒲池浩之, 森耀久, 唐原一郎, 藤田知道, 半場祐子, 日本生態学会大会講演要旨(Web), 65th, 2018

過重力環境で生育したヒメツリガネゴケ茎葉体の力学的特性
新濱梨奈, 森耀久, 小野田雄介, 長嶋寿江, 久米篤, 唐原一郎, 半場祐子, 藤田知道, 蒲池浩之, 日本植物学会大会研究発表記録, 82nd, 2018

1Gとは異なる重力環境におけるヒメツリガネゴケの成長,光合成,遺伝子発現変化
藤田知道, 北島佐紀人, 蒲池浩之, 久米篤, 唐原一郎, 坂田洋一, 半場祐子, 日本植物学会大会研究発表記録, 82nd, 2018

微小重力の宇宙実験下におけるヒメツリガネゴケの栽培環境の検討
安田柚里, 久米篤, 蒲池浩之, 藤田知道, 半場祐子, 日本植物学会大会研究発表記録, 82nd, 2018

Mechanisms for hypergravity response of growth and photosynthesis in Physcomitrella patens - Analysis of gene expression and anatomy -
Hanba Yuko, Yasuda Yuri, Sakaguchi Naoya, Kameishi Ryuji, Takemura Kaori, Watanabe Rina, Kitajima Sakihito, Fujita Tomomichi, Yokoi Maki, Sakata Yoichi, Shinozawa Akihisa, Kume Atsushi, Karahara Ichirou, Kamachi Hiroyuki, Mori Akihisa, 宇宙環境利用シンポジウム 第32回： 平成29年度 = Space Utilization Research, Vol. 32 2017: Proceedings of The Thirty-second Space Utilization Symposium, 32, Jan. 2018
第32回宇宙環境利用シンポジウム (2018年1月15日-16日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県著者人数: 15名資料番号: SA6000117010, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

宇宙における植物の生活環 －茎の組織形成に対する長期過重力影響および根系形態可視化の試み－
唐原 一郎, 篠笥 公隆, 黒金 智文, 村本 雅樹, 玉置 大介, 矢野 幸子, 谷垣 文章, 嶋津 徹, 笠原 春夫, 山内 大輔, 上杉 健太朗, 星野 真人, 峰雪 芳宣, 蒲池 浩之, 久米 篤, 西内 巧, 曽我 康一, 吉田 久美, 半場 祐子, 藤田 知道, 神阪 盛一郎, Karahara Ichirou, Sasaki Kimitaka, Kurogane Tomofumi, Muramoto Masaki, Tamaoki Daisuke, Yano Sachiko, Tanigaki Fumiaki, Shimizu Toru, Kasahara Haruo, Yamauchi Daisuke, Uesugi Kentaro, Hoshino Makoto, Mineyuki Yoshinobu, Kamachi Hiroyuki, Kume Atsushi, Nishiuchi Takumi, Soga Koichi, Yoshida Kumi, Hanba Yuko T., Fujita Tomomichi, Kamisaka Seiichiro, 宇宙環境利用シンポジウム 第32回： 平成29年度 = Space Utilization Research, Vol. 32 2017: Proceedings of The Thirty-second Space Utilization Symposium, 32, Jan. 2018
第32回宇宙環境利用シンポジウム (2018年1月15日-16日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県著者人数: 21名資料番号: SA6000117008, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

Hypergravity of 10g Changes Plant Growth, Anatomy, Chloroplast Size, and Photosynthesis in the Moss Physcomitrella patens
Kaori Takemura, Rina Watanabe, Ryuji Kameishi, Naoya Sakaguchi, Hiroyuki Kamachi, Atsushi Kume, Ichirou Karahara, Yuko T. Hanba, Tomomichi Fujita, MICROGRAVITY SCIENCE AND TECHNOLOGY, 29, 6, 467, 473, Dec. 2017
The photosynthetic and anatomical responses of bryophytes to changes in gravity will provide crucial information for estimating how these plant traits evolved to adapt to changes in gravity in land plant history. We performed long-term hypergravity experiments at 10g for 4 and 8 weeks using the moss Physcomitrella patens with two centrifuges equipped with lighting systems that enable long-term plant growth under hypergravity with irradiance. The aims of this study are (1) to quantify changes in the anatomy and morphology of P. patens, and (2) to analyze the post-effects of hypergravity on photosynthesis by P. patens in relation to these changes. We measured photosynthesis by P. patens for a population of gametophores (e.g., canopy) in Petri dishes and plant culture boxes. Gametophore numbers increased by 9% for a canopy of P. patens, with 24-27% increases in chloroplast sizes (diameter and thickness) in leaf cells. In a canopy of P. patens, the area-based photosynthesis rate (A (canopy)) was increased by 57% at 10g. The increase observed in A (canopy) was associated with greater plant numbers and chloroplast sizes, both of which involved enhanced CO2 diffusion from the atmosphere to chloroplasts in the canopies of P. patens. These results suggest that changes in gravity are important environmental stimuli to induce changes in plant growth and photosynthesis by P. patens, in which an alteration in chloroplast size is one of the key traits. We are now planning an ISS experiment to investigate the responses of P. patens to microgravity., SPRINGER, English

植物栽培における重力環境制御の基礎
唐原一郎, 玉置大介, 久米篤, 蒲池浩之, 半場祐子, 藤田知道, アグリバイオ, 1, 11, 1176‐1179, 1179, 20 Oct. 2017
北隆館, Japanese

過重力下におけるコケ植物の形態変化
亀石隆司, 阪口直哉, 蒲池浩之, 唐原一郎, 久米篤, 藤田知道, 半場祐子, 日本生態学会大会講演要旨(Web), 64th, 2017

モデル植物であるヒメツリガネゴケの過重力応答とそのメカニズム～長期過重力栽培実験の結果から～
阪口 直哉, 亀石 隆司, 竹村 香里, 渡辺 璃那, 久米 篤, 唐原 一郎, 藤田 知道, 蒲池 浩之, 半場 祐子, Sakaguchi Naoya, Kameishi Ryuji, Takemura Kaori, Watanabe Rina, Kume Atsushi, Karahara Ichirou, Fujita Tomomichi, Kamachi Hiroyuki, Hanba Yuko T., 宇宙環境利用シンポジウム 第31回： 平成28年度 = Space Utilization Research, Vol. 31 2016: Proceedings of The Thirty-first Space Utilization Sysmposium, 31, Jan. 2017
第31回宇宙環境利用シンポジウム (2017年1月16日-17日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県資料番号: SA6000061027, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

Effects of Vibration on Plant Growth in a Hypergravity Plant Cultivation System
Mori Akihisa, Kamachi Hiroyuki, Karahara Ichirou, Kume Atsushi, Hanba Yuko T., Takemura Kaori, Fujita Tomomichi, 宇宙環境利用シンポジウム 第31回： 平成28年度 = Space Utilization Research, Vol. 31 2016: Proceedings of The Thirty-first Space Utilization Sysmposium, 31, Jan. 2017
第31回宇宙環境利用シンポジウム (2017年1月16日-17日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県資料番号: SA6000061028, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

Recommendation of [Kameoka H et al., Plant Physiol 2016 172(3):1844-1852]
Fujita T, Kitagawa M, F1000Prime, Oct. 2016

ヒメツリガネゴケCDKAによる光応答の制御
井上夏実, BAO Liang, 石川雅樹, 石川雅樹, 比嘉毅, 日渡祐二, 関根政実, 綿引雅昭, 長谷部光泰, 長谷部光泰, 和田正三, 藤田知道, 日本植物学会大会研究発表記録, 80th, 164, 01 Sep. 2016
Japanese

Recommendation of [Landrein B et al., elife 2015 4:e07811]
Fujita T, Kitagawa M, F1000Prime, May 2016

Effect of hypergravity on growth and photosynthesis of Physcomitrella patens
Hanba Yuko T., Takemura Kaori, Sakaguchi Naoya, Watanabe Rina, Kume Atsushi, Karahara Ichirou, Fujita Tomomichi, Kamachi Hiroyuki, 宇宙環境利用シンポジウム 第30回： 平成27年度 = Space Utilization Research, Vol. 30 2015: Proceedings of The Thirtieth Space Utilization Symposium, 30, Jan. 2016
第30回宇宙環境利用シンポジウム (2016年1月19日-20日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県資料番号: SA6000048007, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

Some considerations on the plant growth environment for variable gravity experiments.
Kume Atsushi, Kamachi Hiroyuki, Karahara Ichirou, Hanba Yuko T, Fujita Tomomichi, 宇宙環境利用シンポジウム 第30回： 平成27年度 = Space Utilization Research, Vol. 30 2015: Proceedings of The Thirtieth Space Utilization Symposium, 30, Jan. 2016
第30回宇宙環境利用シンポジウム (2016年1月19日-20日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県資料番号: SA6000048045, Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS), Japanese

過重力下におけるヒメツリガネゴケの光合成能力と形態変化
竹村香里, 蒲池浩之, 久米篤, 藤田知道, 唐原一郎, 半場祐子, 日本生態学会大会講演要旨(Web), 62nd, 2015

Toward microgravity experiments in moss: The emerging model land plant, Physcomitrella patens on International Space Station and more
Fujita, Tomomichi, Kamachi, Hiroyuki, Karahara, Ichiro, Kume, Atsushi, Sakata, Yoichi, Takabayashi, Atsushi, Tanaka, Ayumi, Nagashima, Hisae, Nishiyama, Tomoaki, Hashimoto, Hirofumi, Hasebe, Mitsuyasu, Hanba, Yuko, Hiwatashi, Yuji, Matsuda, Osamu, Motomura, Taizo, Yano, Sachiko, Proceedings of The Twenty-ninth Space Utilization Sysmposium, 29, 19, 20, 2014
第29回宇宙環境利用シンポジウム (2015年1月24日-25日. 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS)), 相模原市, 神奈川県
Space Utilization Research (January 24-25, 2015. Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency(JAXA)(ISAS)), Sagamihara, Kanagawa Japan
著者人数: 16名
資料番号: SA6000035009
レポート番号: ISAS-SUR29-S04, 宇宙航空研究開発機構宇宙科学研究所(JAXA)(ISAS), Japanese

Toward microgravity experiments in moss: the response of hyper gravity environment of Physcomitrella patens　　　　　　　　　　　　　　　
Kume, Atsushi, Kamachi, Hiroyuki, Hanba, Yuko T, Takemura, Kaori, Karahara, Ichirou, Nagashima, Hisae, Yano, Sachiko, Fujita, Tomomichi, Proceedings of The Twenty-ninth Space Utilization Sysmposium, 29, 21, 22, 2014

過重力下におけるヒメツリガネゴケの光合成能力と形態変化
竹村香里, 蒲池浩之, 久米篤, 藤田知道, 唐原一郎, 半場祐子, 日本植物学会大会研究発表記録, 78th, 2014

過重力下におけるヒメツリガネゴケの光合成能力と形態変化
竹村香里, 蒲池浩之, 久米篤, 藤田知道, 唐原一郎, 半場祐子, 日本生態学会大会講演要旨(Web), 61st, 2014

アブシジン酸依存的な原形質連絡制御
北川宗典, 友井拓実, 佐藤繭子, 豊岡公徳, 坂田洋一, 榊原均, 藤田知道, 日本植物学会大会研究発表記録, 78th, 2014

細胞周期進行と環境応答を同時に制御する植物サイクリン依存性キナーゼCDKAの機能解析
川田慎也, LIANG Bao, 石橋充浩, 巻口勇馬, 野田なつみ, 日渡祐二, 石川雅樹, 石川雅樹, 鈴木穣, 菅野純夫, 長谷部光泰, 長谷部光泰, 藤田知道, 日本分子生物学会年会プログラム・要旨集(Web), 37th, 2014

過重力下におけるヒメツリガネゴケの光合成能力と形態変化
竹村香里, 蒲池浩之, 久米篤, 藤田知道, 唐原一郎, 半場祐子, 日本植物学会大会研究発表記録, 77th, 2013

細胞の分化状態に応じた原形質連絡を介する分子輸送制御
北川宗典, 綾部美晴, 豊岡公徳, 佐藤繭子, 西山智明, 寺内真, 本村泰三, 村田隆, 佐藤良勝, 長谷部光泰, 藤田知道, 日本植物学会大会研究発表記録, 77th, 2013

ヒメツリガネゴケにおけるCDKAの機能解析
石橋充浩, 巻口勇馬, 野田なつみ, 日渡祐二, 日渡祐二, 石川雅樹, 石川雅樹, 鈴木穣, 菅野純夫, 長谷部光泰, 長谷部光泰, 藤田知道, 日本植物生理学会年会要旨集, 54th, 2013

ヒメツリガネゴケ原糸体を用いた細胞レベルでの原形質連絡制御の解析
北川宗典, 松崎潤, 佐藤良勝, 藤田知道, 日本植物生理学会年会要旨集, 51st, 2010

ヒメツリガネゴケにおけるABAによる不等分裂から等分裂への切り替えの解析
仲村康平, 石川雅樹, SAKATA Yoichi, QUATRANO Ralph S., 日渡祐二, 日渡祐二, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本植物生理学会年会要旨集, 51st, 2010

46. Endogenous steroid hormones in Physcomitrella patens
Nomura Takahito, Shibata Kyomi, Fujita Tomomichi, Nakano Takeshi, Yokota Takao, The Janapese Society for Chemical Regulation of Plants, Abstract, 44, 60, 60, 2009
Genetic studies have revealed that brassinosteroids (BRs) are essential plant hormones for growth and development of seed plants. However, little is known about the occurrence and physiological roles of BRs in ferns and moss. The biosynthesis genes of sterols are highly conserved in lower to higher plants. However, the genome of the moss, Physcomitrella patens comprises no genes with high similarity to BR biosynthesis genes of seed plants. In order to know whether P. patens produces BRs, we analyzed endogenous BRs in the protonema using GC-MS. Furthermore, we also analyzed endogenous animal steroid hormones using the same material although their roles and biosynthesis in plants are unknown. As the result, a small amount of castasterone that is a biological active form in seed plants and a series of the precursors such as 6-dexoxocastasterone were identified in P. patens. In addition, animal steroid hormones including progesterone were also identified. These findings may imply that not only BRs but also animal steroid hormones had been utilized as physiologically active steroids before the branching-off from the common ancestor into moss and seed plants., The Janapese Society for Chemical Regulation of Plants, Japanese

原形質連絡による細胞間情報伝達制御と細胞の分化状態との関係性を探る
北川宗典, 佐藤良勝, 藤田知道, 日本植物生理学会年会要旨集, 50th, 2009

不等分裂モデルであるヒメツリガネゴケ頂端幹細胞に局在するタンパク質群の同定および機能解析
丸山剛史, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 久保稔, 小田祥久, 佐野俊夫, 馳澤盛一郎, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本植物生理学会年会要旨集, 50th, 2009

ヒメツリガネゴケにおいて不等分裂制御への関与が推測されるGRASファミリー転写因子の機能解析
一力綾子, 丸山剛史, 今井章裕, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 宮脇香織, 村田隆, 村田隆, 三上浩司, 倉田哲也, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本分子生物学会年会講演要旨集, 32nd, Vol.3, 2009

ヒメツリガネゴケにおけるABAによる不等分裂から等分裂への切り替えの解析
中村康平, 石川雅樹, 坂田洋一, QUATRANO Ralph, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本分子生物学会年会講演要旨集, 32nd, Vol.3, 2009

ヒメツリガネゴケミトコンドリア移行型RecAタンパク質の機能解析
小田原真樹, 井上貴之, 黒岩晴子, 藤田知道, 長谷部光泰, 黒岩常祥, 関根靖彦, 日本植物生理学会年会要旨集, 49th, 278, 15 Mar. 2008
Japanese

植物幹細胞に蓄積するタンパク質群のタイムラプス解析
丸山剛史, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 小田祥久, 佐野俊夫, 馳澤盛一郎, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 生化学, 2008

ヒメツリガネゴケ頂端幹細胞に局在するタンパク質の不等分裂時における動態解析
丸山剛史, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本植物生理学会年会要旨集, 49th, 2008

ヒメツリガネゴケにおける不等分裂に関わる因子の相互作用
一力綾子, 丸山剛史, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 三上浩司, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 日本植物学会大会研究発表記録, 72nd, 2008

陸上植物の発生進化を引き起こしたゲノム進化
長谷部光泰, 西山智明, 棚橋貴子, 青野直樹, 青山剛士, 程朝陽, 藤田知道, 橋本薫, 平井正良, 日渡祐二, 石川雅樹, 岩田美根子, 久保稔, 倉田哲也, 三上浩司, 宮崎さおり, 森長真一, 村田隆, 小原真理, 小栗康子, 小野寺直子, 佐藤良勝, 住川直美, 篠原直貴, 若月幸子, 杉本渚, 日本植物生理学会年会要旨集, 48th, 124, 15 Mar. 2007
Japanese

ヒメツリガネゴケの液胞は微小管によって制御されている
小田祥久, 佐野俊夫, 藤田知道, 日渡祐二, 佐藤良勝, 朽名夏麿, 平田愛子, 長谷部光泰, 長谷部光泰, 長谷部光泰, 馳澤盛一郎, 日本植物生理学会年会要旨集, 48th, 2007

幹細胞に局在する新規タンパク質の同定
藤田知道, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 日本植物生理学会年会要旨集, 48th, 2007

植物幹細胞の不等分裂により非対称に分配されるタンパク質の同定
丸山剛史, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 藤田知道, 生化学, 2007

Evolution of developmental genes in land plants inferred from large-scale phylogenetic analyses 2
TANAHASHI Takako, NISHIYAMA Tomoaki, AONO Naoki, AOYAMA Tsuyoshi, CHENG Chaoyang, FUJITA Tomomichi, HASHIMOTO Kaoru, HIRAI Tadayoshi, HIWATASHI Yuji, ISHIKAWA Masaki, IWATA Mineko, KUBO Minoru, KURATA Tetsuya, MIKAMI Koji, MIYAZAKI Saori, MORINAGA Shinichi, MURATA Takashi, OBARA Mari, OGURI Yasuko, ONODERA Naoko, SATO Yoshikatsu, SHINOHARA Naoki, SUMIKAWA Naomi, WAKAZUKI Sachiko, HASEBE Mitsuyasu, Journal of plant research, 119, 143, 143, 01 Dec. 2006
English

Evolution of developmental genes in land plants inferred from large-scale phylogenetic analyses 1
NISHIYAMA Tomoaki, TANAHASHI Takako, AONO Naoki, AOYAMA Tsuyoshi, CHENG Chaoyang, FUJITA Tomomichi, HASHIMOTO Kaoru, HIRAI Tadayoshi, HIWATASHI Yuji, ISHIKAWA Masaki, IWATA Mineko, KUBO Minoru, KURATA Tetsuya, MIKAMI Koji, MIYAZAKI Saori, MORINAGA Shinichi, MURATA Takashi, OBARA Mari, OGURI Yasuko, ONODERA Naoko, SATO Yoshikatsu, SHINOHARA Naoki, SUMIKAWA Naomi, WAKAZUKI Sachiko, HASEBE Mitsuyasu, Journal of plant research, 119, 142, 143, 01 Dec. 2006
English

Identification of proteins which accumulate preferentially in stem cell daughter rather than in nonstem cell daughter after asymmetric cell division of plant stem cell
FUJITA Tomomichi, HASHIMOTO Kaoru, HIWATASHI Yuji, SATO Yoshikatsu, MURATA Takashi, HASEBE Mitsuyasu, Journal of plant research, 119, 151, 152, 01 Dec. 2006
English

Isolation and functional analysis of PIN-like genes encoding an auxin efflux carrier in the moss Physcomitrella patens
KAWAI Junko, SAKAGUCHI Hisako, FUJITA Tomomichi, HASEBE Mituyasu, ITO Motomi, Journal of plant research, 119, 153, 154, 01 Dec. 2006
English

ヒメツリガネゴケ再生異常を引き起こす因子pphn 36p08の解析
橋本薫, 橋本薫, 藤田知道, 日渡祐二, 日渡祐二, 佐藤良勝, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 日本植物生理学会年会要旨集, 47th, 2006

植物不等分裂に関わるヒメツリガネゴケ遺伝子産物の局在解析
藤田知道, 橋本薫, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 日本植物生理学会年会要旨集, 47th, 2006

In vivo localization of proteins that affect asymmetric cell division by overexpression analysis in Physcomitrella patens
T Fujita, K Hashimoto, Y Hiwatashi, Y Sato, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 47, S117, S117, 2006
OXFORD UNIV PRESS, English, Summary international conference

Regulation of the vacuolar and cytoplasmic localization in protonemata of Physcomitrella patens
Y Oda, N Kutsuna, T Sano, T Fujita, M Hasebe, S Hasezawa, PLANT AND CELL PHYSIOLOGY, 47, S111, S111, 2006
OXFORD UNIV PRESS, English, Summary international conference

Analysis of a novel factor, pphn36p08 that caused abnormal regeneration of protoplast in Physcomitrella patens
K Hashimoto, T Fujita, Y Hiwatashi, Y Sato, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 47, S74, S74, 2006
OXFORD UNIV PRESS, English, Summary international conference

A plant-specific kinesin-like protein, API1, regulates microtubule arrays of spindles and phragmoplasts
Y Hiwatashi, M Obara, T Fujita, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 47, S153, S153, 2006
OXFORD UNIV PRESS, English, Summary international conference

ヒメツリガネゴケミトコンドリア移行型RecAホモログはミトコンドリアゲノムの反復配列間における組換え反応を抑制し,ゲノムの安定化に寄与する
小田原真樹, 井上貴之, 藤田知道, 長谷部光泰, 黒岩晴子, 黒岩常祥, 関根靖彦, 日本分子生物学会年会講演要旨集, 28th, 176, 25 Nov. 2005
Japanese

植物の細胞極性,不等分裂に関わる遺伝子群の細胞内局在解析
藤田知道, 橋本薫, 橋本薫, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 長谷部光泰, 日本分子生物学会年会講演要旨集, 28th, 2005

プロトプラスト再生系を用いた植物細胞極性,不等分裂に関わる遺伝子群
藤田知道, 橋本薫, 佐藤良勝, 日渡祐二, 西山智明, 村田隆, 長谷部光泰, 日本植物生理学会年会要旨集, 46th, 2005

ヒメツリガネゴケ再生異常を引き起こす新規不等分裂因子pphn36p08の解析
橋本薫, 橋本薫, 藤田知道, 日渡祐二, 日渡祐二, 佐藤良勝, 村田隆, 村田隆, 長谷部光泰, 長谷部光泰, 日本分子生物学会年会講演要旨集, 28th, 2005

Visualization of tonoplast and analysis of its structures in Physcomitrella patens
Y Oda, T Fujita, M Hasebe, S Hasezawa, PLANT AND CELL PHYSIOLOGY, 46, S230, S230, 2005
OXFORD UNIV PRESS, English, Summary international conference

Identification of genes involved in cell polarity or asymmetric cell division during protoplast regeneration of Physcomitrella patens
T Fujita, K Hashimoto, Y Sato, Y Hiwatashi, T Nishiyama, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 46, S198, S198, 2005
OXFORD UNIV PRESS, English, Summary international conference

Identification of a novel protein Et21, which is expressed in apical cells of the moss Physcomitrella patens.
Y Hiwatashi, N Sumikawa, T Fujita, M Hasebe, PLANT AND CELL PHYSIOLOGY, 45, S201, S201, 2004
OXFORD UNIV PRESS, English, Summary international conference

Identification of genes involved in cell polarity or asymmetric cell division by overexpression of full-length cDNAs during protoplast regeneration of Physcomitrella patens
T Fujita, M Higuchi, Y Hiwatashi, T Nishiyama, T Murata, K Syono, M Hasebe, PLANT AND CELL PHYSIOLOGY, 45, S200, S200, 2004
OXFORD UNIV PRESS, English, Summary international conference

植物細胞工学シリーズ20、新版植物ホルモンのシグナル伝達 「植物ホルモンの起源を探る－シダ植物，コケ植物，緑色藻類における植物ホルモンの役割から－」　　　　　　　　　　　　　　　
藤田知道, 秀潤社, 2004

Identification of genes involved in cell polarity, asymmetric cell division or cell differentiation by overexpression of full-length cDNAs in the moss Physcomitrella patens
T Fujita, Y Hiwatashi, T Nishiyama, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 44, S128, S128, 2003
OXFORD UNIV PRESS, English, Summary international conference

ヒメツリガネゴケにおけるGFP-tubulinを用いた微小管の可視化
佐藤良勝, 藤田知道, 長井愛, 長谷部光泰, 和田正三, 門田明雄, 日本植物学会大会研究発表記録, 66th, 2002

Comparative analysis of auxin distribution in haploid garactophore of Physcomitrella patens and in diploid shoots of angiosperms
H Sakaguchi, T Fujita, T Sato, M Hasebe, PLANT AND CELL PHYSIOLOGY, 43, S91, S91, 2002
OXFORD UNIV PRESS, English, Summary international conference

Intracellular localization of a kinesin-like protein, API1, in the moss Physcomitrella patens
Y Hiwatashi, T Fujita, T Murata, M Hasebe, PLANT AND CELL PHYSIOLOGY, 43, S212, S212, 2002
OXFORD UNIV PRESS, English, Summary international conference

ANALYSES OF THE FUNCTIONAL REGION OF 6b PROTEIN ON T-DNA OF A.tumefaciens AND THE ROLE OF ITS INTERACTING PROTEINS :
TERAKURA Shinji, KITAKURA Saeko, AZECHI Youko, UENO Yoshihisa, FUJITA Tomomichi, WABIKO Hiroetsu, MACHIDA Yasunori, Plant and cell physiology, 42, s66, 2001
Japanese Society of Plant Physiologists, English

FULL-LENGTH cDNA ACTIVATION SYSTEM OF PHYSCOMITRELLA PATENS TO STUDY AUXIN AND CYTOKININ FUNCTION :
FUJITA Tomomichi, SEKI Motoaki, CARNINCI Piero, HAYASHIZAKI Yoshihide, SHINOZAKI Kazuo, HASEBE Mitsuyasu, Plant and cell physiology, 42, s86, 2001
Japanese Society of Plant Physiologists, English

ANALYSIS OF THE EXPRESSION PATTERN OF GENES WHICH ARE RELATED TO THE LEAF DEVELOPMENT :
UENO Yoshihisa, KITAKURA Saeko, IWAKAWA Hidekazu, SEMIARTI Endang, MACHIDA Chiyoko, FUJITA Tomomichi, MACHIDA Yasunori, Plant and cell physiology, 41, s213, 2000
Japanese Society of Plant Physiologists, English

Agrobacterium tumefaciens T-DNA 6b gene induces plant cell growth and effects on leaf development :
KITAKURA Saeko, FUJITA Tomomichi, UENO Yoshihisa, WABIKOI Hiroetsu, MACHIDA Yasunori, Plant and cell physiology, 41, s213, 2000
Japanese Society of Plant Physiologists, English

Screening of factors which interact with the protein encoded by the Agrobacterium tumefaciens T-DNA gene 6b that stimulates growth of plant cells
KITAKURA Saeko, FUJITA Tomomichi, WABIKO Hiroetsu, MACHIDA Yasunori, 日本分子生物学会年会プログラム・講演要旨集, 21, 507, 507, 01 Dec. 1998
Japanese

コケ分子生物学を用いたSDGsへの取り組み　　　　　　　　　　　　　　　
藤田知道
アグリバイオ 7 (3) 24-28、北隆館, Mar. 2023

植物のサイクリン依存性キナーゼCDKAの光応答における新たな機能　　　　　　　　　　　　　　　
井上夏実, 藤田知道
バイオサイエンスとインダストリー 80巻 pp.414-416、バイオインダストリー協会, 10 Sep. 2022, [Peer-reviewed]

環境ストレスにより新生するコケ植物の幹細胞　　　　　　　　　　　　　　　
神野智世, 藤田知道
アグリバイオ 6 (6) 82-86、北隆館, Jun. 2022

コケ植物の幹細胞新生にみるアブシジン酸を用いたしなやかな生存戦略　　　　　　　　　　　　　　　
神野智世, 藤田知道
細胞 54 (6) 61-64、ニューサイエンス社, May 2022

Tracking intercellular movement of fluorescent proteins in bryophytes.　　　　　　　　　　　　　　　
Takumi Tomoi, Yoan Coudert, Tomomichi Fujita, Plasmodesmata, Methods and Protocols, In Methods in Molecular Biology (MIMB), volume 2457, 321-332; https://doi.org/10.1007/978-1-0716-2132-5_22
v, Mar. 2022, 9781071621318, [Peer-reviewed], [Contributor]

Callose detection and quantification at plasmodesmata in bryophytes.
Arthur Muller, Tomomichi Fujita, Yoan Coudert, Plasmodesmata, Methods and Protocols, In Methods in Molecular Biology (MIMB), volume 2457, 177-187; https://doi.org/10.1007/978-1-0716-2132-5_11
Humana, New York, Mar. 2022, 9781071621318, 177-187, Callose detection and quantification at plasmodesmata in bryophytes., [Peer-reviewed], [Contributor]

An experimental system for examining phototropic response of gametophytic shoots in the moss Physcomitrella patens　　　　　　　　　　　　　　　
Liang Bao, Kotaro Yamamoto, Tomomichi Fujita, Phototropism, In Methods in Molecular Biology, 1924, 45-51. doi: 10.1007/978-1-4939-9015-3_5
Humana Press, New York, NY, 2019, 9781493990153, [Peer-reviewed], [Contributor]

植物栽培における重力環境制御の基礎　　　　　　　　　　　　　　　
唐原一郎, 玉置大介, 久米篤, 蒲池浩之, 半場祐子, 藤田知道
アグリバイオ 10(1), 1172-1175、北隆館, Oct. 2017

植物生理学概論 改訂版　　　　　　　　　　　　　　　
櫻井英博, 柴岡弘郎, 高橋陽介, 小関良宏, 藤田知道
培風館, Sep. 2017, [Contributor]

植物ゲノム科学辞典　　　　　　　　　　　　　　　
藤田知道, 駒嶺穆, 町田泰則, 藤村達人, 田畑哲之, 三位正洋, 斉藤和季
朝倉書店, 2009, [Others]

植物の百科事典「細胞極性、不等分裂」　　　　　　　　　　　　　　　
藤田知道, 石井龍一, 岩槻邦男, 長谷部光泰, 矢澤進, 矢原徹一, 和田正三
朝倉書店, 2009, [Others]

植物の細胞極性形成および不等分裂に関わる遺伝子の機能解析
藤田 知道
[北海道大学理学部], 2008

植物細胞工学シリーズ23、植物の進化 「発生遺伝子の進化」　　　　　　　　　　　　　　　
長谷部光泰, 藤田知道, 倉田哲也, 佐藤良勝, 久保稔, 村田隆, 青山剛士, 三上浩司, 石川雅樹, 日渡祐二, 青野直樹, 篠原直貴, 西山智明, 棚橋貴子
秀潤社, 2007, [Joint work]

植物ホルモンの分子細胞生物学「セン類、タイ類の植物ホルモン」　　　　　　　　　　　　　　　
藤田知道, 小柴共一, 神谷勇治, 勝見允行
講談社サイエンティフィク, 2006

ヒメツリガネゴケを用いた植物幹細胞の不等分裂過程の解析−細胞極性形成から娘細胞の運命決定まで−　　　　　　　　　　　　　　　
藤田知道
植物化学調節学会, 2006

植物の細胞極性および不等分裂に関わる遺伝子の同定と機能解析
藤田 知道
[北海道大学理学部], 2006

Gene tagging, gene- and enhancer-trap systems, and full-length cDNA overexpression in Physcomitrella patens　　　　　　　　　　　　　　　
Fujita, T, Nishiyama, T, Hiwatashi, Y, Hasebe, M
Kluwer Academic Publishers, 2004, [Joint work]

植物細胞工学シリーズ20、新版植物ホルモンのシグナル伝達 「植物ホルモンの起源を探る—シダ植物，コケ植物，緑色藻類における植物ホルモンの役割から—」　　　　　　　　　　　　　　　
藤田 知道
秀潤社, 2004

学術月報「３度のポスドク体験から」　　　　　　　　　　　　　　　
藤田 知道
日本学術振興会, 2000

タバコ遺伝的腫瘍の腫瘍形成に関わる遺伝子の検索　　　　　　　　　　　　　　　
藤田 知道
植物組織培養, 1994

ＤＮＡ複製機構の解明に向けてーＤＮＡ腫瘍ウイルスの手助け　　　　　　　　　　　　　　　
藤田 知道
生物工学会誌, 1994

Habituation as a tumorous state that is interchangeable with a normal state in plant cells　　　　　　　　　　　　　　　
Syono, K, Fujita, T
International Review of Cytology (eds. by Jeon, K. W. and Jarvik J.), 152, pp.265-299, Academic Press,, 1994, [Joint work]

月や火星でのテラフォーミング技術開発を目指す地上研究　　　　　　　　　　　　　　　
2024年度 フロントローディング研究
Apr. 2024 - Feb. 2025
藤田知道
JAXA(宇宙航空研究開発機構)/ISAS(宇宙科学研究所)

植物の多細胞化に重要な構造・原形質連絡の形成に関わる分子基盤の解明とその進化
科学研究費助成事業
01 Apr. 2021 - 31 Mar. 2024
藤田 知道
植物では、細胞壁を貫き細胞間コミュニケーションに不可欠な構造として原形質連絡(PD)の存在が古くより知られている。しかしPDがどのように作られるのか、その分子基盤はいまでもわかっていない。本研究は、コケ植物の変異株で、PDがほぼ消失していたという申請者らの発見を端緒としている。本研究では変異株の責任遺伝子の機能とその標的因子の探索とともに解析を進め、PDを作るための分子基盤とその形成過程を明らかにする。またこの分子基盤が被子植物のシロイヌナズナや陸上植物の祖先群のシャジクモ藻で保存されているかどうかを調べ、シャジクモ藻から陸上植物を含むストレプト植物全体で、PD形成に関わる分子基盤の普遍性を検討する。
このためにモデルコケ植物であるヒメツリガネゴケを用いて①から⑤までを進めることを計画した。①ULMとLMがPD形成に必要十分であるかの遺伝学的検証を行う。② PD形成におけるULM→LM経路の遺伝学的検証を行う。 ③ ULMとLMの細胞内局在および相互作用を調査する。④ LMの下流でPD形成に関わる因子Xの探索とその変異株を作成する。⑤ 小胞体およびPD可視化株を作成する。
次にシロイヌナズナを用いて以下を研究も進めることを計画した。⑥シロイヌナズナでPD形成に関わると予想される変異株の収集および新たな変異株の作成を行う。変異株は致死性を回避するため、薬剤誘導型株を入手する。入手が困難な場合は自分たちで作成する。
その結果、ULM, LMの関与とその経路の重要性についての理解を前進させることができた。またLMの複数の制御因子がPD形成に関わることを明らかにした。さらにストレスホルモンであるアブシジン酸が細胞間コミュニケーション制御に重要であることからPDの形成との関わりを調査したところ、アブシジン酸によりPD形成は抑制されることを新たに見出した。
日本学術振興会, 基盤研究(B), 北海道大学, 21H02516

Elucidation of the unknown molecular basis of the gravitational response in plants and development of growth control technology
Grants-in-Aid for Scientific Research
09 Jul. 2021 - 31 Mar. 2023
久米 篤, 日渡 祐二, 蒲池 浩之, 藤田 知道, 唐原 一郎, 富田 祐子
1）重力環境(G)を変化させた栽培：地上栽培実験で、既存の装置を利用して1g, 2.3g, 10gの３つの異なる過重力環境におけるヒメツリガネゴケの栽培実験を実施した。また、ウキクサ類の栽培実験も実施した。さらに、宇宙ステーション内でコケの長期栽培を実施するための超小型栽培システムの開発を民間企業と協力して開始した。
2）RNA-seqによる遺伝子発現変動解析：地上栽培実験に加えて宇宙栽培実験より回収できたμg, 1gのサンプルを用い、これら全てのサンプルでRNA-seq解析を行い、網羅的に遺伝子の発現変動を調べた。その結果、重力変化に伴う成長制御の鍵となるAP2転写因子を見出した。これらはいずれもこれまでに報告の無いグループであることが判明した。
3）細胞レベルおよび個体レベルにおける重力応答メカニズム解明：転写因子AP2を過剰発現させたヒメツリガネゴケを使用し、過重力をかけて栽培した。その後、形態観察、光合成機能の測定を通して、AP2が植物の過重力応答に果たす役割の詳細を解析した。10gと1gで8週間、ヒメツリガネゴケの野生型とAP2過剰発現体を栽培したところ、すべての系統で10gで栽培したコケコロニーの光合成速度と植物体内CO2コンダクタンスが1gで栽培したコロニーを上回った。また、10gと1gで栽培したコケコロニーの差をそれぞれの系統で比較すると、過剰発現体より野生型での差が大きくなった。転写因子AP2の過剰発現体は過重力栽培により、野生型より葉緑体サイズが大きくなったが、光合成速度の上昇は野生型の光合成速度の上昇に満たなかった。この理由として、過剰発現体の葉緑体サイズは野生型より増大したが、同時に細胞壁が厚くなったため、CO2透過性の上昇が抑制されてしまった可能性が考えられた。
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Kyushu University, 21K19272

スペース・モス創出-テラフォーミングを先導するコケ植物力利用技術開発の挑戦的研究　　　　　　　　　　　　　　　
2023
藤田知道
日本学術振興会 科学研究費助成事業 挑戦的研究（開拓） 2023年4月 - 2027年3月

コケ植物から解き明かす植物幹細胞に特有の動作原理
科学研究費助成事業
01 Apr. 2020 - 31 Mar. 2022
藤田 知道
植物幹細胞が性質の異なる2つの娘細胞になる過程は、不等分裂または非対称分裂と呼ばれ、その分子機構の研究は限られたモデル被子植物で行われてきたものの、知識は断片的でありその大部分はまだよくわかっていない。そこで本研究ではこれまでの成果を発展させ本領域の推進に役立てるため、ヒメツリガネゴケの原糸体頂端幹細胞に着目し、その幹細胞をin vivoの生きた状態で1細胞単位で研究し、植物に特有の分子制御機構を複数明らかにすることを目的とし実施した 。
ヒメツリガネゴケの原糸体は幹細胞が露出しており、このような目的に向いた材料である。また全ゲノム情報が利用でき、遺伝子の機能解析技術も蓄積している。このような材料の利点を生かし、植物幹細胞に特有の特徴を明らかにするために相互に関連の深い次の3つの研究課題に取り組んだ。《1》植物幹細胞の極性・不等分裂の制御機構、《2》植物幹細胞の不等分裂と等分裂を可逆的に制御する機構、《3》植物幹細胞の原形質連絡の制御機構に関する研究をそれぞれ実施した。


その結果、植物特有の転写因子が水チャネルを形成するアクアポリンの発現を誘導し、また並行して液胞形成の局在や形態などを同時に制御していることがわかった。またこの転写因子は細胞壁の構築にも関わり、幹細胞の極性形成制御に重要であると考えられた。次に植物特有のアラビノガラクタンタンパク質に着目し研究を進めた。その結果、この因子はオーキシンの感受性の制御に関わり、オーキシンシグナル伝達系の転写因子を介して細胞壁成分の調整に重要な役割を担っていることがわかってきた。またアブシジン酸シグナル伝達因子の２つに着目し原形質連絡制御の関係を調べ、この両者で働きが違うことを明らかにした。またアブシジン酸により可逆的に新しいストレス耐性の幹細胞の生成が制御されていることを見出した。
日本学術振興会, 新学術領域研究(研究領域提案型), 北海道大学, 20H04878

How do proteoglycans mediate auxin gradient sensing in planar cell polarity?
Grants-in-Aid for Scientific Research
01 Apr. 2019 - 31 Mar. 2022
Teh Ooi Kock
We found that SB plays an inhibitory role in cell wall loosening. SB overexpression inhibits cell expansion while knocking out SB and its functional paralog, SB-like, accelerated gametophore initiation, an indication of loosen cell walls. Mutations analysis on the SB indicated that glycosylations are required for SB functions but has no effect on the SB subcellular localization. We further established a link between SB and a C-type auxin response factor (AFRC) by showing that SB positively regulated the expression level of ARFC. Furthermore, arfc loss-of-function mutants phenocopied sb and repressed the SB overexpressor phenotype, demonstrating that ARFC is acting downstream of SB. We performed RNAseq analyses using an ARFC-inducible line to identify ARFC transcriptional targets. Gene ontology (GO) analyses showed that cell wall-related GO terms were highly enriched. We propose that AGP may act as a cell wall structural protein that senses changes in cell wall compositions.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 19K06701

A novel function of cyclin dependent kinase A in land plants
Grants-in-Aid for Scientific Research
01 Apr. 2018 - 31 Mar. 2021
Fujita Tomomichi
Our results show that PpCDKA is a factor with important functions in various environmental stress responses, in addition to the cell cycle. In particular, we studied in detail the mechanism of molecular regulation of light responses by CDKA and found that CDKA regulates light responses independently of cell cycle regulation, and that this function is conserved not only in Physcomitrella patens but also in the angiosperm Arabidopsis thaliana. We also proposed that CDK1, a CDKA orthologue in humans and animals, may regulate environmental responses in addition to the cell cycle.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 18K06273

コケ植物から解き明かす植物幹細胞に特有の動作原理
科学研究費助成事業
01 Apr. 2018 - 31 Mar. 2020
藤田 知道
植物幹細胞が性質の異なる2つの娘細胞になる過程は、不等分裂または非対称分裂と呼ばれ、その分子機構の研究は限られたモデル被子植物で 行 われてきたものの、知識は断片的でありその大部分はまだよくわかっていない。そこで本研究ではこれまでの成果をさらに発展させ本領域の推進に役立てるため、ヒメツリガネゴケの原糸体頂端幹細胞に着目し、その幹細胞をin vivoの生きた状態で1細胞単位で研究し、植物に特有の分子制御機構を複数明らかにすることを目的とした 。ヒメツリガネゴケの原糸体は幹細胞が露出しており、このような目的に向いた材料である。また全ゲノム情報が利用でき、遺伝子の機能解析技術も蓄積している。このような材料の利点を生かし、植物幹細胞に特有の特徴を明らかにするために相互に関連の深い次の3つの研究課題に取り組んだ。《1》植物幹細胞の極性・不等分裂の制御機構、《2》植物幹細胞の不等分裂と等分裂を可逆的に制御する機構、《3》植物幹細胞の原形質連絡の制御機構に関する研究をそれぞれ実施した。
その結果、植物特有の転写因子が水チャネルを形成するアクアポリンを介して不等分裂の制御に関わることがわかってきた。液胞の形態異常を伴っており液胞が不等分裂の制御に関わる可能性が考えられた。また植物特有のアラビノガラクタンタンパク質に着目し研究を進めた。その結果、このタンパク質の糖鎖修飾部分が原糸体幹細胞の新生に重要である可能性も明らかにすることができた。またアラビノガラクタンタンパク質の下流でオーキシンシグナル伝達系の因子が重要な役割を担っていることを遺伝学的に明らかにできた。さらに原形質連絡を制御するアブシジン酸シグナル伝達因子を２つ同定することができた。今後は、これらの因子による分裂様式の切り替えと原形質連絡の開閉制御の関係をさらに調査する必要があると考えられた。
日本学術振興会, 新学術領域研究(研究領域提案型), 北海道大学, 18H04829

New challenge on planar cell polarity in plants
Grants-in-Aid for Scientific Research
01 Apr. 2016 - 31 Mar. 2019
Fujita Tomomichi, Shindoh Chisato, The Ooi-Kock, Yao Jiawei
How cell polarity is established and maintained is a fundamental question for all organisms. The moss, Physcomitrella patens serves as an excellent model for studying cell polarity due to its easy observation at a cell level, high frequency of gene targeting and available genome information. In order to understand cell polarity regulation in plants, we studied a plant specific, plasma membrane protein in this research by generating overexpression or knock-out mutant lines and performed RNA-seq analysis in these mutants. Thus, we found this gene is, together with auxin signaling factors and microtubule, involved in cell polarity regulation in the moss protonemata.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 16K14747

Identification of the CDKA-target factors that regulate cellular changes during stem cell formation
Grants-in-Aid for Scientific Research
Apr. 2015 - Mar. 2019
Ishikawa Masaki, Hasebe Mitsuyasu, Murata Takashi, Fujita Tomomichi
In the moss Physcomitrella patens, when a leaf is excised from a gametophore and cultivated for a few days on culture medium without phytohormone supplementation, leaf cells facing the cut change into cells that are not distinguishable from chloronema apical cells. During this process, a cell cycle regulator, cyclin-dependent kinase A (CDKA), functions in coordination between cell cycle reactivation and acquisition of new cell-type characteristics. To understand roles of CDKA in the stem cell formation, we performed interactome and phosphoproteomic analyses to identify CDKA-target proteins. As a result, we identified a factor involved in the histone modifications, suggesting that CDKA regulates epigenetic modifications to induce cellular changes. In addition, we developed a sensor to detect the CDKA protein kinase activity at tissue levels.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), National Institute for Basic Biology, 15K07119

Regulation of plasmodesmata in brown algae
Grants-in-Aid for Scientific Research
01 Apr. 2014 - 31 Mar. 2017
NAGASATO Chikako, FUJITA Tomomichi
Brown algae show multicellularity and possess cellular connections called plasmodesmata which enable exchange of molecules between the neighboring cells. Plasmadesmata take tubular structure with 10 - 20 nm in diameter, and vary appearance and distribution with the developmental stages. Some new findings concerning brown algal plasmodesmata were given in this study as follows; 1) plasmodesmata are divided into primary and secondary plasmodesmata as in land plants, 2) plasmodesmatal size exclusion limit are set between 20-40 kDa in multiseriate species, and 3) pit fields in which plasmodesmata concentrate one place on the cross wall pass molecules less than 10 kDa to the next cell.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 26440160

Molecular mechanism of cell-cell communication via plasmodesmata
Grants-in-Aid for Scientific Research
01 Apr. 2013 - 31 Mar. 2016
Fujita Tomomichi, Motomura Taizo
Cell-cell communication via plasmodesmata is important to control growth and environmental response in plants. However, the molecular mechanism is largely unknown.
We developed a quantitative method to analyze cell-cell communication by using a photoconvertible fluorescence protein, Dendra2 in protonemata of the moss, Physcomitrella patens, and studied a regulation of Dendra2 movement. We found two factors that are involved in abscisic acid signaling play a key role to control abscisic acid-dependent suppression of Dendra2 movement.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 25650089

Environmental response and utilization of mosses in space - Space Moss-　　　　　　　　　　　　　　　
Dec. 2015
https://humans-in-space.jaxa.jp/kibouser/subject/life/70682.html
国立研究開発法人宇宙航空研究開発機構JAXA, 北海道大学ほか, Principal investigator

メリステム制御の基盤を支える植物幹細胞の不等分裂の分子機構の解明
科学研究費助成事業
2011 - 2012
藤田 知道, 石川 雅樹, 日渡 祐二
植物の形態形成はメリステムにある幹細胞の増殖と分化のバランスにより制御されている。幹細胞の不等分裂はこのようなバランスを支える基本的しくみであり、メリステムを理解するためには幹細胞の不等分裂の分子制御機構の理解は不可欠である。ヒメツリガネゴケから単離したプロトプラストや原糸体の頂端細胞は露出した幹細胞であり、不等分裂を細胞レベルで研究するのに極めて優れている。そこで本研究はヒメツリガネゴケの幹細胞に着目し、不等分裂による幹細胞の自己複製と細胞分化が細胞周期とともにどのように制御されているかの分子機構を細胞レベルより明らかにすることを目的とした。ヒメツリガネゴケの幹細胞に一過的に過剰発現させることにより、不等分裂異常を引き起こした4種類の因子に着目し、パラログを含めた多重遺伝子破壊体や条件的遺伝子過剰発現体などを作成し、機能解析をすすめた。その結果、GRAS転写因子を条件的に過剰発現することにより、プロトプラストの不等分裂の頻度が下がりかわりに等分裂の頻度が上昇した。また分化した細胞の幹細胞化が促進され、側糸始原細胞の形成頻度が上昇した。一方、転写抑制ドメインを融合した条件的機能抑制体を作成し、機能抑制により細胞増殖が停止し、場合によっては致死となる表現型を得た。これまでの結果とあわせて、この転写因子は幹細胞化を正に制御する上流の転写因子であると考えられた。次に細胞周期制御と極性形成・不等分裂との関係を調べるためにサイクリン依存性キナーゼPpCDKAの機能解析を行った。ゲノム中に2コピー存在するPpCDKAの二重遺伝子破壊体は致死とはならず、この表現型を調べたところ、PpCDKAは細胞周期進行に加え細胞極性や偏光屈性さらにストレス応答の制御に関わることが明らかとなった。またRNA-seqにより二重遺伝子破壊体では多数のストレス制御遺伝子が異常発現していることがわかった。
日本学術振興会, 特定領域研究, 北海道大学, 23012002

Study of molecular mechanism of asymmetry in plant stem cells
Grants-in-Aid for Scientific Research
2011 - 2012
FUJITA Tomomichi
Asymmetric cell division is important for multicellular organisms to control their developmental program. Molecular mechanisms of asymmetric cell division are still largely unknown in plants. The moss, Physcomitrella patens provides a good system for studying the mode of cell division since the whole processes are easily observable at a single cell level.We identified ten of the fluorescent protein-tagged proteins were segregated asymmetrically during cell division, and found distinct types of uneven segregation are underlying during asymmetric cell division. We also found that either under a stress condition or upon abscisic acid treatment, the apical cell changed a cell division mode from asymmetry to symmetry. Then in order to understand a molecular mechanism for how the mode of cell division can be switched, we identified several genes, and further studied their function.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 23657027

メリステム制御の基盤を支える植物幹細胞の不等分裂の分子機構の解明
科学研究費助成事業
2009 - 2011
藤田 知道, 日渡 祐二, 長谷部 光泰
植物の形態形成はメリステムにある幹細胞の増殖と分化のバランスにより制御されている。幹細胞の不等分裂はこのようなバランスを支える基本的しくみであり、メリステムを理解するためには幹細胞の不等分裂の分子制御機構の理解は不可欠である。私達はヒメツリガネゴケから単離したプロトプラストや原糸体の頂端細胞は露出した幹細胞であり、不等分裂を細胞レベルで研究するのに極めて優れていると考えている。そこで本研究はヒメツリガネゴケの幹細胞に着目し、不等分裂による幹細胞の自己複製と細胞分化が細胞周期とともにどのように制御されているかの分子機構を細胞レベルより明らかにすることを目的とし、研究を進めた。その結果、ヒストンシャペロンNAP1とそのパラログの2重遺伝子破壊体では分化した細胞からの幹細胞化が抑制された。また、ヒメツリガネゴケのABAは細胞分裂の分裂様式の切り替え制御に関わることを明らかにし、その制御因子として細胞壁関連タンパク質やABAシグナル因子を同定することができた。それらの機能解析を開始した。また細胞がもつ位置情報は、隣り合う細胞運命の決定や維持に重要であり、このような位置情報が原形質連絡を介してどのように制御されているのか、その時空間的制御を細胞レベルで解析できる実験系を光変換蛍光タンパク質をヒメツリガネゴケの原糸体に応用することで開発できた。その結果、細胞の位置に応じて高分子の移動度が異なることを見出した。またこの違いは原形質連絡のどのような違いが原因で起こりうるのか、電子顕微鏡による超微細構造変化の解析を開始した。
日本学術振興会, 特定領域研究, 北海道大学, 21027002

Isolation and characterization of novel factors that regulate asymmetric cell division in plants
Grants-in-Aid for Scientific Research
2008 - 2010
FUJITA Tomomichi
Since little is known about molecular mechanisms of asymmetric cell division in plants, we undertook to identify genes that are involved in the processes. The moss, Physcomitrella patens provides a good system for the study of asymmetric cell division. We identified more than 60 genes that impaired asymmetric cell division when overexpressed. We explored dynamics of the candidate's proteins during asymmetric cell division by introducing a yellow fluorescent protein as a tag into the c-terminus. We found proteins with unknown function, plant-specific transcription factors, chromatin remodeling factors, small G proteins related to asymmetric cell division. Moreover, putative cell wall proteins are responsible for a suppression of the asymmetric cell division.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 20570030

Characterization of genes for cell polarity and asymmetric cell division in plants
Grants-in-Aid for Scientific Research
2006 - 2007
FUJITA Tomomichi, HASEBE Mitsuyasu
Asymmetric cell division produces two different daughter cells. Although unequal distribution of mRNA or proteins has been known to play a pivotal role to specialize each daughter cell, such molecules in plants remain largely unknown. Protoplasts and apical stem cells of protonemata of the moss, Physcomitrella patens divide asymmetrically to generate two different daughter cells. We have previously reported 59 genes as candidates involved in the asymmetric cell division. For those candidates, we made transgenic plants expressing the citrine-fusion protein by using the gene targeting technique to investigate protein localization under a control of their native promoters. We carried out time-lapse imaging of these proteins during asymmetric cell division and found some proteins accumulated preferentially in the stem cells but not in differentiated protonemal cells. We also showed some of them resumed the accumulation upon dedifferentiation of the protonemal, somatic cells.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 18570031

ポリコーム遺伝子を手がかりとした生活史、特に世代交代進化多様性創出機構の解明
科学研究費助成事業
2005 - 2006
長谷部 光泰, 日渡 祐二, 藤田 知道
ポリコームグループ遺伝子は3種類の遺伝子(C3H2型ジンクフィンガー遺伝子、SETドメイン遺伝子、WD40遺伝子)の総称であり、これらがヒストン脱アセチル化酵素と複合体を作って染色体に結合し、染色体の状態を変化させる。シロイヌナズナでは、3種類の遺伝子のうち前2者は複数個がゲノム内に存在し、組み合わせが変わることで異なった発生プログラムを制御している。昨年度のヒメツリガネゴケゲノム解析によって明らかになったシロイヌナズナのVRN2/FIS2/EMF2/EMF2-like遺伝子オーソログであるPpEMF1,2,3、FIE遺伝子オーソログであるPpFIE、MSI遺伝子オーソログであるPpMSI、CLF/EZA1オーソログであるPpCLF遺伝子の遺伝子破壊体を作出した。1倍体組織である原糸体由来のプロトプラストを用いてPpCLFとPpFIEの各々の遺伝子破壊を行ったところ、両者ともに胞子体様の組織を形成した。形質転換後の原糸体はしばらく成長を続けるが、分岐時に胞子体頂端細胞に酷似した細胞が形成され、その後、胞子体初期発生過程と良く類似した細胞分裂様式を行う。胞子体は頂端分裂細胞が分裂活性を持つ時期と柄分裂組織が形成され胞子嚢形成が起こる時期の2段階に分けることができるが、各遺伝子破壊体では、前者の発生段階のみ観察され、後者の発生段階へは移行しなかった。3つのPpFIS遺伝子それぞれの遺伝子破壊体を作製したどれも野生型と区別できなかった。これは、各遺伝子に機能冗長性があるためではないかと考えられ、機能解析には3重遺伝子破壊体の作出が必要であろう。また、各遺伝子のGUS融合ラインを作製し、1倍体で発現していることがわかった。以上より、本研究によりPcG遺伝子がヒメツリガネゴケにおいて世代交代を制御しており、今後の生活史進化研究の鍵となることがわかった。
日本学術振興会, 萌芽研究, 基礎生物学研究所, 17657033

ヒメツリガネゴケを用いた体軸形成開始点としての植物不等分裂の分子機構の解明
科学研究費助成事業
2005 - 2006
藤田 知道, 長谷部 光泰, 村田 隆, 日渡 祐二
ヒメツリガネゴケプロトプラストにおける完全長cDNAの一過的過剰発現スクリーニング法により得た不等分裂に関わる候補遺伝子58種類について、不等分裂時におけるタンパク質の局在解析を進めた。黄色蛍光タンパク質遺伝子をそれぞれの候補遺伝子に対してノックインした形質転換体を作成し、内在プロモーター制御下における融合タンパク質の局在を観察した。32種類について融合タンパク質の観察を行い、15種類で蛍光シグナルを確認した。これらの中には細胞極性形成部位に蓄積する植物特異的機能未知因子や分化した細胞が再び細胞増殖能を獲得し幹細胞化する過程で発現が誘導される植物特異的転写因子などが含まれていた。
ヒメツリガネゴケ原糸体の頂端幹細胞特異的発現を示す遺伝子トラップラインから同定したユビキチン様タンパク質遺伝子PUBL1、キネシン様タンパク質遺伝子API1の機能解析を進めた。これまでの結果からPUBL1、API1のそれぞれは微小管制御を介して細胞極性や細胞分裂制御に関わることがわかってきた。GFP-チューブリン系統でPUBL1と姉妹遺伝子PUBL2の二重遺伝子破壊を行い細胞分裂時の微小管動態を調べた。その結果、二重遺伝子破壊系統ではフラグモプラスト微小管の崩壊が遅延した。フラグモプラスト赤道面では両極から伸びる微小管のプラス端が交差、架橋されており、その交差領域がフラグモプラストの形成維持に重要であると考えられている。二重遺伝子破壊系統ではこの交差領域が拡大しており、安定化していると考えられた。また、PUBLs-GFP融合タンパク質はフラグモプラスト赤道面の微小管交差領域に局在し、その一部はAPI1タンパク質とも共局在した。API1は交差部位で微小管同士を架橋することでフラグモプラストの制御に関わることを明らかにしており、PUBLsはこの架橋を解除する機能を担っていると考えられた。
日本学術振興会, 特定領域研究, 北海道大学, 17027025

Evolution of proteins in digestive fluid of carnivorous plants
Grants-in-Aid for Scientific Research
2004 - 2006
HASEBE Mitsuyasu, UZAWA Taketoshi, HIWATASHI Yuji, FUJITA Tomomichi, HOSHI Yoshikazu
Activities of various enzymes have been reported in a digestive fluid of carnivorous plants and several genes coding the enzymes were characterized, although their origin and evolution have not been well studied. We analyzed amino acid sequences of secreted proteins in digestive fluids of three independently evolved pitcher plants Nepenthes alata, Sarracenia purpurea, and Cephalotus follicularis as well as a sundew Drosera adelae. Similarity search of the proteins and phylogenetic analyses showed that non-carnivorous plants have homologous proteins to those in the fluid. This suggests that carnivorous enzymes were co-opted from those used in non-carnivorous plants, including pathogenesis and development.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), National Institute for Basic Biology, 16370102

Identification and characterization of genes for cell polarity and asymmetric cell division in plants
Grants-in-Aid for Scientific Research
2004 - 2005
FUJITA Tomomichi, HASEBE Mitsuyasu, MURATA Takashi
Asymmetric cell division generates two different daughter cells. Although unequal distribution of mRNA or proteins has been known to play a pivotal role to specialize each daughter cell, such molecules in plants remain largely unknown. The moss, Physcomitrella patens assures a good system for the study of molecular mechanisms for asymmetric cell division. The protoplast divides asymmetrically to generate apical stem cell and differentiated protonemal, non-stem cell, thereafter the apical stem cell continues to divide asymmetrically to generate a row of differentiated protonemal cells. We have devised a systematic overexpression screening and identified 58 cDNAs as candidates for genes that are involved in asymmetric cell division. For those candidates, we made cDNA-citrine knock-in transgenic plants by using gene targeting technique to investigate protein localization during asymmetric cell division under a control of native promoters. We found several fusion proteins preferentially accumulated in apical stem cells, two of which were intracellularly localized within the apical cell.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), 16570041

多細胞生物進化解明の基盤としての植物不等分裂に関わる遺伝子ネットワークの解明
科学研究費助成事業
2004 - 2004
藤田 知道, 佐藤 良勝, 内山 郁夫, 日渡 祐二
完全長cDNA一過的過剰発現法による植物不等分裂に関わる遺伝子の網羅的単離、機能推定
1、ヒメツリガネゴケから単離したプロトプラストの細胞極性形成開始から不等分裂中の過程で発現している遺伝子群を効率的かつ網羅的に単離するため、プロトプラストを調製後2-3日目の時点で細胞を回収し、RNAを抽出し、完全長cDNAライブラリーを作成した。また、不等分裂細胞を多く含む胞子体2倍体世代の若い胚を集め、RNAを抽出し、完全長cDNAライブラリーを作成した。これら2種類の完全長cDNAライブラリーから、それぞれ約1万クローンのEST解析を行っている。
2、独自に開発したヒメツリガネゴケプロトプラスト一過的過剰発現スクリーニングにより、本年度新たに約600種類のcDNAクローンの一過的過剰発現スクリーニングを行った。その結果等分裂や細胞が巨大化するなど様々な極性形成や不等分裂異常を引き起こす原因遺伝子を見いだした。これまでのスクリーニングとあわせて約200種類について異常頻度を再検討し、不等分裂候補遺伝子を60種類に絞った。これら候補遺伝子の全塩基配列を決定し、配列に基づいた機能推定を行った。
3、候補遺伝子の19種類についてRNA干渉による遺伝子機能抑制による不等分裂の表現型を観察したところ、複数の遺伝子において機能抑制が不等分裂異常を引き起こすことがわかった。さらに、候補遺伝子産物の細胞内局在を調べるため、相同組換えを用いて蛍光タンパク質をノックインした安定形質転換体の作成を開始した。頂端幹細胞で特異的に蓄積するものなどが同定できた。
日本学術振興会, 特定領域研究, 基礎生物学研究所, 16011261

ヒメツリガネゴケとシロイヌナズナで体軸形成開始点としての不等分裂分子機構の解明
科学研究費助成事業
2004 - 2004
藤田 知道, 長谷部 光泰, 村田 隆, 日渡 祐二, 佐藤 良勝
1、植物不等分裂に関わる遺伝子の網羅的単離、機能推定
ヒメツリガネゴケ一過的過剰発現系を用いて、新たに約600種類の完全長cDNAのスクリーニングを行い、様々なプロトプラスト再生異常を引き起こす原因遺伝子を同定した。これまでに同定した再生異常を引き起こす遺伝子と併せて約200種類の再生異常の再現性、頻度を検討し、不等分裂に関わると考えられる候補遺伝子を58種類に絞り込んだ。これら候補遺伝子の全塩基配列を決定した。その中の18種類についてRNA干渉による遺伝子機能抑制による不等分裂の表現型を観察したところ、複数の遺伝子において機能抑制が不等分裂異常を引き起こすことがわかった。さらに、候補遺伝子産物の細胞内局在を調べるため、相同組換えを用いて蛍光タンパク質をノックインした安定形質転換体の作成を開始した。頂端幹細胞で特異的に蓄積するものなどが同定できた。
2、ヒメツリガネゴケ原糸体軸上で不均等に発現する遺伝子の機能解析
II型ユビキチン様タンパク質をコードしているyh78および姉妹遺伝子の二重破壊株の表現型の観察および、YH78-GFP融合タンパク質の細胞内局在を観察した結果、YH78は微小管の制御を通して細胞分裂と細胞極性決定の両方に関わると考えられた。酵母ツーハイブリッド法を用いて相互作用因子を探索し、26Sプロテアソームの構成タンパク質やDnaJ様タンパク質など複数の候補因子を同定することができた。植物特異的キネシンであるAPI1遺伝子の姉妹遺伝子を単離し、二重破壊株を作出した。今後この変異体の表現型の観察を行う。ET21遺伝子と姉妹遺伝子pphn19p09の二重破壊株やシロイヌナズナオーソログ誘導系変異体を作出し表現型解析を行い、機能を検討している。
日本学術振興会, 特定領域研究, 15031225

Analysis on mechanism for microtubule formation in a plant cortical array
Grants-in-Aid for Scientific Research
2003 - 2004
MURATA Takashi, HASEBE Mitsuyasu, FUJITA Tomomichi
Despite the absence of a conspicuous microtubule organizing center, microtubules in interphase plant cells are present in the cell cortex as a well oriented array. By the support of the Grant-in-Aid for Scientific Research, we show that nucleation requires extant cortical microtubules, onto which cytosolic γ-tubulin is recruited. Microtubule-independent nucleation is rarely observed in living tobacco BY-2 cells and nucleation is minimal in the absence of original microtubules in a cell-free system. In both living cells and the cell-free system, microtubules are nucleated as branches on the extant cortical microtubules. The branch points contain γ-tubulin, which is abundant in the cytoplasm, and microtubule nucleation in the cell free system is prevented by inhibiting γ-tubulin function with specific antibodies. When isolated plasma membrane with microtubules is exposed to purified neuro-tubulin, no microtubules are nucleated, but when the membrane is exposed to a cytosolic extract, γ-tubulin binds microtubules on the membrane, and after a subsequent incubation in neuro-tubulin, microtubules are nucleated on the pre-existing microtubules. We propose that a cytoplasmic γ-tubulin complex shuttles between cytoplasm and the side of a cortical microtubule and has nucleation activity only when bound to the microtubule.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), 15570057

多細胞生物進化解明の基盤としての植物不等分裂に関わる遺伝子ネットワークの解明
科学研究費助成事業
2003 - 2003
藤田 知道, 内山 郁夫, 村田 隆
完全長cDNA一過的過剰発現法による植物不等分裂に関わる遺伝子の網羅的単離、機能推定
1、ヒメツリガネゴケから単離したプロトプラストの細胞極性形成開始から不等分裂中の過程で,発現している遺伝子群を効率的かつ網羅的に単離するため、この過程の再生細胞を濃縮して回収する方法を検討した。プロトプラストを単離し、培養開始後2-3日目の時点で細胞を回収し、網目サイズ20μmのフィルターで再生中の細胞と死細胞を分ける操作を行った。回収した再生中の細胞はその後も再生を続けていくことを確認し、この操作が再生細胞の成長に及ぼす影響は排除できると考えられた。このようにして回収した細胞からRNAを抽出し、完全長cDNAライブラリーを作成している。
2、既存の完全長cDNAライブラリーを対象として本年度新たに約1000cDNAクローンの一過的過剰発現スクリーニングを行った。その結果等分裂や細胞が巨大化する、細胞死を起こすなど様々な再生異常を引き起こす原因遺伝子を見いだした。
3、これまでのスクリーニングからあわせて約200cDNAが再生過程に異常を引き起こすことがわかった。これらの表現型の再現性を確認し、細胞極性形成や不等分裂異常に関わると考えられる原因遺伝子を57種類に絞ることができた。これら候補遺伝子の全塩基配列を決定し、配列に基づいて機能の推定を行った。
4、57種類の候補遺伝子についてRNA干渉による表現型観察、ならびにcDNAとGFPの融合タンパク質をプロトプラストに一過的に発現させ不等分裂時の融合タンパク質の細胞内局在を調べるスクリーニングを開始した。RNA干渉、GFP融合コンストラクトを個々の候補遺伝子に対して効率よく作成するためGatewayクローニングを利用できるようにした。これらの方法により候補遺伝子をさらに絞りこむ。
日本学術振興会, 特定領域研究, 岡崎国立共同研究機構, 15011260

ヒメツリガネゴケを用いた細胞の極性形成および不等分裂に関わる遺伝子の単離と解析
科学研究費助成事業
2002 - 2003
藤田 知道
完全長cDNAの一過的過剰発現により、偏光下におけるプロトプラストの極性形成や不等分裂に異常をもたらす原因遺伝子群を同定する。
(1)本年度新たに完全長cDNA約1000種類の一過的過剰発現によるプロトプラスト再生異常のスクリーニングを行い、細胞極性や不等分裂に関わる遺伝子群の一次候補を同定した。
(2)これまでのスクリーニングで得られた一次候補クローンにおいて、表現型の再現性を確認し、不等分裂が等分裂になるもの、不等分裂がおこらず等方位的に細胞が巨大化するものなど細胞極性形成や不等分裂異常に関わると考えられる原因遺伝子を最終的に57種類に絞ることができた。これらcDNAの全長の塩基配列を決定し相同性検索やドメイン解析を行った結果、10種類が既知の極性因子や不等分裂因子と類似していることがわかった。また、機能未知の遺伝子、どの遺伝子とも類似性を示さなかったものも含まれていた。
(3)極性形成や不等分裂は、その過程に関わっている蛋白質自体の極性分布がその作用発現に必須である例が知られている。得られた候補遺伝子とGFP (Green Fluorescence Protein)の融合遺伝子を作成し、プロトプラストで一過的に過剰発現させることにより、融合蛋白質の細胞内局在を調べた。いくつかにおいて原糸体基部に局在するものなどを同定した。
(4)RNA干渉により候補遺伝子の機能抑制時における表現型を調べる。そのためのコンストラクト作りを開始した。このコンストラクトでRNA干渉が実際に働くことをヒメツリガネゴケプロトプラスト再生系において確認した。(3)(4)により、候補遺伝子の不等分裂に関わる機能解析を進めていく。
日本学術振興会, 若手研究(B), 岡崎国立共同研究機構, 14740446

Molecular mechanisms governed the diversity and evolution of flowers
Grants-in-Aid for Scientific Research
2002 - 2003
HASEBE Mistuyasu, FUJITA Tomomichi, UEHARA Kouichi, AOKI Seishiro
Expression patterns of floral homeotic gene homologs in basal angiosperms were examined to infer whether these genes function similarly to those in eudicots. Based on our results, ABC functions of floral homeotic genes were likely established at the stage of basal angiosperms. The intermediate organs between sepals and petals, named as tepals, were formed by the variation of expressions among AP3, P1 and SBP homologs. We also found that AP1 homologs in basal angiosperms were expressed more broadly than eudicot AP1 homologs, and proposed a hypothesis that the broad expression patterns are likely related to the formation of spiral flowers, which often observed in basal angiosperms. Furthermore, we found that petal-like characters of the Houttuynia flower are managed by the heterotopic expression of floral homeotic gene homologs in bracts. Together, this study showed that molecular mechanisms of flower development of eudicots similarly function in basal angiosperms, and the modifications of basic mechanisms are related to the basal-angiosperm specific flowers.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), OKAZAKI NATIONAL RESEARCH INSTITUTES, 14340270

ヒメツリガネゴケとシロイヌナズナで体軸形成開始点としての不等分裂分子機構の解明
科学研究費助成事業
2002 - 2002
長谷部 光泰, 藤田 知道, 村田 隆
(1)ツリガネゴケの約1万5千遺伝子のカタログの中から、代謝、光合成など極性分裂には直接関わりが無いであろうと予想される遺伝子をのぞき、約4000遺伝子を選択した。これらを順次、ヒメツリガネゴケ過剰発現ベクターにサブクローニングし、ヒメツリガネゴケの頂端分裂細胞由来プロトプラストに形質転換し、不等分裂異常をおこす遺伝子を探索している。これまでに約2500のサブクローンが終わり、約1700遺伝子についての形質転換実験を行った。その結果、約100遺伝子が不等分裂に異常を引き起こすことがわかった。これらの中には、不等分裂が等分裂になるもの、分裂細胞が多数できてしまうもの、不等分裂がおこらず1細胞のまま等方位性を保って大きくなったもの、細胞が細胞分裂無しで伸張し続けるものなど様々な表現型に分類できる機能未知の遺伝子および新規遺伝子が含まれている。(2)ヒメツリガネゴケジーントラップライン約2万ラインのスクリーニングの結果、茎頂分裂組織特異的に発現している3ラインのトラップ遺伝子の単離に成功した。これらの遺伝子の機能解析を行った。植物特異的キネシンであるAPI1遺伝子は頂端分裂細胞で発現し、細胞分裂間期に核、終期に細胞板に局在することがGFPとの融合タンパク質の発現解析によりわかった。頂端細胞特異的に発現するユビキチン様タンパク質遺伝子yh78、および姉妹遺伝子であるpph27a22の単独および二重遺伝子破壊株の表現型を観察した。その結果、これらの遺伝子は、細胞分裂と細胞極性決定の両方に関わる機能を持っていると予想された。#21遺伝子は140アミノ酸残基からなるタンパク質をコードしており、そのうちの20アミノ酸はシロイヌナズナのオーソログと高度に類似しているが、それ以外の部分はほとんど類似していないことがわかった。
日本学術振興会, 特定領域研究, 岡崎国立共同研究機構, 14036229

ヒメツリガネゴケで植物ホルモンオーキシン、サイトカイニン機能解明への網羅的研究
科学研究費助成事業
2001 - 2001
長谷部 光泰, 内山 郁夫, 藤田 知道, 村田 隆
本研究は、ヒメツリガネゴケをモデルとして、植物の形態形成に重要な働きを持つ植物ホルモンであるオーキシンとサイトカイニンの作用機作に関わる遺伝子群を網羅的に探索することを目的とした。研究方法は、(1)ホルモン無処理、オーキシン処理、サイトカイニン処理サンプルを用いて完全長cDNAライブラリーを作成、(2)ESTの配列決定、(3)カタログ化したクローンを順次ヒメツリガネゴケで過剰発現させオーキシン処理、サイトカイニン処理したときと同じ表現型を示す遺伝子を網羅的に探索するという3段階ですすめた。その結果、1)予定全ライブラリーについて合計5万ESTの塩基配列を決定しほとんどを一般公開した(遺伝研、新井理、小原雄治との共同研究)。2)本研究で構築したEST配列データと既公開のものを合わせた統合ヒメツリガネゴケESTデータベースを構築した(投稿準備中)。各クローンのBLAST検索結果、contig地図、過剰発現時表現型などが一覧できる。3)ESTの中から興味深いクローンについて全長塩基配列を決定し遺伝子系統樹解析から被子植物ゲノムとヒメツリガネゴケゲノムの比較、その進化について考察した。4)単離プロトプラストにDNAを導入後偏光下で培養することにより第1不等分裂の方向を一方向にそろえる条件を確立した。これにより効果的スクリーニングができるようになった。5)過剰発現用に約1500クローンをサブクローニング、約500cDNAの過剰発現表現型の観察をし、オーキシン、サイトカイニンに対する応答性が異なる10cDNAの単離に成功した。これに加え不等分裂異常など生物学的に興味深い表現型を示すcDNAをカタログ化した。
日本学術振興会, 特定領域研究(C), 岡崎国立共同研究機構, 13202066

ヒメツリガネゴケを用いた茎頂分裂組織形成、維持、器官形成に関する分子機構の解明
科学研究費助成事業
2001 - 2001
長谷部 光泰, 藤田 知道, 村田 隆
(1)PpKNOX1-GUS融合タンパク質は受精前の卵細胞で細胞質に、受精後は受精卵の核で発現が見られ、その後胞子体の分裂組織周辺で発現が維持されることがわかった。GFP-PpKNOX1でも同じ結果が得られた。一方、配偶体世代(原糸体・茎葉体)の頂端分裂組織では発現が検出されなかった。このことから、KNOX遺伝子は胞子体世代の茎頂分裂組織のみで機能していることがわかった。
(2)仮根は茎葉の表皮細胞から分化し、オーキシンによってその形成が誘導されることがわかった。PpHB7遺伝子破壊体は仮根形成数、位置には影響が見られないが、形成される仮根では、色素沈着が少なく、葉緑体数が増え、葉緑体の大きさもおおきくなることがわかった。このことからPpHB7遺伝子は仮根細胞分化を制御していることがわかった。
(3)茎葉体茎頂分裂組織で発現の見られるApi2ラインはユビキチン様遺伝子をトラップしていることがわかった。詳細な発現場所、遺伝子破壊体の表現型を観察中。茎葉体茎頂、原糸体頂端細胞で強い発現のみられる#21ラインは、約130残基のアミノ酸をコードする低分子タンパク質であることがわかった。シロイヌナズナにも1つオーソログがあり、茎頂分裂組織で強く発現していることがわかった。
(4)cDNAアクチベーション系は、全長cDNAライブラリーの5'、3ユ両側からのEST data baseを作成し、順次、各cDNAをヒメツリガネゴケ内で過剰発現させ、変異体を単離する方法である。(1)未処理、(2)オーキシン処理、(3)サイトカイニン処理した原糸体、茎葉体を含むサンプル由来の平均化全長cDNAライブラリーを用いて、ESTクローンを順次プロトプラストで過剰発現させ、分裂異常のおこるcDNAを選抜している。これまで約1000cDNAについて一過的過剰発現を行い、約20の候補cDNAを得た。
日本学術振興会, 特定領域研究(A), 岡崎国立共同研究機構, 13017218

ヒメツリガネゴケを用いたサイトカイニン合成系、情報伝達系関連遺伝子の単離と解析
科学研究費助成事業
2000 - 2001
藤田 知道
1、ヒメツリガネゴケ完全長cDNAライブラリー作成と塩基配列情報のデータベース化
既存の無処理、オーキシン処理に加え、サイトカイニン処理した原糸体由来の完全長ライブラリーを作成した。この3種類から合計4万のcDNAクローンの塩基配列を両末端から決定し、そのほとんどを一般公開した。これらのデータと既公開のものをあわせた統合ヒメツリガネゴケデータベースを構築した。
2、サイトカイニン応答異常変異体の単離
(1)スクリーニングの効率を高めるため、完全長cDNAをヒメツリガネゴケの単離プロトプラストに導入し一過的に過剰発現させ、サイトカイニン応答が異常になる変異体を同定できる系を構築した。また、このための一過的過剰発現用ベクターを新たに作成した。
(2)異なるサイトカイニン、オーキシン濃度を組み合わせた培地におけるヒメツリガネゴケの単離プロトプラストの再生過程を調べた。サイトカイニン濃度が高くなるにつれて、再生5細胞目から多数の頂端細胞からなると思われる細胞塊が誘導された。この基礎データは、過剰発現変異体をスクリーニングする際の1つの指標として用いた。
(3)サイトカイニンとオーキシン添加再生培地下でも正常な形態の原糸体を再生させたものやホルモン無添加再生培地において側枝頂端細胞の形態が異常になったもの、さらには通常オーキシン添加時でのみ見られるカウロネマ細胞が無添加培地において分化してきたものなど、過剰発現によりサイトカイニンやオーキシンに対する応答性に影響を与えた遺伝子を10種類同定することができた。これらのクローンを基にしたより詳細な解析を行っていくことで、サイトカイニンの作用機構に関する制御系の一端が明らかになると考える。
日本学術振興会, 奨励研究(A), 岡崎国立共同研究機構, 12740449

Mechanisms of parallel evolution in plant stems and leaves
Grants-in-Aid for Scientific Research
2000 - 2001
HASEBE Misuyasu, FUJITA Tomomichi, TSUKAYA Hirokazu, MURATA Takashi
1. Class 1 KNOX gene of Physcomitrella patens (PpKNOX 1) was cloned and each GUS and GFP reporter gene was inserted into either 3 and 5 terminal of PpKNOX genomic region using gene targeting technique. The fusion protein was expressed only in the sporophyte generation but not in the gametophyte generation in which stems and leaves develop. This result indicates that molecular mechanisms of stems and leaves between angiosperms and Physcomitrella are different. The ectopic expression of PpKNOX1 gene in Arabidopsis caused the serrated leaves similar to the transformants expressing Arabidopsis class 1 KNOX gene, suggesting that class 1 KNOX function in angiosperms had already established in the common ancestor of angiosperms and Physcomitrella that did not yet have leaves and stems in the sporophyte generation. 2. Polar transport of auxin between gametophores and sporophytes of P. patens, Polytrichum commune, and Bartramia pomiformis was compared. We measured polar auxin transport by a feeding experiment of radiolabelied auxin in either end of the cutting segment of the shoot of mosses, and found that there was little difference of basipetal auxin transport from acropetal transport in gametophores, but obvious polar transport from apical to basal transport was observed in sporophytes. This result also support the hypothesis that leaves and stems of the gametophyte generation in mosses are differently regulated from those of the sporophyte generation in angiosperms.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Okazaki National Research Institutes, 12440242

ヒメツリガネゴケで植物ホルモンオーキシン、サイトカイニン機能解明への網羅的研究
科学研究費助成事業
2000 - 2000
長谷部 光泰, 西山 智明, 藤田 知道
本研究では、ヒメツリガネゴケをモデルとして、オーキシンとサイトカイニンの生合成、受容、情報伝達に関わる遺伝子を誘導cDNAアクチベーション法(全長cDNAを過剰発現プロモーターによって発現させ、変異体を得る方法)により網羅的に単離することを目的としている。
1.mRNAの5'末端に存在するキャップ構造を利用してヒメツリガネゴケ完全長cDNAライブラリーを作成した(理研篠崎研、林崎研との共同研究)。現在、オーキシン処理、サイトカイニン処理したヒメツリガネゴケ由来の完全長cDNAライブラリーを作成中である。
2.作成したヒメツリガネゴケ完全長cDNAライブラリーのうち、1000クローンの塩基配列を決定した結果、90%以上が、完全長cDNAであることがわかった。来年度末までに3万クローンの配列決定、カタログ化を行う予定である。
3.被子植物のオーキシン応答プロモーターであるDR5、GH3プロモーターにGFPをつないだコンストラクトを作成し、ヒメツリガネゴケのニュートラルサイトであるPpHB9サイトに導入した。その結果、GH3プロモーターがヒメツリガネゴケ内で局所的に発現していることがわかり、オーキシンの分布の指標として使える可能性があることがわかった。
4.2で塩基配列決定したクローンを、順次、オーキシン応答プロモーター::GFPを持つヒメツリガネゴケに導入し、過剰発現株の作成を開始した。来年度末までに7千変異体の単離をめざしている。
日本学術振興会, 特定領域研究(C), 岡崎国立共同研究機構, 12202051

ヒメツリガネゴケを用いた茎頂分裂組織形成、維持、器官形成に関する分子機構の解明
科学研究費助成事業
2000 - 2000
長谷部 光泰, 藤田 知道
ヒメツリガネゴケ胞子体由来RNAからヒメツリガネゴケPpKNOX1遺伝子の全長cDNAを単離し、その塩基配列に基づき対応するゲノム領域を単離した。ノザン解析、ジーンターゲティングによりPpKNOX1遺伝子3末にGUS遺伝子を挿入して発現解析を行った。その結果、PpKNOX-GUS融合タンパク質は受精前の卵細胞で細胞質に、受精後は受精卵の核で発現が見られ、その後胞子体の分裂組織周辺で発現が維持される。一方、配偶体世代(原糸体・茎葉体)の頂端分裂組織では発現が検出されなかった。
PpHB遺伝子グループ1と2に属するPpHB3,4,5,6,7,8,9の7遺伝子(Sakakibara et al.投稿中)について恒常的過剰発現形質転換体を作成した。また、相同組換えにより各遺伝子末端にGUS遺伝子を導入した。現在、それぞれについて表現型を観察中である。PHABULOSA遺伝子(PpHBグループ3)ホモログであるPpHB10遺伝子についても同様の実験を進めている。
PpHB7の発現様式、過剰発現体の解析から、この遺伝子は仮根(リゾイド)形成に関与していることがわかった。
シーンターゲティングによりPpMADS1遺伝子末端にGUS遺伝子を挿入し、PpMADS1-GUS融合タンパク質の発現を解析した。発現様式から、PpMADS1遺伝子は卵、精子形成、および胞子体での未知の作用に関与しているのではないかと推察された。
遺伝子トラップ系から得たAPI1はシロイヌナズナの機能未解明なキネシン類似遺伝子とともに、従来報告されたキネシンとは異なった、植物に特異的な新グループを構成することがわかった。
日本学術振興会, 特定領域研究(A), 岡崎国立共同研究機構, 12037218

ヒメツリガネゴケを用いた植物細胞極性、不等分裂、分化全能性の仕組みの解明　　　　　　　　　　　　　　　
Competitive research funding

Molecular characterization of cell polarity, asymmetric cell division and totipotency in plants　　　　　　　　　　　　　　　
Competitive research funding

宇宙環境利用について研究者が語るイベント「宇宙×金融＃12 宇宙実験が拓く！生物・植物の不思議 ─コケ植物培養から火星移住への道筋─」　　　　　　　　　　　　　　　
18 Oct. 2024
Appearance, Panelist, Lecturer
Lecture
[https://digitalblast.co.jp/news/宇宙環境利用について研究者が語るイベント「宇-2/]

Science cafe at Huazhong Agricultural University, Wuhan（華中農業大学、武漢）-"Meet undergraduates"-　　　　　　　　　　　　　　　
13 Oct. 2024
Appearance, Lecturer
Science cafe

第1回 宇宙技術者会議（北大医学部百年記念会館）「“コケ”で宇宙実験！人類を救うプロジェクト！！」　　　　　　　　　　　　　　　
08 Oct. 2024
Lecturer

サイエンスZERO 「人類の未来を変える"吸収力" 小さなコケのミラクルパワー」　　　　　　　　　　　　　　　
08 Sep. 2024
Appearance, Coverage cooperation
NHK Eテレ

BiocK宇宙バイオ実験分科会キックオフイベント「宇宙 x ライフサイエンス in 神戸」　　　　　　　　　　　　　　　
19 Mar. 2024
Appearance, Panelist, Organizing member
Meeting

YAC種子島スペースキャンプ2023夏　　　　　　　　　　　　　　　
30 Jul. 2023 - 01 Aug. 2023
Lecturer, Advisor
YAC 日本宇宙少年団
[https://www.yac-j.com/feature/tanegashima-summer2023/]

千葉県船橋市立西海神小学校「総合：キャリア教育」6年生：ゲストティーチャー　　　　　　　　　　　　　　　
Feb. 2023
Lecturer

北海道大学オープンキャンパス（高校生限定プログラム）(2010〜2019年, 2022〜2023年)　　　　　　　　　　　　　　　
2010 - 2023
Lecturer, Planner
Others

ほっとニュース北海道（報道）「植物の成長を調整するたんぱく質発見」　　　　　　　　　　　　　　　
14 Dec. 2022
Coverage cooperation
NHK札幌
ほっとニュース北海道（2023年1月30日放映）

企画展および企画展記念講演会 こけティッシュ・プレミアムトーク　　　　　　　　　　　　　　　
09 Jan. 2022
Panelist, Coverage cooperation, Lecturer, Advisor, Organizing member
茨城県自然博物館
企画展「こけティッシュ 苔ニューワールド！―地球を包むミクロの森―」

千葉県船橋市立西海神小学校「総合：キャリア教育」6年生：ゲストティーチャー　　　　　　　　　　　　　　　
Feb. 2021
Lecturer

高校生向け 学問新サイト「みらいぶっく・学問・大学なび」掲載　　　　　　　　　　　　　　　
May 2020
Coverage cooperation, Informant, Contribution
河合塾

NASA Media Teleconference（出演）　　　　　　　　　　　　　　　
10 Jul. 2019
Appearance, Coverage cooperation
National Aeronautics and Space Administration (NASA)

ほっとニュース北海道 ：ブーム到来中！宇宙に飛び立つ「コケ」　　　　　　　　　　　　　　　
01 Aug. 2016
Appearance, Coverage cooperation
NHK札幌
ほっとニュース北海道：ほっと中継

文化放送いとうせいこう GREEN FESTA,「スペース・モス」の宇宙実験がJAXAで採用！　　　　　　　　　　　　　　　
16 Feb. 2016
Coverage cooperation
文化放送（2016年2月16日ON AIR）

札幌藻岩高校SPP（サイエンス・パートナーシップ・プロジェクト）　　　　　　　　　　　　　　　
25 Sep. 2009
Lecturer

札幌開成高校（出前授業）　　　　　　　　　　　　　　　
Nov. 2006
Lecturer

広幡小学校 出前授業「科学教室：生き物の遺伝物質をとってみよう」　　　　　　　　　　　　　　　
Feb. 2004
Lecturer
岡崎市立広幡小学校・岡崎ロータリークラブ