2011年09月, 第84回日本生化学会大会, 鈴木紘一メモリアル賞　　　　　　　　　　　　　　　

Taste of microbes: the terroir explained by rhizospheric microbes
Nakano, R.T.
New Phytologist, 243, 5, 1639, 1641, 2024年09月, ［招待有り］, ［筆頭著者, 最終著者, 責任著者］, ［国際誌］
英語, 研究論文（学術雑誌）

Long-Term Consequences of PTI Activation and Its Manipulation by Root-Associated Microbiota
Ryohei Thomas Nakano, Tomohisa Shimasaki
Plant And Cell Physiology, 2024年05月30日, ［査読有り］, ［筆頭著者, 責任著者］, ［国内誌］
英語, 研究論文（学術雑誌）, In nature, plants are constantly colonized by a massive diversity of microbes engaged in mutualistic, pathogenic, or commensal relationships with the host. Molecular patterns present in these microbes activate pattern-triggered immunity (PTI), which detects microbes in the apoplast or at the tissue surface. Whether and how PTI distinguishes among soil-borne pathogens, opportunistic pathogens, and commensal microbes within the soil microbiota remains unclear. PTI is a multi-modal series of molecular events initiated by pattern perception, such as Ca2+ influx, reactive oxygen burst, and extensive transcriptional and metabolic reprogramming. These short-term responses may manifest within minutes to hours, while the long-term consequences of chronic PTI activation persist for days to weeks. Chronic activation of PTI is detrimental to plant growth, so plants need to coordinate growth and defense depending on the surrounding biotic and abiotic environments. Recent studies have demonstrated that root-associated commensal microbes can activate or suppress immune responses to variable extents, clearly pointing to the role of PTI in root-microbiota interactions. However, the molecular mechanisms by which root commensals interfere with root immunity and root immunity modulates microbial behavior remain largely elusive. Here, with a focus on the difference between short-term and long-term PTI responses, we summarize what is known about microbial interference with host PTI, especially in the context of root microbiota. We emphasize some missing pieces that remain to be characterized to promote the ultimate understanding of the role of plant immunity in root-microbiota interactions.

MYB68 regulates radial endodermal differentiation and suberin patterning
Leonie Kraska, Josep Mercadal Melia, Ryohei Thomas Nakano, David Molina, Pau Formosa-Jordan, Laura Ragni, Tonni Grube Andersen
bioRxiv, 2024年05月10日

Apoplastic barrier establishment in roots and nodules of Lotus japonicus is essential for root-shoot signaling and N-fixation
Defeng Shen, Rafael E. Venado, Ulla Neumann, Nadine Dyballa-Rukes, Swati Mahiwal, Sabine Metzger, Ryohei Thomas Nakano, Macarena Marín, Tonni Grube Andersen
bioRxiv, 2023年12月08日

ER body-resident myrosinases and tryptophan specialized metabolism modulate root microbiota assembly.
Basak AK, Piasecka A, Hucklenbroich J, Türksoy GM, Guan R, Zhang P, Getzke F, Garrido-Oter R, Hacquard S, Strzałka K, Bednarek P, Yamada K, Nakano RT
The New phytologist, 2023年09月28日, ［査読有り］, ［最終著者, 責任著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Endoplasmic reticulum (ER) bodies are ER-derived structures that contain a large amount of PYK10 myrosinase, which hydrolyzes tryptophan (Trp)-derived indole glucosinolates (IGs). Given the well-described role of IGs in root-microbe interactions, we hypothesized that ER bodies in roots are important for interaction with soil-borne microbes at the root-soil interface. We used mutants impaired in ER bodies (nai1), ER body-resident myrosinases (pyk10bglu21), IG biosynthesis (myb34/51/122), and Trp specialized metabolism (cyp79b2b3) to profile their root microbiota community in natural soil, evaluate the impact of axenically collected root exudates on soil or synthetic microbial communities, and test their response to fungal endophytes in a mono-association setup. Tested mutants exhibited altered bacterial and fungal communities in rhizoplane and endosphere, respectively. Natural soils and bacterial synthetic communities treated with mutant root exudates exhibited distinctive microbial profiles from those treated with wild-type (WT) exudates. Most tested endophytes severely restricted the growth of cyp79b2b3, a part of which also impaired the growth of pyk10bglu21. Our results suggest that root ER bodies and their resident myrosinases modulate the profile of root-secreted metabolites and thereby influence root-microbiota interactions.

Simultaneous tracking of near-isogenic bacterial strains in synthetic Arabidopsis microbiota by chromosomally-integrated barcodes
Jana Ordon, Julien Thouin, Ryohei Thomas Nakano, Ka-Wai Ma, Pengfan Zhang, Bruno Huettel, Ruben Garrido-Oter, Paul Schulze-Lefert
Nature Microbiology, 2023年04月20日, ［査読有り］
英語, 研究論文（学術雑誌）, AbstractDNA-amplicon-based microbiota profiling can estimate species diversity and abundance but cannot resolve genetic differences within individuals of the same species. Here we report the development of modular bacterial tags (MoBacTags) encoding DNA barcodes that enable tracking of near-isogenic bacterial commensals in an array of complex microbiome communities. Chromosomally integrated DNA barcodes are then co-amplified with endogenous marker genes of the community by integrating corresponding primer binding sites into the barcode. We use this approach to assess the contributions of individual bacterial genes to Arabidopsis thaliana root microbiota establishment with synthetic communities that include MoBacTag-labelled strains of Pseudomonas capeferrum. Results show reduced root colonization for certain mutant strains with defects in gluconic-acid-mediated host immunosuppression, which would not be detected with traditional amplicon sequencing. Our work illustrates how MoBacTags can be applied to assess scaling of individual bacterial genetic determinants in the plant microbiota.

A Friend in Common: A Small GTPase in Independent PTI and ETI Immune Receptor Complexes
Ryohei Thomas Nakano
Plant and Cell Physiology, 62, 11, 1645, 1647, Oxford University Press ({OUP}), 2021年12月10日, ［招待有り］, ［筆頭著者, 最終著者, 責任著者］, ［国内誌］
英語, 研究論文（学術雑誌）

Tryptophan metabolism and bacterial commensals prevent fungal dysbiosis in Arabidopsis roots
Katarzyna W. Wolinska, Nathan Vannier, Thorsten Thiergart, Brigitte Pickel, Sjoerd Gremmen, Anna Piasecka, Mariola Piślewska-Bednarek, Ryohei Thomas Nakano, Youssef Belkhadir, Paweł Bednarek, Stéphane Hacquard
Proceedings of the National Academy of Sciences, 118, 49, Proceedings of the National Academy of Sciences, 2021年12月07日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, In nature, roots of healthy plants are colonized by multikingdom microbial communities that include bacteria, fungi, and oomycetes. A key question is how plants control the assembly of these diverse microbes in roots to maintain host-microbe homeostasis and health. Using microbiota reconstitution experiments with a set of immunocompromised Arabidopsis thaliana mutants and a multikingdom synthetic microbial community (SynCom) representative of the natural A. thaliana root microbiota, we observed that microbiota-mediated plant growth promotion was abolished in most of the tested immunocompromised mutants. Notably, more than 40% of between-genotype variation in these microbiota-induced growth differences was explained by fungal but not bacterial or oomycete load in roots. Extensive fungal overgrowth in roots and altered plant growth was evident at both vegetative and reproductive stages for a mutant impaired in the production of tryptophan-derived, specialized metabolites (cyp79b2/b3). Microbiota manipulation experiments with single- and multikingdom microbial SynComs further demonstrated that 1) the presence of fungi in the multikingdom SynCom was the direct cause of the dysbiotic phenotype in the cyp79b2/b3 mutant and 2) bacterial commensals and host tryptophan metabolism are both necessary to control fungal load, thereby promoting A. thaliana growth and survival. Our results indicate that protective activities of bacterial root commensals are as critical as the host tryptophan metabolic pathway in preventing fungal dysbiosis in the A. thaliana root endosphere.

Retrograde sulfur flow from glucosinolates to cysteine in Arabidopsis thaliana.
Sugiyama R, Li R, Kuwahara A, Nakabayashi R, Sotta N, Mori T, Ito T, Ohkama-Ohtsu N, Fujiwara T, Saito K, Nakano RT, Bednarek P, Hirai MY
Proceedings of the National Academy of Sciences of the United States of America, 118, 22, 2021年06月01日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Specialized (secondary) metabolic pathways in plants have long been considered one-way routes of leading primary metabolite precursors to bioactive end products. Conversely, endogenous degradation of such "end" products in plant tissues has been observed following environmental stimuli, including nutrition stress. Therefore, it is of general interest whether specialized metabolites can be reintegrated into primary metabolism to recover the invested resources, especially in the case of nitrogen- or sulfur-rich compounds. Here, we demonstrate that endogenous glucosinolates (GLs), a class of sulfur-rich plant metabolites, are exploited as a sulfur source by the reallocation of sulfur atoms to primary metabolites such as cysteine in Arabidopsis thaliana Tracer experiments using 34S- or deuterium-labeled GLs depicted the catabolic processing of GL breakdown products in which sulfur is mobilized from the thioglucoside group in GL molecules, potentially accompanied by the release of the sulfate group. Moreover, we reveal that beta-glucosidases BGLU28 and BGLU30 are the major myrosinases that initiate sulfur reallocation by hydrolyzing particular GL species, conferring sulfur deficiency tolerance in A. thaliana, especially during early development. The results delineate the physiological function of GL as a sulfur reservoir, in addition to their well-known functions as defense chemicals. Overall, our findings demonstrate the bidirectional interaction between primary and specialized metabolism, which enhances our understanding of the underlying metabolic mechanisms via which plants adapt to their environments.

Tobacco root endophytic Arthrobacter harbors genomic features enabling the catabolism of host-specific plant specialized metabolites
Shimasaki, T., Masuda, S., Garrido-Oter, R., Kawasaki, T., Aoki, Y., Shibata, A., Shirasu, K., Yazaki, K., Nakano, R.T., Sugiyama, A., Suda, W.
mBio, 12, 3, e0084621, American Society for Microbiology, 2021年05月28日, ［査読有り］, ［責任著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Plant roots constitute the primary interface between plants and soilborne microorganisms and harbor microbial communities called the root microbiota. Recent studies have demonstrated a significant contribution of plant specialized metabolites (PSMs) to the assembly of root microbiota. However, the mechanistic and evolutionary details underlying the PSM-mediated microbiota assembly and its contribution to host specificity remain elusive. Here, we show that the bacterial genus Arthrobacter is predominant specifically in the tobacco endosphere and that its enrichment in the tobacco endosphere is partially mediated by a combination of two unrelated classes of tobacco-specific PSMs, santhopine and nicotine. We isolated and sequenced Arthrobacter strains from tobacco roots as well as soils treated with these PSMs and identified genomic features, including but not limited to genes for santhopine and nicotine catabolism, that are associated with the ability to colonize tobacco roots. Phylogenomic and comparative analyses suggest that these genes were gained in multiple independent acquisition events, each of which was possibly triggered by adaptation to particular soil environments. Taken together, our findings illustrate a cooperative role of a combination of PSMs in mediating plant species-specific root bacterial microbiota assembly and suggest that the observed interaction between tobacco and Arthrobacter may be a consequence of an ecological fitting process. IMPORTANCE Host secondary metabolites have a crucial effect on the taxonomic composition of its associated microbiota. It is estimated that a single plant species produces hundreds of secondary metabolites; however, whether different classes of metabolites have distinctive or common roles in the microbiota assembly remains unclear. Here, we show that two unrelated classes of secondary metabolites in tobacco play a cooperative role in the formation of tobacco-specific compositions of the root bacterial microbiota, which has been established as a consequence of independent evolutionary events in plants and bacteria triggered by different ecological effects. Our findings illustrate mechanistic and evolutionary aspects of the microbiota assembly that are mediated by an arsenal of plant secondary metabolites.

Rhizobiales commensal bacteria promote Arabidopsis thaliana root growth via host sulfated peptide pathway
Jana Hucklenbroich, Tamara Gigolashvili, Anna Koprivova, Philipp Spohr, Mahnaz Nezamiv, Chegini, Gunnar W. Klau, Stanislav Kopriva, Ryohei Thomas Nakano
bioRxiv, Cold Spring Harbor Laboratory, 2021年05月25日, ［最終著者, 責任著者］
研究論文（学術雑誌）

Discovery of a Family of Mixed Lineage Kinase Domain-like Proteins in Plants and Their Role in Innate Immune Signaling.
Mahdi LK, Huang M, Zhang X, Nakano RT, Kopp LB, Saur IML, Jacob F, Kovacova V, Lapin D, Parker JE, Murphy JM, Hofmann K, Schulze-Lefert P, Chai J, Maekawa T
Cell host & microbe, 28, 6, 813, 824, 2020年10月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, HeLo domain-containing mixed lineage kinase domain-like protein (MLKL), a pseudokinase, mediates necroptotic cell death in animals. Here, we report the discovery of a conserved protein family across seed plants that structurally resembles vertebrate MLKL. The Arabidopsis genome encodes three MLKLs (AtMLKLs) with overlapping functions in disease resistance mediated by Toll-interleukin 1-receptor domain intracellular immune receptors (TNLs). The HeLo domain of AtMLKLs confers cell death activity but is dispensable for immunity. Cryo-EM structures reveal a tetrameric configuration, in which the HeLo domain is buried, suggestive of an auto-repressed complex. The mobility of AtMLKL1 along microtubules is reduced by chitin, a fungal immunity-triggering molecule. An AtMLKL1 phosphomimetic variant exhibiting reduced mobility enhances immunity. Coupled with the predicted presence of HeLo domains in plant helper NLRs, our data reveal the importance of HeLo domain proteins for TNL-dependent immunity and argue for a cell death-independent immune mechanism mediated by MLKLs.

Characterization of rhizome transcriptome and identification of a rhizomatous ER body in the clonal plant Cardamine leucantha
Araki, K.S., Nagano, A.J., Nakano, R.T., Kitazume, T., Yamaguchi, K., Hara-Nishimura, I., Shigenobu, S., Kudoh, H.
Scientific Reports, 10, 1, 13291, 13291, 2020年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2020, The Author(s). The rhizome is a plant organ that develops from a shoot apical meristem but penetrates into belowground environments. To characterize the gene expression profile of rhizomes, we compared the rhizome transcriptome with those of the leaves, shoots and roots of a rhizomatous Brassicaceae plant, Cardamine leucantha. Overall, rhizome transcriptomes were characterized by the absence of genes that show rhizome-specific expression and expression profiles intermediate between those of shoots and roots. Our results suggest that both endogenous developmental factors and external environmental factors are important for controlling the rhizome transcriptome. Genes that showed relatively high expression in the rhizome compared to shoots and roots included those related to belowground defense, control of reactive oxygen species and cell elongation under dark conditions. A comparison of transcriptomes further allowed us to identify the presence of an ER body, a defense-related belowground organelle, in epidermal cells of the C. leucantha rhizome, which is the first report of ER bodies in rhizome tissue.

Apoplastic Fluid Preparation from Arabidopsis thaliana Leaves Upon Interaction with a Nonadapted Powdery Mildew Pathogen
Nakano, R.T., Ishihama, N., Wang, Y., Takagi, J., Uemura, T., Schulze-Lefert, P., Nakagami, H.
Methods in Molecular Biology, 2139, 79, 88, Springer US, 2020年01月, ［査読有り］, ［筆頭著者, 責任著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Proteins in the extracellular space (apoplast) play a crucial role at the interface between plant cells and their proximal environment. Consequently, it is not surprising that plants actively control the apoplastic proteomic profile in response to biotic and abiotic cues. Comparative quantitative proteomics of plant apoplastic fluids is therefore of general interest in plant physiology. We here describe an efficient method to isolate apoplastic fluids from Arabidopsis thaliana leaves inoculated with a nonadapted powdery mildew pathogen.

A Golgi-released subpopulation of the trans-Golgi network mediates protein secretion in Arabidopsis
Uemura, T., Nakano, R.T., Takagi, J., Wang, Y., Kramer, K., Finkemeier, I., Nakagami, H., Tsuda, K., Ueda, T., Schulze-Lefert, P., Nakano, A.
Plant Physiology, 179, 2, 519, 532, American Society of Plant Biologists ({ASPB}), 2019年02月, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Spatiotemporal coordination of protein trafficking among organelles is essential for eukaryotic cells. The post-Golgi interface, including the trans-Golgi network (TGN), is a pivotal hub for multiple trafficking pathways. The Golgi-released independent TGN (GI-TGN) is a compartment described only in plant cells, and its cellular and physiological roles remain elusive. In Arabidopsis (Arabidopsis thaliana), the SYNTAXIN OF PLANTS (SYP) 4 group Qa-SNARE (soluble N-ethylmaleimide) membrane fusion proteins are shared components of TGN and GI-TGN and regulate secretory and vacuolar transport. Here we reveal that GI-TGNs mediate the transport of the R-SNARE VESICLE-ASSOCIATED MEMBRANE PROTEIN (VAMP) 721 to the plasma membrane. In interactions with a nonadapted powdery mildew pathogen, the SYP4 group of SNAREs is required for the dynamic relocation of VAMP721 to plant-fungus contact sites via GI-TGNs, thereby facilitating complex formation with its cognate SNARE partner PENETRATION1 to restrict pathogen entry. Furthermore, quantitative proteomic analysis of leaf apoplastic fluid revealed constitutive and pathogen-inducible secretion of cell wall-modification enzymes in a SYP4- and VAMP721-dependent manner. Hence, the GI-TGN acts as a transit compartment between the Golgi apparatus and the plasma membrane. We propose a model in which the GA-TGN matures into the GI-TGN and then into secretory vesicles by increasing the abundance of VAMP721-dependent secretory pathway components.

Modular Traits of the Rhizobiales Root Microbiota and Their Evolutionary Relationship with Symbiotic Rhizobia
Garrido-Oter, R., Nakano, R.T., Dombrowski, N., Ma, K.-W., McHardy, A.C., Schulze-Lefert, P.
Cell Host and Microbe, 24, 1, 155, 167.e5, Elsevier {BV}, 2018年07月, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Rhizobia are a paraphyletic group of soil-borne bacteria that induce nodule organogenesis in legume roots and fix atmospheric nitrogen for plant growth. In non-leguminous plants, species from the Rhizobiales order define a core lineage of the plant microbiota, suggesting additional functional interactions with plant hosts. In this work, genome analyses of 1,314 Rhizobiales isolates along with amplicon studies of the root microbiota reveal the evolutionary history of nitrogen-fixing symbiosis in this bacterial order. Key symbiosis genes were acquired multiple times, and the most recent common ancestor could colonize roots of a broad host range. In addition, root growth promotion is a characteristic trait of Rhizobiales in Arabidopsis thaliana, whereas interference with plant immunity constitutes a separate, strain-specific phenotype of root commensal Alphaproteobacteria. Additional studies with a tripartite gnotobiotic plant system reveal that these traits operate in a modular fashion and thus might be relevant to microbial homeostasis in healthy roots.

Glutathione transferase U13 functions in pathogen-triggered glucosinolate metabolism
Pi?lewska-Bednarek, M., Nakano, R.T., Hiruma, K., Pastorczyk, M., Sanchez-Vallet, A., Singkaravanit-Ogawa, S., Ciesio?ka, D., Takano, Y., Molina, A., Schulze-Lefert, P., Bednarek, P.
Plant Physiology, 176, 1, 538, 551, American Society of Plant Biologists ({ASPB}), 2018年01月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Glutathione (GSH) and indole glucosinolates (IGs) exert key functions in the immune system of the model plant Arabidopsis (Arabidopsis thaliana). Appropriate GSH levels are important for execution of both pre- and postinvasive disease resistance mechanisms to invasive pathogens, whereas an intact PENETRATION2 (PEN2)-pathway for IG metabolism is essential for preinvasive resistance in this species. Earlier indirect evidence suggested that the latter pathway involves conjugation of GSH with unstable products of IG metabolism and further processing of the resulting adducts to biologically active molecules. Here we describe the identification of Glutathione-S-Transferase class-tau member 13 (GSTU13) as an indispensable component of the PEN2 immune pathway for IG metabolism. gstu13 mutant plants are defective in the pathogen-triggered biosynthesis of end products of the PEN2 pathway, including 4-O-β-D-glucosyl-indol-3-yl formamide, indole-3-ylmethyl amine, and raphanusamic acid. In line with this metabolic defect, lack of functional GSTU13 results in enhanced disease susceptibility toward several fungal pathogens including Erysiphe pisi, Colletotrichum gloeosporioides, and Plectosphaerella cucumerina. Seedlings of gstu13 plants fail also to deposit the (1,3)-β-glucan cell wall polymer, callose, after recognition of the bacterial flg22 epitope. We show that GSTU13 mediates specifically the role of GSH in IG metabolism without noticeable impact on other immune functions of this tripeptide. We postulate that GSTU13 connects GSH with the pathogen-triggered PEN2 pathway for IG metabolism to deliver metabolites that may have numerous functions in the innate immune system of Arabidopsis.

PYK10 Myrosinase Reveals a Functional Coordination between ER Bodies and Glucosinolates in Arabidopsis thaliana.
Nakano RT, Piślewska-Bednarek M, Yamada K, Edger PP, Miyahara M, Kondo M, Böttcher C, Mori M, Nishimura M, Schulze-Lefert P, Hara-Nishimura I, Bednarek P
The Plant Journal, 89, 2, 204, 220, WILEY, 2016年09月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, The endoplasmic reticulum body (ER body) is an organelle derived from the ER that occurs in only three families of the order Brassicales and is suggested to be involved in plant defense. ER bodies in Arabidopsis thaliana contain large amounts of b-glucosidases, but the physiological functions of ER bodies and these enzymes remain largely unclear. Here we show that PYK10, the most abundant b-glucosidase in A. thaliana root ER bodies, hydrolyzes indole glucosinolates (IGs) in addition to the previously reported in vitro substrate scopolin. We found a striking co-expression between ER body-related genes (including PYK10), glucosinolate biosynthetic genes and the genes for so-called specifier proteins affecting the terminal products of myrosinase-mediated glucosinolate metabolism, indicating that these systems have been integrated into a common transcriptional network. Consistent with this, comparative metabolite profiling utilizing a number of A. thaliana relatives within Brassicaceae identified a clear phylogenetic co-occurrence between ER bodies and IGs, but not between ER bodies and scopolin. Collectively, our findings suggest a functional link between ER bodies and glucosinolate metabolism in planta. In addition, in silico three-dimensional modeling, combined with phylogenomic analysis, suggests that PYK10 represents a clade of 16 myrosinases that arose independently from the other well-documented class of six thioglucoside glucohydrolases. These findings provide deeper insights into how glucosinolates are metabolized in cruciferous plants and reveal variation of the myrosinase-glucosinolate system within individual plants.

Root Endophyte Colletotrichum tofieldiae Confers Plant Fitness Benefits that Are Phosphate Status Dependent.
Hiruma K, Gerlach N, Sacristán S, Nakano RT, Hacquard S, Kracher B, Neumann U, Ramírez D, Bucher M, O'Connell RJ, Schulze-Lefert P
CELL, 165, 2, 464, 474, CELL PRESS, 2016年04月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, A staggering diversity of endophytic fungi associate with healthy plants in nature, but it is usually unclear whether these represent stochastic encounters or provide host fitness benefits. Although most characterized species of the fungal genus Colletotrichum are destructive pathogens, we show here that C. tofieldiae (Ct) is an endemic endophyte in natural Arabidopsis thaliana populations in central Spain. Colonization by Ct initiates in roots but can also spread systemically into shoots. Ct transfers the macronutrient phosphorus to shoots, promotes plant growth, and increases fertility only under phosphorus-deficient conditions, a nutrient status that might have facilitated the transition from pathogenic to beneficial lifestyles. The host's phosphate starvation response (PSR) system controls Ct root colonization and is needed for plant growth promotion (PGP). PGP also requires PEN2-dependent indole glucosinolate metabolism, a component of innate immune responses, indicating a functional link between innate immunity and the PSR system during beneficial interactions with Ct.

ER bodies in plants of the Brassicales order: Biogenesis and association with innate immunity
Nakano, R.T., Yamada, K., Bednarek, P., Nishimura, M., Hara-Nishimura, I.
Frontiers in Plant Science, 5, MAR, 73, 73, FRONTIERS RESEARCH FOUNDATION, 2014年, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, The endoplasmic reticulum (ER) forms highly organized network structures composed of tubules and cisternae. Many plant species develop additional ER-derived structures, most of which are specific for certain groups of species. In particular, a rod-shaped structure designated as the ER body is produced by plants of the Brassicales order, which includes Arabidopsis thaliana. Genetic analyses and characterization of A. thaliana mutants possessing a disorganized ER morphology or lacking ER bodies have provided insights into the highly organized mechanisms responsible for the formation of these unique ER structures. The accumulation of proteins specific for the ER body within the ER plays an important role in the formation of ER bodies. However, a mutant that exhibits morphological defects of both the ER and ER bodies has not been identified. This suggests that plants in the Brassicales order have evolved novel mechanisms for the development of this unique organelle, which are distinct from those used to maintain generic ER structures. In A. thaliana, ER bodies are ubiquitous in seedlings and roots, but rare in rosette leaves. Wounding of rosette leaves induces de novo formation of ER bodies, suggesting that these structures are associated with resistance against pathogens and/or herbivores. ER bodies accumulate a large amount of beta-glucosidases, which can produce substances that potentially protect against invading pests. Biochemical studies have determined that the enzymatic activities of these beta-glucosidases are enhanced during cell collapse. These results suggest that ER bodies are involved in plant immunity, although there is no direct evidence of this. In this review, we provide recent perspectives of ER and ER body formation in A. thaliana, and discuss clues for the functions of ER bodies. We highlight defense strategies against biotic stress that are unique for the Brassicales order, and discuss how ER structures could contribute to these strategies.

ERMO3/MVP1/GOLD36 Is Involved in a Cell Type-Specific Mechanism for Maintaining ER Morphology in Arabidopsis thaliana
Nakano, R.T., Matsushima, R., Nagano, A.J., Fukao, Y., Fujiwara, M., Kondo, M., Nishimura, M., Hara-Nishimura, I.
PLoS ONE, 7, 11, e49103, PUBLIC LIBRARY SCIENCE, 2012年, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, The endoplasmic reticulum (ER) has a unique, network-like morphology. The ER structures are composed of tubules, cisternae, and three-way junctions. This morphology is highly conserved among eukaryotes, but the molecular mechanism that maintains ER morphology has not yet been elucidated. In addition, certain Brassicaceae plants develop a unique ER-derived organelle called the ER body. This organelle accumulates large amounts of PYK10, a beta-glucosidase, but its physiological functions are still obscure. We aimed to identify a novel factor required for maintaining the morphology of the ER, including ER bodies, and employed a forward-genetic approach using transgenic Arabidopsis thaliana (GFP-h) with fluorescently-labeled ER. We isolated and investigated a mutant (designated endoplasmic reticulum morphology3, ermo3) with huge aggregates and abnormal punctate structures of ER. ERMO3 encodes a GDSL-lipase/esterase family protein, also known as MVP1. Here, we showed that, although ERMO3/MVP1/GOLD36 was expressed ubiquitously, the morphological defects of ermo3 were specifically seen in a certain type of cells where ER bodies developed. Coimmunoprecipitation analysis combined with mass spectrometry revealed that ERMO3/MVP1/GOLD36 interacts with the PYK10 complex, a huge protein complex that is thought to be important for ER body-related defense systems. We also found that the depletion of transcription factor NAI1, a master regulator for ER body formation, suppressed the formation of ER-aggregates in ermo3 cells, suggesting that NAI1 expression plays an important role in the abnormal aggregation of ER. Our results suggest that ERMO3/MVP1/GOLD36 is required for preventing ER and other organelles from abnormal aggregation and for maintaining proper ER morphology in a coordinated manner with NAI1.

Quantitative analysis of ER body morphology in an Arabidopsis mutant
Nagano, A.J., Maekawa, A., Nakano, R.T., Miyahara, M., Higaki, T., Kutsuna, N., Hasezawa, S., Hara-Nishimura, I.
Plant and Cell Physiology, 50, 12, 2015, 2022, OXFORD UNIV PRESS, 2009年, ［査読有り］, ［国内誌］
英語, 研究論文（学術雑誌）, Although fluorescence microscopy screening has proven useful in the identification of genes involved in plant organelle biogenesis and integrity, the quantitative and statistical study of the geometric phenotype is highly limited. This situation could generate unconscious bias in the understanding and presentation of a mutant phenotype. Therefore, we have developed an automated quantification system for green fluorescent protein (GFP) images, which enabled us to easily obtain quantitative data on ER bodies (an endoplasmic reticulum-derived organelle). We isolated an ER body morphology mutant of Arabidopsis thaliana, leb-1 (long ER body). The leb-1 mutant had significantly fewer and larger ER bodies than the wild-type. An amino acid substitution of Cys29 with tyrosine (C29Y) on PYK10, a major component protein of ER bodies, was found in leb-1. Non-reducing SDSPAGE revealed that the electrophoretic mobility of PYK10 in the leb-1 mutant was clearly different from that in the wild type. This difference suggests that the C29Y amino acid substitution caused a tertiary structural change of the PYK10 protein. While the bglu21-1 and pyk10-1 single mutations slightly affected the number and morphology of the ER bodies, a bglu21-1 pyk10-1 double mutant had fewer and larger ER bodies than the wild type. The quantitative ER body phenotypes of leb-1 were similar to those of bglu21-1 pyk10-1 and bglu21-1 leb-1, suggesting that the leb-1 mutation allele acts dominantly to the BGLU21 wild-type allele. The leb-1 type PYK10 protein, which has an abnormal structure, may competitively inhibit interactions between the wild-type BGLU21/PYK10 protein and an unknown partner.

GNOM-LIKE1/ERMO1 and SEC24A/ERMO2 are required for maintenance of endoplasmic reticulum morphology in Arabidopsis thaliana
Nakano, R.T., Matsushima, R., Ueda, H., Tamura, K., Shimada, T., Li, L., Hayashi, Y., Kondo, M., Nishimura, M., Hara-Nishimura, I.
Plant Cell, 21, 11, 3672, 3685, AMER SOC PLANT BIOLOGISTS, 2009年, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, The endoplasmic reticulum (ER) is composed of tubules, sheets, and three-way junctions, resulting in a highly conserved polygonal network in all eukaryotes. The molecular mechanisms responsible for the organization of these structures are obscure. To identify novel factors responsible for ER morphology, we employed a forward genetic approach using a transgenic Arabidopsis thaliana plant (GFP-h) with fluorescently labeled ER. We isolated two mutants with defects in ER morphology and designated them endoplasmic reticulum morphology1 (ermo1) and ermo2. The cells of both mutants developed a number of ER-derived spherical bodies, similar to 1 mu m in diameter, in addition to the typical polygonal network of ER. The spherical bodies were distributed throughout the ermo1 cells, while they formed a large aggregate in ermo2 cells. We identified the responsible gene for ermo1 to be GNOM-LIKE1 (GNL1) and the gene for ermo2 to be SEC24a. Homologs of both GNL1 and SEC24a are involved in membrane trafficking between the ER and Golgi in yeast and animal cells. Our findings, however, suggest that GNL1/ERMO1 and SEC24a/ERMO2 have a novel function in ER morphology in higher plants.

Arthrobacter属細菌におけるニコチン分解遺伝子群の比較ゲノム解析
島崎智久, 増田幸子, 柴田ありさ, 須田亙, 白須賢, 矢崎一史, 市橋泰範, 杉山暁史, 中野亮平, 日本農芸化学会大会講演要旨集(Web), 2022, 2022年

特化代謝産物を介したタバコ-Arthrobacter間相互作用の解析　　　　　　　　　　　　　　　
島崎智久, 島崎智久, 増田幸子, GARRIDO-OTER Ruben, GARRIDO-OTER Ruben, 川崎崇, 青木裕一, 柴田ありさ, 須田亙, 白須賢, 矢崎一史, 中野亮平, 杉山暁史, 植物微生物研究会研究交流会講演要旨集, 2021年

硫酸化ペプチドを介した植物圏コメンサル細菌によるシロイヌナズナ根の発生と免疫の制御
中野亮平, GIGOLASHVILI Tamara, SCHULZE-LEFERT Paul, 日本植物学会大会研究発表記録(CD-ROM), 84th, 2020年

Synthetic community(SynCom)を用いた植物-マイクロバイオータ相互作用の分子生物学的解析　　　　　　　　　　　　　　　
中野トーマス亮平, 日本微生物生態学会大会(Web), 2019年

気孔開閉の制御に対する病原体の選択圧と進化的トレードオフ
峯彰, 峯彰, 福元華織, 中野亮平, 中野亮平, 津田賢一, 日本植物病理学会大会プログラム・講演要旨予稿集, 2018年03月12日
日本語

コロナチンは双子葉植物に広く保存されたアブシジン酸の分解機構を利用して気孔の開口を誘導する
峯彰, 峯彰, 福元華織, 中野亮平, 多田安臣, 津田賢, 日本植物病理学会大会プログラム・講演要旨予稿集, 2016年03月03日
日本語

Colletotrichum tofieldiaeは,シロイヌナズナに無病徴感染しリン不足条件下で宿主の成長を促す
晝間敬, BARBARA Kracher, 中野亮平, SACRISTAN Soledad, O’CONNELL Richard, SCHULZE‐LEFERT Paul, 日本植物病理学会大会プログラム・講演要旨予稿集, 2014年05月10日
日本語

トランスクリプトームからみた地下ストロン(地下茎)メリステムの特性
荒木希和子, 永野惇, 中野亮平, 北爪達也, 山口勝司, 西村いくこ, 重信秀治, 工藤洋, 日本植物生理学会年会要旨集, 2013年03月14日
日本語

ER bodyに蓄積するβ‐グルコシダーゼPYK10の天然基質の同定
中野亮平, BEDNAREK Pawel, 宮原窓, PISLEWSKA‐BEDNAREK Mariola, 森正之, SCHULZE‐LEFERT Paul, 西村いくこ, 日本植物生理学会年会要旨集, 2013年03月14日
日本語

茎か?根か?地下茎メリステムのトランスクリプトーム解析
荒木希和子, 永野惇, 中野亮平, 北爪達也, 山口勝司, 西村いくこ, 重信秀治, 工藤洋, 日本生態学会大会講演要旨集, 2013年03月05日
日本語

ER bodyに蓄積するβ-グルコシダーゼPYK10の天然基質の同定　　　　　　　　　　　　　　　
中野亮平, BEDNAREK Pawel, 宮原窓, PISLEWSKA-BEDNAREK Mariola, 森正之, SCHULZE-LEFERT Paul, 西村いくこ, 日本植物生理学会年会要旨集, 2013年

シロイヌナズナ液胞タンパク質ERMO3は転写因子NAI1と協調して小胞体形態維持に関わる
中野亮平, 松島良, 永野淳, 深尾陽一朗, 藤原正幸, 近藤真紀, 西村幹夫, 西村いくこ, 日本植物生理学会年会要旨集, 2012年03月09日
日本語

内膜系オルガネラの形態制御に関わるGDSLリパーゼERMO3の解析
中野亮平, 松島良, 永野淳, 上田晴子, 田村謙太郎, 嶋田知生, 近藤真紀, 西村幹夫, 西村いくこ, 日本植物生理学会年会要旨集, 2011年03月11日
日本語

液胞タンパク質ERMO3を介した高等植物の小胞体の形態形成・維持機構
中野亮平, 松島良, 永野淳, 深尾陽一朗, 近藤真紀, 西村幹夫, 西村いくこ, 生化学, 2011年
日本語

シロイヌナズナ小胞体の形態維持のための新しい分子機構
中野亮平, 松島良, 上田晴子, 田村謙太郎, 嶋田知生, 李立新, 林八寿子, 近藤真紀, 西村幹夫, 西村いくこ, 日本植物生理学会年会要旨集, 2010年03月12日
日本語

Novel Molecular Pathways Underlying Maintenance of Endoplasmic Reticulum Morphology in Arabidopsis thaliana．　　　　　　　　　　　　　　　
中野亮平, 松島良, 上田晴子, 田村謙太郎, 嶋田知生, 李立新, 林八寿子, 近藤真紀, 西村幹夫, 西村いくこ, 日本植物生理学会年会，熊本，2010年3月18日, 2010年
日本語

ERMO1/GNOM-LIKE1 and ERMO2/SEC24a Are Required for Maintenance of Endoplasmic Reticulum in Arabidopsis thaliana.　　　　　　　　　　　　　　　
R. T. Nakano, R. Matsushima, H. Ueda, K. Tamura, T. Shimada, L. Li, Y. Hayashi, M. Kondo, M. Nishimura, I. Hara-Nishimura, 21st International Conference on Arabidopsis Research 2010, Yokohama (Japan), June 6-10, 2010, 2010年
英語

小胞体の形態と細胞内分布に異常を示すendoplasmic reticulum morphology(ermo)変異体の解析
中野亮平, 松島良, 上田晴子, 林八寿子, 田村謙太郎, 嶋田知生, 西村いくこ, 日本植物生理学会年会要旨集, 2009年03月16日
日本語

小胞体の形態と細胞内分布に異常を示すendoplasmic reticulum morphology (ermo) 変異体の解析
中野 亮平, 松島 良, 上田 晴子, 林 八寿子, 田村 謙太郎, 嶋田 知生, 西村 いくこ, 日本植物生理学会年会およびシンポジウム 講演要旨集, 2009, 0, 35, 35, 2009年, ［査読有り］
小胞体はチューブ状構造とシート状構造が three-way junctionと呼ばれる三叉の接続を介して複雑に組み合わさったネットワーク状の構造をとっている．この形態の形成・維持に寄与する因子を同定するため，小胞体の形態に異常を示す変異体を3系統単離し，endoplasmic reticulum morphology（ermo）と命名した．ermo1とermo2では，小胞体に由来する1 μm前後の球状構造体が数多く観察され，ermo2ではそれらが細胞内の一カ所に凝集していた．一方ermo3では球状の構造体は観察されず，小胞体が一カ所に凝集していた．これらの変異体では異常な構造・凝集体の他に正常なネットワークも観察された．マッピングの結果，ermo1とermo2は小胞体ーゴルジ体間の輸送においてそれぞれCOPI小胞とCOPII小胞の出芽に関わると考えられている遺伝子の変異であることがわかった．しかし，これらの変異体において細胞内輸送に明らかな異常は観察できていない．このことから，ERMO1およびERMO2は小胞体の形態維持に関わる新奇機能をもっている，あるいは変異体における輸送異常は検出できないレベルだが，小胞体ーゴルジ体間の輸送が小胞体形態に重要であることが考えられた．また，ERMO2およびERMO3はオルガネラを細胞内に分布させるのに必要であることが示された．, 日本植物生理学会

研究者、生活を語る 「両立」の舞台裏　　　　　　　　　　　　　　　
中野亮平, 海外で４人の子育てをしながら研究をするということ
岩波書店, 2024年10月, ［分担執筆］

実験医学l：次のパンデミックに備えた新しい感染症研究　　　　　　　　　　　　　　　
UJAだより「帰る！？帰らない！？留学中の心構え」
洋土社, 2024年05月, 9784758125796, ［分担執筆］

現代化学2022年10月号　　　　　　　　　　　　　　　
中野亮平, 「植物と共生・すぐそこにある分子」複雑で魅力的な植物マイクロバイオータ相互作用と、それにかかわる根圏代謝産物群
2022年, ［分担執筆］

アグリバイオ 2021年1月号 持続的農業のための土壌・植物微生物叢の解析と利用　　　　　　　　　　　　　　　
登達也, 中野亮平, 植物微生物叢の学術研究のトレンド
2021年, ［分担執筆］

共生微生物 : 生物と密接に関わるミクロな生命体
大野, 博司, 第14章「根圏と微生物」
化学同人, 2016年10月, 9784759817287, 8, xi, 281p, 日本語, ［分担執筆］

多様な微生物からなるシステムとしての根圏マイクロバイオータによる植物の発生や免疫応答への干渉　　　　　　　　　　　　　　　
Jana Hucklenbroich, 島﨑智久, 中野亮平
第97回日本細菌学会総会 シンポジウム「オミクスのデータ多量解析に基づくシステムバイオロジーは、細菌学の発展にいかに貢献するか」, 2024年08月08日
［招待講演］

根圏常在微生物との相互作用における植物免疫の役割　　　　　　　　　　　　　　　
中野亮平
2024年度 日本農芸化学会北海道支部 第一回学術講演会, 2024年07月13日
［招待講演］

植物免疫・発生・栄養吸収の協調的制御をめぐる植物マイクロバイオータの役割　　　　　　　　　　　　　　　
中野亮平
「植物の栄養研究会」 第8回研究交流会, 2023年09月05日
［招待講演］

ドイツで4人の子どもを育てながら感じたこと　　　　　　　　　　　　　　　
中野亮平
第75回日本細胞生物学会大会 ランチョンセミナー, 2023年06月30日
［招待講演］

植物マイクロバイオータ研究の新時代を創り出すために　　　　　　　　　　　　　　　
中野亮平
植物病理を紡ぐ会, 2023年03月25日
［招待講演］

Glucosinolates mediate root-microbiota interactions　　　　　　　　　　　　　　　
Arpan Kumar Basak, Kenji Yamada, Ryohei Thomas Nakano
International Plant Sulfur Workshop, 2022年07月16日
［招待講演］

植物超個体もリモートで：分泌性物質を介した植物マイクロバイオータ相互作用が根の発生と免疫を制御する　　　　　　　　　　　　　　　
中野亮平
第63回日本植物生理学会年会 シンポジウム「見たい、知りたい、操作したい、植物と微生物の超個体」, 2022年03月23日
［招待講演］

SULFUR BIOLOGY IN PLANT-MICROBIOTA INTERACTIONS　　　　　　　　　　　　　　　
Jana Hucklenbroich, Arpan Kumar Basak, Kenji Yamada, Ryohei Thomas Nakano
Joint Meeting for Plant and Human Sulfur Biology and Glucosinolates, 2021年09月30日
［招待講演］

Toward Understanding the Molecular Dialog between Plants and associated Microbiota　　　　　　　　　　　　　　　
Ryohei Thomas Nakano
University of Bonn-TUAT mini-symposium, 2019年09月16日
［招待講演］

Synthetic community (SynCom) を用いた植物—マイクロバイオータ相互作用の分子生物学的解析　　　　　　　　　　　　　　　
中野亮平
日本微生物生態学会第３３回大会 ＜微生物生態学会-植物生理学会 共催シンポジウム＞ 植物微生物研究で共創する未来, 2019年09月11日
［招待講演］

Ternary interaction assays revealed modular traits of root microbiota that dictate host growth and immune status　　　　　　　　　　　　　　　
Ryohei Thomas Nakano
Frontiers in Rhizosphere Research, 2018年11月20日
［招待講演］