研究者情報

マスター

アカウント（マスター）

• 氏名

  松浦　英幸(マツウラ　ヒデユキ), マツウラ　ヒデユキ

所属（マスター）

• 農学研究院　基盤研究部門　生物機能化学分野

所属（マスター）

• 農学研究院　基盤研究部門　生物機能化学分野

独自項目

syllabus

• 2021, 大学院共通授業科目（教育プログラム）：PARE, Inter-Graduate School Classes(Educational Program):PARE, 修士課程, 大学院共通科目, populations, human activities, environments, resources, food, land, rivers, coasts, atmosphere, soil, water, groundwater, geo-environment, water cycle, element cycles, ecosystems, poverty, sustainability, climate change, erosion, chemical substances, pollution, risk management, green technology
• 2021, 農学フロンティア総論, Advanced Review of Agriculture Frontiers, 修士課程, 農学院, 農学の最先端研究、食品、バイオマス、エネルギー、水土資源、農業経済、持続的生産、生物共生
• 2021, 天然物化学特論, Advanced Natural Product Chemistry, 修士課程, 農学院, 有機化学、天然物化学、化学生物学
• 2021, 環境化学, Environmental Chemistry, 学士課程, 農学部, 大気・河川・湖沼・海洋・地下水・土壌の汚染、人工公害物質、ミスト、バイオレメディエーション、バイオマス廃棄物の資源化・循環、ＬＣＡ、植物ストレス
• 2021, 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), Freshman Seminar, 学士課程, 全学教育, 食料、安全、作物生産、放射能汚染、食品微生物、酵素利用技術、毒物、ガン、アレルギー、メタボリックシンドローム、腸内細菌叢、保健機能食品
• 2021, 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), Freshman Seminar, 学士課程, 全学教育, 天然物化学、生物有機化学、生理活性物質
• 2021, 農産物利用学, Utilization of Agricultural Products, 学士課程, 農学部, デンプン、糖質、タンパク質、脂質、食用加工、貯蔵、風味物質、フレーバー分析
• 2021, 化学実験Ⅰ, Laboratry Work on Chemistry I, 学士課程, 農学部, 天然物化学、有機化学
• 2021, 有機化学Ⅰ, Organic Chemistry I, 学士課程, 農学部, 有機化学、有機分子、化学反応
• 2021, 生物有機化学, Bioorganic Chemistry, 学士課程, 農学部, 酵素、生合成、二次代謝、イソプレノイド経路、ポリケチド経路、シキミ酸経路、アルカロイド

researchmap

プロフィール情報

学位

• 博士（農学）（北海道大学）

プロフィール情報

• プロフィール

  普段、何気なく接している植物ですが、皆さん不思議に思った事はありませんか？春に良く目にする「フキノトウ」、さて、「フキ」との関係は如何に？ジャガイモに「ヘソの緒」がある？おいしそうなスイカ、トマト、果実に種子はあるけれど、果実内で発芽した種子は見た事がありますか？植物は仲間の植物のSOS信号を受け取っている？私はこのような植物の不思議さ、生きる知恵を有機化学的に解明し、これを現代の農業へ応用することを目指しています。

• 姓

  松浦, マツウラ

• 名

  英幸, ヒデユキ

• ID各種

  201301099013531061

対象リソース

http://kaken.nii.ac.jp/d/r/20344492.ja.html

業績リスト

研究キーワード

• セオブロキシド   植物化学生長剤   種子処理   植物生存戦略   アブシシン酸   発芽阻害   低温障害   植物ホルモン   植物化学調節剤   乾燥傷害   根圏   オクタデカノイド経路   花芽形成   有機物シンク形成   亜酸化窒素放出   伝統農法   窒素固定   中強酸性熱帯泥炭土壌   カラマツ・ヒース林   環境負荷緩和   非菌根性植物生育促進真菌類   Lasiodiplodia theobromae   リン・窒素バランス   Xyris complanata   硫酸酸性耐性細菌   ジャスモン酸   酸化還元環境   単生窒素固定細菌   形態分化異常   微生物増殖因子   

研究分野

• ライフサイエンス / 生物有機化学
• 環境・農学 / ランドスケープ科学
• 環境・農学 / 環境農学

経歴

• 2016年10月 - 現在 北海道大学大学院農学研究院 基礎研究部門 教授
• 2007年04月 - 2016年09月 北海道大学 (連合)農学研究科(研究院) 准教授
• 2003年06月 - 2007年03月 北海道大学 (連合)農学研究科(研究院) 助教授
• 2001年11月 - 2003年05月 北海道大学 (連合)農学研究科(研究院) 助手
• 1998年10月 - 2001年10月 北海道科学技術振興財団（現・北海道科学技術総合振興センター） 雇用研究員
• 1996年01月 - 1998年09月 日本学術振興会特別研究員
• 1995年10月 - 1995年12月 北海道大学農学部 農芸化学科 北海道大学農学部農芸化学科博士研究員
• 1993年09月 - 1995年09月 ブリティシュ コロンビア大学 理学部 ブリティシュ コロンビア大学 博士研究員

論文

• Isolation and Identification of the Antibacterial Compounds Produced by Maillard Reaction of Xylose with Phenylalanine or Proline.

  Kazuho Aonishi, Shungo Miyao, Lisa Yokoi, Naoki Kitaoka, Kento Koyama, Hideyuki Matsuura, Shigenobu Koseki

  Journal of agricultural and food chemistry 2024年07月04日 

  Maillard reaction products (MRPs) of xylose with phenylalanine and xylose with proline exhibit high antibacterial activity. However, the active antibacterial compounds in MRPs have not yet been identified or isolated. This study aimed to isolate the active compounds in the two antibacterial MRPs. The organic layer of the MRP solution was separated and purified using silica gel chromatography and high-performance liquid chromatography. The chemical structures of the isolated compounds were determined by mass spectrometry and nuclear magnetic resonance spectroscopy. The compounds inhibited the growth of Bacillus cereus and Salmonella Typhimurium at 25 °C for 7 days at a concentration of 0.25 mM. Furthermore, the isolated compounds inhibited the growth of naturally occurring microflora of lettuce and chicken thighs at 25 °C for 2 days at a concentration of 0.5-1.0 mM. The antibacterial compounds found in MRPs demonstrated a wide range of effectiveness and indicated their potential as alternative preservatives.
• Isolation and Structure Determination of cis-OPDA-α-Monoglyceride from Arabidopsis thaliana.

  Shotaro Hirota, Yusuke Ito, Shiro Inoue, Naoki Kitaoka, Tohru Taniguchi, Kenji Monde, Kosaku Takahashi, Hideyuki Matsuura

  Journal of natural products 2024年04月24日 

  cis-12-oxo-Phytodieneoic acid-α-monoglyceride (1) was isolated from Arabidopsis thaliana. The chemical structure of 1 was elucidated based on exhaustive 1D and 2D NMR spectroscopic measurements and supported by FDMS and HRFDMS data. The absolute configuration of the cis-OPDA moiety in 1 was determined by comparison of 1H NMR spectra and ECD measurements. With respect to the absolute configuration of the β-position of the glycerol backbone, the 2:3 ratio of (S) to (R) was determined by making ester-bonded derivatives with (R)-(+)-α-methoxy-α-trifluoromethylphenylacetyl chloride and comparing 1H NMR spectra. Wounding stress did not increase endogenous levels of 1, and it was revealed 1 had an inhibitory effect of A. thaliana post germination growth. Notably, the endogenous amount of 1 was higher than the amounts of (+)-7-iso-jasmonic acid and (+)-cis-OPDA in intact plants. 1 also showed antimicrobial activity against Gram-positive bacteria, but jasmonic acid did not. It was also found that α-linolenic acid-α-monoglyceride was converted into 1 in the A. thaliana plant, which implied α-linolenic acid-α-monoglyceride was a biosynthetic intermediate of 1.
• Investigation Of The Biosynthesis Pathway That Generates cis-Jasmone.

  Shiro Inoue, Hiromu Tsuzuki, Kazuhiko Matsuda, Naoki Kitaoka, Hideyuki Matsuura

  Chembiochem : a European journal of chemical biology e202300593  2023年11月07日 

  Researchers have established that (+)-7-iso-jasmonic acid ((+)-7-iso-JA) is an intermediate in the production of cis-jasmone (CJ); however, the biosynthetic pathway of CJ has not been fully described. Previous reports stated that CJ, a substructure of pyrethrin II produced by pyrethrum (Tanacetum cinerariifolium), is not biosynthesized through this biosynthetic pathway. To clarify the ambiguity, stable isotope-labelled jasmonates were synthesized, and compounds were applied to apple mint (Mentha suaveolens) via air propagation. The results showed that cis-jasmone is not generated from intermediate (+)-7-iso-JA, and (+)-7-iso-JA is not produced from 3,7-dideydro-JA (3,7-ddh-JA); however, 3,7-didehydro-JA and 4,5-didehydro-7-iso-JA were converted into CJ and JA, respectively.
• Elevated ozone alters long-chain fatty acids in leaves of Japanese white birch saplings

  Noboru Masui, Shiro Inoue, Evgenios Agathokleous, Hideyuki Matsuura, Takayoshi Koike

  Environmental Science and Pollution Research 2023年06月07日
• Airborne Methyl Jasmonate is Metabolized to Jasmonic Acid and 12-Hydroxyjasmonic Acid, and Induces Jasmonate Biosynthesis in Marchantia polymorpha

  Hiromu Tsuzuki, Naoki Kitaoka, Shiro Inoue, Kosaku Takahashi, Hideyuki Matsuura

  CHEMISTRYSELECT 8 21 2023年06月 

  Methyl jasmonate (MeJA) is a volatile jasmonate compound commonly used to induce defense responses in spermatophytes. This study reports that airborne MeJA-d(3), deuterated MeJA, increases the levels of (dinor-)12-oxo-phytodienoic acids [(dn-)OPDAs] and jasmonic acid (JA) as well as JA-d(3) and 12-hydroxyjasmonic acid-d(2) (12-OH-JA-d(2)), MeJA-d(3) metabolites, in the model bryophyte Marchantia polymorpha. Enhancement of JA biosynthesis was substantiated by the expression of JA biosynthetic genes induced by airborne MeJA. Additionally, each of enantiomers, (+)-MeJA and (-)-MeJA, was observed to induce the accumulation of JA and (dn-)OPDAs in M. polymorpha. This study demonstrates that airborne MeJA is metabolized to JA and 12-OH-JA, and induces JA biosynthesis in M. polymorpha. Moreover, the transient increase in endogenous JA level after airborne MeJA treatment provides concrete evidence that M. polymorpha biosynthesizes JA.
• Biological Activity of Anti-bolting Compound, alpha-(7Z,10Z,13Z)-hexadeca-7,10,13-trienoic Acid Monoglyceride to Reduce the Endogenous Amount of Gibberellins

  Tsuyoshi Ogihara, Shunpei Shikama, Akihisa Ishii, Syotaro Hirota, Junichi Kashiwagi, Kaien Fujino, Yuki Mitsui, Takafumi Shimizu, Mitsunori Seo, Naoki Kitaoka, Yasunori Koda, Hideyuki Matsuura

  JOURNAL OF PLANT GROWTH REGULATION 2023年04月 

  A monoacylglycerol, alpha-(7Z,10Z,13Z)-hexadeca-7,10,13-trienoic acid monoglyceride, extracted from leaves of Japanese radish (Raphanus sativus) has been reported as an anti-bolting compound (ABC); however, the mechanism how ABC inhibits the plant bolting has been remained to be elucidated. In this paper, it was found that exogenous applications of ABC led Arabidopsis thaliana seedlings decrease transcriptional levels of AtKO and increase those of AtGA2ox to reduce endogenous levels of gibberellins (GAs) to retard A. thaliana growth, whose physiological phenomena were counteracted by exogenous applications of ent-kaurenoic acid and GA(3), respectively. Furthermore, alpha-oleanolic acid monoglyceride having both activities of supressing KO and enhancing GA2ox inductions retarded A. thaliana growth post germination, although alpha-palmitic acid monoglyceride having only the effect of supressing KO induction was not enough to suppress the growth. These experimental data supported the idea that dual function was essential for ABC to show biological activity.
• Abscisic acid‐mediated sugar responses are essential for vegetative desiccation tolerance in the liverwort Marchantia polymorpha

  Nobiza Khatun, Akihisa Shinozawa, Kosaku Takahashi, Hideyuki Matsuura, Akida Jahan, Mousona Islam, Masudul Karim, Rahul Sk, Mikako Yoshikawa, Kimitsune Ishizaki, Yoichi Sakata, Daisuke Takezawa

  Physiologia Plantarum 175 2 2023年03月
• Pennelliiside D, a New Acyl Glucose from Solanum pennellii and Chemical Synthesis of Pennelliisides

  Rishni Masimbula, Hiroto Kobayashi, Tenki Nakashima, Yurika Nambu, Naoki Kitaoka, Hideyuki Matsuura

  Molecules 27 12 2022年06月09日 

  Acyl glucoses are a group of specialized metabolites produced by Solanaceae. Solanum pennellii, a wild-type tomato plant, produces acyl glucoses in its hair-like epidermal structures known as trichomes. These compounds have been found to be herbicides, microbial growth inhibitors, or allelopathic compounds. However, there are a few reports regarding isolation and investigation of biological activities of acyl glucoses in its pure form due to the difficulty of isolation. Here, we report a new acyl glucose, pennelliiside D, isolated and identified from S. pennellii. Its structure was determined by 1D NMR and 2D NMR, together with FD-MS analysis. To clarify the absolute configuration of the acyl moiety of 2-methylbutyryl in the natural compound, two possible isomers were synthesized starting from β-D-glucose pentaacetate. By comparing the spectroscopic data of natural and synthesized compounds of isomers, the structure of pennelliiside D was confirmed to be 3,4-O-diisobutyryl-2-O-((S)-2-methylbutyryl)-D-glucose. Pennelliiside D and its constituent fatty acid moiety, (S)-2-methylbutanoic acid, did not show root growth-inhibitory activity. Additionally, in this study, chemical synthesis pathways toward pennelliisides A and B were adapted to give 1,6-O-dibenzylpennelliisides A and B.
• Pinellic Acid Isolated from Quercetin-rich Onions has a Peroxisome Proliferator-Activated Receptor-Alpha/Gamma (PPAR-α/γ) Transactivation Activity.

  Mikio Doi, Naoki Morita, Tsugumi Okuzawa, Satoru Ohgiya, Daisaku Okamoto, Kenichi Sato, Yukiya Ito, Hideyuki Matsuura, Yasuyuki Hashidoko

  Planta medica 88 6 440 - 446 2022年05月 

  Quercetin, a flavonol, is a functional compound that is abundant in onions and is known to have antioxidant and anti-inflammatory effects. Quercetin and its glucoside are known to function as peroxisome proliferator-activated receptor (PPAR) ligands and showed high PPAR-α transactivation activity but little PPAR-γ transactivation activity in some reports. In this study, we demonstrated that an aqueous extract of a quercetin-rich onion cultivar increased transactivation activities not only of PPAR-α but also of PPAR-γ. We isolated (9S,12S,13S)-(10E)-9,12,13-trihydroxyoctadec-10-enoic acid (pinellic acid) obtained from the aqueous extract using PPAR-γ transactivation as an index. Furthermore, it was revealed that pinellic acid could transactivate PPAR-α. Our findings are the first report mentioned showing that trihydroxyoctadec-10-enoic acids showed PPAR-α/γ transactivation activities.
• AtGH3.10 is another jasmonic acid-amido synthetase in Arabidopsis thaliana.

  Jay C Delfin, Yuri Kanno, Mitsunori Seo, Naoki Kitaoka, Hideyuki Matsuura, Takayuki Tohge, Takafumi Shimizu

  The Plant journal : for cell and molecular biology 110 4 1082 - 1096 2022年03月04日 

  Jasmonoyl-isoleucine (JA-Ile) is a key signaling molecule that activates jasmonate-regulated flower development and the wound stress response. For years, JASMONATE RESISTANT1 (JAR1) has been the sole jasmonoyl-amino acid synthetase known to conjugate jasmonic acid (JA) to isoleucine, and the source of persisting JA-Ile in jar1 knockout mutants has remained elusive until now. Here we demonstrate through recombinant enzyme assays and loss-of-function mutant analyses that AtGH3.10 functions as a JA-amido synthetase. Recombinant AtGH3.10 could conjugate JA to isoleucine, alanine, leucine, methionine, and valine. The JA-Ile accumulation in the gh3.10-2 jar1-11 double mutant was nearly eliminated in the leaves and flower buds while its catabolism derivative 12OH-JA-Ile was undetected in the flower buds and unwounded leaves. Residual levels of JA-Ile, JA-Ala, and JA-Val were nonetheless detected in gh3.10-2 jar1-11, suggesting the activities of similar promiscuous enzymes. Upon wounding, the accumulation of JA-Ile and 12OH-JA-Ile and the expression of JA-responsive genes OXOPHYTODIENOIC ACID REDUCTASE3 and JASMONATE ZIM-DOMAIN1 observed in WT, gh3.10-1, and jar1-11 leaves were effectively abolished in gh3.10-2 jar1-11. Additionally, an increased proportion of undeveloped siliques associated with retarded stamen development was observed in gh3.10-2 jar1-11. These findings conclusively show that AtGH3.10 contributes to JA-amino acid biosynthesis and functions partially redundantly with AtJAR1 in sustaining flower development and the wound stress response in Arabidopsis.
• Substrate specificity of glycoside hydrolase family 1 β-glucosidase AtBGlu42 from Arabidopsis thaliana and its molecular mechanism.

  Shu Horikoshi, Wataru Saburi, Jian Yu, Hideyuki Matsuura, James R Ketudat Cairns, Min Yao, Haruhide Mori

  Bioscience, biotechnology, and biochemistry 86 2 231 - 245 2022年01月24日 

  Plants possess many glycoside hydrolase family 1 (GH1) β-glucosidases, which physiologically function in cell wall metabolism and activation of bioactive substances, but most remain uncharacterized. One GH1 isoenzyme AtBGlu42 in Arabidopsis thaliana has been identified to hydrolyze scopolin using the gene deficient plants, but no enzymatic properties were obtained. Its sequence similarity to another functionally characterized enzyme Os1BGlu4 in rice suggests that AtBGlu42 also acts on oligosaccharides. Here, we show that the recombinant AtBGlu42 possesses high kcat/Km not only on scopolin, but also on various β-glucosides, cellooligosaccharides, and laminarioligosaccharides. Of the cellooligosaccharides, cellotriose was the most preferred. The crystal structure, determined at 1.7 Å resolution, suggests that Arg342 gives unfavorable binding to cellooligosaccharides at subsite +3. The mutants R342Y and R342A showed the highest preference on cellotetraose or cellopentaose with increased affinities at subsite +3, indicating that the residues at this position have an important role for chain length specificity.
• AtTrxh3, a Thioredoxin, Is Identified as an Abscisic AcidBinding Protein in Arabidopsis thaliana.

  Tomoaki Anabuki, Keisuke Ohashi, Taichi E Takasuka, Hideyuki Matsuura, Kosaku Takahashi

  Molecules (Basel, Switzerland) 27 1 2021年12月28日 

  Abscisic acid (ABA, 1) is a plant hormone that regulates various plant physiological processes such as seed developing and stress responses. The ABA signaling system has been elucidated; binding of ABA with PYL proteins triggers ABA signaling. We have previously reported a new method to isolate a protein targeted with a bioactive small molecule using a biotin linker with alkyne and amino groups, a protein cross-linker, and a bioactive small molecule with an azido group (azido probe). This method was used to identify the unknown ABA binding protein of Arabidopsis thaliana. As a result, AtTrxh3, a thioredoxin, was isolated as an ABA binding protein. Our developed method can be applied to the identification of binding proteins of bioactive compounds.
• Potato tuber-inducing activities of jasmonic acid and related-compounds (II).

  Kanji Miyawaki, Shiro Inoue, Naoki Kitaoka, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 85 12 2378 - 2382 2021年11月24日 

  New information is being accumulated for plant-derived oxylipins, such as jasmonic acid (JA) amino acid conjugates. However, these compounds have not being examined for their activity in promoting potato tuber formation. It was found that (-)-JA had the highest activity followed cis-(-)-OPDA, (+)-4, 5-didehydroJA, cis-(+)-OPDA-l-Ile, and (-)-JA-l-Ile, -Leu, -Phe, -Val, although iso-OPDA and 3,7-didehydroJA did not exhibit activity.
• A Ubiquitously Expressed UDP-Glucosyltransferase, UGT74J1, Controls Basal Salicylic Acid Levels in Rice.

  Daisuke Tezuka, Hideyuki Matsuura, Wataru Saburi, Haruhide Mori, Ryozo Imai

  Plants (Basel, Switzerland) 10 9 2021年09月10日 

  Salicylic acid (SA) is a phytohormone that regulates a variety of physiological and developmental processes, including disease resistance. SA is a key signaling component in the immune response of many plant species. However, the mechanism underlying SA-mediated immunity is obscure in rice (Oryza sativa). Prior analysis revealed a correlation between basal SA level and blast resistance in a range of rice varieties. This suggested that resistance might be improved by increasing basal SA level. Here, we identified a novel UDP-glucosyltransferase gene, UGT74J1, which is expressed ubiquitously throughout plant development. Mutants of UGT74J1 generated by genome editing accumulated high levels of SA under non-stressed conditions, indicating that UGT74J1 is a key enzyme for SA homeostasis in rice. Microarray analysis revealed that the ugt74j1 mutants constitutively overexpressed a set of pathogenesis-related (PR) genes. An inoculation assay demonstrated that these mutants had increased resistance against rice blast, but they also exhibited stunted growth phenotypes. To our knowledge, this is the first report of a rice mutant displaying SA overaccumulation.
• Plant-insect communication in urban forests: Similarities of plant volatile compositions among tree species (host vs. non-host trees) for alder leaf beetle Agelastica coerulea

  Noboru Masui, Evgenios Agathokleous, Akira Tani, Hideyuki Matsuura, Takayoshi Koike

  Environmental Research 204 111996 - 111996 2021年09月
• A promiscuous fatty acid ω-hydroxylase CYP94A90 is likely to be involved in biosynthesis of a floral nitro compound in loquat (Eriobotrya japonica).

  Takuya Yamaguchi, Yumi Matsui, Naoki Kitaoka, Yasumasa Kuwahara, Yasuhisa Asano, Hideyuki Matsuura, Yukari Sunohara, Hiroshi Matsumoto

  The New phytologist 231 3 1157 - 1170 2021年08月 

  Nitro groups are often associated with synthetically manufactured compounds such as medicines and explosives, and rarely with natural products. Loquat emits a nitro compound, (2-nitroethyl)benzene, as a flower scent. The nitro compound exhibits fungistatic activity and is biosynthesised from l-phenylalanine via (E/Z)-phenylacetaldoxime. Although aldoxime-producing CYP79s have been intensively studied, it is unclear what enzymes form nitro groups from aldoximes either in plants or in other organisms. Here, we report the identification of two cytochrome P450s that are likely to be involved in (2-nitroethyl)benzene biosynthesis in loquat through differential gene expression analysis using RNA-seq and functional identification using yeast and tobacco. CYP79D80 and CYP94A90 catalysed the formation of (E/Z)-phenylacetaldoxime from l-phenylalanine and (2-nitroethyl)benzene from the aldoxime, respectively. Expression profiles of CYP79D80 and CYP94A90 were correlated with the emission of (2-nitroethyl)benzene from loquat flowers. CYP94A90 also functioned as a fatty acid ω-hydroxylase as do other CYP94A fatty acid ω-hydroxylases. The CYP94As tested from other plants were all found to catalyse the formation of (2-nitroethyl)benzene from (E/Z)-phenylacetaldoxime. CYP79D80 and CYP94A90 are likely to operate in concert to biosynthesise (2-nitroethyl)benzene in loquat. CYP94A90 and other CYP94As are 'promiscuous fatty acid ω-hydroxylases', catalysing the formation of nitro groups from aldoximes, and are widely distributed in dicot plants.
• Verification of the versatility of the in vitro enzymatic reaction giving (+)-cis-12-Oxo-phytodienoic acid.

  Yusuke Ito, Kento Sasaki, Tsuyoshi Ogihara, Naoki Kitaoka, Kosaku Takahahi, Hideyuki Matsuura

  Bioorganic & medicinal chemistry letters 49 128284 - 128284 2021年07月23日 

  Jasmonic acid (JA) is a plant hormone involved in the defense response against insects and fungi. JA is synthesized from α-linolenic acid (LA) by the octadecanoid pathway in plants. 12-oxo-Phytodienoic acid (OPDA) is one of the biosynthetic intermediates in this pathway. The reported stereo selective total synthesis of cis-(+)-OPDA is not very efficient due to the many steps involved in the reaction as well as the use of water sensitive reactions. Therefore, we developed an enzymatic method for the synthesis of OPDA using acetone powder of flax seed and allene oxide cyclase (PpAOC2) from Physcomitrella patens. From this method, natural cis-(+)-OPDA can be synthesized in the high yield of approximately 40%. In this study, we investigated the substrate specificity of the enzymatic synthesis of other OPDA analogs with successions to afford OPDA amino acid conjugates, dinor-OPDA (dn-OPDA), and OPDA monoglyceride, and it was suggested that the biosynthetic pathway of arabidopsides could occur via MGDG.
• Ozone disrupts the communication between plants and insects in urban and suburban areas: an updated insight on plant volatiles.

  Noboru Masui, Evgenios Agathokleous, Tomoki Mochizuki, Akira Tani, Hideyuki Matsuura, Takayoshi Koike

  Journal of forestry research 1 - 13 2021年01月10日 

  Plant-insect interactions are basic components of biodiversity conservation. To attain the international Sustainable Development Goals (SDGs), the interactions in urban and in suburban systems should be better understood to maintain the health of green infrastructure. The role of ground-level ozone (O3) as an environmental stress disrupting interaction webs is presented. Ozone mixing ratios in suburbs are usually higher than in the center of cities and may reduce photosynthetic productivity at a relatively higher degree. Consequently, carbon-based defense capacities of plants may be suppressed by elevated O3 more in the suburbs. However, contrary to this expectation, grazing damages by leaf beetles have been severe in some urban centers in comparison with the suburbs. To explain differences in grazing damages between urban areas and suburbs, the disruption of atmospheric communication signals by elevated O3 via changes in plant-regulated biogenic volatile organic compounds and long-chain fatty acids are considered. The ecological roles of plant volatiles and the effects of O3 from both a chemical and a biological perspective are presented. Ozone-disrupted plant volatiles should be considered to explain herbivory phenomena in urban and suburban systems. Supplementary information: The online version of this article contains supplementary material available at (10.1007/s11676-020-01287-4) to authorized users.
• Phenazine-1-carboxylic acid (PCA) produced by Paraburkholderia phenazinium CK-PC1 aids postgermination growth of Xyris complanata seedlings with germination induced by Penicillium rolfsii Y-1.

  Masataka Hane, Hanny C Wijaya, Yanetri A Nyon, Yasuko Sakihama, Makoto Hashimoto, Hideyuki Matsuura, Yasuyuki Hashidoko

  Bioscience, biotechnology, and biochemistry 85 1 77 - 84 2021年01月07日 

  Symbiosis of Penicillium rolfsii Y-1 is essential for the seed germination of Hawaii yellow-eyed grass (Xyris complanata). However, the local soil where the plants grow naturally often suppresses the radicle growth of the seedlings. This radicle growth was drastically restored by coinoculation of Paraburkholderia phenazinium isolate CK-PC1, which is a rhizobacterium of X. complanata. It was found that the isolate CK-PC1 produced phenazine-1-carboxylic acid (PCA, 1) as a major metabolite. The biological effects of PCA (1) were investigated using the seeds of X. complanata and Mung bean (Vigna radiata) and it was uncovered that the symbiosis of the isolate CK-PC1was essential for the postgermination growth of X. complanata and the metabolite PCA (1) might partially contribute to promote the growth of the plants.
• Metabolism of airborne methyl salicylate in adjacent plants.

  Rishni Masimbula, Katsunari Oki, Kosaku Takahashi, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 84 9 1780 - 1787 2020年09月 [査読有り][通常論文]
   

  Salicylic acid (SA) and methyl salicylate (MeSA) are synthesized in many plants and are crucial components that establish their disease responses. The metabolism of airborne MeSA to SA has been previously reported. In this report, it was found that SA glucose ester (SAGE), ether (SAG), and salicyloyl-L-aspartic acid (SA-Asp) are metabolites of airborne MeSA. Furthermore, it was found that airborne MeSA was able to increase the endogenous amount of rosmarinic acid in Perilla frutescens, which is known as one of the functional components that contributes to the maintenance of human health.
• Pennelliisides A-C, 2,3,4-Trisubstituted Acyl Glucoses Isolated from Solanum pennellii.

  Tenki Nakashima, Yurika Nambu, Yutaka Inoue, Rishni Masimbula, Hideyuki Matsuura

  Journal of natural products 83 8 2337 - 2346 2020年08月28日 

  Solanum species accumulate a variety of secondary metabolites in their trichomes, and it is well known that acyl sugars are specialized metabolites secreted by the trichomes. However, very few reports provide detailed information on the chemical structure of polyacylated glucose derivatives, due to the α and β isomerization that can occur at the C-1 position. In this study, a strategy was established to isolate polyacylated glucose derivatives. According to the developed strategy, hydroxy groups were derivatized to a benzyloxy group using TriBOT. After isolation of the compounds in pure form and deprotection of the benzyloxy group, the chemical structures of pennelliisides A-C were determined as 2,3,4-O-triisobutyryl-d-glucose, 3-O-(8-methylnonanoyl)-2,4-O-diisobutyryl-d-glucose, and 3-O-decanoyl-2,4-O-diisobutyryl-d-glucose, respectively. Structural elucidation was performed using spectroscopic techniques, including 1D and 2D NMR, FD-MS, and GC-MS. It was also found that the fatty acid moiety contributes to the allelopathic properties of the isolated compounds.
• Jasmonic acid is not a biosynthetic intermediate to produce the pyrethrolone moiety in pyrethrin II.

  Ryo Matsui, Kisumi Takiguchi, Naoshige Kuwata, Katsunari Oki, Kosaku Takahashi, Kazuhiko Matsuda, Hideyuki Matsuura

  Scientific reports 10 1 6366 - 6366 2020年04月14日 [査読有り][通常論文]
   

  Pyrethrum (Tanacetum cinerariifolium) produces insecticidal compounds known as pyrethrins. Pyrethrins are esters; the acid moiety is either trans-chrysanthemic acid or pyrethric acid and the alcohol moiety of pyrethrins is either pyrethrolone, cinerolone, or jasmolone. It was generally accepted that cis-jasmone was biosynthetic intermediate to produce the alcohol moieties of pyrethrin, and the biosynthetic origin of the cis-jasmone was postulated to be jasmonic acid. However, there was no direct evidence to prove this hypothesis. In order to uncover the origin of pyrethrolone moiety in pyrethrin II, feeding experiments were performed employing deuterium- and 13C-labeled compounds as substrates, and the expected labeled compounds were analyzed using UPLC MS/MS system. It was found that the pyrethrolone moiety in pyrethrin II was derived from 12-oxo-phytodienoic acid (OPDA), iso-OPDA and cis-jasmone but not from methyl jasmonate and 3-oxo-2-(2'-[Z]-pentenyl)-cyclopentane-1-hexanoic acid. The results supported that the biosynthesis of the pyrethrolone moiety in pyrethrin II partially used part of the jasmonic acid biosynthetic pathway, but not whole.
• l-Isoleucine (Ile) Promotes Anthocyanin Accumulation in Apples

  Satoru Kondo, Hiroyuki Tomiyama, Peter Opio, Ringo Komiya, Takanori Saito, Katsuya Okawa, Hitoshi Ohara, Hideyuki Matsuura, Yaqin Lang, Daisuke Igarashi

  JOURNAL OF PLANT GROWTH REGULATION 40 2 541 - 549 2020年04月 [査読有り][通常論文]
   

  The functions of l-isoleucine (Ile) in anthocyanin biosynthesis were investigated in apple fruit. Whole trees (Malus x domestica Bokh.) were treated with Ile or n-propyl dihydrojasmonate (PDJ), which is a jasmonic acid (JA) analog, 20 and 30 days before harvest. Both Ile and PDJ treatments stimulated anthocyanin formation at 30 and 35 days after treatment (DAT). The expressions of MdUF3GT and MdMYB1 genes under the Ile and PDJ treatments were higher than those in the untreated control at 19 and 30 DAT. Endogenous JA and jasmonoyl-l-isoleucine (JA-Ile) concentrations in Ile- and PDJ- treated fruit were increased at 19 and 30 DAT. At 30 DAT, the Ile and PDJ treatments stimulated the expressions of MdJAR1, which is related to the conjugation of JA to Ile, and MdMYC2, which is a transcription factor in the JA response. Our study suggests that Ile can induce anthocyanin formation depending on the increase of JA-Ile production and the up-regulation of MdJAR1 and MdMYC2 in apples.
• Role of salicylic acid glucosyltransferase in balancing growth and defence for optimum plant fitness.

  Yudai Kobayashi, Noriho Fukuzawa, Ayaka Hyodo, Hangil Kim, Shota Mashiyama, Tsuyoshi Ogihara, Hirofumi Yoshioka, Hideyuki Matsuura, Chikara Masuta, Takeshi Matsumura, Minoru Takeshita

  Molecular plant pathology 21 3 429 - 442 2020年03月 [査読有り][通常論文]
   

  Salicylic acid (SA), an essential secondary messenger for plant defence responses, plays a role in maintaining a balance (trade-off) between plant growth and resistance induction, but the detailed mechanism has not been explored. Because the SA mimic benzothiadiazole (BTH) is a more stable inducer of plant defence than SA after exogenous application, we analysed expression profiles of defence genes after BTH treatment to better understand SA-mediated immune induction. Transcript levels of the salicylic acid glucosyltransferase (SAGT) gene were significantly lower in BTH-treated Nicotiana tabacum (Nt) plants than in SA-treated Nt control plants, suggesting that SAGT may play an important role in SA-related host defence responses. Treatment with BTH followed by SA suppressed SAGT transcription, indicating that the inhibitory effect of BTH is not reversible. In addition, in BTH-treated Nt and Nicotiana benthamiana (Nb) plants, an early high accumulation of SA and SA 2-O-β-d-glucoside was only transient compared to the control. This observation agreed well with the finding that SAGT-overexpressing (OE) Nb lines contained less SA and jasmonic acid (JA) than in the Nb plants. When inoculated with a virus, the OE Nb plants showed more severe symptoms and accumulated higher levels of virus, while resistance increased in SAGT-silenced (IR) Nb plants. In addition, the IR plants restricted bacterial spread to the inoculated leaves. After the BTH treatment, OE Nb plants were slightly larger than the Nb plants. These results together indicate that SAGT has a pivotal role in the balance between plant growth and SA/JA-mediated defence for optimum plant fitness.
• Prenatal exposure to 11 perfluoroalkyl substances and fetal growth: A large-scale, prospective birth cohort study.

  Ikuko Kashino, Seiko Sasaki, Emiko Okada, Hideyuki Matsuura, Houman Goudarzi, Chihiro Miyashita, Eisaku Okada, Yoichi M Ito, Atsuko Araki, Reiko Kishi

  Environment international 136 105355 - 105355 2020年03月 [査読有り][通常論文]
   

  BACKGROUND: Prenatal maternal exposure to perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) has been reportedly associated with decreased birth weight. Although a majority of epidemiological studies concerning perfluoroalkyl substances (PFAS) have focused on PFOS and PFOA, epidemiological studies of PFAS with longer carbon chains are scarce. In this study, we investigated whether prenatal maternal exposure to 11 PFAS, in particular those with longer carbon chains, is associated with fetal growth. METHODS: The study included 1985 mother-infant pairs (inclusive of preterm and post-term infants), who enrolled in a large-scale, prospective birth cohort study in any of the 37 hospitals in Hokkaido, Japan between 2003 and 2009. The concentration of 11 PFAS was measured in maternal plasma collected during the third trimester of pregnancy, using ultra-performance liquid chromatography in combination with triple quadrupole mass spectrometry. Associations between the measured PFAS values and weight, length, and head circumference of all newborns at birth were examined using multiple regression analyses with adjustment for potential confounders based on data collected from medical records, questionnaires, and those for maternal plasma samples. RESULTS: Of the 11 PFAS analyzed, prenatal perfluorononanoic acid (PFNA) [per log10-unit: regression coefficient (β) = -96.2 g, 95% confidence intervals (95% CI), -165.3 to -27.1] and perfluorodecanoic acid (PFDA) (β = -72.2 g, 95% CI, -138.1 to -6.3) concentrations were inversely associated with birth weight. Furthermore, PFNA concentrations were inversely associated with birth length (per Log10 unit: β = -0.48 cm, 95% CI; - 0.86 to -0.11). Maternal perfluorotridecanoic acid (PFTrDA) exposure showed a significant inverse association with birth weight only for female infants (per Log10 unit: β = -99.8 g, 95% CI, - 193.7 to -6.0) (P for interaction = 0.04). CONCLUSIONS: Our findings suggest that prenatal, maternal exposure to PFAS with longer carbon chains tends to be inversely associated with birth size of newborn infants, which may indicate that these commercially used compounds have an adverse effect on fetal growth.
• Comparative Proteomic Analysis of Wild-Type Physcomitrella Patens and an OPDA-Deficient Physcomitrella Patens Mutant with Disrupted PpAOS1 and PpAOS2 Genes after Wounding.

  Weifeng Luo, Setsuko Komatsu, Tatsuya Abe, Hideyuki Matsuura, Kosaku Takahashi

  International journal of molecular sciences 21 4 2020年02月19日 

  Wounding is a serious environmental stress in plants. Oxylipins such as jasmonic acid play an important role in defense against wounding. Mechanisms to adapt to wounding have been investigated in vascular plants; however, those mechanisms in nonvascular plants remain elusive. To examine the response to wounding in Physcomitrella patens, a model moss, a proteomic analysis of wounded P. patens was conducted. Proteomic analysis showed that wounding increased the abundance of proteins related to protein synthesis, amino acid metabolism, protein folding, photosystem, glycolysis, and energy synthesis. 12-Oxo-phytodienoic acid (OPDA) was induced by wounding and inhibited growth. Therefore, OPDA is considered a signaling molecule in this plant. Proteomic analysis of a P. patens mutant in which the PpAOS1 and PpAOS2 genes, which are involved in OPDA biosynthesis, are disrupted showed accumulation of proteins involved in protein synthesis in response to wounding in a similar way to the wild-type plant. In contrast, the fold-changes of the proteins in the wild-type plant were significantly different from those in the aos mutant. This study suggests that PpAOS gene expression enhances photosynthesis and effective energy utilization in response to wounding in P. patens.
• Does Ozone Alter the Attractiveness of Japanese White Birch Leaves to the Leaf Beetle Agelastica coerulea via Changes in Biogenic Volatile Organic Compounds (BVOCs): An Examination with the Y-Tube Test

  Noboru Masui, Tomoki Mochizuki, Akira Tani, Hideyuki Matsuura, Evgenios Agathokleous, Toshihiro Watanabe, Takayoshi Koike

  Forests 11 1 58 - 58 2020年01月02日 

  Elevated ground-level ozone (O3) reduced C-based defense chemicals; however, severe grazing damages were found in leaves grown in the low O3 condition of a free air O3-concentration enrichment (O3-FACE) system. To explain this phenomenon, this study investigates the role of BVOCs (biogenic volatile organic compounds) as signaling compounds for insect herbivores. BVOCs act as scents for herbivore insects to locate host plants, while some BVOCs show high reactivity to O3, inducing changes in the composition of BVOCs in atmospheres with elevated O3. To assess the aforementioned phenomenon, profiles of BVOCs emitted from birch (Betula platyphylla var. japonica Hara) leaves were analyzed ex situ, and Y-tube insect preference tests were conducted in vitro to study the insect olfactory response. The assays were conducted in June and August or September, according to the life cycle of the adult alder leaf beetle Agelastica coerulea Baly (Coleoptera: Chrysomelidae). The Y-tube tests revealed that the leaf beetles were attracted to BVOCs, and O3 per se had neither an attractant nor a repellent effect. BVOCs became less attractant when mixed with highly concentrated O3 (>80 ppb). About 20% of the total BVOCs emitted were highly O3-reactive compounds, such as β-ocimene. The results suggest that BVOCs emitted from the birch leaves can be altered by elevated O3, thus potentially reducing the attractiveness of leaves to herbivorous insects searching for food.
• Feeding experiment using uniformly 13C-labeled α-linolenic acid supports the involvement of the decarboxylation mechanism to produce cis-jasmone in Lasiodiplodia theobromae.

  Ryo Matsui, Kisumi Takiguchi, Kazuhiko Matsuda, Kosaku Takahashi, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 83 12 2190 - 2193 2019年12月 [査読有り][通常論文]
   

  In our previous report, it was found that Lasiodiplodia theobromae produced cis-jasmone via partially utilizing the biosynthetic pathway of JA. A feeding experiment using uniformly 13C-labeled α-linolenic acid, which was added to the culture media of the fungus, strongly supported that the fungus produced CJ via the decarboxylation step of the biosynthetic pathway.
• Identification of recombinant AtPYL2, an abscisic acid receptor, in E. coli using a substrate-derived bioactive small molecule, a biotin linker with alkyne and amino groups, and a protein cross-linker.

  Tomoaki Anabuki, Yusuke Ito, Keisuke Ohashi, Taichi E Takasuka, Hideyuki Matsuura, Kosaku Takahashi

  Bioorganic & medicinal chemistry letters 29 21 126634 - 126634 2019年11月01日 [査読有り][通常論文]
   

  Target protein identification of bioactive small molecules is one of the most important research in forward chemical genetics. The affinity chromatography technique to use a resin bound with a small molecule is often used for identification of a target protein of a bioactive small molecule. Here we report a new method to isolate a protein targeted with a bioactive small molecule using a biotin linker with alkyne and amino groups, protein cross-linker containing disulfide bond, and a bioactive small molecule with an azido group (azido probe). After an azido probe is associated with a target protein, the complex of a target protein and azido probe is covalently bound through the biotin linker by azide-alkyne Huisgen cycloaddition and protein cross-linker containing disulfide bond. This ternary complex is immobilized on an affinity matrix with streptavidin, and then the target protein is selectively eluted with a buffer containing a reducing agent for cleavage of disulfide bonds. This method uses a probe having an azido group, which a small functional group, and has the possibility to be a solution strategy to overcome the hindrance of a functional group introduced into the probe that reduces association a target protein. The effectiveness of the method in this study was shown using linker 1, 3'-azidoabscisic acid 3, and protein cross-linker containing a disulfide bond (DTSSP 5).
• 12-Hydroxy-Jasmonoyl-l-Isoleucine Is an Active Jasmonate That Signals through CORONATINE INSENSITIVE 1 and Contributes to the Wound Response in Arabidopsis.

  Arati N Poudel, Rebekah E Holtsclaw, Athen Kimberlin, Sidharth Sen, Shuai Zeng, Trupti Joshi, Zhentian Lei, Lloyd W Sumner, Kamlendra Singh, Hideyuki Matsuura, Abraham J Koo

  Plant & cell physiology 60 10 2152 - 2166 2019年10月01日 

  12-hydroxy-jasmonoyl-isoleucine (12OH-JA-Ile) is a metabolite in the catabolic pathway of the plant hormone jasmonate, and is synthesized by the cytochrome P450 subclade 94 enzymes. Contrary to the well-established function of jasmonoyl-isoleucine (JA-Ile) as the endogenous bioactive form of jasmonate, the function of 12OH-JA-Ile is unclear. Here, the potential role of 12OH-JA-Ile in jasmonate signaling and wound response was investigated. Exogenous application of 12OH-JA-Ile mimicked several JA-Ile effects including marker gene expression, anthocyanin accumulation and trichome induction in Arabidopsis thaliana. Genome-wide transcriptomics and untargeted metabolite analyses showed large overlaps between those affected by 12OH-JA-Ile and JA-Ile. 12OH-JA-Ile signaling was blocked by mutation in CORONATINE INSENSITIVE 1. Increased anthocyanin accumulation by 12OH-JA-Ile was additionally observed in tomato and sorghum, and was disrupted by the COI1 defect in tomato jai1 mutant. In silico ligand docking predicted that 12OH-JA-Ile can maintain many of the key interactions with COI1-JAZ1 residues identified earlier by crystal structure studies using JA-Ile as ligand. Genetic alternation of jasmonate metabolic pathways in Arabidopsis to deplete both JA-Ile and 12OH-JA-Ile displayed enhanced jasmonate deficient wound phenotypes and was more susceptible to insect herbivory than that depleted in only JA-Ile. Conversely, mutants overaccumulating 12OH-JA-Ile showed intensified wound responses compared with wild type with similar JA-Ile content. These data are indicative of 12OH-JA-Ile functioning as an active jasmonate signal and contributing to wound and defense response in higher plants.
• 12OHJA, 12OGlcJA, and JA-L-Val as airborne MeJA metabolites.

  Katsunari Oki, Rishni Masimbula, Kanji Miyawaki, Yusuke Takata, Kosaku Takahashi, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 83 9 1709 - 1712 2019年09月 [査読有り][通常論文]
   

  It has been reported that airborne methyl jasmonate (MeJA) was metabolized into jasmonic acid (JA) and jasmonoyl isoleucine (JA-L-Ile). In this report, jasmonoyl valine (JA-L-Val), 12-hydroxy JA (12OHJA), and 12-glucosyloxy JA (12OGlcJA) were identified as metabolites originating from airborne MeJA using tomato (Solanum lycopersicum). Furthermore, the preferable conversion of (-)-MeJA (natural form) into 12OHJA, 12OGlcJA, JA-L-Ile, and JA-L-Val was observed.
• Ability of plant pathogenic fungi Gibberella fujikuroi and Fusarium commune to react with airborne methyl jasmonate.

  Rishni Masimbula, Katsunari Oki, Hiroki Shibata, Hisashi Osawa, Norio Kondo, Kosaku Takahashi, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 83 9 1650 - 1654 2019年09月 [査読有り][通常論文]
   

  The pathogenic fungi Gibberella fujikuroi and Fusarium commune produce jasmonic acid. The application of volatile deuterium-labeled methyl jasmonate increased the amount of nonlabeled JA present in G. fujikuroi and F. commune. These results indicate that the fungi have the ability to react with airborne methyl jasmonate in a manner similar to a plant.
• Jasmonate production through chlorophyll a degradation by Stay-Green in Arabidopsis thaliana.

  Kouhei Ono, Madoka Kimura, Hideyuki Matsuura, Ayumi Tanaka, Hisashi Ito

  Journal of plant physiology 238 53 - 62 2019年07月 [査読有り][通常論文]
   

  Leaf color change through chlorophyll degradation is a characteristic symptom of senescence. Magnesium removal from chlorophyll a is the initial step in chlorophyll a degradation, in a reaction catalyzed by Stay-Green (SGR). Arabidopsis thaliana has three SGR homologs, SGR1, SGR2, and SGR-like. When SGR1 is overexpressed, both chlorophyll a and b are degraded and leaves turn yellow. This process is visually identical to senescence, suggesting that SGR1 overexpression affects various physiological processes in plants. To examine this possibility, gene expression associated with chlorophyll metabolism and senescence was analyzed following dexamethasone-inducible SGR1 introduction into Arabidopsis. When SGR1 was overexpressed following 18 h of dexamethasone treatment, genes involved in chlorophyll degradation were upregulated, as were senescence-associated genes. These observations suggested that chlorophyll a degradation promotes senescence. As jasmonate is the plant hormone responsible for senescence and was expected to be involved in the regulation of gene expression after dexamethasone treatment, the level of jasmonoyl-isoleucine, the active form of jasmonate, was measured. The jasmonoyl-isoleucine level increased slightly after 10 h of SGR1 overexpression, and this increase became significant after 18 h. These observations suggest that jasmonate is produced through chlorophyll a degradation and affects the promotion of senescence.
• Wounding stress induces phenylalanine ammonia lyases, leading to the accumulation of phenylpropanoids in the model liverwort Marchantia polymorpha.

  Mayu Yoshikawa, Weifeng Luo, Genta Tanaka, Yuka Konishi, Hideyuki Matsuura, Kosaku Takahashi

  Phytochemistry 155 30 - 36 2018年11月 [査読有り][通常論文]
   

  Wounding stress induces the biosynthesis of various specialized metabolites in plants. In this study, wounding induced the biosynthesis of luteolin, apigenin, and isoriccardin C, which are biosynthesized through the phenylpropanoid pathway, in the model liverwort Marchantia polymorpha L (Marchantiaceae). Recombinant M. polymorpha phenylalanine ammonia lyases (MpPALs) exhibited PAL activity in vitro and converted phenylalanine into trans-cinnamic acid. Based on semi-quantitative RT-PCR analysis, the expression levels of the MpPAL genes were up-regulated after wounding. α-Aminooxy-β-phenylpropionic acid, a PAL inhibitor, suppressed the production of wounding-induced phenolic compounds, luteolin, apigenin, and isoriccardin C, in M. polymorpha. Thus, PAL is a committed step in the biosynthesis of phenylpropanoids in response to wounding in M. polymorpha. This study suggests that wound-induced specialized metabolites such as phenylpropanoids comprise a conserved defense system in land plants.
• Jasmonic Acid Inhibits Auxin-Induced Lateral Rooting Independently of the CORONATINE INSENSITIVE1 Receptor.

  Yasuhiro Ishimaru, Kengo Hayashi, Takeshi Suzuki, Hidehiro Fukaki, Justyna Prusinska, Christian Meester, Mussa Quareshy, Syusuke Egoshi, Hideyuki Matsuura, Kosaku Takahashi, Nobuki Kato, Erich Kombrink, Richard M Napier, Ken-Ichiro Hayashi, Minoru Ueda

  Plant physiology 177 4 1704 - 1716 2018年08月 [査読有り][通常論文]
   

  Plant root systems are indispensable for water uptake, nutrient acquisition, and anchoring plants in the soil. Previous studies using auxin inhibitors definitively established that auxin plays a central role regulating root growth and development. Most auxin inhibitors affect all auxin signaling at the same time, which obscures an understanding of individual events. Here, we report that jasmonic acid (JA) functions as a lateral root (LR)-preferential auxin inhibitor in Arabidopsis (Arabidopsis thaliana) in a manner that is independent of the JA receptor, CORONATINE INSENSITIVE1 (COI1). Treatment of wild-type Arabidopsis with either (-)-JA or (+)-JA reduced primary root length and LR number; the reduction of LR number was also observed in coi1 mutants. Treatment of seedlings with (-)-JA or (+)-JA suppressed auxin-inducible genes related to LR formation, diminished accumulation of the auxin reporter DR5::GUS, and inhibited auxin-dependent DII-VENUS degradation. A structural mimic of (-)-JA and (+)-coronafacic acid also inhibited LR formation and stabilized DII-VENUS protein. COI1-independent activity was retained in the double mutant of transport inhibitor response1 and auxin signaling f-box protein2 (tir1 afb2) but reduced in the afb5 single mutant. These results reveal JAs and (+)-coronafacic acid to be selective counter-auxins, a finding that could lead to new approaches for studying the mechanisms of LR formation.
• Determinants and Temporal Trends of Perfluoroalkyl Substances in Pregnant Women: The Hokkaido Study on Environment and Children's Health.

  Meng-Shan Tsai, Chihiro Miyashita, Atsuko Araki, Sachiko Itoh, Yu Ait Bamai, Houman Goudarzi, Emiko Okada, Ikuko Kashino, Hideyuki Matsuura, Reiko Kishi

  International journal of environmental research and public health 15 5 2018年05月14日 [査読有り][通常論文]
   

  Perfluoroalkyl substances (PFAS) are persistent bio-accumulative chemicals that impact the health of pregnant women and their children. PFAS derive from environmental and consumer products, which depend on human lifestyle, socioeconomic characteristics, and time variation. Here, we aimed to explore the temporal trends of PFAS in pregnant women and the characteristics related to maternal PFAS concentration. Our study is part of the Hokkaido Study on Environment and Children's Health, the Hokkaido large-scale cohort that recruited pregnant women between 2003 and 2011. Blood samples were acquired from pregnant women during the third trimester to measure PFAS and cotinine concentrations. Maternal basic information was collected with a baseline structured questionnaire. Eleven PFAS were measured from 2123 samples with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry. Eight PFAS were above 80% detection rate and were included in the final analysis. We used multivariable linear regression to analyze the association of pregnant women characteristics with the levels of eight PFAS. The temporal trend of PFAS was observed in two periods (August 2003 to January 2006 and February 2006 to July 2012). The concentration of perfluorooctane sulfonate (PFOS) significantly decreased from August 2003 to January 2006 and from February 2006 to July 2012. The concentrations of perfluorododecanoic acid (PFDoDA), perfluoroundecanoic acid (PFUnDA), and perfluorotridecanoic acid (PFTrDA) increased significantly between August 2003 and January 2006, whereas they decreased significantly between February 2006 and July 2012. Women with pre-pregnancy body mass index (BMI) >25 kg/m² had lower PFUnDA, PFDoDA, and PFTrDA levels than did those with normal BMI (18.5⁻24.9 kg/m²). Pregnant women, who were active smokers (cotinine > 11.49 ng/mL), had higher PFOS than the non-smokers (cotinine < 0.22 ng/mL). Lower levels of PFHxS, PFOS, PFOA, PFNA, and PFDA were observed in women, who had given birth to more than one child. There were also significant positive associations between PFAS levels and annual income or maternal education. PFAS levels varied in women with higher pre-pregnancy BMI, active smoking status, higher education level and annual income. The causes of the individual PFAS differences should be explored in an independent study.
• Biosynthesis and in vitro enzymatic synthesis of the isoleucine conjugate of 12-oxo-phytodienoic acid from the isoleucine conjugate of α-linolenic acid.

  Akira Uchiyama, Takaomi Yaguchi, Hiroyuki Nakagawa, Kento Sasaki, Naoshige Kuwata, Hideyuki Matsuura, Kosaku Takahashi

  Bioorganic & medicinal chemistry letters 28 6 1020 - 1023 2018年04月01日 [査読有り][通常論文]
   

  The isoleucine conjugate of 12-oxo-phytodienoic acid (OPDA-Ile), a new member of the jasmonate family, was recently identified in Arabidopsis thaliana and might be a signaling molecule in plants. However, the biosynthesis and function of OPDA-Ile remains elusive. This study reports an in vitro enzymatic method for synthesizing OPDA-Ile, which is catalyzed by reactions of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC) using isoleucine conjugates of α -linolenic acid (LA-Ile) as the substrate. A. thaliana fed LA-Ile exhibited a marked increase in the OPDA-Ile concentration. LA-Ile was also detected in A. thaliana. Furthermore, stable isotope labelled LA-Ile was incorporated into OPDA-Ile. Thus, OPDA-Ile is biosynthesized via the cyclization of LA-Ile in A. thaliana.
• Novel biotin linker with alkyne and amino groups for chemical labelling of a target protein of a bioactive small molecule.

  Tomoaki Anabuki, Miu Tsukahara, Masanori Okamoto, Hideyuki Matsuura, Kosaku Takahashi

  Bioorganic & medicinal chemistry letters 28 4 783 - 786 2018年02月15日 [査読有り][通常論文]
   

  We synthesized a novel linker (1) with biotin, alkyne and amino groups for the identification of target proteins using a small molecule that contains an azide group (azide probe). The alkyne in the linker bound the azide probe via an azide-alkyne Huisgen cycloaddition. A protein cross-linker effectively bound the conjugate of the linker and an azide probe with a target protein. The covalently bound complex was detected by western blotting. Linker 1 was applied to a model system using an abscisic acid receptor, RCAR/PYR/PYL (PYL). Cross-linked complexes of linker 1, the azide probes and the target proteins were successfully visualized by western blotting. This method of target protein identification was more effective than a previously developed method that uses a second linker with biotin, alkyne, and benzophenone (linker 2) that acts to photo-crosslink target proteins. The system developed in this study is a method for identifying the target proteins of small bioactive molecules and is different from photo-affinity labelling.
• Induction of plant disease resistance upon treatment with yeast cell wall extract.

  Takaomi Yaguchi, Tomohisa Kinami, Tetsuya Ishida, Takaomi Yasuhara, Kosaku Takahashi, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 81 11 2071 - 2078 2017年11月 [査読有り][通常論文]
   

  It has been reported that treatment with yeast cell wall extract (YCWE) induces PDF1 and PR-1 gene expression; these transcripts are important markers of plant disease resistance, though the detailed signaling mechanisms that induce these defense responses are still unknown. In this report, we found that YCWE treatment triggered rice cell suspension cultures to accumulate phenylalanine (Phe), cis-12-oxo-phytodienoic acid (OPDA), 12-hydroxyjasmonoyle isoleucine (12OHJA-Ile), and azelaic acid (AzA). YCWE treatment also reduced endogenous triacylglycerol (TG) content. The addition of 13C-uniform-labeled oleic, linoleic and linolenic acids to the rice cell suspension cultures gave rise to 13C-uniform-labeled AzA. It was also found that YCWE treatment for Arabidopsis thaliana resulted in accumulations of OPDA, AzA, Phe, and camalexin together with enhanced resistance against Botrytis cinerea infection. This suggested that YCWE treatment upon plants may activate JA and AzA signaling systems to induce plant disease resistance.
• N1, N14-diferuloylspermine as an antioxidative phytochemical contained in leaves of Cardamine fauriei.

  Keima Abe, Hideyuki Matsuura, Mitsuko Ukai, Hanako Shimura, Hiroyuki Koshino, Takashi Suzuki

  Bioscience, biotechnology, and biochemistry 81 10 1855 - 1860 2017年10月 [査読有り][通常論文]
   

  Most Brassicaceae vegetables are ideal dietary sources of antioxidants beneficial for human health. Cardamine fauriei (Ezo-wasabi in Japanese) is a wild, edible Brassicaceae herb native to Hokkaido, Japan. To clarify the main antioxidative phytochemical, an 80% methanol extraction from the leaves was fractionated with Diaion® HP-20, Sephadex® LH-20, and Sep-Pak® C18 cartridges, and the fraction with strong antioxidant activity depending on DPPH method was purified by HPLC. Based on the analyses using HRESIMS and MS/MS, the compound might be N1, N14-diferuloylspermine. This rare phenol compound was chemically synthesized, whose data on HPLC, MS and 1H NMR were compared with those of naturally derived compound from C. fauriei. All results indicated they were the same compound. The radical-scavenging properties of diferuloylspermine were evaluated by ORAC and ESR spin trapping methods, with the diferuloylspermine showing high scavenging activities of the ROO·, O2·-, and HO· radicals as was those of conventional antioxidants.
• Elucidation of the biosynthetic pathway of cis-jasmone in Lasiodiplodia theobromae.

  Ryo Matsui, Naruki Amano, Kosaku Takahashi, Yodai Taguchi, Wataru Saburi, Hideharu Mori, Norio Kondo, Kazuhiko Matsuda, Hideyuki Matsuura

  Scientific reports 7 1 6688 - 6688 2017年07月27日 [査読有り][通常論文]
   

  In plants, cis-jasmone (CJ) is synthesized from α-linolenic acid (LA) via two biosynthetic pathways using jasmonic acid (JA) and iso-12-oxo-phytodienoic acid (iso-OPDA) as key intermediates. However, there have been no reports documenting CJ production by microorganisms. In the present study, the production of fungal-derived CJ by Lasiodiplodia theobromae was observed for the first time, although this production was not observed for Botrytis cinerea, Verticillium longisporum, Fusarium oxysporum, Gibberella fujikuroi, and Cochliobolus heterostrophus. To investigate the biosynthetic pathway of CJ in L. theobromae, administration experiments using [18,18,18-2H3, 17,17-2H2]LA (LA-d5), [18,18,18-2H3, 17,17-2H2]12-oxo-phytodienoic acid (cis-OPDA-d5), [5',5',5'-2H3, 4',4'-2H2, 3'-2H1]OPC 8:0 (OPC8-d6), [5',5',5'-2H3, 4',4'-2H2, 3'-2H1]OPC 6:0 (OPC6-d6), [5',5',5'-2H3, 4',4'-2H2, 3'-2H1]OPC 4:0 (OPC4-d6), and [11,11-2H2, 10,10-2H2, 8,8-2H2, 2,2-2H2]methyl iso-12-oxo-phytodienoate (iso-MeOPDA-d8) were carried out, revealing that the fungus produced CJ through a single biosynthetic pathway via iso-OPDA. Interestingly, it was suggested that the previously predicted decarboxylation step of 3,7-didehydroJA to afford CJ might not be involved in CJ biosynthesis in L. theobromae.
• Prenatal exposure to perfluoroalkyl acids and prevalence of infectious diseases up to 4years of age.

  Houman Goudarzi, Chihiro Miyashita, Emiko Okada, Ikuko Kashino, Chi-Jen Chen, Sachiko Ito, Atsuko Araki, Sumitaka Kobayashi, Hideyuki Matsuura, Reiko Kishi

  Environment international 104 132 - 138 2017年07月 [査読有り][通常論文]
   

  Perfluoroalkyl acids (PFAAs) are synthetic chemicals with ability to repel oils and water, and have been widely used in many industrial and household applications such as adhesives and water- and stain-repellent surfaces to nonstick coatings. Animal studies have shown that PFAAs have immunotoxic effects. However, few epidemiological studies have investigated the effects of PFAAs on infectious diseases occurrence. We examined the relationship between prenatal exposure to PFAAs and prevalence of infectious diseases up to 4years of life. A total of 1558 mother-child pairs, who were enrolled in the Hokkaido Study on Environment and Children's Health, were included in this data analysis. Eleven PFAAs were measured in maternal plasma taken at 28-32weeks of gestation using ultra-performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry. Participant characteristics were obtained from medical birth records and self-administered questionnaires during pregnancy and after delivery. Physicians' diagnosis of common infectious diseases including otitis media, pneumonia, respiratory syncytial virus infection, and varicella up to 4years were extracted from the mother-reported questionnaires. The number of children who developed infectious diseases up to 4years of age was as follows: otitis media, 649 (41.4%); pneumonia, 287 (18.4%); respiratory syncytial virus infection, 197 (12.6%); varicella 589 (37.8%). A total of 1046 (67.1%) children had at least one of the diseases defined as total infectious diseases. After adjusting for appropriate confounders, PFOS levels in the highest quartile were associated with increased odds ratios (ORs) of total infectious diseases (Q4 vs. Q1 OR: 1.61; 95% CI: 1.18, 2.21; p for trend=0.008) in all children. In addition, perfluorohexane sulfonate (PFHxS) was associated with a higher risk of total infectious diseases only among girls (Q4 vs. Q1 OR: 1.55, 95% CI: 0.976, 2.45; p for trend=0.045). We found no association between infectious diseases and other examined PFAAs. Our findings suggest that prenatal exposure to PFOS and PFHxS may associated with infectious diseases occurrence in early life. Therefore, prenatal exposure to PFAAs may be immunotoxic for the immune system in offspring.
• Determination of the Absolute Configuration of a Monoglyceride Antibolting Compound and Isolation of Related Compounds from Radish Leaves (Raphanus sativus).

  Tsuyoshi Ogihara, Naruki Amano, Yuki Mitsui, Kaien Fujino, Hiroyuki Ohta, Kosaku Takahashi, Hideyuki Matsuura

  Journal of natural products 80 4 872 - 878 2017年04月28日 [査読有り][通常論文]
   

  A monoglyceride (1) has been reported to possess an antibolting effect in radish (Raphanus sativus), but its absolute configuration at the C-2 position was not determined earlier. In this work, the absolute configuration of 1 was determined to be (2S), and it was also accompanied by one new (2) and two known monoglycerides (3 and 4). The chemical structure of 2 was determined as β-(7'Z,10'Z,13'Z)-hexadecatrienoic acid monoglyceride (β-16:3 monoglyceride). Qualitative and quantitative analytical methods for compounds 1-4 were developed, using two deuterium-labeled compounds (8 and 9) as internal standards. The results revealed a broader range of distribution of 1-4 in several annual winter crops. It was also found that these isolated compounds have an inhibitory effect on the root elongation of Arabidopsis thaliana seedlings at concentrations of 25 and 50 μM in the medium. However, the inhibitory effect of 1 was not dependent on coronatin-insensitive 1 (COI1) protein, which may suggest the involvement of an unidentified signaling system other than jasmonic acid signaling.
• Identification of Jasmonic Acid and Jasmonoyl-Isoleucine, and Characterization of AOS, AOC, OPR and JAR1 in the Model Lycophyte Selaginella moellendorffii.

  Putri Pratiwi, Genta Tanaka, Tomohiro Takahashi, Xiaonan Xie, Koichi Yoneyama, Hideyuki Matsuura, Kosaku Takahashi

  Plant & cell physiology 58 4 789 - 801 2017年04月01日 [査読有り][通常論文]
   

  Jasmonic acid (JA) is involved in a variety of physiological responses in seed plants. However, the detection and role of JA in lycophytes, a group of seedless vascular plants, have remained elusive until recently. This study provides the first evidence of 12-oxo-phytodienoic acid (OPDA), JA and jasmonoyl-isoleucine (JA-Ile) in the model lycophyte Selaginella moellendorffii. Mechanical wounding stimulated the accumulation of OPDA, JA and JA-Ile. These data were corroborated by the detection of enzymatically active allene oxide synthase (AOS), allene oxide cyclase (AOC), 12-oxo-phytodienoic acid reductase 3 (OPR3) and JA-Ile synthase (JAR1) in S. moellendorffii. SmAOS2 is involved in the first committed step of JA biosynthesis. SmAOC1 is a crucial enzyme for generating the basic structure of jasmonates and is actively involved in the formation of OPDA. SmOPR5, a functionally active OPR3-like enzyme, is also vital for the reduction of (+)-cis-OPDA, the only isomer of the JA precursor. The conjugation of JA to Ile by SmJAR1 demonstrates that S. moellendorffii produces JA-Ile. Thus, the four active enzymes have characteristics similar to those in seed plants. Wounding and JA treatment induced the expression of SmAOC1 and SmOPR5. Furthermore, JA inhibited the growth of shoots in S. moellendorffii, which suggests that JA functions as a signaling molecule in S. moellendorffii. This study proposes that JA evolved as a plant hormone for stress adaptation, beginning with the emergence of vascular plants.
• GTR1 is a jasmonic acid and jasmonoyl-l-isoleucine transporter in Arabidopsis thaliana.

  Yasuhiro Ishimaru, Takaya Oikawa, Takeshi Suzuki, Syohei Takeishi, Hideyuki Matsuura, Kosaku Takahashi, Shin Hamamoto, Nobuyuki Uozumi, Takafumi Shimizu, Mitsunori Seo, Hiroyuki Ohta, Minoru Ueda

  Bioscience, biotechnology, and biochemistry 81 2 249 - 255 2017年02月 [査読有り][通常論文]
   

  Jasmonates are major plant hormones involved in wounding responses. Systemic wounding responses are induced by an electrical signal derived from damaged leaves. After the signaling, jasmonic acid (JA) and jasmonoyl-l-isoleucine (JA-Ile) are translocated from wounded to undamaged leaves, but the molecular mechanism of the transport remains unclear. Here, we found that a JA-Ile transporter, GTR1, contributed to these translocations in Arabidopsis thaliana. GTR1 was expressed in and surrounding the leaf veins both of wounded and undamaged leaves. Less accumulations and translocation of JA and JA-Ile were observed in undamaged leaves of gtr1 at 30 min after wounding. Expressions of some genes related to wound responses were induced systemically in undamaged leaves of gtr1. These results suggested that GTR1 would be involved in the translocation of JA and JA-Ile in plant and may be contributed to correct positioning of JA and JA-Ile to attenuate an excessive wound response in undamaged leaves.
• Proteomics of Physcomitrella patens protonemata subjected to treatment with 12-oxo-phytodienoic acid.

  Weifeng Luo, Yohei Nanjo, Setsuko Komatsu, Hideyuki Matsuura, Kosaku Takahashi

  Bioscience, biotechnology, and biochemistry 80 12 2357 - 2364 2016年12月 [査読有り][通常論文]
   

  12-Oxo-phytodienoic acid (OPDA) is induced by mechanical wounding and suppresses the growth of Physcomitrella patens; OPDA is considered as a signal compound in this moss species. In this study, a proteomic analysis of P. patens protonemata treated with OPDA was performed. The abundance levels of 41 proteins were significantly altered by OPDA, with decreased levels for 40 proteins. The proteins for which abundance decreased in response to OPDA at the protonema developmental stage were mainly involved in the metabolism of proteins and carbohydrates. The effects of inhibition on protein abundance are likely a major physiological function of OPDA in P. patens. OPDA also suppressed the expression of histones at the protein level and gene transcription level. Suppression of histone expression might be an OPDA-specific function in P. patens protonemata. In P. patens, a subset of the physiological responses caused by OPDA is shown to differ between protonema and gametophore developmental stages.
• Effects of prenatal exposure to perfluoroalkyl acids on prevalence ofallergic diseases among 4-year-old children.

  Houman Goudarzi, Chihiro Miyashita, Emiko Okada, Ikuko Kashino, Sumitaka Kobayashi, Chi-Jen Chen, Sachiko Ito, Atsuko Araki, Hideyuki Matsuura, Yoichi M Ito, Reiko Kishi

  Environment international 94 124 - 132 2016年09月 [査読有り][通常論文]
   

  Perfluoroalkyl acids (PFAAs) are ubiquitous chemicals extremely resistant and widespread throughout the environment, frequently being detected in human blood samples. Animal studies have revealed that exposure to PFAAs results in immunotoxicity. However, the association between PFAAs, especially long-chain PFAAs, and allergies in humans is not well established. We examined whether prenatal exposure to PFAAs is associated with allergic diseases among 4-year-old children in a large-scale prospective birth cohort in Hokkaido, Japan. In total, 1558 mother-child pairs were included in this study and prenatal levels of eleven PFAAs were measured in maternal plasma samples obtained between 28 and 32weeks of pregnancy by using ultra-performance liquid chromatography-tandem mass spectrometry. Participant demographic and characteristic information were obtained from self-administered pre- and postnatal questionnaires and medical birth records. Infant allergies were assessed using the Japanese version of the International Study of Asthma and Allergies in Childhood (ISAAC) Phase Three questionnaire, which was administered 4years post-delivery. Symptoms included eczema, wheezing and rhinoconjunctivitis with a prevalence of 19.0%, 18.7%, and 5.4%, respectively. Associations of PFAA quartiles with allergic outcomes were examined using logistic models. Adjusted odds ratios (ORs) in the 4th quartile vs. 1st quartile (Q4 vs. Q1) for total allergic diseases (including at least one allergic outcome) significantly decreased for perfluorododecanoic acid (PFDoDa) (Q4 vs. Q1 OR: 0.621; 95% confidence interval (CI): 0.454, 0.847) and perfluorotridecanoic acid (PFTrDA) (Q4 vs. Q1 OR: 0.712; 95% CI: 0.524, 0.966) in all children. We obtained similar results when examining the association between PFAAs and eczema. The adjusted OR (Q4 vs. Q1) for wheezing in relation to higher maternal PFHxS levels was 0.728 (95% CI: 0.497, 1.06) in all children. In conclusion, prenatal exposure to long-chain PFAAs, such as PFDoDa and PFTrDA may have an immunosuppressive effect on allergic diseases in 4-year-old children.
• RNA-seq-based evaluation of bicolor tepal pigmentation in Asiatic hybrid lilies (Lilium spp.).

  Kazuma Suzuki, Tomohiro Suzuki, Takashi Nakatsuka, Hideo Dohra, Masumi Yamagishi, Kohei Matsuyama, Hideyuki Matsuura

  BMC genomics 17 1 611 - 611 2016年08月11日 [査読有り][通常論文]
   

  BACKGROUND: Color patterns in angiosperm flowers are produced by spatially and temporally restricted deposition of pigments. Identifying the mechanisms responsible for restricted pigment deposition is a topic of broad interest. Some dicots species develop bicolor petals, which are often caused by the post-transcriptional gene silencing (PTGS) of chalcone synthase (CHS) genes. An Asiatic hybrid lily (Lilium spp.) cultivar Lollypop develops bicolor tepals with pigmented tips and white bases. Here, we analyzed the global transcription of pigmented and non-pigmented tepal parts from Lollypop, to determine the main transcriptomic differences. RESULTS: De novo assembly of RNA-seq data yielded 49,239 contigs (39,426 unigenes), which included a variety of novel transcripts, such as those involved in flavonoid-glycosylation and sequestration and in regulation of anthocyanin biosynthesis. Additionally, 1258 of the unigenes exhibited significantly differential expression between the tepal parts (false discovery rates <0.05). The pigmented tepal parts accumulated more anthocyanins, and unigenes annotated as anthocyanin biosynthesis genes (e.g., CHS, dihydroflavonol 4-reductase, and anthocyanidin synthase) were expressed 7-30-fold higher than those in non-pigmented parts. These results indicate that the transcriptional regulation of biosynthesis genes is more likely involved in the development of bicolor lily tepals rather than the PTGS of CHS genes. In addition, the expression level of a unigene homologous to LhMYB12, which often regulates full-tepal anthocyanin pigmentation in lilies, was >2-fold higher in the pigmented parts. Thus, LhMYB12 should be involved in the transcriptional regulation of the biosynthesis genes in bicolor tepals. Other factors that potentially suppress or enhance the expression of anthocyanin biosynthesis genes, including a WD40 gene, were identified, and their involvement in bicolor development is discussed. CONCLUSIONS: Our results indicate that the bicolor trait of Lollypop tepals is caused by the transcriptional regulation of anthocyanin biosynthesis genes and that the transcription profile of LhMYB12 provides a clue for elucidating the mechanisms of the trait. The tepal transcriptome constructed in this study will accelerate investigations of the genetic controls of anthocyanin color patterns, including the bicolor patterns, of Lilium spp.
• Ascorbic acid accumulates as a defense response to Turnip mosaic virus in resistant Brassica rapa cultivars.

  Ayaka Fujiwara, Satoko Togawa, Takahiro Hikawa, Hideyuki Matsuura, Chikara Masuta, Tsuyoshi Inukai

  Journal of experimental botany 67 14 4391 - 402 2016年07月 [査読有り][通常論文]
   

  We initially observed that Brassica rapa cultivars containing the Turnip mosaic virus (TuMV) resistance gene, Rnt1-1, accumulated a high level of endogenous ascorbic acid (AS) and dehydroascobic acid (DHA) when infected with TuMV. We here hypothesized a possible contribution of an elevated level of AS+DHA (TAA) to the Rnt1-1-mediated resistance, and conducted a series of experiments using B. rapa and Arabidopsis plants. The application of l-galactose (the key substrate in AS synthesis) to a susceptible cultivar could increase the TAA level ~2-fold, and simultaneously lead to some degree of enhanced viral resistance. To confirm some positive correlation between TAA levels and viral resistance, we analyzed two Arabidopsis knockout mutants (ao and vtc1) in the AS pathways; the TAA levels were significantly increased and decreased in ao and vtc1 plants, respectively. While the ao plants showed enhanced resistance to TuMV, vtc1 plants were more susceptible than the control, supporting our hypothesis. When we analyzed the expression profiles of the genes involved in the AS pathways upon TuMV infection, we found that the observed TAA increase was mainly brought about by the reduction of AS oxidation and activation of AS recycling. We then investigated the secondary signals that regulate endogenous TAA levels in response to viral infection, and found that jasmonic acid (JA) might play an important role in TAA accumulation. In conclusion, we reason that the elevated TAA accumulation in B. rapa plants would be at least partly mediated by the JA-dependent signaling pathway and may significantly contribute to viral resistance.
• A chloroplast-localized protein LESION AND LAMINA BENDING affects defence and growth responses in rice.

  Muluneh Tamiru, Hiroki Takagi, Akira Abe, Takao Yokota, Hiroyuki Kanzaki, Haruko Okamoto, Hiromasa Saitoh, Hideyuki Takahashi, Koki Fujisaki, Kaori Oikawa, Aiko Uemura, Satoshi Natsume, Yusuke Jikumaru, Hideyuki Matsuura, Kenji Umemura, Matthew J Terry, Ryohei Terauchi

  The New phytologist 210 4 1282 - 97 2016年06月 [査読有り][通常論文]
   

  Understanding how plants allocate their resources to growth or defence is of long-term importance to the development of new and improved varieties of different crops. Using molecular genetics, plant physiology, hormone analysis and Next-Generation Sequencing (NGS)-based transcript profiling, we have isolated and characterized the rice (Oryza sativa) LESION AND LAMINA BENDING (LLB) gene that encodes a chloroplast-targeted putative leucine carboxyl methyltransferase. Loss of LLB function results in reduced growth and yield, hypersensitive response (HR)-like lesions, accumulation of the antimicrobial compounds momilactones and phytocassanes, and constitutive expression of pathogenesis-related genes. Consistent with these defence-associated responses, llb shows enhanced resistance to rice blast (Magnaporthe oryzae) and bacterial blight (Xanthomonas oryzae pv. oryzae). The lesion and resistance phenotypes are likely to be caused by the over-accumulation of jasmonates (JAs) in the llb mutant including the JA precursor 12-oxo-phytodienoic acid. Additionally, llb shows an increased lamina inclination and enhanced early seedling growth due to elevated brassinosteroid (BR) synthesis and/or signalling. These findings show that LLB functions in the chloroplast to either directly or indirectly repress both JA- and BR-mediated responses, revealing a possible mechanism for controlling how plants allocate resources for defence and growth.
• 3-O-β-D-Glucopyranosyltheobroxide from Aerial Parts of Cowpea (Vigna unguiculata).

  Arata Aoi, Yudai Yamashita, Xiquan Gao, Makoto Uematsu, Maremichi Ota, Kosaku Takahashi, Teruhiko Yoshihara, Hideyuki Matsuura

  Natural product communications 11 5 673 - 6 2016年05月 [査読有り][通常論文]
   

  Theobroxide has been isolated from culture filtrates of Lasiodiplodia theobromae as a potato tuber-inducing compound. In this study, the metabolism of theobroxide was investigated using cowpea as an experimental model and [2H3-7]theobroxide as a substrate for analyzing a metabolite, which revealed that theobroxide applied exogenously to the roots was converted into 3-O-β-D-glucopyranosyltheobroxide.
• Chemical Promotion of Endogenous Amounts of ABA in Arabidopsis thaliana by a Natural Product, Theobroxide.

  Yudai Yamashita, Maremichi Ota, Yutaka Inoue, Youko Hasebe, Masanori Okamoto, Tsuyoshi Inukai, Chikra Masuta, Yasuko Sakihama, Yasuyuki Hashidoko, Mikiko Kojima, Hitoshi Sakakibara, Yasuyuki Inage, Kosaku Takahashi, Teruhiko Yoshihara, Hideyuki Matsuura

  Plant & cell physiology 57 5 986 - 99 2016年05月 [査読有り][通常論文]
   

  Plant hormones are a group of structurally diverse small compounds that orchestrate the cellular processes governing proper plant growth and environmental adaptation. To understand the details of hormonal activity, we must study not only their inherent activities but also the cross-talk among plant hormones. In addition to their use in agriculture, plant chemical activators, such as probenazole and uniconazole, have made great contributions to understand hormonal cross-talk. However, the use of plant chemical activators is limited due to the lack of activators for certain hormones. For example, to the best of our knowledge, there are only a few chemical activators previously known to stimulate the accumulation of ABA in plants, such as absinazoles and proanthocyanidins. In many cases, antagonistic effects have been examined in experiments using exogenously applied ABA, although these studies did not account for biologically relevant concentrations. In this report, it was found that a natural product, theobroxide, had potential as a plant chemical activator for stimulating the accumulation of ABA. Using theobroxide, the antagonistic effect of ABA against GAs was proved without exogenously applying ABA or using mutant plants. Our results suggest that ABA levels could be chemically controlled to elicit ABA-dependent biological phenomena.
• Metabolomics approach in soybean

  Takuji Nakamura, Keiki Okazaki, Noureddine Benkeblia, Jun Wasaki, Toshihiro Watanabe, Hideyuki Matsuura, Hirofumi Uchimiya, Setsuko Komatsu, Takuro Shinano

  Genetics, Genomics, and Breeding of Soybean 313 - 330 2016年04月19日 

  Profiling plant metabolites is able to serve a new technology for understanding metabolic pathway and it may also help picking up the difference beneath plant phenotypes. CE-MS is one of the suitable tools to analyze a large number of plant metabolites, while GC-MS is also a very powerful especially when focusing on primary metabolites, and LC-MS(/MS) is suitable for the detection of secondary metabolites. Several applications of metabolomics approach by using these analytical tools with fractionation of cellular organelles and extraction procedures are demonstrated in this chapter.
• Hormone crosstalk in wound stress response: wound-inducible amidohydrolases can simultaneously regulate jasmonate and auxin homeostasis in Arabidopsis thaliana.

  Tong Zhang, Arati N Poudel, Jeremy B Jewell, Naoki Kitaoka, Paul Staswick, Hideyuki Matsuura, Abraham J Koo

  Journal of experimental botany 67 7 2107 - 20 2016年03月 [査読有り][通常論文]
   

  Jasmonate (JA) and auxin are essential hormones in plant development and stress responses. While the two govern distinct physiological processes, their signaling pathways interact at various levels. Recently, members of the Arabidopsis indole-3-acetic acid (IAA) amidohydrolase (IAH) family were reported to metabolize jasmonoyl-isoleucine (JA-Ile), a bioactive form of JA. Here, we characterized three IAH members, ILR1, ILL6, and IAR3, for their function in JA and IAA metabolism and signaling. Expression of all three genes in leaves was up-regulated by wounding or JA, but not by IAA. Purified recombinant proteins showed overlapping but distinct substrate specificities for diverse amino acid conjugates of JA and IAA. Perturbed patterns of the endogenous JA profile in plants overexpressing or knocked-out for the three genes were consistent with ILL6 and IAR3, but not ILR1, being the JA amidohydrolases. Increased turnover of JA-Ile in the ILL6- and IAR3-overexpressing plants created symptoms of JA deficiency whereas increased free IAA by overexpression of ILR1 and IAR3 made plants hypersensitive to exogenous IAA conjugates. Surprisingly, ILL6 overexpression rendered plants highly resistant to exogenous IAA conjugates, indicating its interference with IAA conjugate hydrolysis. Fluorescent protein-tagged IAR3 and ILL6 co-localized with the endoplasmic reticulum-localized JA-Ile 12-hydroxylase, CYP94B3. Together, these results demonstrate that in wounded leaves JA-inducible amidohydrolases contribute to regulate active IAA and JA-Ile levels, promoting auxin signaling while attenuating JA signaling. This mechanism represents an example of a metabolic-level crosstalk between the auxin and JA signaling pathways.
• Selective regulation of pyrethrin biosynthesis by the specific blend of wound induced volatiles in Tanacetum cinerariifolium.

  Koji Sakamori, Naoaki Ono, Makoto Ihara, Hideyuki Suzuki, Hideyuki Matsuura, Ken Tanaka, Daisaku Ohta, Shigehiko Kanaya, Kazuhiko Matsuda

  Plant signaling & behavior 11 4 e1149675  2016年 

  Natural pyrethrins are used to control household and agricultural pests, and it is of value to understand biosynthesis in Tanacetum cinerariifolium for enhanced production. We previously found that a blend of four green leaf volatiles (GLVs) and (E)-β-farnesene emitted by T. cinerariifolium seedlings enhanced gene expressions of certain biosynthetic enzymes in unwounded seedlings; however, the extent to which such a regulation facilitates pyrethrin biosynthesis remains unknown. Here we have investigated the effects of the blend of the volatile organic compounds (VOCs) on gene expressions of seven biosynthetic enzymes. VOC treatment resulted in enhanced chrysanthemyl diphosphate synthase (CDS), chrysanthemic acid synthase (CAS), Tanacetum cinerariifolium GDSL lipase (TcGLIP) and acyl-Coenzyme A oxidase 1 (ACX1) gene expressions that reached a peak at a 12 h VOC treatment, whereas the treatment minimally influenced the expressions of other biosynthetic genes. In undifferentiated Tanacetum tissues, such VOC-induced amplification of CDS, CAS, TcGLIP and ACX1 gene expressions were markedly reduced, suggesting that a high-resolution, VOC-mediated communication is an event selective to differentiated plants.
• Tandem photoaffinity labeling of a target protein using a linker with biotin, alkyne and benzophenone groups and a bioactive small molecule with an azide group.

  Tomoaki Anabuki, Miu Tsukahara, Hideyuki Matsuura, Kosaku Takahashi

  Bioscience, biotechnology, and biochemistry 80 3 432 - 9 2016年 [査読有り][通常論文]
   

  A novel linker containing biotin, alkyne and benzophenone groups (1) was synthesized to identify target proteins using a small molecule probe. This small molecule probe contains an azide group (azide probe) that reacts with an alkyne in 1 via an azide-alkyne Huisgen cycloaddition. Cross-linking of benzophenone to the target protein formed a covalently bound complex consisting of the azide probe and the target protein via 1. The biotin was utilized via biotin-avidin binding to identify the cross-linked complex. To evaluate the effectiveness of 1, it was applied in a model system using an allene oxide synthase (AOS) from the model moss Physcomitrella patens (PpAOS1) and an AOS inhibitor that contained azide group (3). The cross-linked complex consisting of PpAOS1, 1 and 3 was resolved via SDS-PAGE and visualized using a chemiluminescent system. The method that was developed in this study enables the effective identification of target proteins.
• Changes in Polysome Association of mRNA Throughout Growth and Development in Arabidopsis thaliana

  Shotaro Yamasaki, Hideyuki Matsuura, Taku Demura, Ko Kato

  Plant and Cell Physiology 56 11 2169 - 2180 2015年11月01日 [査読有り][通常論文]
   

  Translational control is a key regulatory step in the expression of genes as proteins. In plant cells, the translational efficiency of mRNAs differs for different mRNA species, and the efficiency dynamically changes in various conditions. To gain a global view of translational control throughout growth and development, we performed genome-wide analysis of polysome association of mRNA during growth and leaf development in Arabidopsis thaliana by subjecting the mRNAs in polysomes to DNA microarray. This analysis revealed that the degree of polysome association of mRNA was different depending on the mRNA species, and the polysome association changed greatly throughout growth and development for each. In the growth stage, transcripts showed varying changes in polysome association from strongly depressed to unchanged, with the majority of transcripts showing dissociation from ribosomes. On the other hand, during leaf development, the polysome association of transcripts showed a normal distribution from repressed to activated mRNAs when comparing expanding and expanded leaves. In addition, functional category analysis of the microarray data suggested that translational control has a physiological significance in the plant growth and development process, especially in the categories of signaling and protein synthesis. In addition to this, we compared changes in polysome association of mRNAs between various conditions and characterized translational controls in each. This result suggested that mRNA translation might be controlled by complicated mechanisms for response to each condition. Our results highlight the importance of dynamic changes in mRNA translation in plant development and growth.
• Identification of rice Os4BGlu13 as a β-glucosidase which hydrolyzes gibberellin A4 1-O-β-d-glucosyl ester, in addition to tuberonic acid glucoside and salicylic acid derivative glucosides.

  Hua Y, Ekkhara W, Sansenya S, Srisomsap C, Roytrakul S, Saburi W, Takeda R, Matsuura H, Mori H, Ketudat Cairns JR

  Archives of biochemistry and biophysics 583 36 - 46 2015年10月01日 [査読有り][通常論文]
   

  Gibberellin 1-O-β-d-glucose ester hydrolysis activity has been detected in rice seedling extracts, but no enzyme responsible for this activity has ever been purified and identified. Therefore, gibberellin A4 glucosyl ester (GA4-GE) β-d-glucosidase activity was purified from ten-day rice seedling stems and leaves. The family 1 glycoside hydrolase Os4BGlu13 was identified in the final purification fraction. The Os4BGlu13 cDNA was amplified from rice seedlings and expressed as an N-terminal thioredoxin-tagged fusion protein in Escherichia coli. The purified recombinant Os4BGlu13 protein (rOs4BGlu13) had an optimum pH of 4.5, for hydrolysis of p-nitrophenyl β-d-glucopyranoside (pNPGlc), which was the best substrate identified, with a kcat/Km of 637 mM(-1) s(-1). rOs4BGlu13 hydrolyzed helicin best among natural glycosides tested (kcat/Km of 74.4 mM(-1) s(-1)). Os4BGlu13 was previously designated tuberonic acid glucoside (TAG) β-glucosidase (TAGG), and here the kcat/Km of rOsBGlu13 for TAG was 6.68 mM(-1) s(-1), while that for GA4-GE was 3.63 mM(-1) s(-1) and for salicylic acid glucoside (SAG) is 0.88 mM(-1) s(-1). rOs4BGlu13 also hydrolyzed oligosaccharides, with preference for short β-(1 → 3)-linked over β-(1 → 4)-linked glucooligosaccharides. The enzymatic data suggests that Os4BGlu13 may contribute to TAG, SAG, oligosaccharide and GA4-GE hydrolysis in the rice plant, although helicin or a similar compound may be its primary target.
• Abscisic acid induces biosynthesis of bisbibenzyls and tolerance to UV-C in the liverwort Marchantia polymorpha.

  Akito Kageyama, Kimitsune Ishizaki, Takayuki Kohchi, Hideyuki Matsuura, Kosaku Takahashi

  Phytochemistry 117 547 - 553 2015年09月 [査読有り][通常論文]
   

  Environmental stresses are effective triggers for the biosynthesis of various secondary metabolites in plants, and phytohormones such as jasmonic acid and abscisic acid are known to mediate such responses in flowering plants. However, the detailed mechanism underlying the regulation of secondary metabolism in bryophytes remains unclear. In this study, the induction mechanism of secondary metabolites in the model liverwort Marchantia polymorpha was investigated. Abscisic acid (ABA) and ultraviolet irradiation (UV-C) were found to induce the biosynthesis of isoriccardin C, marchantin C, and riccardin F, which are categorized as bisbibenzyls, characteristic metabolites of liverworts. UV-C led to the significant accumulation of ABA. Overexpression of MpABI1, which encodes protein phosphatase 2C (PP2C) as a negative regulator of ABA signaling, suppressed accumulation of bisbibenzyls in response to ABA and UV-C irradiation and conferred susceptibility to UV-C irradiation. These data show that ABA plays a significant role in the induction of bisbibenzyl biosynthesis, which might confer tolerance against UV-C irradiation in M. polymorpha.
• Glucosinolate profiles in Cardamine fauriei and effect of light quality on glucosinolate concentration

  Keima Abe, Shigenori Kido, Tomoo Maeda, Daisuke Kami, Hideyuki Matsuura, Hanako Shimura, Takashi Suzuki

  SCIENTIA HORTICULTURAE 189 12 - 16 2015年06月 [査読有り][通常論文]
   

  Most Brassicaceae plants produce glucosinolates (GSLs), which are hydrolyzed to form isothiocyanates (ITCs) that contribute to a stimulative flavor and antibiotic activity beneficial to human health. Three glucosinolates (GSLs), gluconapin, glucoibarin and glucobrassicin, were isolated from plants of Cardamine fauriei Maxim. (Brassicaceae; Ezo-wasabi in Japanese), a wild, edible herb originating from Hokkaido, Japan. These GSLs comprise two types: aliphatic (gluconapin and glucoibarin) and indolic (glucobrassicin). Total GSL concentration in the leaf blade, petiole and root of the plant grown in natural light were 62.81, 43.94 and 17.63 mg g(-1) dry mass, respectively. When the plants were grown under various wavelengths of light supplied by LEDs, total GSL content in the leaf blade was highest when plants were irradiated by red + blue light. In addition, the ratio of the content of the indolic GSL to that of aliphatic GSL was greater after irradiation with by red or green alone rather than blue alone, red + blue, or white (red + blue + green). Because the last three types of irradiation include blue wavelengths, aliphatic GSL content can be increased by irradiation with blue light wavelengths, whereas the indolic GSL levels decreased. The results indicate that it is possible to control the GSL component ratio in C. fauriei by irradiating plants with particular wavelengths. (C) 2015 Elsevier B.V. All rights reserved.
• UDP-Glc非依存性配糖体化酵素による新規なジャスモン酸の配糖体化と その生理的意義

  竹松 知紀, 瀬戸 義哉, 宮澤 吉郎, 和久田 真司, 荻原 毅, 佐分利 亘, 森 春英, 高橋 公咲, 松浦 英幸

  天然有機化合物討論会講演要旨集 57 Oral14  天然有機化合物討論会実行委員会 2015年 

  【目 的】

  植物は大地に根を張り移動が困難である事から、自己の被る様々な環境要因，ストレスに適応して生活環を終結させなくてはならない。よって、植物は独自の環境応答機構を有している。この応答機構を司る生理活性物質として植物ホルモンと呼称される一群の化合物が知られ、多くの研究がなされてきた。我々の研究室では長年、植物ホルモンの一種であるジャスモン酸(JA)について、鋭意研究を行っている。JA類は植物の傷害応答シグナル物質として必須であり、他の興味ある生理活性としてバレイショ塊茎の誘導活性、植物の就眠運動における就眠誘導、花粉管伸長促進などが知られている。近年、JA類の生理活性の制御の観点から、配糖化、チトクロームP450による酸化などに注目した報告がある。

  我々はJA類の配糖体化に関して研究を進め、イネに含まれるOsSGTを12-hydroxyJA(12-OHJA)配糖化酵素として報告した[1]。OsSGT は糖供与体としてUDP-Glcを用いるが、12-OHJAよりサリチル酸(SA)に対してより高い糖転位活性を示した。しかしながら、本酵素の探索過程において、SAに対してほとんど配糖化活性を示さず、12-OHJAに特異性の高いUDP-Glc非依存性の配糖化酵素の存在も明らかとした。現在のところ、UDP-Glc非依存性の植物2次代謝産物を標的とした配糖化酵素に関しては松葉らの報告[2]があるのみであり、植物ホルモンの配糖体化に関してUDP-Glc非依存性の糖転位酵素が関与しているとの報告は一切無い。

  上記の興味ある発見の詳細を明らかとすべく我々は、1）イネカルスに含まれるUDP-Glc非依存性の配糖化酵素の精製、2）イネカルスに含まれる本配糖化酵素の糖供与体の単離精製、構造決定,3）本配糖化酵素の諸性質の究明を進めたところ、本配糖化酵素はb−glucosydase, OsBGlu1 （accession number: AK100165）であると突き止め、イネにおいてサリチル酸グルコシド(SAG)が糖供与体として働くことを明らかとした(図1)。本酵素の生理学的意義は水酸化JA類を配糖化しJAの活性調節に寄与していると示唆できた事から、本成果について報告する。

  【研究方法および結果】

  1. UDP-Glc非依存性配糖化酵素の発見

  イネカルス粗酵素溶液にUDP-Glcを糖供与体としない12-OHJA 選択的な配糖化酵素が存在する事を、市販の糖供与体を用い、以下の実験より発見した。糖転位反応液としてイネカルスより抽出した粗酵素溶液、糖受容体としてJA水酸化体の一種である12-OHJA、糖供与体として市販のGlc 1-phosphate, TDP-Glc, octyl-Glc, UDP-Glcを用いて実験を行った。その結果、octyl-Glc はUDP-Glcよりも12-OHJAにより選択的に糖を供与し、SAはほとんど配糖化されなかった。

  2. イネ由来のUDP-Glc非依存性配糖化酵素の同定

  12-OHJA選択的糖転位活性を指標に、イネカルス粗酵素溶液に含まれる糖転位酵素の同定を試みた。糖供与体としてoctyl-Glc、糖受容体として12-OHJAを用い、種々の精製操作の後、得られた精製タンパク質はペプチドマスフィンガープ

  (View PDFfor the rest of the abstract.)

• Overexpression of RelA/SpoT homologs, PpRSH2a and PpRSH2b, induces the growth suppression of the moss Physcomitrella patens.

  Michio Sato, Tomohiro Takahashi, Kozo Ochi, Hideyuki Matsuura, Kensuke Nabeta, Kosaku Takahashi

  Bioscience, biotechnology, and biochemistry 79 1 36 - 44 2015年 [査読有り][通常論文]
   

  Two genes encoding RelA/SpoT homologs, PpRSH2a and PpRSH2b, which are involved in the synthesis of bacterial alarmone guanosine 5'-diphosphate 3'-diphosphate (ppGpp) for the stringent response, were isolated from the moss, Physcomitrella patens. A complementary analysis of PpRSH2a and PpRSH2b in Escherichia coli showed that these genes had ppGpp biosynthetic activity. The recombinant PpRSH2a and PpRSH2b were also shown to synthesize ppGpp in vitro. Both proteins were localized to the chloroplasts of P. patens. Expression of the PpRSH genes was induced upon treatment with abscisic acid or abiotic stresses, such as dehydration and UV irradiation. Overexpression of PpRSH2a and PpRSH2b caused suppression of the growth in response to 1% (w/v) of glucose. The present study suggests the existence of a mechanism to regulate the growth of P. patens, which is governed by plant RSH in chloroplasts.
• Endoplasmic reticulum-associated inactivation of the hormone jasmonoyl-L-isoleucine by multiple members of the cytochrome P450 94 family in Arabidopsis.

  Abraham J Koo, Caitlin Thireault, Starla Zemelis, Arati N Poudel, Tong Zhang, Naoki Kitaoka, Federica Brandizzi, Hideyuki Matsuura, Gregg A Howe

  The Journal of biological chemistry 289 43 29728 - 38 2014年10月24日 [査読有り][通常論文]
   

  The plant hormone jasmonate (JA) controls diverse aspects of plant immunity, growth, and development. The amplitude and duration of JA responses are controlled in large part by the intracellular level of jasmonoyl-L-isoleucine (JA-Ile). In contrast to detailed knowledge of the JA-Ile biosynthetic pathway, little is known about enzymes involved in JA-Ile metabolism and turnover. Cytochromes P450 (CYP) 94B3 and 94C1 were recently shown to sequentially oxidize JA-Ile to hydroxy (12OH-JA-Ile) and dicarboxy (12COOH-JA-Ile) derivatives. Here, we report that a third member (CYP94B1) of the CYP94 family also participates in oxidative turnover of JA-Ile in Arabidopsis. In vitro studies showed that recombinant CYP94B1 converts JA-Ile to 12OH-JA-Ile and lesser amounts of 12COOH-JA-Ile. Consistent with this finding, metabolic and physiological characterization of CYP94B1 loss-of-function and overexpressing plants demonstrated that CYP94B1 and CYP94B3 coordinately govern the majority (>95%) of 12-hydroxylation of JA-Ile in wounded leaves. Analysis of CYP94-promoter-GUS reporter lines indicated that CYP94B1 and CYP94B3 serve unique and overlapping spatio-temporal roles in JA-Ile homeostasis. Subcellular localization studies showed that CYP94s involved in conversion of JA-Ile to 12COOH-JA-Ile reside on endoplasmic reticulum (ER). In vitro studies further showed that 12COOH-JA-Ile, unlike JA-Ile, fails to promote assembly of COI1-JAZ co-receptor complexes. The double loss-of-function mutant of CYP94B3 and ILL6, a JA-Ile amidohydrolase, displayed a JA profile consistent with the collaborative action of the oxidative and the hydrolytic pathways in JA-Ile turnover. Collectively, our results provide an integrated view of how multiple ER-localized CYP94 and JA amidohydrolase enzymes attenuate JA signaling during stress responses.
• Prenatal exposure to perfluoroalkyl acids and allergic diseases in early childhood.

  Emiko Okada, Seiko Sasaki, Ikuko Kashino, Hideyuki Matsuura, Chihiro Miyashita, Sumitaka Kobayashi, Kumiko Itoh, Tamiko Ikeno, Akiko Tamakoshi, Reiko Kishi

  Environment international 65 127 - 34 2014年04月 [査読有り][通常論文]
   

  Perfluoroalkyl acids (PFAAs) are persistent organic pollutants that are detected in humans worldwide. Laboratory animal studies have shown that PFAAs are associated with immunotoxic effects. However, epidemiological studies investigating the role of PFAAs, in particular PFAAs with longer chains than perfluorooctanoic acid, are scarce. We investigated associations between prenatal exposure to PFAAs, including long-chain compounds, and infant allergic diseases at 12 and 24months in a large study population. The participants included mothers and their infants who enrolled in the Hokkaido Study on Environment and Children's Health 2003-2009. Eleven PFAAs were measured in maternal plasma taken at 28-32weeks of gestation using ultra-performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry. Characteristics of participants and information on infant allergic diseases were obtained from self-administered questionnaires and medical records. At 24months, the adjusted odds ratio (OR) (first vs. fourth quartiles) for eczema in association with higher maternal perfluorotridecanoic acid (PFTrDA) levels was 0.62 (95% confidence interval (CI) 0.45, 0.86). After stratification by gender, the adjusted ORs in female infants from mothers with higher maternal perfluoroundecanoic acid (PFUnDA) and PFTrDA levels were also statistically significant (PFUnDA: OR=0.50; 95% CI, 0.30, 0.81; PFTrDA: OR=0.39; 95% CI, 0.23, 0.64). Our findings suggest that lower prenatal exposure to PFTrDA may decrease the risk of developing eczema in early childhood, only in female infants.
• CYP94B3 activity against jasmonic acid amino acid conjugates and the elucidation of 12-O-β-glucopyranosyl-jasmonoyl-L-isoleucine as an additional metabolite.

  Naoki Kitaoka, Hiroshi Kawaide, Naruki Amano, Takuya Matsubara, Kensuke Nabeta, Kosaku Takahashi, Hideyuki Matsuura

  Phytochemistry 99 6 - 13 2014年03月 [査読有り][通常論文]
   

  The hormonal action of jasmonate in plants is controlled by the precise balance between its biosynthesis and inactivation. Oxidation of jasmonoyl-L-isoleucine at the C-12 position, which is catalyzed by cytochrome P450s CYP94B3 and CYP94C1, is thought to be one of the main inactivation pathways. In this study, an additional function of CYP94B3 was elucidated, as well additional jasmonoyl-L-isoleucine metabolites being investigated. It was found that CYP94B3 also catalyzes the hydroxylation of jasmonoyl-L-valine and jasmonoyl-L-phenylalanine, and that these hydroxyl compounds accumulated after wounding and possessed lower activity than non-hydroxylated compounds. Additionally, 12-O-β-glucopyranosyl-jasmonoyl-L-isoleucine accumulated after wounding, suggesting that it is a metabolite of jasmonoyl-L-isoleucine.
• Proteomic analysis of Physcomitrella patens treated with 12-oxo-phytodienoic acid, an important oxylipin in plants.

  Erika Toshima, Yohei Nanjo, Setsuko Komatsu, Tatsuya Abe, Hideyuki Matsuura, Kosaku Takahashi

  Bioscience, biotechnology, and biochemistry 78 6 946 - 53 2014年 [査読有り][通常論文]
   

  12-Oxo-phytodienoic acid (OPDA) is biosynthesized in the octadecanoid pathway and is considered to be a signaling molecule in plants. In Physcomitrella patens, OPDA is induced by bacterial infection and mechanical stress and is known to suppress growth; however, the functional mechanism of OPDA signaling remains elusive. In this study, we performed a proteomic analysis of P. patens treated with OPDA and found that the expression of 82 proteins was significantly altered, with approximately 80% of these proteins being downregulated by OPDA. The identified proteins were mainly categorized as being involved in photosynthesis, metabolism, and protein synthesis, and most of the proteins that were upregulated by OPDA are involved in light-dependent reactions, suggesting that OPDA regulates a function in chloroplasts. Additionally, OPDA induced the expression of an allene oxide cyclase (PpAOC1) in the octadecanoid pathway, demonstrating positive feedback regulation by OPDA in P. patens.
• Production of 1,5-anhydro-d-fructose by an α-glucosidase belonging to glycoside hydrolase family 31.

  Maneesan J, Matsuura H, Tagami T, Mori H, Kimura A

  Bioscience, biotechnology, and biochemistry 78 12 2064 - 8 2014年 [査読有り][通常論文]
   

  α-1,4-Glucan lyases [glycoside hydrolase family (GH) 31] catalyze an elimination reaction to form 1,5-anhydro-d-fructose (AF), while GH31 α-glucosidases normally catalyze a hydrolytic reaction. We determined that a small amount of AF was produced by GH31 Aspergillus niger α-glucosidase from maltooligosaccharides by elimination reaction, likely via an oxocarbenium ion intermediate.
• Temporal trends of perfluoroalkyl acids in plasma samples of pregnant women in Hokkaido, Japan, 2003-2011.

  Emiko Okada, Ikuko Kashino, Hideyuki Matsuura, Seiko Sasaki, Chihiro Miyashita, Jun Yamamoto, Tamiko Ikeno, Yoichi M Ito, Toru Matsumura, Akiko Tamakoshi, Reiko Kishi

  Environment international 60 89 - 96 2013年10月 [査読有り][通常論文]
   

  Perfluoroalkyl acids (PFAAs) are persistent organic pollutants that are used in a wide range of consumer products. Recent epidemiological studies have shown that prenatal exposure to toxic levels of PFAAs in the environment may adversely affect fetal growth and humoral immune response in infants and children. Here we have characterized levels of prenatal exposure to PFAA between 2003 and 2011 in Hokkaido, Japan, by measuring PFAA concentrations in plasma samples from pregnant women. The study population comprised 150 women who enrolled in a prospective birth cohort study conducted in Hokkaido. Eleven PFAAs were measured in maternal plasma samples using simultaneous analysis by ultra-performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry. At the end of the study, in 2011, age- and parity-adjusted mean concentrations of perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnDA), perfluorododecanoic acid (PFDoDA), perfluorotridecanoic acid (PFTrDA), perfluorohexane sulfonate (PFHxS), and perfluorooctane sulfonate (PFOS) were 1.35ng/mL, 1.26ng/mL, 0.66ng/mL, 1.29ng/mL, 0.25ng/mL, 0.33ng/mL, 0.28ng/mL, and 3.86ng/mL, respectively. Whereas PFOS and PFOA concentrations declined 8.4%/y and 3.1%/y, respectively, PFNA and PFDA levels increased 4.7%/y and 2.4%/y, respectively, between 2003 and 2011. PFUnDA, PFDoDA, and PFTrDA were detected in the vast majority of maternal samples, but no significant temporal trend was apparent. Future studies must involve a larger population of pregnant women and their children to determine the effects of prenatal exposure to PFAA on health outcomes in infants and children.
• Isolation, structural elucidation, and biological evaluation of a 5-hydroxymethyl-2-furfural derivative, asfural, from enzyme-treated asparagus extract

  Tomohiro Ito, Atsuya Sato, Tomoko Ono, Kazunori Goto, Takahiro Maeda, Jun Takanari, Hiroshi Nishioka, Kenichi Komatsu, Hideyuki Matsuura

  Journal of Agricultural and Food Chemistry 61 38 9155 - 9159 2013年09月25日 [査読有り][通常論文]
   

  A novel 5-hydroxymethyl-2-furfural (HMF 1) derivative, which is named asfural (compound 2), was isolated from enzyme-treated asparagus extract (ETAS) along with HMF (1) as a heat shock protein 70 (HSP70) inducible compound. The structure of compound 2 was elucidated on the basis of its spectroscopic data from HREIMS and NMR, whereas the absolute configuration was determined using chiral HPLC analysis, compared to two synthesized compounds, (S)- and (R)-asfural. As a result, compound 2 derived from ETAS was assigned as (S)-(2-formylfuran-5-yl)methyl 5-oxopyrrolidine-2-carboxylate. When compound 2, synthesized (S)- and (R)-asfural, and HMF (1) were evaluated in terms of HSP70 mRNA expression-enhancing activity in HL-60 cells, compound 2 and (S)-asfural significantly increased the expression level in a concentration-dependent manner. HMF (1) also showed significant activity at 0.25 mg/mL. © 2013 American Chemical Society.
• Response of rice to insect elicitors and the role of OsJAR1 in wound and herbivory-induced JA-Ile accumulation.

  Kaori Fukumoto, Kabir Alamgir, Yuko Yamashita, Izumi C Mori, Hideyuki Matsuura, Ivan Galis

  Journal of integrative plant biology 55 8 775 - 84 2013年08月 [査読有り][通常論文]
   

  Plants produce jasmonic acid (JA) and its amino acid conjugate, jasmonoyl-L-isoleucine (JA-Ile) as major defense signals in response to wounding and herbivory. In rice (Oryza sativa), JA and JA-Ile rapidly increased after mechanical damage, and this increase was further amplified when the wounds were treated with oral secretions from generalist herbivore larvae, lawn armyworms (Spodoptera mauritia), revealing for the first time active perception mechanisms of herbivore-associated elicitor(s) in rice. In the rice genome, two OsJAR genes can conjugate JA and Ile and form JA-Ile in vitro; however, their function in herbivory-induced accumulation of JA-Ile has not been investigated. By functional characterization of TOS17 retrotransposon-tagged Osjar1 plants and their response to simulated herbivory, we show that OsJAR1 is essential for JA-Ile production in herbivore-attacked, field-grown plants. In addition, OsJAR1 was required for normal seed development in rice under field conditions. Our results suggest that OsJAR1 possesses at least two major functions in rice defense and development that cannot be complemented by the additional OsJAR2 gene function, although this gene previously showed overlapping enzyme activity in vitro.
• Ushikulide C, a New Immunosuppressant from Streptomyces sp IUK-102

  Takahashi Kosaku, Fukushi Eri, Kawabata Jun, Matsuura Hideyuki, Kurosawa Kazuhiko

  NATURAL PRODUCT COMMUNICATIONS 8 6 815 - 816 2013年06月 [査読有り][通常論文]
  
• A Computational and Experimental Approach Reveals that the 5 '-Proximal Region of the 5 '-UTR has a Cis-Regulatory Signature Responsible for Heat Stress-Regulated mRNA Translation in Arabidopsis

  Hideyuki Matsuura, Shinya Takenami, Yuki Kubo, Kiyotaka Ueda, Aiko Ueda, Masatoshi Yamaguchi, Kazumasa Hirata, Taku Demura, Shigehiko Kanaya, Ko Kato

  PLANT AND CELL PHYSIOLOGY 54 4 474 - 483 2013年04月 [査読有り][通常論文]
   

  Translation of specific plant mRNAs is differentially regulated under certain abiotic stress conditions such as heat, oxygen deprivation and dehydration. The majority of transcripts exhibit varying degrees of translational repression, whereas a subset of transcripts escape such repression and remain actively translated. The underlying mechanisms that mediate this control, and in particular the identities of the regulatory RNA elements involved, remain poorly understood. Using a combined computational and experimental approach, we identified a novel cis-regulatory element in the 5'-untranslated region (5'-UTR) that affects differential translation in response to heat stress (HS) in Arabidopsis thaliana. First, we selected a set of genes with distinct translational responses to HS, based on our previously reported genome-wide data regarding changes in polysome loading induced by HS in A. thaliana cultured cells. We evaluated the 5'-UTRs of these messages for their ability to mediate expression, when fused to reporter mRNAs, in protoplasts under HS. The data from the reporter assay and the nucleotide sequences of the 5'-UTRs tested were used to define regulatory elements in the 5'-UTRs, with the help of a partial least square (PLS) regression model. The computational analysis using PLS and subsequent experimental characterization of a series of 5'-UTR mutants provided evidence that the 5'-proximal sequence of the 5'-UTR is a primary and position-dependent determinant of 5'-UTR-mediated differential translation in response to HS. Finally, we discuss the possible mechanism underlying HS regulation of differential mRNA translation.
• 傷害応答シグナルとしてのJA-Ile

  佐藤 千鶴, 松浦 英幸

  植物の生長調節 48 1 93 - 101 一般社団法人植物化学調節学会 2013年 

  Determining the mobile signal used by plants to defend against biotic and abiotic stresses has proved elusive, but jasmonic acid (JA) and its derivatives appear to be involved. Using deuterium-labeled analogs, we investigated the systemic transport of jasmonoyl isoleucine (JA-Ile) in response to leaf wounding in tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum) plants. It was found that de novo synthesized JA-Ile responded to wound damages was transported to induce JA-mediated wound response in healthy systemic organ.
• Profiling of jasmonic acid-related metabolites and hormones in wounded leaves.

  Yusuke Jikumaru, Mitsunori Seo, Hideyuki Matsuura, Yuji Kamiya

  Methods in molecular biology (Clifton, N.J.) 1011 113 - 22 2013年 [査読有り][通常論文]
   

  The endogenous concentration of N-jasmonoyl-L-isoleucine (JA-Ile) is regulated by the balance between biosynthesis and deactivation and controls plant developmental processes and stress responses. Therefore, profiling of its precursors and metabolites is required to understand the mechanism by which the JA-Ile concentration is regulated. Also, other hormones, such as indole-3-acetic acid, abscisic acid, salicylic acid, and ethylene, have been suggested to interact with JA-Ile signaling. Profiling of these hormones and their metabolites should give us insights into their interaction mode. Liquid chromatography-electrospray ionization-tandem mass spectrometry has enabled us to develop a highly sensitive and high-throughput comprehensive quantification analysis of phytohormones.
• Identification of rice β-glucosidase with high hydrolytic activity towards salicylic acid β-d-glucoside

  Nami Himeno, Wataru Saburi, Shinji Wakuta, Ryosuke Takeda, Hideyuki Matsuura, Kensuke Nabeta, Sompong Sansenya, James R, Ketudat Cairns, Haruhide Mori, Ryozo Imai, Hirokazu Matsui

  Bioscience, Biotechnology and Biochemistry 77 5 934 - 9 2013年 [査読有り][通常論文]
   

  β-Glucosidases (EC 3.2.1.21) split β-glucosidic linkages at the non-reducing end of glucosides and oligosaccharides to release β-D-glucose. One of the important functions of plant β-glucosidase is deglucosylation of inactive glucosides of phytohormones to regulate levels of active hormones. Tuberonic acid is a jasmonaterelated compound that shows tuber-inducing activity in the potato. We have identified two enzymes, OsTAGG1 and OsTAGG2, that have hydrolytic activity towards tuberonic acid β-D-glucoside in rice (Oryza sativa L.). The expression of OsTAGG2 is upregulated by wounding and by methyl jasmonate, suggesting that this isozyme is involved in responses to biotic stresses and wounding, but the physiological substrate of OsTAGG2 remains ambiguous. In this study, we produced recombinant OsTAGG2 in Pichia pastoris (rOsTAGG2P), and investigated its substrate specificity in detail. From 1L of culture medium, 2.1mg of purified recombinant enzyme was obtained by ammonium sulfate precipitation and Ni-chelating column chromatography. The specific activity of rOsTAGG2P (182 U/mg) was close to that of the native enzyme (171 U/mg), unlike recombinant OsTAGG2 produced in Escherichia coli, which had approximately 3-fold lower specific activity than the native enzyme. The optimum pH and temperature for rOsTAGG2P were pH 3.4 and 60 βC. After pH and heat treatments, the enzyme retained its original activity in a pH range of 3.4-9.8 and below 55 βC. Native OsTAGG2 and rOsTAGG2P showed 4.5-4.7-fold higher activities towards salicylic acid β-D-glucoside, an inactive storageform of salicylic acid, than towards tuberonic acid β-Dglucoside (TAG), although OsTAGG2 was originally isolated from rice based on TAG-hydrolytic activity.
• Transportation of de novo synthesized jasmonoyl isoleucine in tomato.

  Hideyuki Matsuura, Syohei Takeishi, Naoki Kiatoka, Chizuru Sato, Kae Sueda, Chikara Masuta, Kensuke Nabeta

  Phytochemistry 83 25 - 33 2012年11月 [査読有り][通常論文]
   

  In plants, jasmonic acid (JA) and its derivatives are thought to be involved in mobile forms of defense against biotic and abiotic stresses. In this study, the distal transport of JA-isoleucine (JA-Ile) that is synthesized de novo in response to leaf wounding in tomato (Solanum lycopersicum) plants was investigated. JA-[¹³C₆]Ile was recovered in distal untreated leaves after wounded leaves were treated with [¹³C₆]Ile. However, as [¹³C₆]Ile was also recovered in the distal untreated leaves, whether JA-Ile was synthesized in the wounded or in the untreated leaves was unclear. Hence, stem exudates were analyzed to obtain more detailed information. When [¹³C₆]Ile and [²H₆]JA were applied separately into the wounds on two different leaves, JA-[¹³C₆]Ile and [²H₆]JA-Ile were detected in the stem exudates but [²H₆]JA-[¹³C₆]Ile was not, indicating that JA was conjugated with Ile in the wounded leaf and that the resulting JA-Ile was then transported into systemic tissues. The [²H₃]JA-Ile that was applied exogenously to the wounded tissues reached distal untreated leaves within 10 min. Additionally, applying [²H₃]JA-Ile to the wounded leaves at concentrations of 10 and 60 nmol/two leaves induced the accumulation of PIN II, LAP A, and JAZ3 mRNA in the distal untreated leaves of the spr2 mutant S. lycopersicum plants. These results demonstrate the transportation of de novo synthesized JA-Ile and suggest that JA-Ile may be a mobile signal.
• Genome-Wide Analyses of Changes in Translation State Caused by Elevated Temperature in Oryza sativa

  Kiyotaka Ueda, Hideyuki Matsuura, Masatoshi Yamaguchi, Taku Demura, Ko Kato

  PLANT AND CELL PHYSIOLOGY 53 8 1481 - 1491 2012年08月 [査読有り][通常論文]
   

  It has been reported that the translational status of mRNAs responds dramatically to abiotic stresses. While many useful results have demonstrated translational control in dicotyledonous model plants, little is known about changes in the translation state in response to abiotic stresses in monocotyledonous plants. To understand global changes in translation of mRNAs, we performed genome-wide analyses using Oryza sativa treated with heat stress (HS). These analyses showed that most mRNAs were translationally repressed, while the translation of some mRNAs was maintained. In addition to other regulatory steps in gene expression, including transcription and processing, it is thought that translational regulation is a critical step in adaptation to new conditions because of the functional tendencies of proteins that are either translationally maintained or highly repressed upon HS. When we compared the functional tendencies of translationally regulated proteins in rice with those in Arabidopsis thaliana cells exposed to HS, some showed similar regulation, arguing for both common and different features of translational regulation in the two plants.
• Isolation of antitrypanosomal compounds from Vitis repens, a medicinal plant of Myanmar.

  Khine Swe Nyunt, Ahmed Elkhateeb, Yusuke Tosa, Kensuke Nabata, Ken Katakura, Hideyuki Matsuura

  Natural product communications 7 5 609 - 10 2012年05月 [査読有り][通常論文]
   

  Bioactivity-guided fractionation of an ethanolic extract of Vitis repens led to the isolation of resveratrol (1), 11-O-acetyl bergenin (2), and stigmast-4-en-3-one (3). The compounds were examined for their in vitro antitrypanosomal activities against trypomastigotes of Trypanosoma evansi. Resveratrol showed antitrypanosomal activity with an IC50 value of 0.13 microM, whereas 11-O-acetyl bergenin and stigmast-4-en-3-one exhibited IC50 values of 0.17 and 0.15 microM, respectively.
• Biological activity and tissue specific accumulation of fluorescently labeled methyl jasmonate.

  Naoki Kitaoka, Yuzou Sano, Seizo Fujikawa, Kensuke Nabeta, Hideyuki Matsuura

  Natural product communications 7 2 215 - 8 2012年02月 [査読有り][通常論文]
   

  The C-7 position of jasmonate is practical for synthesis of a probe to use for chemical biological studies. To confirm the utility, we synthesized fluorescent-labeled methyl jasmonate. The synthesized compound exhibited Arabidopsis thaliana root growth inhibitory and meandering activity, and potent fluorescence was observed inside the root and root hairs.
• Antitrypanosomal activities of acetylated bruceines A and C; a structure-activity relationship study.

  Ahmed Elkhateeb, Yusuke Tosa, Hideyuki Matsuura, Kensuke Nabeta, Ken Katakura

  Journal of natural medicines 66 1 233 - 40 2012年01月 [査読有り][通常論文]
   

  The crude extract of Brucea javanica showed strong in vitro inhibitory activity against Trypanosoma evansi. Among the isolated quassinoids, bruceines A, C, and bruceantinol were found to be the most potent compounds against T. evansi. To gain a deeper understanding of the relationship between the free hydroxyl groups and the activity, several O-acetylated derivatives of bruceines A and C were synthesized and their in vitro antitrypanosomal activities against trypomastigotes of T. evansi were examined and compared with those of the original compounds. The following structure-activity relationships were observed: (1) the free hydroxyl groups at positions C-3, C-11, and C-12 are essential for antitrypanosomal activity; (2) the C-11 and C-12 hydroxyl groups are more important for the activity than the enolic hydroxyl group at C-3, and; (3) the free hydroxyl group at C-4' of bruceine C does not have any significant effect on the activity.
• Infection dynamics in viral spread and interference under the synergism between Cucumber mosaic virus and Turnip mosaic virus.

  Minoru Takeshita, Emiko Koizumi, Makiko Noguchi, Kae Sueda, Hanako Shimura, Noriko Ishikawa, Hideyuki Matsuura, Kazusato Ohshima, Tomohide Natsuaki, Shigeru Kuwata, Naruto Furuya, Kenichi Tsuchiya, Chikara Masuta

  Molecular plant-microbe interactions : MPMI 25 1 18 - 27 2012年01月 [査読有り][通常論文]
   

  Mixed infection of Cucumber mosaic virus (CMV) and Turnip mosaic virus (TuMV) induced more severe symptoms on Nicotiana benthamiana than single infection. To dissect the relationships between spatial infection patterns and the 2b protein (2b) of CMV in single or mixed infections, the CMV vectors expressing enhanced green fluorescent or Discosoma sp. red fluorescent proteins (EGFP [EG] or DsRed2 [Ds], respectively were constructed from the same wild-type CMV-Y and used for inoculation onto N. benthamiana. CMV2-A1 vector (C2-A1 [A1]) has a functional 2b while CMV-H1 vector (C2-H1 [H1]) is 2b deficient. As we expected from the 2b function as an RNA silencing suppressor (RSS), in a single infection, A1Ds retained a high level of accumulation at initial infection sites and showed extensive fluorescence in upper, noninoculated leaves, whereas H1Ds disappeared rapidly at initial infection sites and could not spread efficiently in upper, noninoculated leaf tissues. In various mixed infections, we found two phenomena providing novel insights into the relationships among RSS, viral synergism, and interference. First, H1Ds could not spread efficiently from vasculature into nonvascular tissues with or without TuMV, suggesting that RNA silencing was not involved in CMV unloading from vasculature. These results indicated that 2b could promote CMV to unload from vasculature into nonvascular tissues, and that this 2b function might be independent of its RSS activity. Second, we detected spatial interference (local interference) between A1Ds and A1EG in mixed infection with TuMV, between A1Ds (or H1Ds) and TuMV, and between H1Ds and H1EG. This observation suggested that local interference between two viruses was established even in the synergism between CMV and TuMV and, again, RNA silencing did not seem to contribute greatly to this phenomenon.
• Efficient synthesis of (+)-cis-12-oxo-phytodienoic acid by an in vitro enzymatic reaction.

  Akiyuki Kajiwara, Tatsuya Abe, Takahiro Hashimoto, Hideyuki Matsuura, Kosaku Takahashi

  Bioscience, biotechnology, and biochemistry 76 12 2325 - 8 2012年 [査読有り][通常論文]
   

  12-Oxo-phytodienoic acid (OPDA) is an important metabolite on the octadecanoid pathway of plants. This study shows an efficient in vitro synthesis of (+)-cis-OPDA by using a flaxseed extract and an allene oxide cyclase. The OPDA yield of the reaction in this study was almost 7-fold higher than that in the conventional reaction with the flaxseed extract.
• High level expression of transgenes by use of 5′-untranslated region of the Arabidopsis thaliana arabinogalactan-protein 21 gene in dicotyledons

  Takeshi Matsui, Hideyuki Matsuura, Kazutoshi Sawada, Eiji Takita, Satoko Kinjo, Shinya Takenami, Kiyotaka Ueda, Naoya Nishigaki, Shotaro Yamasaki, Kensuke Hata, Masatoshi Yamaguchi, Taku Demura, Ko Kato

  Plant Biotechnology 29 3 319 - 322 2012年 [査読有り][通常論文]
   

  For higher expression of a foreign gene in plant cells, it is important to optimize nucleotide sequences corresponding to 5′-untranslated region (5′-UTR), because it usually has great impacts on the expression of the gene mainly at the translational level. In this study, with an aim to find useful 5′-UTRs, thirty nine 5′-UTRs derived from Arabidopsis thaliana genes were tested by transient expression of firefly luciferase (Fluc), and that of A. thaliana arabinogalactanprotein 21 (AtAGP21) gene was selected for further analyses. Its activity was either equaling or surpassing that of known translational enhancer, A. thaliana alchol dehydrogenase (AtADH) 5′-UTR in dicotyledons, and was further improved by the optimizing sequence context of the initiating codon (-3 to -1 of AUG). Finally, we also found that the modified AtAGP21 5′-UTR was useful in recombinant expression of horseradish peroxidase (HRP) in tobacco cultured cells, and the yield was as much as 23 mg l -1 culture medium in seven days. © 2012 The Japanese Society for Plant Cell and Molecular Biology.
• Isolation of polyphenol oxidase genes from Portulaca oleracea and evaluation of their ability to metabolize endocrine-disrupting chemicals

  Hirokazu Kaneda, Takeshi Matsui, Ryosuke Tomiyasu, Yukako Kuroda, Yuka Higashimoto, Tomoka Oda, Hitoshi Miyasaka, Hiroshi Okuhata, Satoshi Tanaka, Kazuo Harada, Hideyuki Matsuura, Hideki Nakayama, Ko Kato, Kazumasa Hirata

  Plant Biotechnology 29 4 351 - 357 2012年 [査読有り][通常論文]
   

  Portulaca (Portulaca oleracea cv.), a garden plant, efficiently removes endocrine-disrupting chemicals (EDCs) including bisphenol A (BPA) from hydroponic solution. We hypothesized that polyphenol oxidase (PPO) was involved in the initial steps of detoxifying EDCs in portulaca roots. In order to elucidate the molecular basis of portulaca's ability to metabolize EDCs, we first isolated five PPO genes (PoPPO1-5) that were expressed mainly in portulaca roots. Among these genes, PoPPO2, PoPPO4 and PoPPO5 were introduced into cultured tobacco cells and expressed as active forms. We found that crude extracts from the cells expressing PoPPO2, PoPPO5, and to a lesser extent PoPPO4, could metabolize BPA. In addition, we found that the BPA metabolites from crude extracts of cells expressing PoPPO2, PoPPO4 and PoPPO5 were identical to those of portulaca. Moreover, PoPPO2 and PoPPO5 also caused hydroxylation of octylphenol, nonylphenol and 17β-estradiol. Therefore, these results strongly suggest that PoPPOs significantly contribute to the superior ability of portulaca to metabolize EDCs. © 2012 The Japanese Society for Plant Cell and Molecular Biology. © 2012 The Japanese Society for Plant Cell and Molecular Biology.
• Bioconversion of proposed precursors into theobroxide and related compounds.

  Peng Li, Kosaku Takahashi, Ahmed Elkhateeb, Hideyuki Matsuura, Teruhiko Yoshihara, Kensuke Nabeta

  Natural product communications 6 12 1801 - 4 2011年12月 [査読有り][通常論文]
   

  We have previously reported a tetraketide origin for theobroxide and its related compound. In the present study, bioconversion of natural and deuterium-labeled precursors of this proposed biosynthetic pathway by Lasiodipoldia theobromae was investigated. Theobroxide was quantified after bioconversion from each proposed precursor. The transformation of the isotopically labeled precursor to products was tracked by 2H NMR measurement.
• Toxicological characterization of N-methyl-N-nitrosourea-induced cataract in rats by LC/MS-based metabonomic analysis

  Yuko Miyazono, Kazuo Harada, Koji Sugiyama, Motonobu Ueno, Mikinori Torii, Ikuo Kato, Hideyuki Matsuura, Kazumasa Hirata

  JOURNAL OF APPLIED TOXICOLOGY 31 7 655 - 662 2011年10月 [査読有り][通常論文]
   

  Cataract is one of the most serious drug-induced side effects that can terminate the development of drug candidates, and pharmaceutical companies consider it important to evaluate cataract-inducing potential in the early phases. Metabonomics is expected to be a powerful approach for the safety evaluation of drug candidates. In this study, we conducted a toxicological characterization of N-methyl-N-nitrosourea (MNU)-induced cataract in rats by LC/MS-based metabonomic analysis. MNU was intraperitoneally administered once to 15-day old rats at 70 mg kg(-1). After that, animals were kept for 3 weeks waiting for cataract formation. Lens samples for metabonomic analysis were collected on 7, 14 and 21 days after MNU administration. Comprehensive analyses of lens metabolites were conducted using an LC/MS system, and multivariate data for each sample were compared by principal component analysis (PCA) to find any changes in lens metabolites. Lens opacity was confirmed by ophthalmoscopy 14 days after dosing, and even by gross observation 21 days after dosing. PCA of the lens samples for the control and MNU-treated groups revealed that the metabolite profiles of lens differed from each other, and several lens metabolites, such as lots of a-amino acids and gluthathione, decreased after MNU treatment. In conclusion, metabonomic analysis enabled us to identify new marker candidates for cataract and provided a better understanding of the mechanism related to MNU-induced cataract. It was considered that metabonomics is a useful approach for the characterization of drug-induced toxicity. Copyright (C) 2011 John Wiley & Sons, Ltd.
• Arabidopsis CYP94B3 encodes jasmonyl-L-isoleucine 12-hydroxylase, a key enzyme in the oxidative catabolism of jasmonate.

  Naoki Kitaoka, Takuya Matsubara, Michio Sato, Kosaku Takahashi, Shinji Wakuta, Hiroshi Kawaide, Hirokazu Matsui, Kensuke Nabeta, Hideyuki Matsuura

  Plant & cell physiology 52 10 1757 - 65 2011年10月 [査読有り][通常論文]
   

  The hormonal action of jasmonate in plants is controlled by the precise balance between its biosynthesis and catabolism. It has been shown that jasmonyl-L-isoleucine (JA-Ile) is the bioactive form involved in the jasmonate-mediated signaling pathway. However, the catabolism of JA-Ile is poorly understood. Although a metabolite, 12-hydroxyJA-Ile, has been characterized, detailed functional studies of the compound and the enzyme that produces it have not been conducted. In this report, the kinetics of wound-induced accumulation of 12-hydroxyJA-Ile in plants were examined, and its involvement in the plant wound response is described. Candidate genes for the catabolic enzyme were narrowed down from 272 Arabidopsis Cyt P450 genes using Arabidopsis mutants. The candidate gene was functionally expressed in Pichia pastoris to reveal that CYP94B3 encodes JA-Ile 12-hydroxylase. Expression analyses demonstrate that expression of CYP94B3 is induced by wounding and shows specific activity toward JA-Ile. Plants grown in medium containing JA-Ile show higher sensitivity to JA-Ile in cyp94b3 mutants than in wild-type plants. These results demonstrate that CYP94B3 plays a major regulatory role in controlling the level of JA-Ile in plants.
• P-67 Jasmonoyl-L-isoleucineの代謝に関する研究(ポスター発表の部)

  北岡 直樹, 松原 卓也, 佐藤 道大, 高橋 公咲, 和久田 真司, 川出 洋, 松井 博和, 鍋田 憲助, 松浦 英幸

  天然有機化合物討論会講演要旨集 53 469 - 474 天然有機化合物討論会 2011年09月02日 

  The hormonal action of jasmonate in plants is controlled by the precise balance between its biosynthesis and catabolism. It has been shown that jasmonyl-L-isoleucine (JA-Ile, 6) is the bioactive form involved in the jasmonate-mediated signaling pathway. However, the catabolism of JA-Ile (6) is poorly understood. Although a metabolite, TA-Ile (7), has been characterized, detailed functional studies of the compound and the enzyme that produces it have not been conducted. In our results, the kinetics of wound-induced accumulation of TA-Ile (7) in plants was examined, and its involvement in plant wound response is described. Candidate genes for the catabolic enzyme were narrowed down from 272 Arabidopsis cytochrome P450 genes using Arabidopsis mutants. The candidate gene was functionally expressed in Pichia pastoris to reveal that CYP94B3 encodes JA-Ile (6) 12-hydroxylase. Expression analyses demonstrate that expression of CYP94B3 is induced by wounding and shows the activity toward JA-Ile (6). Plants grown in medium containing JA-Ile (6) show higher sensitivity to JA-Ile (6) in cyp94b3 mutants than in wild-type plants. These results demonstrate that CYP94B3 plays a major regulatory role in controlling the level of JA-Ile (6) in plants. CYP94B3 can use other JA-amino acid conjugates as substrates. Our other results uncovered that JA amino acid conjugates induced by wounding, JA-Phe and JA-Val, possessed biological activities same as JA-Ile (6).
• Isolation and partial characterization of the Chlamydomonas reinhardtii γ-glutamyl kinase.

  Miyoshi K, Matsuura H, Kumon T, Matsuda K, Yoshikawa T, Bamba T, Harada K, Hirata K

  Journal of bioscience and bioengineering 112 1 75 - 78 1 2011年07月 [査読有り][通常論文]
   

  We isolated a Chlamydomonas gene encoding putative gamma-glutamyl kinase (GK), an enzyme that catalyzes the first step of proline biosynthesis. Using an Escherichia coli auxotroph and a purified recombinant protein, we show that Chlamydomonas GK is a functional GK. The sensitivity of this kinase to feedback inhibition by proline was lower than in that of microbial GKs previously reported. (C) 2011, The Society for Biotechnology, Japan. All rights reserved.
• OsJAR1 and OsJAR2 are jasmonyl-L-isoleucine synthases involved in wound- and pathogen-induced jasmonic acid signalling.

  Shinji Wakuta, Erika Suzuki, Wataru Saburi, Hideyuki Matsuura, Kensuke Nabeta, Ryozo Imai, Hirokazu Matsui

  Biochemical and biophysical research communications 409 4 634 - 9 2011年06月17日 [査読有り][通常論文]
   

  The synthesis of JA-Ile was catalysed by JA-Ile synthase, which is a member of the group I GH3 family of proteins. Here, we showed evidence that OsGH3.5 (OsJAR1) and OsGH3.3 (OsJAR2) are the functional JA-Ile synthases in rice, using recombinant proteins. The expression levels of OsJAR1 and OsJAR2 were induced in response to wounding with the concomitant accumulation of JA-Ile. In contrast, only the expression of OsJAR1 was associated with the accumulation of JA-Ile after blast infection. Our data suggest that these two JA-Ile synthases are differentially involved in the activation of JA signalling in response to wounding and pathogen challenge in rice.
• Molecular Cloning and Partial Characterization of a Peroxidase Gene Expressed in the Roots of Portulaca oleracea cv., One Potentially Useful in the Remediation of Phenolic Pollutants

  Takeshi Matsui, Yuki Nomura, Mai Takano, Sofue Imai, Hideki Nakayama, Hitoshi Miyasaka, Hiroshi Okuhata, Satoshi Tanaka, Hideyuki Matsuura, Kazuo Harada, Takeshi Bamba, Kazumasa Hirata, Ko Kato

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 75 5 882 - 890 2011年05月 [査読有り][通常論文]
   

  Portulaca (Portulaca oleracea cv.) efficiently removes phenolic pollutants from hydroponic solution. In plant roots, peroxidase (PRX) is thought to be involved in the removal of phenolic pollutants by the cross-linking them to cell wall polysaccharides or proteins at the expense of reduction of hydrogen peroxide (H2O2). In this study, we found that portulaca roots secreted an acidic PRX isozyme that had relatively high H2O2 affinity. We isolated five PRX genes, and the recombinant PRX proteins produced in cultured tobacco cells were partially characterized. Among these genes, PoPRX2 probably encoded the acidic PRX isozyme. PoPRX2 had an extra N-terminal region which has not been reported for other PRX proteins. We found that PoPRX2 oxidized phenolic pollutants, including bisphenol A, octylphenol, nonylphenol, and 17 beta-estradiol. In addition, we found that the Cys261 residue of PoPRX2 played an important role in the determination of affinity for H2O2 and stability toward H2O2.
• Distal transport of exogenously applied jasmonoyl-isoleucine with wounding stress.

  Chizuru Sato, Kensuke Aikawa, Shunpei Sugiyama, Kensuke Nabeta, Chikara Masuta, Hideyuki Matsuura

  Plant & cell physiology 52 3 509 - 17 2011年03月 [査読有り][通常論文]
   

  Determining the mobile signal used by plants to defend against biotic and abiotic stresses has proved elusive, but jasmonic acid (JA) and its derivatives appear to be involved. Using deuterium-labeled analogs, we investigated the distal transport of JA and jasmonoyl-isoleucine (JA-Ile) in response to leaf wounding in tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum) plants. We recovered [(2)H(2)-2]JA ([(2)H(2)]JA) and [(2)H(3)-12]JA-Ile ([(2)H(3)]JA-Ile) in distal leaves of N. tabacum and S. lycopersicum after treating wounded leaves with [(2)H(2)]JA or [(2)H(3)]JA-Ile. We found that JA-Ile had a greater mobility than JA, despite its lower polarity, and that application of exogenous JA-Ile to wounded leaves of N. tabacum led to a higher accumulation of JA and JA-Ile in distal leaves compared with wounded control plants. We also found that exudates from the stem of S. lycopersicum plants with damaged leaflets contained JA and JA-Ile at higher levels than in an undamaged plant, and a significant difference in the levels of JA-Ile was observed 30 min after wounding. Based on these results, it was found that JA-Ile is a transportable compound, which suggests that JA-Ile is a signaling cue involved in the resistance to biotic and abiotic stresses in plants.
• Cellulase applied to the leaves of sweet pepper (Capsicum annuum L. var. grossum) upregulates the production of salicylic and azelaic acids.

  Chizuru Sato, Norikuni Oka, Kensuke Nabeta, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 75 4 761 - 3 2011年 [査読有り][通常論文]
   

  Treating the leaves of sweet pepper (Capsicum annuum L. var. grossum) with an aqueous solution of cellulase resulted in a four-fold increase in the salicylic acid level compared to a control plant. The level of endogenous azelaic acid was also elevated by the cellulase treatment. Azelaic acid has recently been reported to act as a mobile "priming" agent to arm plants against pathogenic attack. Our results are consistent with this and that the cellulase treatment enhanced the ability of sweet pepper to withstand viral attack.
• Tobacco salicylic acid glucosyltransferase is active toward tuberonic acid (12-hydroxyjasmonic acid) and is induced by mechanical wounding stress.

  Yoshiya Seto, Shigeki Hamada, Hiroyuki Ito, Chikara Masuta, Hirokazu Matsui, Kensuke Nabeta, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 75 12 2316 - 20 2011年 [査読有り][通常論文]
   

  Recently we reported that rice salicylic acid (SA) glucosyltransferase (OsSGT) is active toward 12-hydroxyjasmonic acid (tuberonic acid, TA) and that OsSGT gene expression is induced by wounding stress. Here we report that tobacco SA glucosyltransferase (NtSGT), which is thought to be an ortholog of OsSGT, is also active toward TA. Although NtSGT expression is known to be induced by biotrophic stress, it was also induced by wounding stress in the same manner as OsSGT. These results indicate that this glucosyltransferase is important not only in biotrophic stress but also for wounding stress. It was found that this enzyme is dually functional, with activity both toward TA and SA.
• Comparison of enzymatic properties and gene expression profiles of two tuberonic acid glucoside β-glucosidases from Oriza sativa L.

  Wakuta, S, Hamada, S, Ito, H, Imai, R, Mori, H, Matsuura, H, Nabeta, K, Matsui, H

  Journal of Applied Glycoscience 58 2 67 - 70 Japanese Society of Applied Glycoscience 2011年 [査読有り][通常論文]
  
• A Short Period of Mannitol Stress but Not LiCl Stress Led to Global Translational Repression in Plants

  Hideyuki Matsuura, Ueda Kiyotaka, Yu Ishibashi, Yuki Kubo, Masatoshi Yamaguchi, Kazumasa Hirata, Taku Demura, Ko Kato

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 74 10 2110 - 2112 2010年10月 [査読有り][通常論文]
   

  In plant cells, high salinity stress induces rapid inhibition of general protein synthesis. In this study, we found that treatment with mannitol, but not lithium stress, led to rapid global translational repression, suggesting that a rapid response at the level of translation might be induced by the osmotic but not the ionic components of salinity stress.
• Chemical inhibitors of viviparous germination in the fruit of watermelon.

  Yoshiki Kobayashi, Kensuke Nabeta, Hideyuki Matsuura

  Plant & cell physiology 51 9 1594 - 8 2010年09月 [査読有り][通常論文]
   

  It is well known that the seeds of watermelon [Citrullus lanatus (Thunb.) Matsum and Nakai] have a high potential to germinate when the fruit has ripened. When removed from the mature fruit, the seeds can germinate under appropriate conditions. However, it is unclear why they cannot germinate in the flesh of the fruit. Here, we show that cis-ABA and its β-D-glucopyranosyl ester (ABA-β-GE) accumulate in the flesh of the fruit at levels high enough to inhibit seed germination. This result indicates the existence of chemical factors that inhibit viviparous seed germination of watermelon.
• Resistant and susceptible responses in tomato to cyst nematode are differentially regulated by salicylic acid.

  Taketo Uehara, Shunpei Sugiyama, Hideyuki Matsuura, Tsutomu Arie, Chikara Masuta

  Plant & cell physiology 51 9 1524 - 36 2010年09月 [査読有り][通常論文]
   

  To understand the machinery underlying a tomato cultivar harboring the Hero A gene against cyst nematode using microarrays, we first analyzed tomato gene expression in response to potato cyst nematode (PCN; Globodera rostochiensis) during the early incompatible and compatible interactions at 3 and 7 days post-inoculation (dpi). Transcript levels of the phenylalanine ammonia lyase (PAL) and Myb-related genes were up-regulated at 3 dpi in the incompatible interaction. Transcription of the genes encoding pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) was also up-regulated at 3 dpi in the incompatible interaction. On the other hand, the four genes (PAL, Myb, PDC and ADH) were down-regulated in the compatible interaction at 3 dpi. When the expression levels of several pathogenesis-related (PR) protein genes in tomato roots were compared between the incompatible and compatible interactions, the salicylic acid (SA)-dependent PR genes were found to be induced in the incompatible interaction at 3 dpi. The PR-1(P4) transcript increased to an exceptionally high level at 3 dpi in the cyst nematode-infected resistant plants compared with the uninoculated controls. The free SA levels were elevated to similar levels in both incompatible and compatible interactions. We then confirmed that PR-1(P4) was not significantly induced in the NahG tomato harboring the Hero A gene, compared with the resistant cultivar. We thus found that PR-1(P4) was a hallmark for the cultivar resistance conferred by Hero A against PCN and that nematode parasitism resulted in the inhibition of the SA signaling pathway in the susceptible cultivars.
• Identification of a beta-glucosidase hydrolyzing tuberonic acid glucoside in rice (Oryza sativa L.)

  Shinji Wakuta, Shigeki Hamada, Hiroyuki Ito, Hideyuki Matsuura, Kensuke Nabeta, Hirokazu Matsui

  PHYTOCHEMISTRY 71 11-12 1280 - 1288 2010年08月 [査読有り][通常論文]
   

  Tuberonic acid (TA) and its glucoside (TAG) have been isolated from potato (Solanum tuberosum L) leaflets and shown to exhibit tuber-inducing properties. These compounds were reported to be biosynthesized from jasmonic acid (JA) by hydroxylation and subsequent glycosylation, and to be contained in various plant species. Here we describe the in vivo hydrolytic activity of TAG in rice. In this study, the TA resulting from TAG was not converted into JA. Tuberonic acid glucoside (TAG)-hydrolyzing beta-glucosidase, designated OsTAGG1, was purified from rice by six purification steps with an similar to 4300-fold purification. The purified enzyme migrated as a single band on native PAGE, but as two bands with molecular masses of 42 and 26 kDa on SDS-PAGE. Results from N-terminal sequencing and peptide mass fingerprinting of both polypeptides suggested that both bands were derived from a single polypeptide, which is a member of the glycosyl hydrolase family 1. In the native enzyme, the K(m) and V(max) values of TAG were 31.7 mu M and 0.25 mu katal/mg protein, OsTAGG1 preferentially hydrolyzed TAG and methyl TAG. Here we report that OsTAGG1 is a specific beta-glucosidase hydrolyzing TAG, which releases the physiologically active TA. (C) 2010 Elsevier Ltd. All rights reserved.
• Evaluation of Myanmar medicinal plant extracts for antitrypanosomal and cytotoxic activities.

  Saw Bawm, Saruda Tiwananthagorn, Kyaw San Lin, Junichi Hirota, Takao Irie, Lat Lat Htun, Ni Ni Maw, Tin Tin Myaing, Nyunt Phay, Satoshi Miyazaki, Tatsuya Sakurai, Yuzaburo Oku, Hideyuki Matsuura, Ken Katakura

  The Journal of veterinary medical science 72 4 525 - 8 2010年04月 [査読有り][通常論文]
   

  Current chemotherapeutic options for African trypanosomiasis in humans and livestock are very limited. In the present study, a total of 71 medicinal plant specimens from 60 plant species collected in Myanmar were screened for antitrypanosomal activity against trypomastigotes of Trypanosoma evansi and cytotoxicity against MRC-5 cells in vitro. The methanol extract of dried rootbark of Vitis repens showed the highest antitrypanosomal activity with IC(50) value of 8.6 +/- 1.5 microg/ml and the highest selectivity index of 24.4. The extracts of Brucea javanica, Vitex arborea, Eucalyptus globulus and Jatropha podagrica had also remarkable activity with IC(50) values and selectivity indices in the range of 27.2-52.6 microg/ml and 11.4-15.1 respectively.
• Genome-Wide Analyses of Early Translational Responses to Elevated Temperature and High Salinity in Arabidopsis thaliana

  Hideyuki Matsuura, Yu Ishibashi, Atsuhiko Shinmyo, Shigehiko Kanaya, Ko Kato

  PLANT AND CELL PHYSIOLOGY 51 3 448 - 462 2010年03月 [査読有り][通常論文]
   

  Abiotic stress influences the translation of mRNAs in plants. To gain a global view of the early translational response to abiotic stress, we investigated genome-wide changes in mRNA translation in Arabidopsis thaliana suspension cell cultures exposed to brief periods of two types of stress: elevated temperature (37C) and high salinity (200mM NaCl). Microarray analyses revealed that polysome association of most transcripts, which were monitored by using polysomal- and non-polysomal-associated RNA pools, was variably depressed by both stresses within 10min. We also inspected coordination of changes in translational profiles with transcriptional profiles, and found no simple correlations between the changes in these two processes under both stresses. In addition, we uncovered that the 10min heat- and salt-inducible changes in polysome association of individual transcripts affected specific biological functions differently; some functional classes were recalcitrant to the overall depression, while others were hypersensitive to it. Heat and salt stresses imposed similar, but not identical, changes in polysome association of individual transcripts, and the functional categories with differential responses from all other genes (i.e. recalcitrant or hypersensitive functional categories) displayed some overlap between the two stresses, suggesting similar underlying mechanisms. Our results highlight the importance of dynamic changes in mRNA translation, which include selective translation and extensive repression of a subset of transcripts, in plant abiotic stress responses.
• Effect of the sequence context of the AUG initiation codon on the rate of translation in dicotyledonous and monocotyledonous plant cells

  Tadatoshi Sugio, Hideyuki Matsuura, Takeshi Matsui, Manabu Matsunaga, Tatsuya Nosho, Shigehiko Kanaya, Atsuhiko Shinmyo, Ko Kato

  JOURNAL OF BIOSCIENCE AND BIOENGINEERING 109 2 170 - 173 2010年02月 [査読有り][通常論文]
   

  The sequence context around the AUG initiation codon strongly contributes to the translation initiation step in mammalian and plant cells. Here, we investigated the effect of the three nucleotides immediately upstream of the initiating AUG (positions -3 to -1) on the translation efficiency of a reporter gene, beta-glucuronidase, in dicotyledonous and monocotyledonous plant cells. 2009, The Society for Biotechnology, Japan. All rights reserved.
• ツベロン酸グルコシドグルコシダーゼアイソザイム間の酵素特性および発現特性の比較解析

  和久田 真司, 濱田 茂樹, 伊藤 浩之, 松浦 英幸, 鍋田 憲介, 今井 亮三, 松井 博和

  Journal of Applied Glycoscience Supplement 2010 72 - 72 日本応用糖質科学会 2010年
• Biosynthesis of resorcylic acid lactone (5S)-5-hydroxylasiodiplodin in Lasiodiplodia theobromae.

  Takasumi Kashima, Kosaku Takahashi, Hideyuki Matsuura, Kensuke Nabeta

  Bioscience, biotechnology, and biochemistry 73 11 2522 - 4 2009年11月 [査読有り][通常論文]
   

  An administration study of (2)H-labeled precursors showed that the 9-hydroxydecanoyl unit, the acyl intermediate of lasiodiplodin (1), was also the intermediate of (5S)-5-hydroxylasiodiplodin (2) in Lasiodiplodia theobromae. The incorporation of [O-methyl-(2)H(3)]-lasiodiplodin (6) into 2 indicated that hydroxylation at C-5 occurred after cyclization.
• Cloning and functional analysis of an allene oxide synthase in Physcomitrella patens.

  P K G S S Bandara, Kosaku Takahashi, Michio Sato, Hideyuki Matsuura, Kensuke Nabeta

  Bioscience, biotechnology, and biochemistry 73 10 2356 - 9 2009年10月 [査読有り][通常論文]
   

  Jasmonic acid (JA) is a plant hormone that plays important roles in a large number of processes in stress adaptation and development in flowering plants. A search of genome database indicated the existence of allene oxide synthase (AOS), an enzyme of JA biosynthesis, in Physcomitrella patens, a model plant among mosses. In this study, the presence of JA was detected in P. patens. The recombinant AOS of P. patens, which was overexpressed in Escherichia coli, showed AOS activity. These data suggest that the octadecanoid pathway also exists in P. patens.
• Kinetics of the accumulation of jasmonic acid and its derivatives in systemic leaves of tobacco (Nicotiana tabacum cv. Xanthi nc) and translocation of deuterium-labeled jasmonic acid from the wounding site to the systemic site.

  Chizuru Sato, Yoshiya Seto, Kensuke Nabeta, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 73 9 1962 - 70 2009年09月 [査読有り][通常論文]
   

  In plants, the mobile signal needed for wound-induced systemic acquired resistance (WSR) has been elusive. The signal compound involved in WSR is supposed to be JA or its derivatives. On the basis of kinetic study of the accumulation of JA or its derivatives, it was discovered that JA, JA-Ile, tuberonic acid (TA, 12-OH epi-JA), and tuberonic acid glucoside (TAG) accumulated in systemic tissues in response to mechanical wounding stress in the tobacco plant (Nicotiana tabacum). Attempts to recover deuterium-labeled JA in systemic leaves after feeding the wounded leaves with deuterium-labeled JA were successfully done. It was also found that the translocated deuterium-labeled JA was metabolized to TA in systemic leaves under feeding of deuterium-labeled JA to the wounding leaves.
• Potato tuber cell expansion-inducing activity of stereochemically restricted JA analogs.

  Naoki Kitaoka, Eri Fukushi, Yasunori Koda, Kensuke Nabeta, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 73 8 1872 - 6 2009年08月 [査読有り][通常論文]
   

  Stereochemically restricted analogues of C-7 substituted 7-epi-jasmonate, together with 12-hydroxy jasmonic acid, 12-hydroxy jasmonic acid glucoside, and jasmonic acid conjugated with L-isoleucine (JA-Ile), were synthesized and then tested for potato tuber cell expansion-inducing activity. JA-Ile showed almost the same activity as JA, while the C-7 substituted 7-epi-jasmonates exhibited weaker activity than JA and showed an antagonist effect against JA.
• Isolation and structural elucidation of a new cyclohexenone compound from Lasiodiplodia theobromae.

  Naoki Kitaoka, Kensuke Nabeta, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 73 8 1890 - 2 2009年08月 [査読有り][通常論文]
   

  A new cyclohexenone compound was isolated as a mixture of enantiomers from a culture filtrate of Lasiodiplodia theobromae. The relative structure was determined to be 4,5-dihydroxy-3-methyl-cyclohex-2-enone on the basis of MS, (1)H-NMR, and (13)C-NMR spectroscopic analyses, including 2D-NMR experiments. Resolution of the enantiomers was conducted by a coupling reaction with (S)-MTPA-Cl followed by HPLC separation.
• Biosynthesis of resorcylic acid lactone lasiodiplodin in Lasiodiplodia theobromae.

  Takasumi Kashima, Kosaku Takahashi, Hideyuki Matsuura, Kensuke Nabeta

  Bioscience, biotechnology, and biochemistry 73 5 1118 - 22 2009年05月 [査読有り][通常論文]
   

  The biosynthesis of lasiodiplodin (1) and its (5S)-5-hydroxylated derivative (2) were investigated by the administration of (13)C-labeled acetates to Lasiodiplodia theobromae. The labeling patterns of biosynthetically (13)C-labeled 1 and 2 were determined by (13)C-NMR and INADEQUATE spectra, demonstrating the octaketide origins of 1 and 2. Taking into account the biosynthetic study of resorcylic acid lactones, the involvement of highly reduced acyl intermediates in the biosynthesis of lasiodiplodins was presumed; thus, we synthesized (2)H-labeled hypothetical acyl intermediates of 1, 9-hydroxydecanoic acid (4) and its N-acetylcysteamine thioester (SNAC, 5). When L. theobromae was incubated with 5 mM of a (2)H-labeled intermediate, the (2)H-label from the intermediate was incorporated at the expected position of 1. These incorporation studies revealed that 1 was produced via a pathway which closely resembles that of resorcylic acid lactone biosynthesis.
• Expression of allene oxide cyclase from Pharbitis nil upon theobroxide treatment.

  Fan-Jiang Kong, Ying Li, Jun Abe, Baohui Liu, Florian Schaller, Markus Piotrowski, Shungo Otagaki, Kosaku Takahashi, Hideyuki Matsuura, Teruhiko Yoshihara, Kensuke Nabeta

  Bioscience, biotechnology, and biochemistry 73 5 1007 - 13 2009年05月 [査読有り][通常論文]
   

  In previous reports we have reported that theobroxide induces characteristic accumulation of allene oxide cyclase (AOC; EC 5.3.99.6) protein and jasmonic acid (JA) in Pharbitis nil. In the present study, PnAOC, an AOC gene from Pharbitis nil was cloned. Immunofluorescence assays indicated that the AOC protein is located in the chloroplast of vascular bundles in Pharbitis nil leaves. The PnAOC cDNA sequence lacking the chloroplast signal peptide was successfully expressed in Escherichia coli, and a gas chromatography-mass spectrum assay suggested the relative AOC activity of the recombinant PnAOC protein in comparison with Arabidopsis AOC2. Interestingly, a biphasic expression of PnAOC was induced by theobroxide, which is consistent with the accumulation patterns of AOC protein and JA. All these results indicate that AOC is the primary target of theobroxide regulation and suggest that feedback regulation of PnAOC by JA occurs upon theobroxide treatment in Pharbitis nil.
• Simultaneous UPLC MS/MS analysis of endogenous jasmonic acid, salicylic acid, and their related compounds

  Hideyuki Matsuura, Arata Aoi, Chizuru Satou, Mino Nakaya, Chikara Masuta, Kensuke Nabeta

  Plant Growth Regulation 57 3 293 - 301 2009年04月 [査読有り][通常論文]
   

  Jasmonic acid (JA) and salicylic acid (SA) are plant hormones involved in plant growth and development. Recent studies demonstrated that presence of a complex interplay between JA and SA signaling pathways to response to pathogenesis attack and biotic stresses. To our best knowledge, no method has existed for simultaneous analyses of JA, SA, and their related compounds. Especially, the glucosides are thought to be the storages or the inactivated compounds, but their contribution should be considered for elucidating the amount of the aglycons. It is also valuable for measuring the endogenous amount of phenylalanine, cinnamic acid, and benzoic acid that are the biosynthetic intermediates of SA due to the existence of isochorismate pathway to synthesize SA. We established this method using deuterium labeled compounds as internal standards. This is the first report of simultaneous analysis of endogenous JA, SA, and their related compounds. Measuring the endogenous JA, SA, and their related compounds that had been accumulated in tobacco plants proved the practicality of the newly developed method. It was demonstrated that accumulation of JA, SA and their related compounds were induced in both case of TMV infection and abiotic stresses. © 2008 Springer Science+Business Media B.V.
• Isolation of antibabesial compounds from Brucea javanica, Curcuma xanthorrhiza, and Excoecaria cochinchinensis.

  Kazuaki Yamada, Subeki, Kensuke Nabeta, Masahiro Yamasaki, Ken Katakura, Hideyuki Matsuura

  Bioscience, biotechnology, and biochemistry 73 3 776 - 80 2009年03月23日 [査読有り][通常論文]
   

  One new curcuminoid, 3'-demethoxycyclocurcumin (1), was isolated from Curcuma xanthorrhiza as an antibabesial compound, together with p-hydroxybenzaldehyde (2) and cleomiscosin A (3) from Brucea javanica and (+)-epiloliolide (4) from Excoecaria cochinchinensis. The antibabesial activities were examined in vitro, and compounds 1-4, and diminazene aceturate were studied with IC(50) values of 16.6, 7.6, 15.6, 10.0, and 0.6 microg/ml, respectively.
• Purification and cDNA cloning of a wound inducible glucosyltransferase active toward 12-hydroxy jasmonic acid.

  Yoshiya Seto, Shigeki Hamada, Hideyuki Matsuura, Mana Matsushige, Chizuru Satou, Kosaku Takahashi, Chikara Masuta, Hiroyuki Ito, Hirokazu Matsui, Kensuke Nabeta

  Phytochemistry 70 3 370 - 9 2009年02月 [査読有り][通常論文]
   

  Tuberonic acid (12-hydroxy epi-jasmonic acid, TA) and its glucoside (TAG) were isolated from potato leaflets (Solanumtuberosum L.) and shown to have tuber-inducing properties. The metabolism of jasmonic acid (JA) to TAG in plant leaflets, and translocation of the resulting TAG to the distal parts, was demonstrated in a previous study. It is thought that TAG generated from JA transmits a signal from the damaged parts to the undamaged parts by this mechanism. In this report, the metabolism of TA in higher plants was demonstrated using [12-(3)H]TA, and a glucosyltransferase active toward TA was purified from the rice cell cultures. The purified protein was shown to be a putative salicylic acid (SA) glucosyltransferase (OsSGT) by MALDI-TOF-MS analysis. Recombinant OsSGT obtained by overexpression in Escherichia coli was active not only toward TA but also toward SA. The OsSGT characterized in this research was not specific, but this is the first report of a glucosyltransferase active toward TA. mRNA expressional analysis of OsSGT and quantification of TA, TAG, SA and SAG after mechanical wounding indicated that OsSGT is involved in the wounding response. These results demonstrated a crucial role for TAG not only in potato tuber formation, but also in the stress response in plants and that the SA glucosyltransferase can work for TA glucosylation.
• Down-regulation of flavonoid 3'-hydroxylase gene expression by virus-induced gene silencing in soybean reveals the presence of a threshold mRNA level associated with pigmentation in pubescence.

  Atsushi Nagamatsu, Chikara Masuta, Hideyuki Matsuura, Keisuke Kitamura, Jun Abe, Akira Kanazawa

  Journal of plant physiology 166 1 32 - 9 2009年01月01日 [査読有り][通常論文]
   

  Changes in flavonoid content are often manifested as altered pigmentation in plant tissues. Two loci have been identified as controlling pigmentation in soybean pubescence. Of these, the T locus appears to encode flavonoid 3'-hydroxylase (F3'H) protein: the T and t alleles are associated with tawny and gray colors, respectively, in pubescence. We previously down-regulated F3'H gene expression by virus-induced gene silencing (VIGS) in soybean. Despite this successful VIGS, the tawny pubescence pigmentation proved to be unchanged in greenhouse-grown plants. We hypothesized that the reduced mRNA level of the F3'H gene resulting from VIGS remained high enough to induce pigmentation. To verify this hypothesis, in the present study, we performed F3'H VIGS on plants grown under controlled conditions, in which the steady-state mRNA level of the F3'H gene was reduced to approximately 5% of that of greenhouse-grown plants. This VIGS treatment resulted in the loss of tawny pigmentation in pubescence, suggesting that the sf3'h1 gene is involved in the control of pigmentation in pubescence. We detected a marked decrease in target mRNA, an accumulation of short interfering RNAs (siRNAs), and a decrease in quercetin content relative to kaempferol in leaf tissues, indicating that sequence-specific mRNA degradation of the F3'H gene was induced. These results suggest that leaf tissues have a threshold mRNA level of the F3'H gene, which is associated with the occurrence of tawny pigmentation in pubescence. The estimated threshold mRNA level for pigmentation in pubescence was approximately 3% of the steady-state mRNA level of the F3'H gene in greenhouse-grown plants.
• Evaluation of efficacy of bruceine A, a natural quassinoid compound extracted from a medicinal plant, Brucea javanica, for canine babesiosis.

  Ryo Nakao, Chiaki Mizukami, Yuta Kawamura, Subeki, Saw Bawm, Masahiro Yamasaki, Yoshimitsu Maede, Hideyuki Matsuura, Kensuke Nabeta, Nariaki Nonaka, Yuzaburo Oku, Ken Katakura

  The Journal of veterinary medical science 71 1 33 - 41 2009年01月 [査読有り][通常論文]
   

  Bruceine A, a natural quassinoid compound extracted from the dried fruits of Brucea javanica (L.) Merr., was evaluated for its antibabesial activity in vitro and in vivo. Bruceine A inhibited the in vitro growth of Babesia gibsoni in canine erythrocytes at lower concentration compared with the standard antibabesial drug diminazene aceturate and killed the parasites within 24 hr at a concentration of 25 nM. Oral administration of bruceine A at a dosage of 6.4 mg/kg/day for 5 days resulted in no clinical findings in a dog with normal ranges of hematological and biochemical values in the blood. Three dogs were infected with B. gibsoni and two of them were treated with bruceine A at a dosage of 6.4 mg/kg/day for 6 days from day 5 post-infection. An untreated dog developed typical acute babesiosis symptoms including severe anemia, high fever, and complete loss of appetite and movement. However, the two bruceine A-treated dogs maintained their healthy conditions throughout the experimental period of 4 weeks although complete elimination of parasites from the peripheral blood was not achieved and decreases in the packed cell volume and the erythrocyte and platelet counts were observed. Since natural quassinoid compounds have been used as traditional medicines for the treatment of various ailments including cancer and malaria, the present results suggest that bruceine A or other related compounds are potential candidates for the treatment of canine babesiosis.
• ツベロン酸グルコシドグルコシダーゼの酵素学的諸性質の解明および遺伝子の同定

  和久田 真司, 濱田 茂樹, 伊藤 浩之, 松浦 英幸, 鍋田 憲助, 松井 博和

  Journal of Applied Glycoscience Supplement 2009 41 - 41 日本応用糖質科学会 2009年
• In vitro antitrypanosomal activities of quassinoid compounds from the fruits of a medicinal plant, Brucea javanica.

  Saw Bawm, Hideyuki Matsuura, Ahmed Elkhateeb, Kensuke Nabeta, Subeki, Nariaki Nonaka, Yuzaburo Oku, Ken Katakura

  Veterinary parasitology 158 4 288 - 94 2008年12月20日 [査読有り][通常論文]
   

  The medicinal plant Brucea javanica (L.) Merr. (Simaroubaceae) is widely distributed throughout Asia where its bitter fruits have been used in traditional medicine for various ailments. Fifteen C-20 quassinoids were isolated from the fruits of B. javanica and examined for their in vitro antitrypanosomal activities against trypomastigotes of Trypanosoma evansi. Bruceine A, bruceantinol, bruceine C, brusatol, and bruceine B showed strong antitrypanosomal activities with IC(50) values in the range of 2.9-17.8nM, which compared well with the standard trypanocidal drugs diminazene aceturate (IC(50)=8.8nM) and suramin (IC(50)=43.2nM). However, dehydrobruceine A, dehydrobruceine B, and dehydrobrusatol were about 2100, 900, and 1200 times less active, respectively, than bruceine A, bruceine B, and brusatol. The relationship of the structure and antitrypanosomal activity of these quassinoid compounds suggested that the presence of a diosphenol moiety in ring A and the nature of the C-15 side chain are important for their activities against T. evansi. This is the first report on the antitrypanosomal activity of isolated quassinoids.
• RNA silencing of a gene involved in flavonoid synthesis reveals the presence of a threshold mRNA level associated with pigmentation in plant tissues

  Nagamatsu Atsushi, Masuta Chikara, Matsuura Hideyuki, Kitamura Keisuke, Abe Jun, Kanazawa Akira

  GENES & GENETIC SYSTEMS 83 6 515  2008年12月 [査読有り][通常論文]
  
• New potato micro-tuber-inducing cyclohexene compounds related to theobroxide from Lasiodiplodia theobromae.

  Ryo Takei, Kosaku Takahashi, Hideyuki Matsuura, Kensuke Nabeta

  Bioscience, biotechnology, and biochemistry 72 8 2069 - 73 2008年08月 [査読有り][通常論文]
   

  Two new cyclohexene compounds related to theobroxide (3) were isolated from the mycelia of Lasiodiplodia theobromae OCS71. The structures of these compounds were determined to be (4S,5S)-4,5-dihydroxy-2-methyl-cyclohex-2-enone (1) and (3aS,4R,5S,7aR)-4,5-dihydroxy-7-methyl-3a,4,5,7a-tetrahydrobenzo[1,3]dioxol-2-one (2) by means of spectroscopic analyses and chemical correlation to 3. Compound 2 was shown to take up the carbonate ion to form a carbonic acid ester non-enzymatically. The compounds also showed potato micro-tuber-inducing activities at a concentration of 10(-3) M, using a culture of single-node segments of potato stems in vitro.
• The 5 '-untranslated region of the Oryza sativa alcohol dehydrogenase gene functions as a translational enhancer in monocotyledonous plant cells

  Tadatoshi Sugio, Junko Satoh, Hideyuki Matsuura, Atsuhiko Shinmyo, Ko Kato

  JOURNAL OF BIOSCIENCE AND BIOENGINEERING 105 3 300 - 302 2008年03月 [査読有り][通常論文]
   

  The F-untranslated region (5'-UTR) functions as a translational enhancer in monocotyledonous plant cells is necessary to express a foreign gene efficiently. Here, we show that the 5'-UTR of the rice alcohol dehydrogenase gene contributes to efficient translation in not only dicotyledonous plant cells, but also in monocotyledonous rice cells.
• Temperature regulates tuber-inducing lipoxygenase-derived metabolites in potato (Solanum tuberosum)

  Kyong-Hee Nam, Fanjiang Kong, Hideyuki Matsuura, Kosaku Takahashi, Kensuke Nabeta, Teruhiko Yoshihara

  JOURNAL OF PLANT PHYSIOLOGY 165 2 233 - 238 2008年02月 [査読有り][通常論文]
   

  Temperature is one of the major environmental factors affecting potato tuberization. It has been suggested that lipoxygenase (LOX) mediates between temperature and tuber induction. In this study, the contents of the LOX-derived metabolites hydroperoxylinolenic acid (HPOT), jasmonic acid (JA), tuberonic acid (TA) and tuberonic acid glucoside (TAG) were analyzed in leaves of potatoes growing at different temperatures. At low, tuber-inducing temperature, endogenous levels of JA, TA and TAG rise, indicating their crucial role in tuber induction. The concentration of 13(S)-HPOT seems not to be directly affected by temperature. Instead, the molecule has only a short half-life in leaves and is readily metabolized. (c) 2007 Elsevier GmbH. All rights reserved.
• Veinal necrosis induced by turnip mosaic virus infection in Arabidopsis is a form of defense response accompanying HR-like cell death.

  Bomin Kim, Chikara Masuta, Hideyuki Matsuura, Hideki Takahashi, Tsuyoshi Inukai

  Molecular plant-microbe interactions : MPMI 21 2 260 - 8 2008年02月 [査読有り][通常論文]
   

  In the pathosystems of Turnip mosaic virus (TuMV) with Brassicaceae crops, various symptoms, including mosaic and necrosis, are observed. We previously reported a necrosis-inducing factor TuNI in Arabidopsis thaliana, a model species. In this study, we show that the necrotic symptom induced by TuNI, observed along the veins, was actually a form of defense response accompanying a hypersensitive reaction (HR)-like cell death in the veinal area. The virus is often localized in the necrotic region. The necrotic response is associated with the production of H2O2, accumulation of salicylic acid (SA), emission of ethylene, and subsequent expression of defense-related genes. Additionally, this HR-like cell death is eased or erased by a shading treatment. These features are similar to the HR-associated resistance reaction to pathogens. However, unlike HR, two phytohormones--SA and ethylene--are involved in the necrosis induction, and both SA- and ethylene-dependent pathogenesis-related genes are activated. We concluded that the veinal necrosis induced by TuMV is regulated by a complex and unique network of at least two signaling pathways, which differs from the signal transduction for the known HR-associated resistance.
• Preferential translation mediated by Hsp81-3 5 '-UTR during heat shock involves ribosome entry at the 5 '-end rather than an internal site in Arabidopsis suspension cells

  Hideyuki Matsuura, Atsuhiko Shinmyo, Ko Kato

  JOURNAL OF BIOSCIENCE AND BIOENGINEERING 105 1 39 - 47 2008年01月 [査読有り][通常論文]
   

  Translational inhibition of most mRNAs and preferential translation of mRNAs coding heat shock proteins (Hsps) occur in most cells under heat shock stress. For most Hsp mRNAs, preferential translation in heat-shocked cells is conferred by their 5'-untranslated regions (5'-UTRs). However, the preferential translation directed by 5'-UTRs during heat shock remains mostly unknown in plants. Here, we found that the mRNA of Hsp81-3, which is an Arabidopsis Hsp90 family gene, continued to be associated with polysomes in heat-shocked Arabidopsis suspension-cultured cells. The Hsp81-3 5'-UTR was found to contribute to the efficient translation of capped reporter mRNAs in heat-shocked Arabidopsis protoplasts using a transient expression assay. Further characterization of the Hsp81-3 5'-UTR revealed that the anterior half of the 5'-UTR is important for the efficient translation in heat-shocked protoplasts. Moreover, the Hsp81-3 5'-UTR was highly capable of enhancing translation from uncapped reporter mRNAs relative to the 5'-UTR of a housekeeping gene in both normal and heat-shocked protoplasts. These Hsp81-3 5'-UTR-directed translations both in capped and uncapped reporter mRNAs were substantially reduced by the insertion of an upstream AUG at the 5'-end of the 5'-UTR, indicating that ribosomes are recruited to the 5'-end of the Hsp81-3 5'-UTR regardless of temperature and the presence or absence of the cap structure. These results suggest that the preferential translation of Hsp81-3 mRNA in heat-shocked Arabidopsis cells involves a ribosome scanning from the 5'-end of the 5'-UTR rather than ribosome entry to the internal site.
• Germination stimulant from root exudates of Vigna unguiculata

  Hideyuki Matsuura, Kumiko Ohashi, Hiroshi Sasako, Noriko Tagawa, Yuuko Takano, Yumiko Ioka, Kensuke Nabeta, Teruhiko Yoshihara

  Plant Growth Regulation 54 1 31 - 36 2008年01月 [査読有り][通常論文]
   

  Root exudate of Vigna unguiculata was extracted from a soil system consisting of charcoal and vermiculite. Germination stimulating activity for Striga gesnerioides was found in extracts of the soil system, and an active compound was isolated. The chemical structure of the active ingredient was determined to be (+)-4-O-acetylorobanchol, based on analysis of the spectral data of 1-D and 2-D NMR together with nuclear Overhauser effect (NOE) experiments. Application of the active compound to the seeds of S. gesnerioides at a concentration of 0.35 × 10-9 mol/disk led to 69% germination. The germination observed with application of GR-24, a positive control, at 0.57 × 10-10 mol/disk was 80%. © 2007 Springer Science+Business Media B.V.
• Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing

  Atsushi Nagamatsu, Chikara Masuta, Mineo Senda, Hideyuki Matsuura, Atsushi Kasai, Sung Hong, Keisuke Kitamura, Jun Abe, Akira Kanazawa

  PLANT BIOTECHNOLOGY JOURNAL 5 6 778 - 790 2007年11月 [査読有り][通常論文]
   

  Virus-induced gene silencing (VIGS) is a powerful tool for functional analysis of genes in plants. A wide-host-range VIGS vector, which was developed based on the Cucumber mosaic virus (CMV), was tested for its ability to silence endogenous genes involved in flavonoid biosynthesis in soybean. Symptomless infection was established using a pseudorecombinant virus, which enabled detection of specific changes in metabolite content by VIGS. It has been demonstrated that the yellow seed coat phenotype of various cultivated soybean lines that lack anthocyanin pigmentation is induced by natural degradation of chalcone synthase (CHS) mRNA. When soybean plants with brown seed coats were infected with a virus that contains the CHS gene sequence, the colour of the seed coats changed to yellow, which indicates that the naturally occurring RNA silencing is reproduced by VIGS. In addition, CHS VIGS consequently led to a decrease in isoflavone content in seeds. VIGS was also tested on the putative flavonoid 3'-hydroxylase (F3'H) gene in the pathway. This experiment resulted in a decrease in the content of quercetin relative to kaempferol in the upper leaves after viral infection, which suggests that the putative gene actually encodes the F3'H protein. In both experiments, a marked decrease in the target mRNA and accumulation of short interfering RNAs were detected, indicating that sequence-specific mRNA degradation was induced. The present report is a successful demonstration of the application of VIGS for genes involved in flavonoid biosynthesis in plants; the CMV-based VIGS system provides an efficient tool for functional analysis of soybean genes.
• Screening of Indonesian medicinal plant extracts for antibabesial activity and isolation of new quassinoids from Brucea javanica

  Subeki, Hideyuki Matsuura, Kosaku Takahashi, Kensuke Nabeta, Masahiro Yamasaki, Yoshimitsu Maede, Ken Katakura

  JOURNAL OF NATURAL PRODUCTS 70 10 1654 - 1657 2007年10月 [査読有り][通常論文]
   

  Boiled extracts derived from 28 Indonesian medicinal plants were screened for their antibabesial activity against Babesia gibsoni in vitro. Of these extracts, the fruit of Brucea javanica was the most active in inhibiting parasite growth at a concentration of 10 mu g/mL. Bioassay-guided fractionation of the fruit extract of Br. javanica led to the isolation of two new quassinoids, bruceantinol B and bruceine J, and the structures of these compounds were elucidated on the basis of their spectroscopic data and by chemical transformation to known compounds. In addition, the known quassinoids bruceines A-D, bruceantinol, and yadanziolide A were isolated. Antibabesial activities were also examined in vitro, and bruceine A and bruceantinol were shown to be more potent than diminazene aceturate, a drug IC50 = 103 ng/mL) used clinically against B. gibsoni, with IC50 values of 4 and 12 ng/mL, respectively.
• Novel cyclic peptide, epichlicin, from the endophytic fungus, Epichloe typhina

  Yoshiya Seto, Kosaku Takahashi, Hideyuki Matsuura, Yasunori Kogami, Hiroshi Yada, Teruhiko Yoshihara, Kensuke Nabeta

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 71 6 1470 - 1475 2007年06月 [査読有り][通常論文]
   

  The novel cyclic peptide, epichlicin, was isolated from Epichloe typhina, an endophytic fungus of the timothy plant (Phleum pretense L.). Its structure was determined by NMR studies and by mass spectrometry. Enantiomers of 3-amino tetradecanoic acid, a constituent amino acid of epichlicin, were synthesized as authentic standards. The stereochemistry of each amino acid was elucidated through a combination of the advanced Marfey method and chemical manipulation. Epichlicin showed inhibitory activity toward the spore germination of Cladosporium phlei, a pathogenic fungus of the timothy plant at an IC50 value of 22 nM.
• Anti-babesial compounds from Curcuma xanthorrhiza

  Hideyuki Matsuura, Shinkichi Nomura, Subeki, Kazuaki Yamada, Masahiro Yamasaki, Osamu Yamato, Yoshimitsu Maede, Ken Katakura, Trimurningsih, Chairul, Teruhiko Yoshihara, Kensuke Nabeta

  NATURAL PRODUCT RESEARCH 21 4 328 - 333 2007年 [査読有り][通常論文]
   

  Anti-babesial ingredients, (12R)- and (12S)-12,13-dihydro-12,13-dihydroxyxanthorrhizols,were isolated from Curcuma xanthorrhiza. The structures were established by the extensive NMR techniques. The assignments of H-1 NMR data of (12R)-12,13-dihydro-12,13-dihydroxyxanthorrhizol was revised, and (12S)-12,13-dihydro-12,13-dihydroxyxanthorrhizol was isolated as a pure form for the first time. The IC50 of the active compounds were compared with that of commercial drug, diminazene aceturate (Ganaseg). IC50 values of Ganaseg, (12R)-, and (12S)- 12,13-dihydro-12,13-dihydroxyxanthorrhizols were 0.6 mu g mL(-1), 8.3 mu g mL(-1) and 11.6 mu g mL(-1), respectively.
• Anti-babesial compounds from Berberis vulgaris

  A. Elkhateeb, K. Yamada, K. Takahashi, H. Matsuura, M. Yamasaki, Y. Maede, K. Katakura, K. Nabeta

  NATURAL PRODUCT COMMUNICATIONS 2 2 173 - 176 2007年 [査読有り][通常論文]
   

  Extracts of nine North African medicinal plants have been evaluated for their anti-babesial properties in vitro. The extracts of Berberis vulgaris and Rosa damascena showed more than 90% inhibition of the parasite growth at a test concentration of 100 mu g/mL. In addition to berberine alkaloids, we isolated nine active ingredients from B. vulgaris: E-coniferyl alcohol, (-)-simulanol, p-hydroxybenzaldehyde, 3-hydroxy-4,5-dimethoxybenzoic acid, trans-ferulic acid, syringic acid, vanillic acid, cis-ferulic acid, and syringaresinol-p-D-glucoside. These compounds are all known, but are newly reported for B. vulgaris. Among them, trans-ferulic acid showed the lowest IC50 value (7.33 mu g/mL), not much above that of the standard drug, diminazene aceturate (0.60 mu g/mL).
• Anti-babesial compounds from Rosa damascena mill.

  Ahmed Elkhateeb, Hideyuki Matsuura, Masahiro Yamasaki, Yoshimitsu Maede, Ken Katakura, Kensuke Nabeta

  NATURAL PRODUCT COMMUNICATIONS 2 7 765 - 769 2007年 [査読無し][通常論文]
   

  Bioassay-guided investigation of extracts of the flowers of Rosa damascena Mill. led to the isolation of four anti-babesial compounds, 3,4-dihydroxy benzoic acid (1), gallic acid (2), 2-phenylethyl 6-O-galloyl-beta-D-glucopyranoside (3), and quercetin 3-O-beta-D-(6-O-acetyl)-glucopyranosyl-(1 -> 4)-alpha-L-rhamnopyranoside (9), in addition to five flavonoid compounds which possessed weak anti-babesial activity, kaempferol 3-O-beta-D-glucopyranoside (4), kaempferol 3-O-beta-D-Xylopyranoside (5), kaempferol 3-O-alpha-L-rhamnopyranoside (6), quercetin 3-O-beta-D-glucopyranoside (7), and quercetin 3-O-alpha-L-rhamnopyranoside (8). Among the isolated compounds, 3 possessed the greatest anti-babesial activity with an IC50 value of 11.78 mu g/mL. Although compound 3 is known, no published reports list detailed spectroscopic data for the molecule. Therefore, the first complete structural elucidation of 3 using 1D and 2D NMR spectroscopic analysis is reported here. This is the first report of the isolation of compounds 1, 5, and 9 from R. damascena.
• Inhibitory rote of gibberellins in theobroxide-induced flowering of Pharbitis nil

  XQ Gao, FJ Kong, F Wang, H Matsuura, T Yoshihara

  JOURNAL OF PLANT PHYSIOLOGY 163 4 398 - 404 2006年03月 [査読有り][通常論文]
   

  Theobroxide, a novel active compound isolated from a fungus, has been reported previously to induce potato tuberization and flower bud formation in Pharbitis nil under non-inductive long-day conditions. Up to date, the action mechanism of theobroxide on flower-bud induction of P nil, however, is stilt unknown. In the present study, we observed a reduction of the stem length, along with the induction of flower buds, in theobroxide-treated and short-day-grown P nil plants. Also, the results showed that flower bud formation was delayed markedly in P nil seedlings with removal of cotyledons or exposure to night break. The suppression effect of night-break and cotyledon-rem ova L, however, was abolished completely by spraying theobroxide. Endogenous gibberellin(1/3) contents in P nil plants treated with theobroxide or grown under short-day conditions were relatively lower, suggesting that gibberellins probably are negatively involved in theobroxide- and short-day-induced flower-bud formation of P nil. (c) 2005 Elsevier GmbH. All. rights reserved.
• Inhibition of stem elongation in spinach by theobroxide

  FJ Kong, XQ Gao, KH Nam, K Takahashi, H Matsuura, T Yoshihara

  JOURNAL OF PLANT PHYSIOLOGY 163 5 557 - 561 2006年03月 [査読有り][通常論文]
   

  In the current study, we investigated the influences of theobroxide on stem elongation in spinach (Spinacia oleracea). Our results showed that stem elongation and flower formation were inhibited by spraying spinach plants with theobroxide under inductive, long day conditions (16 h light/8 h dark), white application of exogenous applied GA(3) prevented the effect of theobroxide. Quantitative analysis showed that theobroxide suppressed GA(1) biosynthesis, whereas the endogenous content of jasmonic acid was unchanged. However, under short day conditions (10 h light/14 h dark), there were no differences in stem length between treated and untreated plants. These results suggest that the inhibition of stem elongation by theobroxide is probably due to the suppression of gibberellin biosynthesis. (C) 2005 Elsevier GmbH. All rights reserved.
• Allene oxide cyclase is essential for theobroxide-induced jasmonic acid biosynthesis in Pharbitis nil

  FJ Kong, J Abe, K Takahashi, H Matsuura, T Yoshihara, K Nabeta

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 336 4 1150 - 1156 2005年11月 [査読有り][通常論文]
   

  Theobroxide, a natural product, strongly stimulates the biosyndthesis of jasmonic acid (JA) in Pharbitis nil. In this study, we investigated the accumulation of protein by the immunoblot analysis of lipoxygenase (LOX), allene oxide synthase (AOS), and. allene oxide cyclase (AOC), key enzymes in JA biosynthesis, and how the endogenous levels of JA in P. nil are affected by theobroxide. The effect of JA on the accumulations of these proteins was monitored simultaneously. The results show that theobroxide treatment led to a high level accumulation of JA, which is due to high accumulations of LOX, AOS, and AOC proteins induced by theobroxide treatment both under short day (SD) and long day (LD) conditions. However, under SD conditions AOS and AOC proteins are not enhanced by JA treatment. Kinetic analysis of protein levels shows that a biphasic activation of AOC protein by theobroxide is displayed and the first activation of AOC protein together with elevated JA levels is observed within 30 min after treatment. Meanwhile, AOS and LOX proteins are activated by theobroxide later than AOC protein, suggesting that AOC plays an essential role in the initial JA formation induced by theobroxide. Since theobroxide-increased JA levels also show a biphasic manner similar to AOC activation and AOS, LOX proteins are activated later than AOC, and thus we propose a positive JA feedback regulation. Interestingly, AOS protein, which is also the enzyme for the biosynthesis of 9,10-ketol-octadecadienoic acid (KODA, a flowering inducing factor), accumulates markedly due to the simultaneous involvement of theobroxide and SID conditions, suggesting that AOS probably plays a role in flower bud formation in P. nil. (c) 2005 Elsevier Inc. All rights reserved.
• Anti-babesial ellagic acid rhamnosides from the bark of Elaeocarpus parvifolius

  A Elkhateeb, Subeki, K Takahashi, H Matsuura, M Yamasaki, O Yamato, Y Maede, K Katakura, T Yoshihara, K Nabeta

  PHYTOCHEMISTRY 66 21 2577 - 2580 2005年11月 [査読有り][通常論文]
   

  Bioassay-guided investigation of the bark of Elaeocarpus parvifolius led to the isolation of three new ellagic acid derivatives, 4-O-methylellagic acid 3'-alpha-rhamnoside (2), 4-O-methylellagic acid 3'-(3"-O-acetyl)-alpha-rhamnoside (3), and 4-O-methylellagic acid 3'-(4"-0acetyl) -alpha-rhamnoside (4) in addition to the known ellagic acid derivative, 4-O-methylellagic acid 3'-(2", 3"-di-O-acetyl)-alpha-rhamnoside (1). Their structures were elucidated on the basis of analysis of H-1 NMR, C-13 NMR, HMQC, HMBC and MS spectroscopic data. Compounds 1-4 were evaluated for their growth-inhibitory effect on Babesia gibsoni in vitro. Compounds 2 and 4 showed very weak activity, while compounds I and 3 showed moderate activity, with IC50 values of 28.5 and 52.1 mu g/ml, respectively. (c) 2005 Elsevier Ltd. All rights reserved.
• Theobroxide inhibits stem elongation in Pharbitis nil by regulating jasmonic acid and gibberellin biosynthesis

  FJ Kong, XQ Gao, KH Nam, K Takahashi, H Matsuura, T Yoshihara

  PLANT SCIENCE 169 4 721 - 725 2005年10月 [査読有り][通常論文]
   

  In this study, exogenous factors affecting the elongation growth in the short day plant, Pharbitis nil, was investigated. Theobroxide inhibited stem elongation in P. nil both under short day (SD) and long day (LD) conditions. Salicy1hydroxamic acid (SHAM), an inhibitor of jasmonic acid (JA) biosynthesis, and GA(3) recovered the inhibitory effect of theobroxide on stem elongation. Quantitative analysis of JA showed that the level of endogenous JA increased significantly in theobroxide treated plants, while exogenously applied GA(3) and SHAM suppressed JA biosynthesis stimulated by theobroxide. The activity of lipoxygenase (LOX, the key enzyme of JA biosynthesis) also was stimulated by theobroxide and this stimulation was nullified by SHAM and GA(3). Quantitative analysis of GA(1) showed that theobroxide suppressed GA(1) biosynthesis. In non theobroxide treated R nil, SD conditions stimulated JA biosynthesis and LOX activity, while GA(1) biosynthesis was suppressed. All these results suggest that JA probably is involved negatively in the control of stem elongation, and the balance between JA and gibberellin might determine the stem growth in P. nil. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
• Theobroxide triggers jasmonic acid production to induce potato tuberization in vitro

  XQ Gao, F Wang, Q Yang, H Matsuura, T Yoshihara

  PLANT GROWTH REGULATION 47 1 39 - 45 2005年09月 [査読有り][通常論文]
   

  Theobroxide, a novel compound isolated from fungus Lasiodiplodia theobromae culture, stimulates potato (Solanum tuberosum L.) tuber formation in vitro and in vivo, and induces flowering of Japanese morning glory (Pharbitis nil) under non-inductive long day conditions. To assess the mode of action of theobroxide in the tuberization process we measured endogenous levels of jasmonic acid and lipoxygenase activity in the cultures after the treatment with theobroxide. The results showed that theobroxide not only stimulated microtuber formation alone, but also enhanced the inductive effect of jasmonic acid when they were used in combination. The endogenous JA content increased in response to theobroxide in both old and new tissues. Moreover, theobroxide increased lipoxygenase activity in the cultures, at 2 and 3 week after culture initiation. Additionally, histological observations indicated that theobroxide might play a role in the swelling of micro-tubers formed in vitro in a similar manner as that of jasmonic acid. These results suggest that the inductive effect of theobroxide on potato micro-tuber formation might be achieved through triggering jasmonic acid production.
• Production of phleichrome by Cladosporium phlei as stimulated by diketopiperadines of Epichloe typhina

  Y Seto, Y Kogami, T Shimanuki, K Takahashi, H Matsuura, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 69 8 1515 - 1519 2005年08月 [査読有り][通常論文]
   

  Epichloe typhina is an endophytic fungus, while Cladosporium phlei is a pathogenic fungus of the timothy plant (Phleum pretense L.). We found two activities in the culture filtrate of E. typhina: one stimulated the pathogenic fungus, C phlei, to produce phleichrome and the other inhibited its growth. The active ingredients that stimulated the production of phleichrome and inhibited the growth of C. phlei were isolated and characterized. The isolated compounds were identified as cyclo-(L-Pro-L-Leu) and cyclo-(L-Pro-L-Phe), which were stimulatory compounds, and p-hydroxybenzaldehyde, which was the growth inhibitory compound, based on an analysis of their spectral data. Of the two stimulatory compounds, cyclo-(L-Pro-L-Phe) showed higher activity. However, when 500 Kg Of cyclo-(L-Pro-L-Phe) was spotted on the TLC plate for bio-autography, a growth inhibitory zone was identified in the central red region, which contained phleichrome. On the other hand, phleichrome showed antifungal activity against E. typhina in the light, so it is assumed that there might be antagonism between the endophytic fungus, E. typhina, and the pathogenic fungus, C. phlei.
• Novel potato micro-tuber-inducing compound, (3R,6S)-6-hydroxylasiodiplodin, from a strain of Lasiodiplodia theobromae

  P Li, K Takahashi, H Matsuura, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 69 8 1610 - 1612 2005年08月 [査読有り][通常論文]
   

  A novel potato micro-tuber-inducing compound was isolated from the culture broth of Lasiodiplodia theobromae Shimokita 2. The structure of the isolated compound was determined as (3R,6S)-6-hydroxylasiodiplodin by means of spectroscopic analyses, the modified Mosher method, and chemical conversion. The compound showed potato micro-tuber-inducing activity at a concentration of 10(-4) M, using the culture of single-node segments of potato stems in vitro.
• Evaluation of the inhibitory activities of the extracts of Indonesian traditional medicinal plants against Plasmodium falciparum and Babesia gibsoni

  T Murnigsih, Subeki, H Matsuura, K Takahashi, M Yamasaki, O Yamato, Y Maede, K Katakura, M Suzuki, S Kobayashi, Chainul, T Yoshihara

  JOURNAL OF VETERINARY MEDICAL SCIENCE 67 8 829 - 831 2005年08月 [査読有り][通常論文]
   

  Twenty-four kinds of water extracts derived from 22 plants that are traditionally used for the treatment of malaria on Java Island, Indonesia, were screened for their antibabesial and antimalarial activities. Among the extracts, 8 extracts displayed strong antimalarial activity, with an inhibition range from 89.6 to 100%, and 15 showed strong antibabesial activity, with an inhibition range from 84.2 to 98.1%. The extracts of Achillea millefolium, Baeckea frutenscens, Brucea javanica, Curcuma xanthorrhiza, Strychnos lucida and Swietenia macrophylla showed both strong antibabesial and antimalarial activities. The antimalarial activities paralleled the antibabesial activities, but the converse was not true.
• Relation between environmental factors and the LOX activities upon potato tuber formation and flower-bud formation in morning glory

  KH Nam, C Minami, FJ Kong, H Matsuura, K Takahashi, T Yoshihara

  PLANT GROWTH REGULATION 46 3 253 - 260 2005年07月 [査読有り][通常論文]
   

  Life cycles of plants including tuberization and flowering are strongly related to environmental factors such as photoperiod and temperature. Theobroxide induces potato tuber formation and flower bud formation of morning glory under non-inductive conditions and stimulates the activity of lipoxygenase (LOX). In this study, to understand the LOX activity more systematically, the relationships between LOX activity and light and temperature, which effects potato tuber and flower-bud formation, have been investigated. The results showed that LOX activity in morning glory was greatly enhanced up to 30 min and then declined after switching from the light to the dark condition, while the activity did not vary when switching from the dark to the light condition. In addition, the temperature profile of measured LOX activity in the potato and morning glory plants was nearly consistent with the time taken to form potato tubers and flower buds in morning glory, respectively, at different growing temperatures. These results strongly suggest that LOX activity is directly connected with light and temperature to regulate the formation of tubers and flower-buds.
• The Arabidopsis gene CAD1 controls programmed cell death in the plant immune system and encodes a protein containing a MACPF domain

  C Morita-Yamamuro, T Tsutsui, M Sato, H Yoshioka, M Tamaoki, D Ogawa, H Matsuura, T Yoshihara, A Ikeda, Uyeda, I, J Yamaguchi

  PLANT AND CELL PHYSIOLOGY 46 6 902 - 912 2005年06月 [査読有り][通常論文]
   

  To clarify the processes involved in plant immunity, we have isolated and characterized a single recessive Arabidopsis mutant, cad1 (constitutively activated cell death 1), which shows a phenotype that mimics the lesions seen in the hypersensitive response (HR). This mutant shows spontaneously activated expression of pathogenesis-related (PR) genes, and leading to a 32-fold increase in salicylic acid (SA). Inoculation of cad1 mutant plants with Pseudomonas syringae pv tomato DC3000 shows that the cad1 mutation results in the restriction of bacterial growth. Cloning of CAD1 reveals that this gene encodes a protein containing a domain with significant homology to the MACPF (membrane attack complex and perforin) domain of complement components and perforin proteins that are involved in innate immunity in animals. Furthermore, cell death is suppressed in transgenic cad1 plants expressing nahG, which encodes an SA-degrading enzyme. We therefore conclude that the CAD1 protein negatively controls the SA-mediated pathway of programmed cell death in plant immunity.
• Anti-babesial activity of some Central Kalimantan plant extracts and active oligostilbenoids from Shorea balageran

  Subeki, S Nomura, H Matsuura, M Yamasaki, O Yamato, Y Maede, K Katakura, M Suzuki, Trimurningsih, Chairul, T Yoshihara

  PLANTA MEDICA 71 5 420 - 423 2005年05月 [査読有り][通常論文]
   

  Bark extracts from a total of 22 species of Central Kalimantan plants were evaluated for their anti-babesial activity against Babesia gibsoni in vitro. Of these plant species, extracts of Calophyllum tetrapterum, Garcinia rigida, Lithocarpus sp., Sandoricum emarginatum, and Shorea balangeran showed more than 90 % inhibition of the parasite growth at a test concentration of 1000 mu g/mL. Activity-guided fractionation of the bark of S. balangeran (Dipterocarpaceae) led to the reisolation of oligostil-benoids, vaticanol A (1), B (2), and G (3). The structures were determined on the basis of spectral evidence. Compounds 1 and 3 showed complete inhibition on the growth of Babesia gibsoni in vitro at a concentration of 25 mu g/mL, and compound 2 at concentration of 50 mu g/mL.
• Anti-babesial compounds from Curcuma zedoaria

  K Kasahara, S Nomura, Subecki, H Matsuura, M Yamasaki, O Yamato, Y Maede, K Katakura, M Suzuki, Trimurningsih, Chairul, T Yoshihara

  PLANTA MEDICA 71 5 482 - 484 2005年05月 [査読有り][通常論文]
   

  Anti-babesial activity was confirmed in an extract of the bark of Curcuma zedoaria. The active ingredients were isolated, and their chemical structures were determined to be zecloalactones A, B, and C based on spectral data. Zedoalactone C is a hitherto unreported compound. The IC50 vales of these active compounds against Babesia gibsoni were compared with a standard drug, diminazene aceturate. The IC50 value of diminazene aceturate was 0.6 mu g/mL, while those of zedoalactones A, B, and C were 16.5,1.6 and 4.2 mu g/mL, respectively.
• Anti-babesial and anti-plasmodial compounds from Phyllanthus niruri

  Subeki, H Matsuura, K Takahashi, M Yamasaki, O Yamato, Y Maede, K Katakura, S Kobayashi, Trimurningsih, Chairul, T Yoshihara

  JOURNAL OF NATURAL PRODUCTS 68 4 537 - 539 2005年04月 [査読有り][通常論文]
   

  Bioassay-guided fractionation of boiled aqueous extracts from the whole plant of Phyllanthus niruri led to the isolation of 1-O-galloyl-6-O-luteoyl-a-D-glucose (1), with IC50 values of 4.7 mu g/mL against Babesia gibsoni and 1.4 mu g/mL against Plasmodium falciparum in vitro. The known compounds P-glucogallin (2), quercetin 3-O-beta-D-glucopyranosyl-(2 -> 1)-O-beta-D-xylopyranoside (3), beta-sitosterol, and gallic acid were also isolated. Structures of these compounds were elucidated on the basis of their chemical and spectroscopic data.
• Antibabesial activity of protoberberine alkaloids and 20-hydroxyecdysone from Arcangelisia flava against Babesia gibsoni in culture

  Subeki, H Matsuura, K Takahashi, M Yamasaki, O Yamato, Y Maede, K Katakura, M Suzuki, Trimurningsih, Chairul, T Yoshihara

  JOURNAL OF VETERINARY MEDICAL SCIENCE 67 2 223 - 227 2005年02月 [査読有り][通常論文]
   

  Bioassay-guided fractionation of the boiled extract from the stems of Arcangelisia flava led to the isolation of palmatine (1), berberine (2), jatrorrhizine (3), dihydroberberine (4) and 20-hydroxyecdysone (5). The chemical structures of these compounds were elucidated on the basis of their chemical and spectral evidence. The isolated compounds were evaluated for their growth inhibiting effects on Babesia gibsoni in culture for a week. Compounds (1-4) showed significant inhibitions at concentrations from 100 to 1.0 mug/ml, while compound 5 at a concentration of 100 mug/ml, only.
• The Arabidopsis gene CAD1 controls programmed cell death in the plant immune system and encodes a protein containing a MACPF domain.

  C Yamamuro, T Tsutsui, Y Asada, H Yoshioka, M Tamaoki, D Ogawa, H Matsuura, T Yoshihara, A Ikeda, J Yamaguchi

  PLANT AND CELL PHYSIOLOGY 46 6 S120 - S120 2005年 [査読有り][通常論文]
  
• Anti-fungal sesquiterpenoid from the root exudate of Solanum abutiloides

  T Yokose, K Katamoto, S Park, H Matsuura, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 68 12 2640 - 2642 2004年12月 [査読有り][通常論文]
   

  The Solanum abutiloides plant is highly resistant to soil-borne pathogens such as Fusarium oxysporum f. sp. melongenae, Verticillium dahliae, and Ralstonia solanacearum. This species is utilized as a mating source of resistant cultivars and is also used as a rootstock. The root exudate of Solanum abutiloides was extracted from a soil system composed of charcoal and vermiculite. Anti-fungal activity was found in the extract, and an active ingredient was isolated. The chemical structure of the active compound was determined to be 3-beta-acetoxysolavetivone, a new sesquiterpenoid. The anti-fungal activity of 3-beta-acetoxysolavetivone examined by the inhibition of spore germination of Fusarium oxysporum was close to that of lubimin, and higher than that of solavetivone.
• Effects of theobroxide, a natural product, on the level of endogenous jasmonoids

  Q Yang, XQ Gao, Y Fujino, H Matsuura, T Yoshihara

  ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES 59 11-12 828 - 834 2004年11月 [査読有り][通常論文]
   

  The natural potato microtuber inducing substance, theobroxide, strongly induces the formation of tuber of potato (Solanum tuberosum L.) and flower bud of morning glory (Pharbitis nil) plants under non-inducing conditions (long days) (Yoshihara et al., 2000). In the present study, theobroxide was evaluated for its effect on the level of endogenous jasmonoids in different tissues of such two plants. An in vitro bioassay using cultures of single-node segments of potato stems was performed with the supplement of theobroxide in the medium. The endogenous jasmonic acid (JA) and its analogue tuberonic acid (TA, 12-hydroxyjasmonic acid) in segments and microtubers were quantitatively analyzed. The increase in the endogenous JA level caused by theobroxide was observed in both segments and microtubers. Endogenous TA was only detected in segments, and the content increased with the concentration of theobroxide. As for morning glory, the whole plant was sprayed with theobroxide for 1 5 weeks under different photoperiods and endogenous JA in the leaves was quantitatively analyzed. Theobroxide spraying increased the level of endogenous JA in the leaves of the plants grown under both long and short days.
• Isolation of an alpha-methylene-7-butyrolactone derivative, a toxin from the plant pathogen Lasiodiplodia theobromae

  GC He, H Matsuura, T Yoshihara

  PHYTOCHEMISTRY 65 20 2803 - 2807 2004年10月 [査読有り][通常論文]
   

  Lasiodiplodia theobromae is known as a multi-infectious microorganism that causes considerable crop damage, particularly to tropical fruits. When the fruits are infected by L. theobromae, the typical symptom is the appearance of black spots on the surface of the infected fruit. When injected in to the peel of banana, the culture filtrate of L. theobromae induced formation of black spots. The structure of the isolated compound responsible for this effect was determined to be (3S,4R)-3-carboxy-2-methylene-heptan-4-olide on the basis of analysis of MS, IR, and H-1 and C-13 NMR spectroscopic data, including HMQC, HMBC, and H-1-H-1 COSY experiments. The active compound was not only isolated from the culture filtrate derived from potato dextrose medium, but also from the extract of infected peels of bananas. (C) 2004 Elsevier Ltd. All rights reserved.
• A new antifungal metabolite from Penicillium expansum

  GC He, H Matsuura, T Takushi, S Kawano, T Yoshihara

  JOURNAL OF NATURAL PRODUCTS 67 7 1084 - 1087 2004年07月 [査読有り][通常論文]
   

  A new antifungal compound, (3S)-4,6-dihydro-8-methoxy-3,5-dimethyl-6-oxo-3H-2-benzopyran (4), was isolated from Penicillium expansum. During the isolation procedure 4 was determined to be unstable and readily reacted with methanol, ethanol, and water, forming three new isochromans, (1S,3S)-6-hydroxy-1,8-dimethoxy-3,5-dimethylisochroman (1), 1-ethoxy-6-hydroxy-8-methoxy-3,5-dimethylisochroman (2), and 1,6-dihydroxy-8-methoxy-3,5-dimethylisochroman (3), respectively. (3S)-6-Hydroxy-8-methoxy-3,5-dimethylisochroman (5) was reisolated from P. expansum. In fungicide disk assays, compounds 1, 2, and 4 inhibited the mycelial growth of Lasiodiplodia theobromae at 100 mug/mL by 76%, 74%, and 69%, respectively.
• Effects of Central Kalimantan plant extracts on intraerythrocytic Babesia gibsoni in culture

  Subeki, H Matsuura, M Yamasaki, O Yamato, Y Maede, K Katakura, M Suzuki, Trimurningish, Chairul, T Yoshihara

  JOURNAL OF VETERINARY MEDICAL SCIENCE 66 7 871 - 874 2004年07月 [査読有り][通常論文]
   

  The inhibitory effects of 45 plant extracts selected from Central Kalimantan, Indonesia on Babesia gibsoni in vitro and their acute toxicity to mice were evaluated. Of these plant extracts studied, Arcangelisia flava, Curcuma zedoaria, Garcinia benthamiana, Lansium domesticum and Peronema canescens were found to have appreciable antibabesial activity with IC50 values from 5.3 to 49.3,ug/ml without acute toxicity in mice at the intraperitoneal dose of 0.7 g/kg of body weight.
• Single-molecule imaging of the dynamic interactions between macromolecules

  H Yokota, K Kaseda, H Matsuura, Y Arai, A Iwane, Y Ishii, T Kodama, T Yanagida

  JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY 4 6 616 - 621 2004年07月 [査読有り][通常論文]
   

  In recent years, the development of single-molecule detection techniques has allowed the dynamic properties of biomolecules, which are normally obscured in conventional ensemble measurements, to be measured. One of these single-molecule detection techniques allows the measurement of dissociation and association events of individual molecules to be measured. This technique is based on the unique premise that the mobility between molecules that are bound and the mobility between those that are free in solution are different. The binding of ATP at the beginning and its dissociation at the end of the hydrolysis reaction were detected at the single-molecule level in real time. In this study, we extended this technique to image the dynamic interactions between large biomolecules (protein/protein and protein/polysaccharide). The binding and dissociation of fluorescently labeled macromolecules to partner molecules fixed on a glass surface were visualized by total internal reflection fluorescence microscopy. The dynamic interactions between the proteins in two energy conversion systems, that is, signaling proteins and enzyme molecules moving on dextran, have been measured. In these systems, the dynamic interactions were sensitive to the factors determining the chemical reactions. Thus, the dynamic interactions monitored in the single-molecule measurements provided useful information to further the understanding of the underlying mechanisms of energy conversion systems.
• Antifungal compounds from the root and root exudate of Zea mays

  S Park, Y Takano, H Matsuura, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 68 6 1366 - 1368 2004年06月 [査読有り][通常論文]
   

  A maize plant (Zea mays) planted in a test tube was found to inhibit the growth of the soil-borne plant pathogen, Fusarium oxysporum C sp. melongenae. The antifungal compounds, 6-methoxybenzoxazolinone and 6,7-dimethoxybenzoxazolinone, were isolated from an ethanol extract of Zea mays roots, and (6R)-7,8-dihydro-3-oxo-alpha-ionone and (6R, 9R)-7,8-dihydro-3-oxo-alpha-ionol were isolated from the root exudate.
• Isolation of alpha-glusosidase inhibitors from hyssop (Hyssopus officinalis)

  H Matsuura, H Miyazaki, C Asakawa, M Amano, T Yoshihara, J Mizutani

  PHYTOCHEMISTRY 65 1 91 - 97 2004年01月 [査読有り][通常論文]
   

  alpha-Glucosidase inhibitory activity was found in aqueous methanol extracts of dried hyssop (Hyssopus officinalis) leaves. Active principles against alpha-glucosidase, prepared from rat small intestine acetone powders, were isolated and characterized. The structures of these isolated compounds were determined to be (7S, 8S)-syringoylglycerol-9-O-(6'-O-cinnamoyl)-beta-D-glucopyranoside and (7S, 8S)-syringoylglycerol 9-O-beta-D-glucopyranoside by analysis of physical and spectroscopic data (FDMS, H-1 NMR, C-13 NMR, HMQC. and HMBC experiments) together with chemical syntheses. (C) 2003 Elsevier Ltd. All rights, reserved.
• DPPH radical scavengers from dried leaves of oregano (Origanum vulgare)

  H Matsuura, H Chiji, C Asakawa, M Amano, T Yoshihara, J Mizutani

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 67 11 2311 - 2316 2003年11月 [査読有り][通常論文]
   

  1,1-Dipehnyl-2-picrylhydrazyl (DPPH) radical scavenging activities were found in the extract of dried leaves of oregano (Origanum vulgare). The water-soluble active ingredients were isolated, and their structures were determined to be 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybenzyl protocatechuate and 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybenzyI 4-O-methyl-protocatechuate by H-1-, C-13-NMR, DEPT, HMQC, and HMBC spectral analyses, and by NOE experiments. The DPPH radical scavenging activities of these compounds were compared with those of rutin, quercetin and rosmarinic acid at a concentration of 2 x 10(-5) M. The scavenging activity of 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybenzyl protocatechuate was almost the same as that of quercetin and rosmarinic acid, but that of 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybennzyl, 4-O-methylprotocatechuate was less than that of quercetin, rosmarinic acid and 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybenzyl protocatechuate. The amount of 4'-O-beta-D-glucopyranosyl-3',4'-dihydroxybenzyI protocatechuate was estimated to be 3.8 mg/1 g of dried leaves by an HPLC analysis.
• Inhibitory effect of salicylhydroxamic acid on theobroxide-induced potato tuber formation

  Xiquan Gao, Qing Yang, Chisato Minami, Hideyuki Matsuura, Atsuo Kimura, Teruhiko Yoshihara

  Plant Science 165 5 993 - 999 2003年11月01日 [査読無し][通常論文]
   

  Theobroxide is a natural compound from the fungus Lasiodiplodia theobromae and has a significantly inductive effect on potato tuber formation in vitro and in vivo. Considering the similar roles of theobroxide and jasmonic acid (JA) in tuberization, we investigated the influence of theobroxide on endogenous levels of JA and its analogue tuberonic acid (TA), as well as on lipoxygenase (LOX) activity in potato leaves. Salicylhydroxamic acid (SHAM), the JA biosynthesis inhibitor, was used to test its effect on theobroxide-induced tuber formation, on endogenous JA and TA levels, and on LOX activity. The results showed that theobroxide increased endogenous levels of JA and TA, and LOX activity. However, this inductive effect was inhibited by SHAM, associated with a reduction in tuber formation. These results suggest that the inductive effect of theobroxide on tuber formation is probably achieved by stimulating JA and TA synthesis. © 2003 Elsevier Ireland Ltd. All rights reserved.
• Inhibitory effects of hyssop (Hyssopus officinalis) extracts on intestinal alpha-glucosidase activity and postprandial hyperglycemia

  H Miyazaki, H Matsuura, C Yanagiya, J Mizutani, M Tsuji, C Ishihara

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 49 5 346 - 349 2003年10月 [査読有り][通常論文]
   

  It has been known that Hyssopus officinalis (hyssop) is a herb that grows in the wild and is a source of natural antioxidants. We previously reported that alpha-glucosidase inhibitors, (2S,3S)1-O-beta-D-6'-O-cinnamoylglucopyranosyl-3-(3",5"-dimethoxy-4"-hydroxyphenyl)-1,2,3-propanetriol and (2S,3S)1-O-beta-D-glucopranosyl-3-(3",5"-dimethoxy-4"-hydroxyphenyl)-1,2,3-propanetriol, from the dry leaves of hyssop, were isolated. This study examined the a-glucosidase inhibitory effects of hyssop extracts on intestinal carbohydrate absorption in rat everted gut sac and carbohydrate-loaded hyperglycemia in mice. In the everted gut sac experiment, 10 mM sucrose- and 5 mm maltose-treated increases in glucose concentration in the serosal compartment were inhibited in the presence of 0.5 and 1.0 mg/mL hyssop extracts, although a 10 mm glucose-induced increase in serosal glucose was not inhibited by the extracts. Additionally, hyperglycemia in sucrose- and maltose-loaded mice was significantly suppressed at an early stage, within 30 to 60 min by oral pre-administration of 300 and 100 mg/kg hyssop extracts, respectively. These findings suggest that hyssop extracts inhibited the digestion of complex carbohydrates, but not that of absorbable monosaccharide, and might be a useful supplemental food for hyperglycemia.
• Metabolism of deuterium-labeled jasmonic acid and OPC 8 : 0 in the potato plant (Solanum tuberosum L.)

  H Matsuura, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 67 9 1903 - 1907 2003年09月 [査読有り][通常論文]
   

  The metabolism of deuterium-labeled (+/-)-jasmonic acid and 3-oxo-2-[(Z)pent-2'-enyl]-cyclopentan-1-octanoic acid in potato (Solanum tuberosum L.) was examined by using cultures of potato single-node stems. Deuterium-labeled (+/-)-jasmonic acid and 3-oxo-2-[(Z) pent-2'-enyl]cyclopentan-1-octanoic acid, which had been prepared from commercially available methyl (+/-)-jasmonate, were fed to the cultures, and the metabolites were extracted from the plants and analyzed by a liquid chromatography-selected ion monitoring system. The metabolism of deuterium-labeled (+/-)-jasmonic acid and 3-oxo-2-[(Z)-2'-pentenyl]cyclopentan-1-octanoic acid to 5' and 4'-O-glucopyranosyloxyjasmoic acids was strongly suggested.
• Isolation and identification of sodium 2-propenyl thiosulfatei from boiled garlic (Allium sativum) that oxidizes canine erythrocytes

  O Yamato, Y Sugiyama, H Matsuura, KW Lee, K Goto, MA Hossain, Y Maede, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 67 7 1594 - 1596 2003年07月 [査読有り][通常論文]
   

  Sodium 2-propenyl thiosulfate was identified in boiled garlic (Allium sativum). When canine erythrocytes were incubated with sodium 2-propenyl thiosulfate, the methemoglobin concentration and Heinz body percentage in erythrocytes were both increased, indicating that the compound induced oxidative damage in canine erythrocytes. It seems that this compound is one of the causative agents of garlic-induced hemolysis in dogs.
• Isolation and measurement of quercetin glucosides in flower buds of Japanese butterbur (Petasites japonicus subsp gigantea Kitam.)

  H Matsuura, M Amano, J Kawabata, J Mizutani

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 66 7 1571 - 1575 2002年07月 [査読有り][通常論文]
   

  Three quercetin glucosides were isolated from flower buds of Japanese butterbur (Petasites japonicus subsp. gigantea Kitam.) together with caffeic acid as the ingredients that had DPPH radical scavenging activity, using the DPPH-HPLC method for measuring the radical scavenging activity. These quercetin glucosides were identified as quercetin 3-O-beta-D-glucoside, quercetin 3-O-beta-D-6"-O-acetylglucoside, and rutin, and the amounts of the glucosides in flower buds were also examined by HPLC. The flower buds were harvested from four different sites, the total amount of quercetin glucosides in each site was 100-170 mg/100 g fr. wt., and there were no great differences of the amounts between growing fields
• alpha-glucosidase inhibitor from the seeds of balsam pear (Momordica charantia) and the fruit bodies of Grifola frondosa

  H Matsuura, C Asakawa, M Kurimoto, J Mizutani

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 66 7 1576 - 1578 2002年07月 [査読有り][通常論文]
   

  alpha-Glucosidase inhibitory activities were found in aqueous methanol extracts of the seeds of Momordica charantia and the fruit bodies of Grifola frondosa. An active principle against the enzyme prepared from rat small intestine acetone powders was isolated and characterized. The structure of the isolated compound was identified as D-(+)-trehalose by FDMS, H-1-, C-13-NMR, and [alpha](D) measurements. The inhibitory activity of trehalose was compared with 1-deoxynojirimycin. Trehalose showed 45% inhibitory activity at the concentration of 2 x 10(-3) M, but 1-deoxynojirimycin had 52% inhibitory activity at 1 x 10(-7) M.
• Occurrence of 11-hydroxyjasmonic acid glucoside in leaflets of potato plants (Solanum tuberosum L.)

  H Matsuura, Y Ohkubo, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 65 2 378 - 382 2001年02月 [査読有り][通常論文]
   

  In order to examine the occurrence of 11-hydroxyjasmonic acid glucoside in potato plants, a synthesis of 11-hydroxyjasmonic acid was accomplished, and the synthetic compound was employed as a standard for an LC-SIM analysis. The existence of 11-hydroxyjasmonic acid glucoside was proved by the LC-SIM analysis.
• 36.バレイショ茎断片とアサガオ葉に投与したセオブロキシドの内生ジャスモン酸類生成への影響(第36回大会研究発表抄録)

  楊 青, 松浦 英幸, 藤野 由美子, 吉原 照彦

  植物の生長調節 36 2 238 - 238 一般社団法人植物化学調節学会 2001年
• Qualitative and quantitative analysis of endogenous jasmonoids in potato plant (Solanum tuberosum L.)

  H Matsuura, F Ohmori, M Kobayashi, A Sakurai, T Yoshihara

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 64 11 2380 - 2387 2000年11月 [査読有り][通常論文]
   

  Qualitative and quantitative analyses of endogenous jasmonoids were done by liquid chromatography/ selected ion monitoring (LC-SIM) using deuterium-labeled compounds as internal standards. To prove the practicality of this way of analyzing the contents of endogenous jasmonoids in plants, the method was used for estimating jasmonoids in potato plants.
• Potato micro-tuber inducing hydroxylasiodiplodins from Lasiodiplodia theobromae

  Q Yang, M Asai, H Matsuura, T Yoshihara

  PHYTOCHEMISTRY 54 5 489 - 494 2000年06月 [査読有り][通常論文]
   

  Three hydroxylasiodiplodins were isolated from the mycelium extracts of Lasiodiplodia theobromae IFO 31059; and their structures were identified as (3R),(4S)-4-hydroxylasiodiplodin, (3R),(6R)-6-hydroxy-de-O-methyllasiodiplodin and (3R),(5R)-5-hydroxy-de-O-methyllasiodiplodin. All showed potato micro-tuber inducing activity. (C) 2000 Elsevier Science Ltd. All rights reserved.
• Induction of plant tubers and flower buds under noninducing photoperiod conditions by a natural product, theobroxide

  Teruhiko Yoshihara, Fumihiro Ohmori, Kaoru Nakamori, Michiko Amanuma, Takashi Tsutsumi, Akitami Ichihara, Hideyuki Matsuura

  Journal of Plant Growth Regulation 19 4 457 - 461 2000年 [査読有り][通常論文]
   

  Theobroxide is an epoxy cyclohexene compound isolated from the culture filtrate of the fungus Lasiodiplodia theobromae that induces potato microtuber formation in vitro (Nakamori and others 1994). When sprayed on potato (Solanum tuberosum L.) and morning glory (Pharbitis nil) plants, which require short days to induce tubers and flower buds, respectively, potato plants kept in noninducing conditions (long days) produced tubers. Theobroxide spray treatment also produced flower buds in morning glory plants kept under noninducing conditions (long days). Furthermore, under inducing conditions (short days), the number of flowers of seedlings sprayed with theobroxide was about 1.5 times that of controls.
• An antifungal compound from roots of Welsh onion

  N Phay, T Higashiyama, M Tsuji, H Matsuura, Y Fukushi, A Yokota, F Tomita

  PHYTOCHEMISTRY 52 2 271 - 274 1999年09月 [査読無し][通常論文]
   

  A novel antifungal compound, fistulosin (octadecyl 3-hydroxyindole), was isolated from roots of Welsh onion (Allium fistulosum L.), and its structure was elucidated by spectroscopic means. This compound showed high activity against Fusarium oxysporum primarily inhibiting protein synthesis. (C) 1999 Elsevier Science Ltd. All rights reserved.
• Three lasiodiplodins from Lasiodiplodia theobromae IFO 31059

  Hideyuki Matsuura, Kaoru Nakamori, E. A. Omer, Chiaki Hatakeyama, Teruhiko Yoshihara, Akitami Ichihara

  Phytochemistry 49 2 579 - 584 1998年09月28日 [査読有り][通常論文]
   

  Three new fungal metabolites were isolated from the culture filtrate of the fungus, Lasiodiplodia theobromae IFO 31059 and identified as (5R) and (5S) 5-hydroxylasiodiplodins and 5-oxolasiodiplodin. The first two showed weak potato micro-tuber inducing activities, although only at very high dosage levels.
• 55(P13) 重水素ラベルジャスモン酸類、ツベロン酸、ツベロン酸グルコシドの合成とその定量分析、生合成研究への応用(ポスター発表の部)

  松浦 英幸, 小原 直美, 吉原 照彦

  天然有機化合物討論会講演要旨集 40 323 - 328 天然有機化合物討論会 1998年08月31日 

  Tuber development of potato plants (Solanum tuberosum L.) was controlled by environmental factors, mainly by photoperiods. Short days stimulate this process, but long days have opposite effect against this process. This means potato plants distinguish the day length. But it is still unknown the relationships between day length and potato tuber inducing compounds, jasmonic acid (JA), 12-hydroxyjasmonic acid (tuberonic acid, TA), and tuberonic acid glucoside (TAG), since there is no qualitative and quantitative analysis method for these compounds. The synthesis of deuterium labeled TAG, TA, JA, (1R, 2R) 3-ox-2-(2'-pentenyl)-cyclopentanebutanoic acid (OPC 4: 0), (1R 2R) 3-oxo-2-(2'-pentenyl)-cyclopentanehexanoic acid (OPC 6: 0), and (1R, 2R) 3-oxo-2-(2'-pentenyl)-cyclopentaneoctanoic acid (OPC 8: 0) were accomplished. The qualitative and quantitative analysis for the endogenous jasmonides was carried out using these compounds as internal standards and monitoring by liquid chromatography/mass spectrometry (LC-SIM). This methodology was applied to estimate the endogenous jasmonides, TAG, TA, JA, OPC 4: 0, OPC 6: 0, and OPC 8: 0, in potato plant. The contents of TAG, TA, JA in the leaflets of potato (Solanum tuberosum L. cv. Irish Cobbler) which were harvested in July, Hokkaido, were estimated to be 7-5μg/1g leaflets, 170-100 ng/1g leaflets and 20-10ng/1g leaflets, respectively, and those of OPC 4: 0, OPC 6: 0, OPC 8: 0 to be below ng level/1g leaflets. The contents of TAG of the leaflets of potato which grown under long and short day conditions were analyzed, respectively, by means of this developed methodology. The contents of TAG were higher in the leaflets grown under long days than in those grown in short days. It is suggested that the short day condition might stimulate the transportation of jasmonids from leaflets to the other parts. The bio-synthetic pathway for TA and TAG was established in the first time because the deuterium leveled JA and OP 8: 0 were converted to TAG in the cultures on single-node segments of potato stem in vitro.
• Novel cyclohexene compound from lasiodiplodia theobromae IFO 31059

  Hideyuki Matsuura, Naomi Obara, Noriko Chisaka, Akitami Ichihara, Teruhiko Yoshihara

  Bioscience, Biotechnology and Biochemistry 62 12 2460 - 2462 1998年 [査読有り][通常論文]
   

  A novel cyclohexene compound (1), which is structurally related to theobroxide (2), was isolated from a culture filtrate of the fungus, Lasiodiplodia theobromae IFO 31059. The potato micro-tuber-inducing activity of this compound was observed at a concentration of 10-3 M in the medium, whereas theobroxide (2) showed its activity at 10-5 M. © 1998, Taylor & Francis Group, LLC. All rights reserved.
• Metabolism and transport of [2-14C](±) jasmonic acid in the potato plant

  Teruhiko Yoshihara, Michiko Amanuma, Takashi Tsutsumi, Yukihiro Okumura, Hideyuki Matsuura, Akitami Ichihara

  Plant and Cell Physiology 37 5 586 - 590 1996年 [査読有り][通常論文]
   

  Metabolism and transport of [2-14C](±) jasmonic acid (JA) were studied using 2-week-old potato plants. Plants grown under tuber inducing conditions (short days) and non-inducing conditions (long days) were assayed 1, 5, 10, 15, 20, and 25 days after substance application. The incorporated [2-14C](±)JA was metabolized to tuberonic acid glucoside (TAG) within 2 weeks. The radioactivity of the ethanol extract from each plant part was measured. More than half of the total radioactivity was still associated with the treated leaves up to 25 days after the application of [214C](±)JA. In the other parts of the plants, the upper leaves contained the most total radioactivity, followed by the stems and lower leaves. Under long days no tubers were produced, and the radioactivity of the stolons was observed. In contrast, under short days tuber were produced, and the radioactivities were present in stolons and tubers. Flower buds and flowers were produced more under long days and accumulated high radioactivity. It is concluded from the combined data that JA is metabolized to TAG and transported to all parts of the plant. A high accumulation of TAG in tubers and flower buds may induce the formation of tubers and flower buds.
• Potato micro-tuber inducing substances from Lasiodiplodia theobromae

  Kaoru Nakamori, Hideyuki Matsuura, Teruhiko Yoshihara, Akitami Ichihara, Yasunori Koda

  Phytochemistry 35 4 835 - 839 1994年 [査読有り][通常論文]
   

  Three potato-tuber inducing substances were isolated from Lasiodiplodia theobromae IFO 31059, and their structures identified as mellein, jasmonic acid and a previously unrecorded cyclohexene named theobroxide. © 1994.
• 4 NEW POLYACETYLENIC GLUCOSIDES, METHYL BETA-D-GLUCOPYRANOSYL HELIANTHENATE C-F, FROM JERUSALEM-ARTICHOKE (HELIANTHUS-TUBEROSUS L)

  H MATSUURA, T YOSHIHARA, A ICHIHARA

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 57 9 1492 - 1498 1993年09月 [査読有り][通常論文]
   

  Four new polyacetylenic glucosides isolated from the leaf of Jerusalem artichoke (Helianthus tuberosus L.) were characterized as methyl beta-D-glucopyranosyl helianthenate C (5), D (6), E (7), and F (8). The absolute stereochemistry of the glucosyloxymethine was also determined to be of R configuration by preparing the relative compounds with Sharpless asymmetric epoxidation as the kev step and source of optical activity.
• Tuber-forming Substances in Jerusalem Artichoke (Helianthus tuberosus L.)

  Hideyuki Matsuura, Teruhiko Yoshihara, Akitami Ichihara, Yoshio Kikuta, Yasunori Koda

  Bioscience, Biotechnology and Biochemistry 57 8 1253 - 1256 1993年 [査読有り][通常論文]
   

  Four tuber-forming substances in Jerusalem artichoke were isolated from the leaves. The structures were established by spectroscopic methods as jasmonic acid (2), methyl β-d-glucopyranosyl tuberonate (3), and two new Polyacetylene compounds, methyl β-d-glucopyranosyl helianthenate A (4, C19H24O8) and B (5, C17H22O8). © 1993, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.
• 53 植物及び微生物起源のバレイショ塊茎形成物質(口頭発表の部)

  松浦 英幸, 中森 薫, 吉原 照彦, 市原 耿民, 幸田 泰則, 喜久田 嘉郎

  天然有機化合物討論会講演要旨集 33 409 - 416 天然有機化合物討論会 1991年09月07日 

  Tuber forming substances from plants and microorgansms were surveyed using a bioassay consisting single-node stem segment culture. From leaves of Jerusalem artichoke (Helianthus tuberosus. L), two cyclopentanoid fatty acids, jasmonic acid(2) and methyl tuberonic acid glucoside(3), and six polyacetylene compounds(4-9) were isolated and the chemical structures were determinded. From the fungi, Lasiodiplodia theobromae IFO 31059, a cyclohexene epoxide(10) and mellein(11) were isolated. Cyclopentanoid fatty acids(2,3) have revealed tuber forming activity at more than 10^<-5>M and cyclohexene epoxide(10) at 10^<-4>M. Polyacetylene compounds(4-9) and mellein(11) have displayed weak activity.
• 21 バレイショ塊茎形成誘導物質に関する化学的研究(ポスター発表の部)

  吉原 照彦, オメール エルサエド, 柴田 治樹, 越野 広雪, 松浦 英幸, 坂村 貞雄, 幸田 泰則, 喜久田 嘉郎, 岡澤 養三

  天然有機化合物討論会講演要旨集 30 158 - 164 天然有機化合物討論会 1988年09月26日 

  The concept of tuber forming stimulus of potato has been suggested by grafting experiments. Recently the occurrence of the substances has been evidenced by our experiment using a bioassay of a potato single-node stem segment culture. The object of this study was to isolate and determine the chemical structure of the substance from potato leaves by using of above-mentioned bioassay. The ethanol extracts of potato leaves (100kg) was separated by means of the subsequent liquid chromatographies and HPLCs. Purification processes resulted in 2.7mg of the active compound (1) which was able to induce tuberization in vitro even in lower concentration of 0.01 ppm. The FD mass spectrum showed the molecular ion peak at m/z 388 and that of the acetate at m/z 556. These data with that of GLC suggested that the active compound (1) is a monoglucoside. The PMR spectrum (δ 4.26, d, J=8.0Hz) and CMR spectrum (δ 104.4) suggested the presence of β-D-glucose moiety. EI-HR MS of the acetate (2) showed the molecular formula, C_<26>H_<36>O_<13>. The ^1H-NMR COSY experiment revealed the partial structures, I and II. These data suggested that the active compound (1) was similar to jasmonic acid (3). The chemical structure of the active compound (1) was determined to be 3-oxo-2- (5-β-D-glucopyranosyloxy-2-cis-pentenyl)-cyclopentane-1-acetic acid. A (cis)-double bond and a carboxyl group were essential for the activity. The active compound (1) and/or similar compound(s)(jasmonic acid?) were presented in all kinds of plants.

MISC

• 植物病原菌が生産するストリゴラクトン様活性分子の探索

  鈴木泰輝, 松浦英幸, 北岡直樹, 都築玄武, 来馬道生, 西山康太郎, 瀬戸義哉 植物の生長調節 58 (Supplement) 2023年
• The bHLH transcription factor MpHYPNOS regulates gemma dormancy in the liverwort Marchantia polymorpha

  Hirotaka Kato, Nami Yoshimura, Mikako Yoshikawa, Hideyuki Matsuura, Kosaku Takahashi, Daisuke Takezawa, Tomoyuki Furuya, Yuki Kondo, Hidehiro Fukaki, Tetsuro Mimura, Kimitsune Ishizaki bioRxiv 488978 2022年04月25日 [査読無し]
  
• ダイズの柱頭上花粉発芽に関わるFTオルソログの機能

  平田万季, 針谷康平, 竹島亮馬, 松浦英幸, 藤野介延, XU M., 山田哲也, 阿部純 育種学研究 21 2019年
• 胎児期の有機フッ素化合物曝露が7歳のアレルギーに及ぼす影響 北海道スタディ

  アイツバマイ ゆふ, 荒木 敦子, 宮下 ちひろ, 岡田 恵美子, 樫野 郁子, グウダルジ・ホウマヌ, 松浦 英幸, 岸 玲子 日本衛生学雑誌 73 (Suppl.) S225 -S225 2018年03月 [査読無し][通常論文]
  
• 胎児期の有機フッ素化合物曝露が7歳のアレルギーに及ぼす影響 北海道スタディ

  アイツバマイ ゆふ, 荒木 敦子, 宮下 ちひろ, 岡田 恵美子, 樫野 郁子, グウダルジ・ホウマヌ, 松浦 英幸, 岸 玲子 日本衛生学雑誌 73 (Suppl.) S225 -S225 2018年03月 [査読無し][通常論文]
  
• 胎児期の有機フッ素化合物曝露が7歳のアレルギーに及ぼす影響-北海道スタディ

  アイツバマイ ゆふ, 荒木敦子, 宮下ちひろ, 岡田恵美子, 岡田恵美子, 樫野郁子, 樫野郁子, グウダルジ ホウマヌ, グウダルジ ホウマヌ, 松浦英幸, 岸玲子 日本衛生学雑誌(Web) 73 (Supplement) 2018年
• ゲノム編集を用いたサリチル酸高蓄積イネ変異体の作出

  手塚大介, 手塚大介, 佐分利亘, 森春英, 松浦英幸, 今井亮三 植物の生長調節 53 (Supplement) 2018年
• 前向きコーホート研究に基づく先天異常,免疫アレルギーおよび小児発達障害のリスク評価と環境物質に対する遺伝的感受性の解明 DNAメチル化を介する有機フッ素化合物胎児期曝露の児への影響

  岸玲子, 石塚真由美, 松浦英幸, 三浦りゅう, 小林祥子, 小林澄貴 前向きコーホート研究に基づく先天異常、免疫アレルギーおよび小児発達障害のリスク評価と環境化学物質に対する遺伝的感受性の解明 平成28年度 総括・分担研究報告書 127‐134 2017年 [査読無し][通常論文]
  
• S07 バレイショ塊茎形成におけるジャスモン酸類の働き(オーラル発表,植物化学調節学会第50回大会)

  井上 悠敬, 清水 崇史, 高橋 公咲, 崎浜 靖子, 瀬尾 光範, 松浦 英幸 植物化学調節学会研究発表記録集 50 29 -29 2015年10月01日
• P075 GTR1は傷害葉から非傷害葉へのJA-Ile移行に関与し、過剰なJA応答を抑制する(ポスター発表,植物化学調節学会第50回大会)

  及川 貴也, 石丸 泰寛, 鈴木 健史, 武石 翔平, 松浦 英幸, 高橋 公咲, 浜本 晋, 魚住 信之, 清水 崇史, 瀬尾 光範, 太田 啓之, 上田 実 植物化学調節学会研究発表記録集 50 93 -93 2015年10月01日
• P076 ジャスモン酸はAux/IAAの分解を部位特異的に抑制し側根形成を阻害する(ポスター発表,植物化学調節学会第50回大会)

  鈴木 健史, 石丸 泰寛, Meesters Christian, Kombrink Erich, 知念 拓実, 臼井 健郎, 高橋 公咲, 松浦 英幸, 林 謙一郎, 深城 英弘, 上田 実 植物化学調節学会研究発表記録集 50 94 -94 2015年10月01日
• P055 Trehalose-induced Systemic Resistance(TSR)の分子基盤(ポスター発表,植物化学調節学会第50回大会)

  手塚 大介, 和久田 真司, 加藤 英樹, 松浦 英幸, 佐分 利亘, 森 春英, 松井 博和, 今井 亮三 植物化学調節学会研究発表記録集 50 (0) 73 -73 2015年10月01日 [査読無し][通常論文]
  
• Trehalose‐induced Systemic Resistance(TSR)の分子基盤

  手塚大介, 手塚大介, 和久田真司, 加藤英樹, 松浦英幸, 佐分利亘, 森春英, 松井博和, 今井亮三, 今井亮三 植物の生長調節 50 (Supplement) 73 2015年10月01日 [査読無し][通常論文]
  
• ゼニゴケのT‐DNA挿入変異株を用いたOPDA作用機序に関する研究

  大鹿遵, 西浜竜一, 河内孝之, 松浦英幸, 高橋公咲 植物の生長調節 50 (Supplement) 95 2015年10月01日 [査読無し][通常論文]
  
• P077 ゼニゴケのT-DNA挿入変異株を用いたOPDA作用機序に関する研究(ポスター発表,植物化学調節学会第50回大会)

  大鹿 遵, 西浜 竜一, 河内 孝之, 松浦 英幸, 高橋 公咲 植物化学調節学会 研究発表記録集 50 (0) 95 -95 2015年 [査読無し][通常論文]
  
• 42. ヒメツリガネゴケの12-オキソファイトジエン酸の代謝に関わるグルタチオン-S-トランスフェラーゼの機能解析(口頭発表,植物化学調節学会第49回大会)

  梶原 章志, 戸島 江里香, 南条 洋平, 小松 節子, 松浦 英幸, 高橋 公咲 植物化学調節学会研究発表記録集 49 60 -60 2014年10月01日
• 19. 低温に応答したGA量の抑制によりイネの耐性を高めることができる(口頭発表,植物化学調節学会第49回大会)

  高橋 直希, 川又 彩, 手塚 大介, 佐分利 亘, 松浦 英幸, 森 春英, 今井 亮三 植物化学調節学会研究発表記録集 49 (0) 37 -37 2014年10月01日 [査読無し][通常論文]
  
• 43. 傷害時のイネにおけるサリチル酸グルコシドを糖ドナーとするツベロン酸への糖転移反応(口頭発表,植物化学調節学会第49回大会)

  竹松 知紀, 瀬戸 義哉, 宮澤 吉郎, 和久田 真司, 佐分利 亘, 森 春英, 高橋 公咲, 松浦 英幸 植物化学調節学会研究発表記録集 49 (0) 61 -61 2014年10月01日 [査読無し][通常論文]
  
• 52. イネのトレハロースにより誘導されるシステミックな病害抵抗性にはジャスモン酸が関与する(口頭発表,植物化学調節学会第49回大会)

  手塚 大介, 坂井 志帆, 和久田 真司, 加藤 英樹, 松浦 英幸, 佐分利 亘, 森 春英, 松井 博和, 今井 亮三 植物化学調節学会研究発表記録集 49 (0) 70 -70 2014年10月01日 [査読無し][通常論文]
  
• 62. イネの傷害応答におけるサリチル酸グルコシド加水分解酵素の機能(口頭発表,植物化学調節学会第49回大会)

  武田 遼介, 佐分利 亘, 姫野 奈美, 和久田 真司, 松浦 英幸, 今井 亮三, 松井 博和, 森 春英 植物化学調節学会研究発表記録集 49 (0) 80 -80 2014年10月01日 [査読無し][通常論文]
  
• イネの傷害応答におけるサリチル酸グルコシド加水分解酵素の機能

  武田遼介, 佐分利亘, 姫野奈美, 和久田真司, 松浦英幸, 今井亮三, 松井博和, 森春英 植物の生長調節 49 (Supplement) 80 2014年10月01日 [査読無し][通常論文]
  
• イネのトレハロースにより誘導されるシステミックな病害抵抗性にはジャスモン酸が関与する

  手塚大介, 坂井志帆, 和久田真司, 加藤英樹, 松浦英幸, 佐分利亘, 森春英, 松井博和, 今井亮三 植物の生長調節 49 (Supplement) 70 2014年10月01日 [査読無し][通常論文]
  
• 傷害時のイネにおけるサリチル酸グリコシドを糖ドナーとするツベロン酸への糖転移反応

  竹松知紀, 瀬戸義哉, 宮澤吉郎, 和久田真司, 佐分利亘, 森春英, 高橋公咲, 松浦英幸 植物の生長調節 49 (Supplement) 61 2014年10月01日 [査読無し][通常論文]
  
• 低温に応答したGA量の抑制によりイネの耐性を高めることができる

  高橋直希, 川又彩, 手塚大介, 佐分利亘, 松浦英幸, 森春英, 今井亮三 植物の生長調節 49 (Supplement) 37 2014年10月01日 [査読無し][通常論文]
  
• Brassica rapaにおける抗ウイルス反応としてのアスコルビン酸蓄積の誘導メカニズム

  十川聡子, 藤原綾香, 松浦英幸, 犬飼剛, 増田税 日本植物病理学会大会 プログラム・講演要旨予稿集 134 2014年06月 [査読無し][通常論文]
  
• セオブロキシド処理によるウイルス抵抗性の低下機構の解明

  山下雄大, 松浦英幸, 犬飼剛, 増田税, 吉原照彦 日本植物病理学会大会 プログラム・講演要旨予稿集 101 2014年06月 [査読無し][通常論文]
  
• 有機フッ素化合物の胎児曝露が乳幼児期のアレルギー症状に及ぼす影響

  岡田恵美子, 佐々木成子, 樫野いく子, 松浦英幸, 宮下ちひろ, 小林澄貴, 伊藤久美子, 池野多美子, 玉腰暁子, 岸玲子 北海道医学雑誌 89 (1) 70 2014年 [査読無し][通常論文]
  
• シロイヌナズナβ‐グルコシダーゼ様タンパク質の生化学的機能解析

  谷口沙希, 佐分利亘, 松浦英幸, 今井亮三, 松井博和, 森春英 日本農芸化学会北海道支部講演会講演要旨 2013 36 2013年11月27日 [査読無し][通常論文]
  
• シロイヌナズナβ‐グルコシダーゼ様タンパク質の生化学的機能解析

  谷口沙希, 佐分利亘, 松浦英幸, 今井亮三, 松井博和, 森春英 日本農芸化学会北海道支部講演会講演要旨 2013 23 2013年11月27日 [査読無し][通常論文]
  
• 25. ゼニゴケにおけるアブシシン酸シグナル伝達と二次代謝産物の変化(口頭発表,植物化学調節学会第48回大会)

  蔭山 暁人, 石崎 公庸, 河内 孝之, 松浦 英幸, 高橋 公咲 植物化学調節学会研究発表記録集 48 40 -40 2013年10月04日
• 19. Theobroxide処理に対する植物応答機構の解明(口頭発表,植物化学調節学会第48回大会)

  山下 雄大, 犬飼 剛, 高橋 公咲, 増田 税, 松浦 英幸 植物化学調節学会研究発表記録集 48 34 -34 2013年10月04日
• 38. 傷害応答時における移動性シグナルとしてのジャスモン酸イソロイシンの機能(口頭発表,植物化学調節学会第48回大会)

  武石 翔平, 佐藤 千鶴, 増田 税, 松浦 英幸 植物化学調節学会研究発表記録集 48 53 -53 2013年10月04日
• 12‐オキソファイトジエン酸で処理したヒメツリガネゴケのプロテオーム解析

  戸島江里香, 阿部達也, 南條洋平, 小松節子, 松浦英幸, 高橋公咲 日本農芸化学会大会講演要旨集(Web) 2013 3A44A12 (WEB ONLY) 2013年03月05日 [査読無し][通常論文]
  
• イネ由来ツベロン酸β‐グルコシドグルコシダーゼのアグリコン認識メカニズムの解析

  姫野奈美, 佐分利亘, 武田遼介, 和久田真司, 松浦英幸, 鍋田憲助, 森春英, 今井亮三, 松井博和 日本農芸化学会大会講演要旨集(Web) 2013 3C16A13 (WEB ONLY) 2013年03月05日 [査読無し][通常論文]
  
• シロイヌナズナにおけるCYP94B2の機能解析

  天野就基, 北岡直樹, 川出洋, 谷口沙希, 佐分利亘, 松井博和, 松浦英幸 日本農芸化学会大会講演要旨集(Web) 2013 4A44A10 (WEB ONLY) 2013年03月05日 [査読無し][通常論文]
  
• イネ由来ツベロン酸β‐グルコシドグルコシダーゼアイソザイム1の機能解析

  武田遼介, 姫野奈美, 佐分利亘, 和久田真司, 森春英, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 日本農芸化学会大会講演要旨集(Web) 2013 3C16A12 (WEB ONLY) 2013年03月05日 [査読無し][通常論文]
  
• 有機フッ素化合物(11種類)の胎児期曝露による出生時体格への影響

  樫野 いく子, 佐々木 成子, 岡田 恵美子, 宮下 ちひろ, 山本 潤, 松浦 英幸, 伊藤 陽一, 松村 徹, 玉腰 暁子, 岸 玲子 日本衛生学雑誌 68 (Suppl.) S160 -S160 2013年03月 [査読無し][通常論文]
  
• 有機フッ素化合物の胎児期曝露による1歳までのアレルギー症状との関連

  岡田 恵美子, 樫野 いく子, 佐々木 成子, 宮下 ちひろ, 山本 潤, 伊藤 陽一, 松浦 英幸, 松村 徹, 玉腰 暁子, 岸 玲子 日本衛生学雑誌 68 (Suppl.) S161 -S161 2013年03月 [査読無し][通常論文]
  
• ツベロン酸グルコシド加水分解酵素の基質認識機構の解明

  姫野奈美, 和久田真司, 武田遼介, 佐分利亘, 森春英, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 日本農芸化学会北海道支部講演会講演要旨 2012 24 2012年11月01日 [査読無し][通常論文]
  
• 組換えツベロン酸グルコシドグルコシダーゼの酵素化学的諸性質の解析

  武田遼介, 姫野奈美, 佐分利亘, 和久田真司, 森春英, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 日本農芸化学会北海道支部講演会講演要旨 2012 23 2012年11月01日 [査読無し][通常論文]
  
• イネの傷害時のSAGをdonorとしたTAに対する糖転移反応

  竹松知紀, 宮澤吉郎, 瀬戸義哉, 和久田真司, 佐分利亘, 鍋田憲助, 松井博和, 松浦英幸 日本農芸化学会北海道支部講演会講演要旨 2012 15 2012年11月01日 [査読無し][通常論文]
  
• 37. ゼニゴケにおけるアレンオキシドシクラーゼの機能解析(口頭発表,植物化学調節学会第47回大会)

  山本 祐介, 石崎 公庸, 河内 孝之, 高橋 知寛, 松浦 英幸, 高橋 公咲 植物化学調節学会研究発表記録集 47 54 -54 2012年10月05日
• ジャスモン酸メチルの植物移行性及び代謝に関する有機化学的研究

  松原卓也, 北岡直樹, 松浦英幸 植物の生長調節 47 (Supplement) 103 2012年10月05日 [査読無し][通常論文]
  
• ゼニゴケにおけるアレンオキシドシクラーゼの機能解析

  山本祐介, 石崎公庸, 河内孝之, 高橋知寛, 松浦英幸, 高橋公咲 植物の生長調節 47 (Supplement) 54 2012年10月05日 [査読無し][通常論文]
  
• 86. ジャスモン酸メチルの植物移行性及び代謝に関する有機化学的研究(口頭発表,植物化学調節学会第47回大会)

  松原 卓也, 北岡 直樹, 松浦 英幸 植物化学調節学会研究発表記録集 47 (0) 103 -103 2012年10月05日 [査読無し][通常論文]
  
• イネの根脂質改変とアルミニウム耐性

  前島恵理子, 渡部敏裕, 松浦英幸, 黒田昌治, 豊増知伸, 岡咲洋三, 斉藤和季, 大崎満, 我妻忠雄 日本土壌肥料学会講演要旨集 58 (58) 74 -74 2012年09月04日 [査読無し][通常論文]
  
• 妊婦の血中PFOS/PFOAおよび類縁化合物の定量法の確立

  樫野いく子, 岡田恵美子, 松浦英幸, 山本潤, 佐々木成子, 宮下ちひろ, 松村徹, 岸玲子 環境化学討論会要旨集(CD-ROM) 21st ROMBUNNO.P-163 2012年07月10日 [査読無し][通常論文]
  
• 北海道における妊婦の血中PFOS/PFOAおよび類縁化合物濃度

  岡田恵美子, 樫野いく子, 松浦英幸, 山本潤, 佐々木成子, 宮下ちひろ, 松村徹, 岸玲子 環境化学討論会要旨集(CD-ROM) 21st ROMBUNNO.P-169 2012年07月10日 [査読無し][通常論文]
  
• シロイヌナズナ由来シトクロムP450 CYP94C1の機能解析

  谷口沙希, 和久田真司, 北岡直樹, 佐分利亘, 川出洋, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 日本農芸化学会大会講演要旨集(Web) 2012 3A30P18 (WEB ONLY) 2012年03月05日 [査読無し][通常論文]
  
• 植物ホルモン・ジャスモン酸の主要な不活性化経路 ジャスモン酸イソロイシン12位水酸化酵素CYP94B3の発見

  北岡直樹, 松浦英幸 化学と生物 50 (3) 156 -157 2012年03月01日 [査読無し][通常論文]
  
• 植物ホルモン・ジャスモン酸の主要な不活性化経路 : ジャスモン酸イソロイシン12位水酸化酵素CYP94B3の発見

  北岡 直樹, 松浦 英幸 化学と生物 50 (3) 156 -157 2012年03月01日 [査読無し][通常論文]
  
• 11-5 イネの根脂質改変とアルミニウム耐性(11.植物の有害元素,2012年度鳥取大会)

  前島 恵理子, 渡部 敏裕, 松浦 英幸, 黒田 昌治, 豊増 知伸, 岡咲 洋三, 斉藤 和季, 大崎 満, 我妻 忠雄 日土肥講要 58 (0) 74 -74 2012年 [査読無し][通常論文]
  
• 85. 12位の水酸化から始まるジャスモン酸イソロイシンンの代謝に関する研究(口頭発表,植物化学調節学会第46回大会)

  北岡 直樹, 松原 卓也, 宮澤 吉郎, 佐藤 道大, 瀬戸 義哉, 高橋 公咲, 和久田 真司, 川出 洋, 松井 博和, 鍋田 憲助, 松浦 英幸 植物化学調節学会研究発表記録集 (46) 101 -101 2011年10月03日 

  Jasmonate is plant hormone that plays crucial roles in development and defense response. It has been shown that 7-iso-jasmonyl-L-isoleucine (JA-Ile) is the bioactive form involved in the jasmonate-mediated signaling pathway. The hormonal action of jasmonate in plants is controlled by the precise balance between its biosynthesis and catabolism. Recently, CYP94B3, JA-Ile 12-hydroxylase was considered one of important enzymes that played fine tuning system of active form. Here, we studied the metabolic pathway of JA-Ile started with 12-hydroxylation. Firstly, we synthesized 12-carboxy1JA-Ile and 12-O-D-β-glucopyranosy1JA-Ile, which were expected metabolites of 12-hydroxyJA-Ile. Secondly, the endogenous levels of the compounds in wounded leaves of wild type or cyp94b3 mutant were analyzed by UPLC-MS/MS. Both compounds accumulated and the amount of 12-O-D-β-glucopyranosy1JA-Ile in cyp94b3 mutant was lower than that in wild type. In other hands, the 12-carboxy1JA-Ile level was same in wildtype and cyp94b3 mutant. This result suggests 12-carboxy1JA-Ile is synthesized by enzyme different from CYP94B3. Finally, we investigated 12-hydroxyJA-Ile glucosyltransferase. The enzyme which converted 12-hydroxyJA to 12-O-D-P-glucopyranosy1JA also had activity when 12-hydroxyJA-Ile was used as substrate. Currently, we survey the biological activity of 12-carboxy1JA-Ile and 12-O-D-13-glucopyranosy1JA-Ile.
• 84. UDP-glucose非依存性イネ由来糖転移酵素に関わる糖供与体の探索(口頭発表,植物化学調節学会第46回大会)

  宮澤 吉郎, 瀬戸 義哉, 小野 祐太朗, 佐藤 道大, 高橋 公咲, 和久田 真司, 松井 博和, 鍋田 憲助, 松浦 英幸 植物化学調節学会研究発表記録集 (46) 100 -100 2011年10月03日 

  Plant hormone, jasmonic acid (JA), has been recognized as a signal molecule involved in the wounding response. But two metabolites of JA, tuberonic acid (TA; 12-hydroxyjasmonic acid) and its β-glucoside (TAG) have not been reported to show any clear physiological function on the wounding response. In order to establish functions of these compounds, we previously purified and characterized a glucosyltransferase in rice, that catalyzes the glucosylation from TA to TAG using UDPG as a glucose donor [1]. Moreover, we found out the existence of UDPG-independent one in the crude enzyme, that is more specific to TA than that one. In order to understand this glucosylation, we investigated a glucose donor except UDPG in this study. To test an activity of glucosyltranslation, we carried out TA glucosyltransferase assay using a crude enzyme in the rice cell culture. We used TA as a substrate and analyzed resulting TAG by UPLC-MS/MS. Using this assay system, an 80% ethanol extract derived from rice cell cultures (376.3 g) was purified by several series of column chromatography, and finally, compound 1 (1.5 mg) was obtained. The chemical structure of compound 1 is determined by NMR analysis that is to be 3-(4-hydroxyphenyl)prop-2-enyl β-D-glucopyranoside. This was synthesized from p-coumaric acid as starting compound, and the synthesized compound also transferred glucose to TA. Therefore, we concluded that compound 1 is a natural glucose donor in the rice cell culture.
• 葉部において生合成されたジャスモノイルイソロイシンの移動と生物活性

  松浦英幸, 佐藤千鶴, 末田香恵, 増田税, 鍋田憲助 植物の生長調節 46 (Supplement) 102 2011年10月03日 [査読無し][通常論文]
  
• 12位の水酸化から始まるジャスモン酸イソロイシンの代謝に関する研究

  北岡直樹, 松原卓也, 宮澤吉郎, 佐藤道大, 瀬戸義哉, 高橋公咲, 和久田真司, 川出洋, 松井博和, 鍋田憲助, 松浦英幸 植物の生長調節 46 (Supplement) 101 2011年10月03日 [査読無し][通常論文]
  
• UDP‐glucose非依存性イネ由来糖転移酵素に関わる糖供与体の探索

  宮澤吉郎, 瀬戸義哉, 小野祐太朗, 佐藤道大, 高橋公咲, 和久田真司, 松井博和, 鍋田憲助, 松浦英幸 植物の生長調節 46 (Supplement) 100 2011年10月03日 [査読無し][通常論文]
  
• 86. 葉部において生合成されたジャスモノイルイソロイシンの移動と生物活性(口頭発表,植物化学調節学会第46回大会)

  松浦 英幸, 佐藤 千鶴, 末田 香恵, 増田 税, 鍋田 憲助 植物化学調節学会研究発表記録集 0 (46) 102 -102 2011年10月03日 [査読無し][通常論文]
   

  Jasmonic acid (JA) and its derivatives are involved in mobile signaling in plants during defense against biotic and abiotic stress. In our recent paper, it was found that exogenously applied jasmonoyl isoleucine (JA-Ile) in biological concentration is transported to distal sites in wounded tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum). Here we investigated the distal transport of de novo-synthesized JA-Ile in response to leaf wounding in tobacco and tomato using [^2H_1-10, ^2H_2-11, ^2H_3-12] JA ([^2H_6]JA) and [U-^<13>C]L-Ile ([^<13>C_6]Ile). [^2H_6]JA-[^<13>C_6]Ile, [^2H_6...
• キュウリモザイクウイルスとカブモザイクウイルスの移行に関する相互作用

  竹下稔, 小泉恵美子, 野口眞季子, 末田香恵, 志村華子, 石川典子, 松浦英幸, 大島一里, 夏秋知英, 桑田茂, 古屋成人, 土屋健一, 増田税 日本植物細胞分子生物学会大会・シンポジウム講演要旨集 29th 276 2011年09月06日 [査読無し][通常論文]
  
• Pichia pastorisによる組換えツベロン酸グルコシドβ‐glucosidaseの諸性質の解明

  姫野奈美, 和久田真司, 佐分利亘, 森春英, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 応用糖質科学 1 (3) (47) 2011年07月20日 [査読無し][通常論文]
  
• 前向きコホート研究による先天異常モニタリング,特に尿道下裂,停留精巣のリスク要因と環境化学物質に対する感受性の解明 有機フッ素化合物(PFCs)曝露による小児の体格および疾患への影響に関する疫学研究のレビュー

  岸玲子, 吉岡英治, 岡田恵美子, 樫野いく子, 坂晋, 金澤文子, 鷲野考揚, 小西香苗, 馬場俊明, THAMAR Yila, 宮下ちひろ, TITILOLA Braimoh, 小林澄貴, 大竹裕子, 伊藤久美子, LIMPAR Mariko, 松浦英幸 前向きコホート研究による先天異常モニタリング、特に尿道下裂、停留精巣のリスク要因と環境化学物質に対する感受性の解明 平成20-22年度 総合研究報告書 186 -198 2011年 [査読無し][通常論文]
  
• 前向きコホート研究による先天異常モニタリング,特に尿道下裂,停留精巣のリスク要因と環境化学物質に対する感受性の解明 有機フッ素化合物(PFCs)曝露による小児の体格および疾患への影響に関する疫学研究のレビュー

  岸玲子, 吉岡英治, 岡田恵美子, 樫野いく子, 坂晋, 金澤文子, 鷲野考揚, 小西香苗, 馬場俊明, THAMAR Yila, 宮下ちひろ, TITILOLA Braimoh, 小林澄貴, 大竹裕子, 伊藤久美子, LIMPAR Mariko, 松浦英幸 前向きコホート研究による先天異常モニタリング、特に尿道下裂、停留精巣のリスク要因と環境化学物質に対する感受性の解明 平成22年度 総括・分担研究報告書 62 -74 2011年 [査読無し][通常論文]
  
• イネの病傷害ストレス応答におけるOsJAR1およびOsJAR2の機能

  鈴木絵里香, 和久田真司, 佐分利亘, 松浦英幸, 鍋田憲助, 今井亮三, 松井博和 日本農芸化学会北海道支部講演会講演要旨 2011 5 2011年 [査読無し][通常論文]
  
• 54.植物の傷害応答伝達シグナルとしてのJA-Ileの植物内における移動性の検討(口頭発表)

  佐藤 千鶴, 相川 健介, 杉山 俊平, 鍋田 憲助, 増田 税, 松浦 英幸 植物化学調節学会研究発表記録集 (45) 71 -71 2010年10月01日 

  Determining the mobile signal used by plants to defend against biotic and abiotic stresses has proved elusive, but jasmonic acid (JA) and its derivatives appear to be involved. Using deuterium-labeled analogs, we investigated the systemic transport of JA and jasmonoyl isoleucine (JA-Ile) in response to leaf wounding in tobacco (Nicotiana tabacum) plants. We recovered [2H2-2]JA (JA d2) and ([2H3-12]JA-Ile (JA-Ile d3) in systemic leaves of tobacco after treating wounded leaves with JA d2 or JA-Ile d3. We found that JA-Ile had a greater mobility than JA, despite its lower polarity, and that application of exogenous JA-Ile to wounded leaves led to a higher accumulation of JA and JA-Ile in systemic leaves compared to wounded control plants. We also found that exudates from the stem of tomato (Solanum lycopersicum) plants with damaged leaflets contained JA and JA-Ile in higher levels than in an undamaged plant, and a significant difference in the levels of JA-Ile was observed 30min after wounding. These results suggest that JA-Ile might be transportable compound involved in the resistance to biotic and abiotic stresses in plants.
• 抗トリパノソーマ活性を有するミャンマー産薬用植物の探索

  片倉賢, BAWM Saw, TIWANANTHAGORN, Saruda, LIN Kyaw San, HTUN Lat Lat, MAW Ni Ni, MYAING Tin Tin, PHAY Nyunt, 宮崎智史, 櫻井達也, 奥祐三郎, 松浦英幸 日本寄生虫学会大会プログラム・抄録集 79th 77 2010年05月 [査読無し][通常論文]
  
• ミャンマー産薬用植物の抗トリパノソーマ活性と細胞毒性の評価(Evaluation of Myanmar medicinal plants for antitrypanosomal and cytotoxic activities)

  Bawm S, 松浦 英幸, 宮崎 智史, Lin K.S, 廣田 淳一, 入江 隆夫, Htun L.L, Maw N.N, Tiwananthagorn S, 櫻井 達也, 奥 祐三郎, 片倉 賢 獣医寄生虫学会誌 8 (2) 126 -126 2010年03月 [査読無し][通常論文]
  
• Cloning and Functional Analysis of an Allene Oxide Synthase in Physcomitrella patens (vol 73, pg 2356, 2009)

  P. K. G. S. S. Bandara, K. Takahashi, M. Sato, H. Matsuura, K. Nabeta BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 73 (11) 2009年11月 [査読無し][通常論文]
  
• 植物の傷害応答時におけるジャスモン酸類の代謝と移動

  松浦 英幸 化学と生物 47 (11) 756 -763 2009年11月01日 [査読無し][通常論文]
   

  生理活性物質の代謝，移動，到達点での生理活性発現解析のために様々な手法が用いられ，研究が行なわれてきた．特に植物内での移動実験に関しては 14C などの放射活性なラベル体を用いた実験が多く見受けられるが，移動先での検出は多くの場合，放射活性を測定するのみであり，実際の化合物の化学構造については議論がなされていない．しかし，放射活性を有したまま化合物が構造変換を受け，移動した場合，上記の手法には限界がある．一方，昨今の質量分析機器の性能の向上は目覚ましく，迅速な微量物質の分析が可能になった．この技術を安定同位体ラベルされた目的化合物と組み合わせ，上記の問題解決を試みたので本稿で紹介したい．また，この技術を利用してジャスモン酸(JA)類の代謝経路に関わる酵素の単離精製に至った経緯もあわせ紹介する．
• 70.スイカ果肉に由来する種子発芽阻害物質の探索(口頭発表)

  小林 義氣, 鍋田 憲助, 松浦 英幸 植物化学調節学会研究発表記録集 (44) 84 -84 2009年10月06日 

  Seed germination is regulated by many environmental signals to complete this critical event exactly. We haven't seen germinated seeds in a fruit. This may be due to seed dormancy, lack of oxygen or available water. But it is assumed that chemical(s) which exist around seeds are also involved. Using extracts of watermelon [Citrullus lanatus (Thunb.) Matsum and Nakai)] flesh, bioassay-guided purification led to three fractions which possess inhibitory activity. Structural analysis following more separation identified six compounds: abscisic acid, 4-hydroxybenzoic acid, indole-3-carboxylic acid, abscisic acid glucose ester, abscisic alcohol glucoside and dehydrodiconiferyl alcohol glucoside. Among these, abscisic acid has very strong activity at a concentration equal to that of watermelon flesh.
• 90.ヒメツリガネゴケにおけるジャスモン酸の同定およびアレンオキシドシンターゼの機能解析(口頭発表)

  高橋 公咲, Bandara P. K. G. S. S, 佐藤 道大, 松浦 英幸, 鍋田 憲助 植物化学調節学会研究発表記録集 (44) 104 -104 2009年10月06日 

  Jasmonic acid (JA) is a plant hormone that plays important roles for a large number of processes in stress adaptation and development in vascular plants. The functions and signal transduction of JA in vascular plants have been studied extensively however those in non-vascular plants have been overlooked. The UPLC-MS/MS analysis of JA showed the presence of JA in Physcomitrella patens that is a model plant in mosses. The genomic database and protein homology searches predicted that P. patens would possess two allene oxide synthases, PpAOS1 and PpAOS2, evolved in JA biosynthesis. Recombinant PpAOSs were tried to obtain by overexpression in Escherichia coli. Only recombinant PpAOS1 was obtained as a soluble form. PpAOS1 had AOS activity and showed a pH optimum of approximately pH 6.0 using 13-HPOT as a substrate. PpAOS1 had substrate specificity for 13-HPOT, showing that it was 13-AOS. Semi-quantitative RT-PCR of PpAOSs showed that the expression of PpAOS1 was higher than that of PpAOS2 this indicated that PpAOS1 might operate JA biosynthesis in P. patens predominantly. These data suggest that the octadecanoid pathway also exists in P. patens.
• 薬用植物Brucea javanicaから抽出したクアシノイドのin vitroにおける抗エバンス.トリパノソーマ活性(Quassinoids from a medicinal plant Brucea javanica: their in vitro activities against Trypanosoma evansi)

  Bawm S, 松浦 英幸, Subeki, Elkhateeb A, 鍋田 憲助, 野中 成晃, 奥 祐三郎, 片倉 賢 獣医寄生虫学会誌 8 (1) 65 -65 2009年09月 [査読無し][通常論文]
  
• 抗寄生虫性と細胞毒性活性のミャンマー薬用植物の評価(Evaluation of Myanmar medicinal plants for antiparasitic and cytotoxic activities)

  Saw Bawm, 松浦 英幸, 宮崎 智史, Lin Kyaw San, 広田 淳一, 入江 隆夫, Htun Lat Lat, Maw Ni Ni, Tiwananthagorn Saruda, 櫻井 達也, 奥 祐三郎, 片倉 賢 日本獣医学会学術集会講演要旨集 148回 182 -182 2009年09月 [査読無し][通常論文]
  
• Isolation of quassinoids from a medicinal plant, Brucea javanica and their in vitro activities against Trypanosoma evansi

  S. Bawm, H. Matsuura, A. Elkhateeb, K. Nabeta, Subeki, N. Nonaka, Y. Oku, K. Katakura JOURNAL OF VETERINARY PHARMACOLOGY AND THERAPEUTICS 32 160 -161 2009年08月 [査読無し][通常論文]
  
• 薬用植物Brucea javanicaから抽出したカシノイド Trypanosoma evansiに対するin vitroでの作用(Quassinoids from a medicinal plant Brucea javanica: their in vitro activities against Trypanosoma evansi)

  Saw Bawm, 松浦 英幸, Subeki Subeki, Elkhateeb Ahmed, 鍋田 憲助, 野中 成晃, 奥 祐三郎, 片倉 賢 日本獣医学会学術集会講演要旨集 147回 233 -233 2009年03月 [査読無し][通常論文]
  
• 63.傷害応答時におけるジャスモン酸の代謝および移動に関する研究(口頭発表)

  佐藤 千鶴, 瀬戸 義哉, 松重 真奈, 葵 新, 鍋田 憲助, 松浦 英幸 植物化学調節学会研究発表記録集 (43) 77 -77 2008年10月06日 

  Plants respond to wounding and herbivore attack with a series of defense responses, which are put in motions locally and systemically. When the plants are suffered from wounding stress, a mobile signal is translocated from wounded sites to undamaged distal sites. This phenomenon is classified as wound-induced systemic acquired resistance (WSR). The key compound involved in this systemic signal transduction is supposed to be JA or its derivatives. Based on the kinetic study of the accumulation of JA and its derivatives responding to wounding stress, it was uncovered that JA was accumulated in undamaged systemic tissues within 0.5h after wounding and then, metabolized to JA-Ile, tuberonic acid (12-OH JA), and tuberonic acid glucoside. Attempts to recover deuterium and radio isotope labeled JA, which were fed to wounding leaves, were successfully accomplished. Therefore, it was demonstrated that JA could be transported from the damaged site to the systemic leaves.
• 62.ツベロン酸グルコシルトランスフェラーゼの精製とその生理的作用に関する研究(口頭発表)

  瀬戸 義哉, 浜田 茂樹, 松浦 英幸, 松重 真奈, 佐藤 千鶴, 高橋 公咲, 増田 税, 伊藤 浩之, 松井 博和, 鍋田 憲助 植物化学調節学会研究発表記録集 0 (43) 76 -76 2008年10月06日 [査読無し][通常論文]
   

  Tuberonic acid (12-hydroxy epi-jasmonic acid, TA) and its glucoside (TAG) were isolated from potato leaflets (Solanum tuberosum L.) and shown to have tuber-inducing properties. These compounds are known to be biosynthesized by an octadecanoid pathway. The metabolism of jasmonic acid (JA) to TAG in plant leaflets and translocation of the resulting TAG to the distal parts was demonstrated. It is thought that TAG generated from JA transmits a wounded signal from damaged parts to undamaged parts by this mechanism. The metabolism of TA in higher plants was demonstrated using [12-^3H] TA, and TA ...
• (401) カブモザイクウイルスに感染したアラビドプシスにおけるえそ誘導機構(平成20年度日本植物病理学会大会講演要旨)

  犬飼 剛, 金 甫〓, 増田 税, 松浦 英幸, 高橋 英樹 日本植物病理學會報 74 (3) 274 -274 2008年08月25日 [査読無し][通常論文]
  
• フラボノイド合成系遺伝子のRNAサイレンシングにより明らかになった植物組織の着色に必要なmRNA量の閾値の存在

  永松敦, 増田税, 松浦英幸, 喜多村啓介, 阿部純, 金澤章 日本遺伝学会大会プログラム・予稿集 80th 113 2008年08月08日 [査読無し][通常論文]
  
• 植物の根に関する諸問題(179)食糧難を引き起こす寄生雑草,ストライガ

  松浦 英幸, 吉原 照彦 農業および園芸 83 (7) 799 -805 2008年07月
• 3種のモデル植物における低リン適応機構の比較解析

  崔祥子, 和崎淳, 信濃卓郎, 松浦英幸, 大崎満 日本植物生理学会年会要旨集 49th 109 2008年03月15日 [査読無し][通常論文]
  
• 薬用植物由来天然化合物bruceine Aのイヌ・バベシア症に対する治療効果

  中尾 亮, 水上 智秋, 川村 悠太, スベキ, S.・ボン, 山崎 真大, 前出 吉光, 松浦 英幸, 高橋 公咲, 鍋田 憲助, 野中 成晃, 奥 祐三郎, 片倉 賢 獣医寄生虫学会誌 6 (2) 92 -92 2008年03月 [査読無し][通常論文]
  
• Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing (vol 5, pg 778, 2007)

  A. Nagamatsu, C. Masuta, M. Senda, H. Matsuura, A. Kasai, J. -S. Hong, K. Kitamura, J. Abe, A. Kanazawa PLANT BIOTECHNOLOGY JOURNAL 6 (1) 103 -103 2008年01月 [査読無し][通常論文]
  
• Anti-babesial quassinoids from the fruits of Brucea javanica

  Ahmed Elkhateeb, Masahiro Yamasaki, Yoshimitsu Maede, Ken Katakura, Kensuke Nabeta, Hideyuki Matsuura NATURAL PRODUCT COMMUNICATIONS 3 (2) 145 -148 2008年 [査読無し][通常論文]
   

  The medicinal plant Brucea javanica (L.) Merr. (Simaroubaceae), grown in China, was examined for anti-babesial properties. The anti-babesial activity of the fruit was found to be attributed to its quassinoid constituents. Ten active compounds were isolated and purified from a chloroform extract. The identities of these compounds were confirmed from NMR spectroscopic and mass spectral data as brusatol (1), bruceantin (2), bruceine A (3), bruceantinol (4), dehydrobruceine B (5), dehydrobrusatol (6), dehydrobruceine A (7), bruceine D (8), bruceoside A (9), and yadanzioside G (10). When tested in vitro against Babesia gibsoni, compounds 1-10 had IC50 values of 0.74, 13.4, 4.0, 12.0, 308.2, 10.5, 835.0, >1000, and >1000 ng/mL, respectively. Compounds 1-4, 6 and 7 had far higher activity than the commercial anti-babesial drug diminazene aceturate, which possesses an IC50 value of 70.5 ng/mL. Except for bruceine A (3), bruceantinol (4) and bruceine D (8), this is the first report of the anti-babesial activity of these isolated quassinoids.
• ダイズにおいてF3'H mRNA蓄積量が毛茸の着色に関与する

  永松 敦, 増田 税, 松浦 英幸, 喜多村 啓介, 阿部 純, 金澤 章 日本育種学会・日本作物学会北海道談話会会報 (48) 103 -104 2007年12月 [査読無し][通常論文]
  
• 薬用植物由来天然化合物bruceine Aのイヌ・バベシア症に対する治療効果

  中尾亮, 水上智秋, 川村悠太, SUBEKI Subeki, BAWM Saw, 山崎真大, 前出吉光, 松浦英幸, 高橋公咲, 鍋田憲助, 片倉賢 日本獣医学会学術集会講演要旨集 144th 61 2007年08月27日 [査読無し][通常論文]
  
• 2‐Benzoxazolinone類縁体の抗バベシア活性

  山田一明, 野村信吉, 松浦英幸, SU Beki, 山崎真大, 前出吉光, 片倉賢, 吉原照彦, 鍋田憲助 日本農芸化学会大会講演要旨集 2007 250 2007年03月05日 [査読無し][通常論文]
  
• インドネシア産薬用植物から抽出したbrucein Aのイヌ・バベシア症に対する治療効果

  中尾亮, 水上智秋, 川村悠太, SUBEKI Subeki, SAW Bawm, 山崎真大, 前出吉光, 松浦英幸, 高橋公咲, 鍋田憲助, 片倉賢 日本寄生虫学会大会プログラム・抄録集 76th 64 2007年02月23日 [査読無し][通常論文]
  
• Virus‐induced gene silencingを利用したダイズにおけるフラボノイドの生合成に関わる遺伝子の機能解析と組成の改変

  永松敦, 千田峰生, 松浦英幸, 葛西厚史, HONG Jin‐Sung, 喜多村啓介, 阿部純, 増田税, 金澤章 生化学 3P-0751 2007年 [査読無し][通常論文]
  
• Curcuma xanthorrhizaからの抗バベシア化合物の単離

  山田一明, 野村信吉, 松浦英幸, SUBEKI, 山崎真大, 大和修, 前出吉光, 片倉賢, TRIMURNINGSIH, CHAIRUL, 吉原照彦, 鍋田憲助 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨 2006 14 2006年 [査読無し][通常論文]
  
• インドネシア産民間薬用植物抽出物における抗マラリア, 抗バベシア活性の相関(寄生虫病学)

  MURNIGSIH Tri, Subeki, 松浦 英幸, 高橋 公咲, 山崎 真大, 大和 修, 前出 吉光, 片倉 賢, 鈴木 守, 小林 寿美子, Chairul, 吉原 照彦 The journal of veterinary medical science 67 (8) 829 -831 2005年08月 [査読無し][通常論文]
   

  宿主や媒介昆虫は異なるが, バベシア, マラリア原虫の生活環は酷似している事から, インドネシアで抗マラリア薬として用いられている民間伝承薬用植物の抗バベシア活性に関して検討した.活性の評価にはイヌバベシア原虫(Babesia gibsoni)ならびに熱帯マラリア原虫(Plasmodium falciparum)を用いた.その結果, 抗マラリア活性を有する植物は抗バベシア活性を有するが, 抗バベシア活性を示す植物が必ずしも抗マラリア活性を示すことは無いと評価できた.
• 培養中のBabesia gibsoniに対するArcangelisia flavaから得られたprotoberberine alkaloidsと20-hydroxyecdysoneの抗バベシア活性(短報)(内科学)

  スベキ, 松浦 英幸, 高橋 公咲, 山崎 真大, 大和 修, 前出 吉光, 片倉 賢, 鈴木 守, トリムルニンシ, カイルー, 吉原 照彦 The journal of veterinary medical science 67 (2) 223 -227 2005年02月 [査読無し][通常論文]
   

  抗バベシア活性を指標として,Arcangelisia flaveから5活性化合物が単離された.planatube, berberine, jatororrhizine及びdihydroberbrineはIn vitroにおいて1.0μg/mlでBabesia gibsoniに対する成長を抑制した.20-hydroxyecdysonの成長抑制濃度は100μg/mlという低い感受性であった.
• シロイヌナズナ恒常的細胞死変異体cad1の単離と解析

  山室千鶴子, 筒井友和, 浅田裕, 吉岡博文, 玉置雅紀, 小川大輔, 松浦英幸, 吉原照彦, 池田亮 日本植物生理学会年会要旨集 46th 2005年
• 赤血球内バベシア原虫に対する中央カリマンタン植物抽出物の効果(短報)(内科学)

  スベキ スベキ, 松浦 英幸, 山崎 真大, 大和 修, 前出 吉光, 片倉 賢, 鈴木 守, トリムルニシン トリムルニシン, カイルル カイルル, 吉原 照彦 The journal of veterinary medical science 66 (7) 871 -874 2004年07月 [査読無し][通常論文]
   

  インドネシア,中央カリマンタンより採取した45種類の植物から熱湯抽出物を得た.これらの抽出物について,in vitroにおけるBabesia gibsoniの増殖抑制効果を測定し,また,マウスに対する急性毒性作用を観察した.これら45種類の植物から得られた抽出物のうち,Arcangelisia flava, Curcuma zedoaria, Garcinia benthamiana, Lansium domesticum,ならびにPeronema canescensに非常に高い増殖抑制が観察され,IC_<50>は5.3μg/mlから49.3μg/mlであると判明した.また,上記植物の抽出物を体重1kgにつき0.7gを腹膜投与したがマウスに対する急性毒性は観察されなかった.
• インドネシア産植物における抗Babesia gibson活性物質の探索

  野村信吉, SUBEKI, 松浦英幸, 山崎真大, 大和修, 片倉賢, 鈴木守, TRIMURNINGSIH, CHAIRUL 日本農芸化学会大会講演要旨集 2004 283 2004年03月05日 [査読無し][通常論文]
  
• インドネシア産薬用植物Curcuma zedoariaにおける抗バベシア・マラリア活性化合物の探索

  笠原堅, 野村信吉, SUBEKI, 松浦英幸, 山崎真大, 大和修, 前出吉光, 片倉賢, 鈴木守, 小林寿美子, TRIMURNINGSIH, CHAIRUL, 吉原照彦 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨 2004 27 2004年 [査読無し][通常論文]
  
• 36 Infuluence of theoroXide,a natural Product, on the accumulation of endogegenous jasmonoids in in vitro cultures of potato single-node segment and morning glory leaves

  Qing Yang, Matsuura Hideyuki, Fujino Yumiko, Yoshihara Teruhiko 植物化学調節学会研究発表記録集 (36) 83 -84 2001年10月09日 

  Theobroxide is an epoxy cyclohexene compound isolated from the fungus Lasiodiplodia theobromae and induces potato microtuber formation in vitro as low as 5x10^<-6>M (Nakamori et al., 1994). When sprayed on potato (Solanum tuberosum L.) and morning glory (Pharbitis nil) plants, which require short days to induce tubers and flower buds, respectively, these plants kept in non-inducing conditions (long days) produced corresponded tubers and flower buds (Yoshihara et al., 2000). However, the action mechanism of theobroxide is still not clear. The examination of the hypothesis that theobroxide may play an important role in the accumulation of endogenous jasmonoids was carried out in this study. In vitro bioassay using cultures of single-node segment of potato stems was performed with the supplement of theobroxide in the medium at different concentrations, the endogenous jasmonic acid (JA), tuberonic acid (TA, 12-hydroxy jasmonic acid) and its glucoside (TAG) in segments, stolons and microtubers were respectively quantitative analyzed. As concern as morning glory, plant seedlings were sprayed with theobroxide solution at a two-day interval, the endogenous JA and IA in the leaves were analyzed. An increase in the content of endogenous JA correlated with the concentration of theobroxide was observed in both segments and microtubers. Endogenous TA was only detected in segments, and the content increased with the concentration of theobroxide. The increase in the content of endogenous TAG caused by the treatment of theobroxide was also observed in microtubers. However, this trend was not found in stolons and segments. It was also found that spraying theobroxide on morning glory plants for 2-3 weeks under short and long days respectively increased 1.6 times and 2.3 times of JA in leaves. The theobroxide spray under short days caused 1.5 times increase in TA level in all treatments.
• フキノトウ(Petasites japonicus)からの抗酸化成分の単離

  松浦英幸, 天野みどり, 川端潤, 水谷純也 日本農芸化学会誌 74 (11) 1314 -1315 2000年11月01日 [査読無し][通常論文]
  
• ニガウリ(Momordica charantia L.)からのα‐Glucosidase阻害物質の単離

  松浦英幸, 浅川智香子, 栗本成敬, 川端潤, 水谷純也 日本農芸化学会誌 74 (11) 1314 2000年11月01日 [査読無し][通常論文]
  
• ニガウリ(Momordica charantia L.)からのα‐glucosidase阻害物質の単離

  松浦英幸, 浅川智香子, 栗本成敬, 川端潤, 水谷純也 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨 2000 12 2000年07月21日 [査読無し][通常論文]
  
• フキノトウ(Petasites japonicus)からの抗酸化成分の単離

  松浦英幸, 天野みどり, 川端潤, 水谷純也 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨 2000 13 2000年07月21日 [査読無し][通常論文]
  
• 23 バレイショ葉におけるジャスモン酸の代謝

  松浦 英幸, 大久保 泰宏, 吉原 照彦, 市原 耿民 植物化学調節学会研究発表記録集 0 (32) 51 -52 1997年11月28日 [査読無し][通常論文]
  
• 40 セオブロキシドは長日条件下アサガオの花芽形成を誘導する

  吉原 照彦, 松浦 英幸, 中森 薫, 大森 史裕, 天沼 美知子, 堤 崇史, 市原 耿民 植物化学調節学会研究発表記録集 (31) 81 -81 1996年10月04日
• 微生物由来のバレイショマイクロチューバー形成物質の検索 : 有機化学・天然物化学

  Omer E.A., 畠山 千明, 堤 崇史, 天沼 美和子, 松浦 英幸, 吉原 昭彦, 市原 耿民, 佐藤 豊三 日本農藝化學會誌 70 60 -60 1996年03月05日
• 38 植物体に於ける標識ジャスモン酸の代謝・移動

  吉原 照彦, 天沼 美和子, 松浦 英幸, 堤 崇史, 市原 耿民 植物化学調節学会研究発表記録集 (28) 71 -71 1993年10月15日
• A NOVEL SESQUITERPENE, TUBERONONE, FROM SOLANUM-TUBEROSUM L

  H MATSUURA, T YOSHIHARA, A ICHIHARA BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 56 (11) 1890 -1891 1992年11月 [査読無し][通常論文]
  
• 16.Lasiodiplodia theobromaeの生産するバレイショ塊茎形成誘導物質 (第26回大会研究発表抄録)

  吉原 照彦, 中森 薫, 松浦 英幸, 市原 耿民, 幸田 泰則, 喜久田 嘉郎 植物の化学調節 26 (2) 221 -221 1991年12月25日
• 16 Lasiodiplodia theobromaeの生産するバレイショ塊茎形成誘導物質

  吉原 照彦, 中森 薫, 松浦 英幸, 市原 耿民, 幸田 泰則, 喜久田 嘉郎 植物化学調節学会研究発表記録集 (3) 33 -34 1991年10月14日

所属学協会

• 植物化学調節学会   日本農芸化学会   

共同研究・競争的資金等の研究課題

• シロイヌナズナ、ビート、ダイコン、の抽台抑制機構の解明と抽台抑制剤の開発

  日本学術振興会：科学研究費助成事業

  研究期間 : 2022年04月 -2025年03月 

  代表者 : 松浦 英幸, 北岡 直樹, 久保 友彦, 黒田 洋輔, 柏木 純一, 北崎 一義, 橋本 誠
• 植物内生シグナル物質としての脂肪酸ーアミノ酸縮合体の機能解析

  日本学術振興会：科学研究費助成事業

  研究期間 : 2021年04月 -2024年03月 

  代表者 : 高橋 公咲, 松浦 英幸

   

  シロイヌナズナ、ナス、トマト、トウモロコシおよびダイズを80％メタノールで抽出し、固相抽出カラムで前処理し夾雑物を除去した。本前処理で得られたサンプルをUPLC-MS/MSで分析し、代表的な脂肪酸-アミノ酸縮合体であるN-リノレノイルグルタミンが植物内に存在するのか否かを検討した。その結果、分析したいずれの植物にもN-リノレノイルグルタミンが存在することが判明した。N-リノレノイルグルタミンは、幼虫の唾液中だけではなく植物にも普遍的に存在することが示唆された。しかし、その内生量は植物により大きく異なっていた。 GH3タンパク質ファミリーの中には、カルボキシル基を持つ植物ホルモンの3-インドール酢酸、ジャスモン酸およびサリチル酸とアミノ酸を縮合するタンパク質が存在する。グルタミンと3-インドール酪酸を縮合することが報告されているシロイヌナズナのGH3.15タンパク質の組換えタンパク質を大腸菌で生産した。本タンパク質のα-リノレン酸とグルタミンの縮合活性を評価した。その結果、GH3.15タンパク質はα-リノレン酸とグルタミンの縮合活性を示した。この結果は、植物にN-リノレノイルグルタミンが存在することを支持する結果であった。さらに、α-リノレン酸を一方の基質とし、GH3.15タンパク質に対する20種のアミノ酸の基質特異性を調べた。その結果、20種すべてのアミノ酸が本反応の基質となりうることが明らかとなった。それらの中でグルタミンは4番目に高い基質特異性を示すアミノ酸であった。 N-リノレノイルグルタミンのシロイヌナズナの生育に対する影響を調べたところ、本化合物は100μMの濃度でシロイヌナズナの根の伸長を抑制するとともに、根を垂直ではなく、斜めに伸長させることが明らかとなった。
• なぜ植物は共生菌を受容するか？相利共生構築に寄与する化学コミュニケーションの解析

  日本学術振興会：科学研究費助成事業 新学術領域研究(研究領域提案型)

  研究期間 : 2020年04月 -2022年03月 

  代表者 : 松浦 英幸

   

  植物の成長に有益な効果をもたらし、植物と共生関係にある内生菌（エンドファイト、endophyte）の存在が明らかとされ、様々な研究がなされてきた。その一例として、共生菌の一種、Veronaeopsis simplexと共生関係を結んだトマトは、本来トマト幼苗が生育できない低pH条件でも通常の生育を示す。本菌は培地上で暗色(Dark)のコロニーを形成し比較的生育が遅く、菌糸に隔壁(Septa)があることから、Dark Septate Endophytic fungus (DSE)に属する菌として知られており、根部に感染し植物に対して有益な効果を示す。他のDSE菌、 Heteroconium chaetosnira, Phialocephala fortiniiの存在も知られ、その効果が実証されている。しかしながら、『どうしてエンドファイトは植物に排除されずに感染できるか？』、『共生したエンドファイトは植物に如何にして劣悪環境突破力をあたえるのか？』については未だ不明なところが多い。本研究ではP. fortiniiの培養濾液より新規な化合物、8-hydroxy-6-methoxy-3,7-dimethyl-1H-2-benzopyrpyran-1-oneを単離し、植物に対する活性として内生のABA含量を増加させる生物活性を見出した。また、我々が主題として取り組む糸状菌由来の生理活性物質、セオブロキシドに、内生菌の感染量を増加させる活性を見出した。感染量を増加させる生理活性の発現には、内生のABA含量増加に伴う、PR1遺伝子の転写量の低下が起因しているであろうとの結果を得た。また、植物と内生菌が共生関係を結ぶ際に重要な役割が想定されているジャスモン酸の類縁体の生合成に関して、新たな知見を得た。
• 新たな農資源としての醸造後の酵母資源の利用とその生理活性発現機構の解明

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2019年07月 -2022年03月 

  代表者 : 松浦 英幸, 柏木 純一

   

  地球上の埋蔵エネルギーには限りがあり、未利用バイオマス資源の有効活用は人類永続の一助となる。本資源の有効活用の観点から、これらを原料とした多く農業資材が市販され、利用されている。しかし、学術的な裏付けが乏しく、その使用が一部に限られている。大量に産出される未利用バイオマスの一事例として、ビール、日本酒、ブドウ酒、焼酎などの醸造過程の副産物があげられる。本課題ではワイン酒醸造後の残渣種子を中心に研究を行い、バレイショ増収効果に期待がもたれる資材であるとこを報告する。本研究の成果は資源循環型の社会構築に向けた研究事例のモデルケースとなる。
• 葉部より転流する生理活性物質による植物の抽だい抑制機構の解明と抽だい抑制剤の開発

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2018年04月 -2021年03月 

  代表者 : 松浦 英幸, 三井 裕樹, 柏木 純一

   

  植物の生態型には一年生、冬一年生および多年生などがある。冬一年生植物は夏に発芽し、越冬後の春に抽だい、花成誘導を行う。よって、夏から秋の栄養成長期間中の抽だいは厳密に抑制されなければならない。植物の抽だい抑制機構に関しては低分子生理活性物質の関与が示唆されているが、長距離シグナルとしての実態や移動先での作用メカニズムは不明な点が多い。本研究では葉部で生合成された生理活性物質の茎頂点への移動と移動先での生理活性を明らかとし、環境条件によって制御される植物の抽だい抑制機構を明らかとすることを目的とした。 ダイコン葉部より、2,3-dihydroxypropyl (7Z,10Z,13Z)-hexadeca-7,10,13-trienoateが抑制物質として単離されている。活性に必要な官能基を探るために構造活性相関研究を行なった。材料にはシロイヌナズナを用い、播種後の植物の伸長抑制効果を指標とした。この結果、母骨格に結合する炭素鎖に関して、1箇所でも二重結合があれば活性を有し、cis,trans体のいずれでも活性を有することが明らかとなった。他器官への移動を検証するための重水素ラベル体についてはラベルパターン異なる化合物を合成できた。 また、ジベレリンに対する生合成抑制効果についてはその内生量をUPLC MS/MSによって測定した。この結果、抽台抑制物質に晒されたシロイヌナズナでは全体的なジベレリン含量が下がっていることが明らかとなった。本結果は抽台抑制化合物のターゲットはジベレリン生合成の初期段階を抑制していることが示唆された。さらにプランターで育てた大根（品種:耐病そう太り）に2,3-dihydroxypropyl (7Z,10Z,13Z)-hexadeca-7,10,13-trienoateを一定期間葉面散布したところ、無処理の植物に比べて、抽台時期の遅延効果が示された。
• 褐藻切断組織の組織癒合過程におけるオーキシン調節作用の解明

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2017年04月 -2020年03月 

  代表者 : 田中 厚子, 松浦 英幸

   

  本研究は、動物や陸上植物とは異なる進化の過程で，独自に多細胞体制を獲得した褐藻類の傷害応答機構に注目し，傷害応答における植物ホルモンの役割を明ら かにすることを目標として掲げている。具体的には、褐藻アミジグサで既にオーキシンによる促進作用が確認されている傷害組織の癒合に焦点を当て、オーキシ ンの合成から組織内の移動、作用機序までの各段階を形態学的・分子生物学的アプローチを用いて解明する予定である。 本年度は３つのサブプロジェクトを進めた。一つ目は傷害の形状や位置における生長促進作用と側芽形成数の比較から、傷害シグナルが作用する組織と傷害との位置関係を考察した。傷害の形状は２型を比較したところ、その生長促進作用に違いが見られた。さらに傷害応答の際には、生長促進作用と側芽形成との間にトレードオフの関係が生じることが明らかとなった。この結果は日本藻類学会第42回大会（東北大学）にて発表された。二つ目は傷害応答におけるトランスクリプトームの再解析を行い、傷害で活性化する遺伝子は抑制される遺伝子の10分の１であることが判明した。さらに遺伝子オントロジー解析から特に傷害によって発現が促進される遺伝子群についての機能同定を試みた。三つ目として、オーキシンを含む植物ホルモンが潜在的に褐藻類、特に本研究の研究対象であるアミジグサ類に含まれるか否かについて検証した。アミジグサ類は沖縄を中心に亜熱帯地域に多くの種類が生息するため、26個体の天然アミジグサ類藻体についてUPLC-MS/MSを用いた解析を行った。その結果、特定の植物ホルモンについてその存在が示された。さらに分子系統解析を用いて正確なアミジグサ類の種同定を行ったところ、日本新産種と思われる数種を発見した。現在はこれらの知見を投稿論文としてまとめている。
• 新規標的タンパク質同定法を用いた植物シグナル伝達物質受容体の同定

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2017年04月 -2020年03月 

  代表者 : 高橋 公咲, 松浦 英幸

   

  モデル実験により、生理活性物質にアジド基を有するプローブ（アジドプローブ）と、アミノ基、アルキンおよびビオチンを有するリンカーを用いて標的タンパクを同定する方法が有効であることが示された。そこで本手法を用いて、植物生長促進活性を有するアザヒポキサンチン（AHX）の結合タンパク質の同定を試みた。 モデル植物であるイネのタンパク質抽出液にアジド基を導入したAHX（N3-AHX）を添加し、タンパク質とN3-AHXを十分に相互作用させた。タンパク質とN3-AHXの複合体は、リンカーを介してストレプトアビジン固定化レジンに結合させた。非特異的にレジンに結合するタンパク質を十分に洗浄した後に、N3-AHXと結合するタンパク質を選択的に溶出させた画分（Fr.1）が得られた。イネのタンパク質抽出液に過剰のAHXを添加し、Fr.1を得た方法と同様な方法で得られた画分をFr.2とした。イネのタンパク質抽出液を何も結合させていないストレプトアビジン固定化レジンに供した後、本レジンに非特異的に結合するタンパク質が溶出がされた画分をFr.3とした。この3つの画分をそれぞれnano-LC-MS/MSに供し、各画分に含まれるタンパク質を検出した。Fr.1にのみ含まれ、Fr.2およびFr.3には含まれないタンパク質が14個あり、これらのタンパク質がAHXに結合する候補タンパク質と推定された。その中で、Os07t0112800（Similar to Eukaryotic translation initiation factor 5A-4 (eIF-5A-4)）が、ウェスタンブロッッティングによりAHXとの結合が確認された。本タンパク質はAHX結合タンパク質として有望であることが示された。
• 『トマトの生きる知恵』の生物有機化学的解明

  日本学術振興会：科学研究費助成事業 挑戦的萌芽研究

  研究期間 : 2015年04月 -2017年03月 

  代表者 : 松浦 英幸

   

  申請者は研究期間内に、下記の2点を明らかにすることを目的とした。1】トマト果実内に存在する胎生発芽抑制物質の化学構造とその生理機能を明らかとする。2】トマトの原種、Solanum pennelliiの植物体を覆う、粘性物質の化学構造および展着剤としての可能性を明らかとする。項目1に関しては、種子を含むロキールの抽出物に発芽抑制活性は確認できたものの、活性物質の単離、同定には至らなかった。項目2に関しては、粘性物質の本体であるacylglucoseの単離精製方法を考案し、その化学構造を明らかとすることができた。また、その粘性を利用した農業資材の利用法に関して言及できた。
• 前向き出生コホート研究による化学物質の次世代影響並びに環境遺伝交互作用の解明

  日本学術振興会：科学研究費助成事業 基盤研究(A)

  研究期間 : 2013年04月 -2016年03月 

  代表者 : 岸 玲子, 池野 多美子, 荒木 敦子, 宮下 ちひろ, 中島 そのみ, 佐々木 成子, 花岡 知之, 梶原 淳睦, 松浦 英幸, 室橋 春光, 白石 秀明, 佐田 文宏, 西條 泰明, 吉岡 英治, 安住 薫, 安住 薫

   

  母児20000組を対象とした北海道内40産院コホートで、妊娠中の化学物質、特に喫煙・ダイオキシン類・有機フッ素化合物に対する曝露と、学童期までの児の発育、神経行動発達および免疫アレルギーとの関連を検討した。妊娠中の化学物質曝露が(1) PPARβ遺伝子多型を介して血清脂質に、(2) 母のDNA修復（XRCC1）遺伝子多型を介して児の出生体重に、(3) 児のIGF2のDNAメチル化に、そして(4) 1.5歳児の神経行動発達に影響することを明らかにした。加えて、学童期のADHDに関するリスク要因が検討され、妊娠中の飲酒や喫煙がリスクとなることが明らかにされた。
• 植物ホルモンの新規な代謝酵素の同定と機能解析

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2012年04月 -2015年03月 

  代表者 : 松井 博和, 佐分 利亘, 松浦 英幸

   

  ジャスモン酸 (JA) は病傷害ストレス応答，生長などに関与する植物ホルモンである．本研究ではJAシグナルの不活性化に働くと推定したCYP94D1，CYP94D2およびILL6の機能解析を行った．CYP94D1については遺伝子発現量の調整により表現型の変化が認められた．すなわち，遺伝子欠損株ではJAへの感受性の低下，過剰発現体ではJAの水酸化物の誘導体ツベロン酸グルコシドの蓄積が見られた．
• イネ由来新規糖転移酵素の探索とジャスモン酸の不活性化機構の解明

  日本学術振興会：科学研究費助成事業 挑戦的萌芽研究

  研究期間 : 2012年04月 -2015年03月 

  代表者 : 松浦 英幸

   

  植物の傷害応答機構、特にジャスモン酸(JA)シグナル伝達機構の不活性化に寄与すると想定される配糖化酵素として、OsBGlu1を同定した。種々のJA類を用いてイネ幼苗の根の伸長阻害試験を行った。この結果、JAが最も伸長阻害を示し、12-hydroxyjasmonic acid および12-hydrohyjasmonoyl L-isoleucine配糖体がほとんど伸長阻害を示さなかった。以上の事から生理活性を示すJAの重要な代謝経路の一つとして、OsBGlu1が介する糖転位反応を証明できたと考える。
• ミャンマーとタイ北部における家畜寄生虫症の分子疫学と抗寄生虫薬用植物の探索

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2010年04月 -2014年03月 

  代表者 : 片倉 賢, 櫻井 達也, 宮崎 智史, 加藤 大智, 松浦 英幸, 板垣 匡, 佐々木 均

   

  家畜の寄生虫としてバベシア、タイレリア、トリパノソーマ、トキソプラズマ、ネオスポラおよび肝蛭に焦点をあてた疫学調査をミャンマーの各都市で実施した。その結果、これらの寄生虫の分布、生活環、進化に関する新知見がえられた。また、アジア産薬用植物を寄生虫症の予防や治療に地産地消的観点から有効活用することを目的として、ミャンマー産薬用植物から抗トリパノソーマ活性物質の分離・精製を試みた。本研究の成果は、ミャンマーにおいて家畜寄生虫症対策を立案するうえで有用な基礎情報を提供し、同国の農畜産業の発展に寄与することが期待される。
• スイカ果実に含まれるスイカ種子発芽抑制物質の有機化学的研究

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2009年 -2011年 

  代表者 : 松浦 英幸

   

  植物は大地に根を張って生きねばならない故、環境要因が悪化したからといって逃げ出す事はできない。よって、植物は生育環境に応じて発芽、生育，開花、結実の行程を踏み、環境要因に呼応して次の生育ステージへ移行する。植物の生活環制御は興味に尽きない。ここで、植物に含まれる低分子生理活性物質に目を向けた場合、植物ホルモンと呼ばれる一群の生理活性物質が知られている。種子発芽の抑制もしくは促進、栄養生長時に於ける大盛な生長、栄養生長にブレーキを踏み生殖生長(老化)に切り替える働き等、植物ホルモンが関与している事例が多く知られ、鋭意研究が行われている。しかしながら多くの研究者が植物の生活環制御の不思議さ，巧妙さに興味を抱きつつ、研究がなされてきたが、「種子の発芽は果実中に存在する限り起こりえない」と云う事例に目を向けた研究は未だに行われていない。この事象についてスイカ果実を実験材料として用い、植物ホルモンの一種であるアブシシン酸が主たる活性物質である事を明らかとした。本年度は他の植物を用いて上記の生物現象を検討した。数種の植物を用いて検証したところ、果実と種子が直接接しているような植物においては化学物質を介した果実中での発芽抑制機構が存在する事が示唆された。そこで、発芽抑制の確認されたトマト(Solanum lycopericum)を用いて、果実中の発芽抑制物質の単離を試みた。今の所、単離、精製を試みている段階であり、生理活性物質の実態は不明である。しかしながら、精製を進めた段階に於いてもトマト種子発芽抑制活性を有する事から数ステップの精製操作により目的化合物の取得が可能と思われる。
• コケ植物におけるジャスモン酸の生合成及び生理機能の解明

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2009年 -2011年 

  代表者 : 高橋 公咲, 松浦 英幸, 鍋田 憲助

   

  ヒメツリガネゴケ(Physcomitrella patens)におけるオクタデカノイド経路中(ジャスモン酸生合成)の各種酵素の活性を調べた。その結果、いずれの酵素も高等植物と同様の酵素活性が示された。ヒメツリガネゴケの生育においては、ジャスモン酸(JA)がほとんど影響を与えなかったのに対して、その生合成中間体である12-オキソファイトジエン酸(OPDA)は生育を阻害した。オクタデカノイド経路に関わる酵素遺伝子の変異株を作製したが、それらの表現型は野生株と大きな差異は無かった。本研究の結果から、ヒメツリガネゴケではJA ではなくOPDA が活性化合物として機能していることが示唆された。
• 根圏有機物シンク形成に連動する根圏生物複合系の高度負荷環境緩和機構の検証と利用

  日本学術振興会：科学研究費助成事業 基盤研究(A)

  研究期間 : 2008年 -2011年 

  代表者 : 橋床 泰之, 原口 昭, 木村 淳夫, 崎浜 靖子, 玉井 裕, 松浦 英幸

   

  インドネシア・カリマンタンの負荷土壌に対処できる根圏微生物コンソーシアムについて検討した。中強酸性泥炭湿地に自生するチガヤ根面から分離したBurkholderia属細菌A-KA株にSphingomonas属など根圏微生物の増殖を促進する活性が認められ,また,酸性硫酸塩土壌耐性イネ根から分離したBurkholderia mimosarumにはイネ苗立枯細菌病を引き起こすBurkholderia plantarii々のトロポロン産生を抑え,種子付着菌を根圏で増殖させる能力を示した。熱帯のポリフェノールリッチな土壌とフタバガキ幼木根圏から分離した細菌群からは,ピロガロール基をもつポリフェノール存在でインドール分解活性が急激に活性化されるBurkholderia属細菌を見いだした。共生系では,ソテツ根と窒素固定性シアノバクテリアNostoc punchiformeとのサンゴ状根共生において,シアノバクテリア誘引に必要な運動性付与がジアシルグリセロールの一分子種で起こることを明らかにした。また,宿主の土壌伝播病原菌感染を抑制する根圏細菌Psuedomonas jessenii EC-S101株やTrichoderma sp. PS1-7株の放出するシグナル物質の化学構造とその機能性を解明した。根圏での脱窒制御に関しては,熱帯泥炭地の亜酸化窒素生成菌では亜酸化窒素還元酵素(N2OR)遺伝子が欠損しており,また,様々な植物二次代謝産物が根圏での脱窒能に影響を与えることが明らかにされた。
• 高度負可土壌耐性植物がもつ根圏複合系の機能性解析とアグロテクノロジーへの応用研究

  日本学術振興会：科学研究費助成事業 基盤研究(A)

  研究期間 : 2004年 -2007年 

  代表者 : 橋床 泰之, 玉井 裕, 原口 昭, 松浦 英幸, 岩橋 均, 俵谷 圭太郎, 佐野 雄三

   

  インドネシア熱帯泥炭林荒廃地の滅菌現地土壤(ペースト状の木質泥炭)におけるX.comp1anataの種子発芽はPenicillium属糸状菌Y-1株によってのみ強力に促進されたが、発芽種子は子葉の伸長以降は成長できず、1ヶ月後には真菌がまとわりついた様に立ち枯れて死に絶えてしまった。とこ芽種子は極めて良好な生育を6ヶ月以上持続したため、土壌生/種子付着性真菌のX.comp1anata発芽ステージでの発芽亢進機能とそれに引き続く土壌細菌のX.comp1anata芽生え生育ほう助機能とが明確に分担されていることが強く示唆された。 シベリア永久凍土のタイガ林林床土壌では、低栄養要求性の単生窒素固定性Burkholderia属ならびにPseudomonas属細菌が主要な菌相をなし、細菌群集は0.02〜O.05%のマンニトールで十分に1%炭素源以上のアセチレン還元活性を示すことが分かった。また、表層に近い腐植成分を多く含む層から得た土壌微生物群集の窒素固定能力は10〜15℃で最大限に発揮されること、中一アルカリ性(pH 6.0以上)では極端に抑えられ、弱酸性(pH 5.0付近)では極めて高い能力が発揮されることから、現地負荷環境条件の再現が当該微生物コンソーシアムの機能性を最大限に発揮させると結論付けた。 2000年に噴火した有珠山の火山性荒廃地における先駆性植物の外生菌根(ECM)およびアーバスキュラー菌根(AM)共生について調査した。荒廃地に優占的に更新するヤナギ類3種、ドロノキ(Populusmaximowiczii)、エゾノバッコヤナギ(Salix hultenii var. angustifolia)およびオノエヤナギ(Salix sachalinensis)は定常的にECMを形成しており、全体での外生菌根形成率は、17〜42%であった。草本ではスギナとオオイタドリ以外の10種は全てAMを形成していた。植生再生の初期過程においては、菌根共生が植生再生と根圏土壌環境負荷緩和に先駆的役割を果たしていることが示された。
• 種子へのセオブロキシド処理による低温障害抑制効果の有機化学的・分子生物学的解析

  日本学術振興会：科学研究費助成事業 萌芽研究

  研究期間 : 2005年 -2006年 

  代表者 : 松浦 英幸, 高橋 公咲

   

  Theobroxide(1mM水溶液、1粒/40ml)をカウピー(Vigna unguiculata)播種時の苗床に施し、播種約10日後、傷害応答シグナルとして認知されているsalicyhc acid(SA),salicylic acid glucoside(SAG)の幼植物内での含有量を測定した。Theobroxide処理の植物においてこれらの化合物の有意な蓄積量の上昇が確認された。傷害耐性付与が期待できたので、断水処理、キュウリモザイクウィルスの接種ならびにダイズ茎疫病菌の接種、それぞれ3つの処理を行った。これらの3種のストレスに対してtheobroxide処理の植物は無処理の植物と比較し、有意に耐性効果が観察された。以上の結果からtheobroxideの病傷害耐性付与効果を有する農業資材としての可能性が示唆された。また、ATP生産に関わる遺伝子のうち、ATP消費を促進する遺伝子(AOX)について検討したところ、theobroxide処理の植物ではコントロールに比べ有意に抑制されていることが解り、エネルギー節約型の植物、倹約型を導いているのではないかと示唆された。また、アサガオやホウレンソウを用いてtheobroxide効果を検討したところ植物ホルモンとして知られているジベレリンに対して何らかの阻害作用を示しているデータが得られた。
• 花芽形成調節剤の開発研究

  日本学術振興会：科学研究費助成事業 基盤研究(A)

  研究期間 : 2001年 -2003年 

  代表者 : 吉原 照彦, 松浦 英幸, 田原 哲士

   

  1.セオブロキシド処理による花芽形成誘導 すでに効果が明らかになっているアサガオ以外の植物の花芽形成誘導について調べた。1mMセオブロキシド水溶液を2日に1回植物葉に散布した。その結果、短日性植物であるイネ、イチゴ、ダイズ、ポインセチアの花芽形成誘導に効果があった。しかし、イチゴ、ダイズでは品種間の差異が大きかった。 2.セオブロキシドの処理による塊茎形成誘導 実験室レベルでバレイショにセオブロキシド水溶液を散布することにより塊茎形成が誘導されるが、圃場実験でも20%の収量の増大が認められた。さらに、バレイショの病害抵抗性の付与にも関与することが明らかになり、病害防除とともに植物化学調節剤としての実用化の可能性が大きくなった。 3.セオブロキシド処理による抽苔抑制 長日条件で抽苔するホウレンソウにセオブロキシド水溶液を散布したところ、長日で抽苔を抑制する効果が認められた。植物化学調節剤としての実用化とともに抽苔現象解明のプローブとして重要な役割を持つ化合物であることが明らかとなった。 4.セオブロキシドの作用機作 リポキシゲナーゼ阻害剤サリシルヒドロキサミン酸を用いた実験により、セオブロキシドの作用機作はリポキシゲナーゼ活性を高めることが明らかになった。このことによりセオブロキシドによる塊茎と花芽形成は、リノレン酸酸化物の生成が促進され、その酸化物により両者の誘導が行われるとの結論に達した。 5.セオブロキシド製造特許 醗酵法および合成法によるセオブロキシドの製造法を開発し、特許を申請した。
• 植物の日長応答と内生機能物質の生理・有機化学的研究

  日本学術振興会：科学研究費助成事業 特別研究員奨励費

  研究期間 : 1998年 -1998年 

  代表者 : 松浦 英幸

産業財産権

• 特開2008-211993:形質転換されたダイズ及びその製造方法。  2008年09月18日

  金澤 章, 松浦 英幸, 喜多村 啓介, 阿部 純, 増田 税  国立大学法人 北海道大学  200903043089823192
• 特開2007-223962:抗バベシア剤および新規クアシノイド誘導体  2007年09月06日

  スベキ, 前出 吉光, 片倉 賢, 松浦 英幸, 鍋田 憲助  国立大学法人 北海道大学  200903090213774845
• 特許第3990387号:ヒソップ抽出物の血液流動性改善剤、血流改善剤およびその利用  2007年07月27日

  山岸 喬, 船木 稔, 松浦 英幸, 石原 智明, 宮崎 浩之, 金澤 勉  独立行政法人科学技術振興機構  200903055622549217
• 特開2007-051068:植物用矮化剤および植物の矮化方法  2007年03月01日

  吉原 照彦, 松浦 英幸, 高橋 公咲  国立大学法人 北海道大学  200903013800644739
• 特開2006-327995:植物用自己防御物質誘導剤および自己防御物質の生成誘導方法  2006年12月07日

  吉原 照彦, 松浦 英幸, 高橋 公咲  国立大学法人 北海道大学  200903098147387437
• 特開2006-219456:抗バベシア剤および新規ベンズオキサゾリノン誘導体  2006年08月24日

  吉原 照彦, 前出 吉光, 片倉 賢, 松浦 英幸  国立大学法人 北海道大学  200903069907900270
• 特開2006-028046:植物の抽だい誘導抑制及び/又は花芽分化抑制剤  2006年02月02日

  吉原 照彦, 松浦 英幸, 高橋 公咲  国立大学法人 北海道大学  200903072376529823
• 特開2006-008555:オレガノ抽出物のα−グルコシダーゼ阻害剤およびその利用  2006年01月12日

  山岸 喬, 船木 稔, 松浦 英幸, 石原 智明, 金澤 勉, 宮崎 浩之  独立行政法人科学技術振興機構  200903099934377941
• 特開2005-314345:植物抽出抗バベシア剤  2005年11月10日

  吉原 照彦, 松浦 英幸, スベキ, 前出 吉光, 片倉 賢, 野村 信吉  独立行政法人科学技術振興機構  200903079153984418