Nomura T

References (28)

Title : Synthesis of Carlactone Derivatives to Develop a Novel Inhibitor of Strigolactone Biosynthesis - Kawada_2023_ACS.Omega_8_13855
Author(s) : Kawada K , Saito T , Onoda S , Inayama T , Takahashi I , Seto Y , Nomura T , Sasaki Y , Asami T , Yajima S , Ito S
Ref : ACS Omega , 8 :13855 , 2023
Abstract : Strigolactones (SLs), phytohormones that inhibit shoot branching in plants, promote the germination of root-parasitic plants, such as Striga spp. and Orobanche spp., which drastically reduces the crop yield. Therefore, reducing SL production via chemical treatment may increase the crop yield. To design specific inhibitors, it is valid to utilize the substrate structure of the target proteins as lead compounds. In this study, we focused on Os900, a rice enzyme that oxidizes the SL precursor carlactone (CL) to 4-deoxyorobanchol (4DO), and synthesized 10 CL derivatives. The effects of the synthesized CL derivatives on SL biosynthesis were evaluated by the Os900 enzyme assay in vitro and by measuring 4DO levels in rice root exudates. We identified some CL derivatives that inhibited SL biosynthesis in vitro and in vivo.
ESTHER : Kawada_2023_ACS.Omega_8_13855
PubMedSearch : Kawada_2023_ACS.Omega_8_13855
PubMedID: 37091382

Title : Unveiling the complexity of strigolactones: Exploring structural diversity, biosynthesis pathways and signaling mechanisms - Nomura_2023_J.Exp.Bot__
Author(s) : Nomura T , Seto Y , Kyozuka J
Ref : J Exp Bot , : , 2023
Abstract : Strigolactone (SL) is the collective name for compounds containing a butenolide as a part of their structure, first discovered as compounds that induce seed germination of root parasitic plants. They were later found to be rhizosphere signaling molecules that induce hyphal branching of arbuscular mycorrhizal (AM) fungi, and finally, they emerged as a class of plant hormones. SLs are found in root exudates, where they display a great variability in their chemical structure. Their structure varies among plant species, and multiple SLs can exist in one species. Over 30 SLs have been identified, yet the chemical structure of the SL that functions as an endogenous hormone and is found in the above-ground parts of plants remains unknown. We discuss our current knowledge of the synthetic pathways of diverse SLs and their regulation as well as recent progresses in identifying SLs as plant hormones. SL is perceived by the D14 receptor, an alpha/beta hydrolase which originated by gene duplication of KARRIKIN INSENSITIVE 2 (KAI2). D14 and KAI2 signaling pathways are partially overlapping paralogous pathways. Progress in understanding the signaling mechanisms mediated by two alpha/beta hydrolase receptors as well as remaining challenges in the field of SL research are reviewed.
ESTHER : Nomura_2023_J.Exp.Bot__
PubMedSearch : Nomura_2023_J.Exp.Bot__
PubMedID: 37877933

Title : An ancestral function of strigolactones as symbiotic rhizosphere signals - Kodama_2022_Nat.Commun_13_3974
Author(s) : Kodama K , Rich MK , Yoda A , Shimazaki S , Xie X , Akiyama K , Mizuno Y , Komatsu A , Luo Y , Suzuki H , Kameoka H , Libourel C , Keller J , Sakakibara K , Nishiyama T , Nakagawa T , Mashiguchi K , Uchida K , Yoneyama K , Tanaka Y , Yamaguchi S , Shimamura M , Delaux PM , Nomura T , Kyozuka J
Ref : Nat Commun , 13 :3974 , 2022
Abstract : In flowering plants, strigolactones (SLs) have dual functions as hormones that regulate growth and development, and as rhizosphere signaling molecules that induce symbiosis with arbuscular mycorrhizal (AM) fungi. Here, we report the identification of bryosymbiol (BSB), an SL from the bryophyte Marchantia paleacea. BSB is also found in vascular plants, indicating its origin in the common ancestor of land plants. BSB synthesis is enhanced at AM symbiosis permissive conditions and BSB deficient mutants are impaired in AM symbiosis. In contrast, the absence of BSB synthesis has little effect on the growth and gene expression. We show that the introduction of the SL receptor of Arabidopsis renders M. paleacea cells BSB-responsive. These results suggest that BSB is not perceived by M. paleacea cells due to the lack of cognate SL receptors. We propose that SLs originated as AM symbiosis-inducing rhizosphere signaling molecules and were later recruited as plant hormone.
ESTHER : Kodama_2022_Nat.Commun_13_3974
PubMedSearch : Kodama_2022_Nat.Commun_13_3974
PubMedID: 35803942

Title : Canonical strigolactones are not the major determinant of tillering but important rhizospheric signals in rice - Ito_2022_Sci.Adv_8_eadd1278
Author(s) : Ito S , Braguy J , Wang JY , Yoda A , Fiorilli V , Takahashi I , Jamil M , Felemban A , Miyazaki S , Mazzarella T , Chen GE , Shinozawa A , Balakrishna A , Berqdar L , Rajan C , Ali S , Haider I , Sasaki Y , Yajima S , Akiyama K , Lanfranco L , Zurbriggen MD , Nomura T , Asami T , Al-Babili S
Ref : Sci Adv , 8 :eadd1278 , 2022
Abstract : Strigolactones (SLs) are a plant hormone inhibiting shoot branching/tillering and a rhizospheric, chemical signal that triggers seed germination of the noxious root parasitic plant Striga and mediates symbiosis with beneficial arbuscular mycorrhizal fungi. Identifying specific roles of canonical and noncanonical SLs, the two SL subfamilies, is important for developing Striga-resistant cereals and for engineering plant architecture. Here, we report that rice mutants lacking canonical SLs do not show the shoot phenotypes known for SL-deficient plants, exhibiting only a delay in establishing arbuscular mycorrhizal symbiosis, but release exudates with a significantly decreased Striga seed-germinating activity. Blocking the biosynthesis of canonical SLs by TIS108, a specific enzyme inhibitor, significantly lowered Striga infestation without affecting rice growth. These results indicate that canonical SLs are not the determinant of shoot architecture and pave the way for increasing crop resistance by gene editing or chemical treatment.
ESTHER : Ito_2022_Sci.Adv_8_eadd1278
PubMedSearch : Ito_2022_Sci.Adv_8_eadd1278
PubMedID: 36322663

Title : Strigolactone biosynthesis catalyzed by cytochrome P450 and sulfotransferase in sorghum - Yoda_2021_New.Phytol_232_1999
Author(s) : Yoda A , Mori N , Akiyama K , Kikuchi M , Xie X , Miura K , Yoneyama K , Sato-Izawa K , Yamaguchi S , Nelson DC , Nomura T
Ref : New Phytol , 232 :1999 , 2021
Abstract : Root parasitic plants such as Striga, Orobanche, and Phelipanche spp. cause serious damage to crop production world-wide. Deletion of the Low Germination Stimulant 1 (LGS1) gene gives a Striga-resistance trait in sorghum (Sorghum bicolor). The LGS1 gene encodes a sulfotransferase-like protein, but its function has not been elucidated. Since the profile of strigolactones (SLs) that induce seed germination in root parasitic plants is altered in the lgs1 mutant, LGS1 is thought to be an SL biosynthetic enzyme. In order to clarify the enzymatic function of LGS1, we looked for candidate SL substrates that accumulate in the lgs1 mutants and performed in vivo and in vitro metabolism experiments. We found the SL precursor 18-hydroxycarlactonoic acid (18-OH-CLA) is a substrate for LGS1. CYP711A cytochrome P450 enzymes (SbMAX1 proteins) in sorghum produce 18-OH-CLA. When LGS1 and SbMAX1 coding sequences were co-expressed in Nicotiana benthamiana with the upstream SL biosynthesis genes from sorghum, the canonical SLs 5-deoxystrigol and 4-deoxyorobanchol were produced. This finding showed that LGS1 in sorghum uses a sulfo group to catalyze leaving of a hydroxyl group and cyclization of 18-OH-CLA. A similar SL biosynthetic pathway has not been found in other plant species.
ESTHER : Yoda_2021_New.Phytol_232_1999
PubMedSearch : Yoda_2021_New.Phytol_232_1999
PubMedID: 34525227

Title : Evaluation and Quantification of Natural Strigolactones from Root Exudates - Xie_2021_Methods.Mol.Biol_2309_3
Author(s) : Xie X , Yoneyama K , Nomura T
Ref : Methods Mol Biol , 2309 :3 , 2021
Abstract : Strigolactones (SLs) in the root exudates can be detected by germination assays with root parasitic weed seeds, but precise and accurate evaluation and quantification are possible only by chemical analysis with the liquid chromatography-tandem mass spectrometry (LC-MS/MS). Here we describe methods for root exudate collection, sample preparation, and LC-MS/MS analysis of SLs.
ESTHER : Xie_2021_Methods.Mol.Biol_2309_3
PubMedSearch : Xie_2021_Methods.Mol.Biol_2309_3
PubMedID: 34028674

Title : Identification of two oxygenase genes involved in the respective biosynthetic pathways of canonical and non-canonical strigolactones in Lotus japonicus - Mori_2020_Planta_251_40
Author(s) : Mori N , Nomura T , Akiyama K
Ref : Planta , 251 :40 , 2020
Abstract : A cytochrome P450 and a 2-oxoglutarate-dependent dioxygenase genes responsible, respectively, for the biosyntheses of canonical and non-canonical strigolactones in Lotus japonicus were identified by transcriptome profiling and mutant screening. Strigolactones (SLs) are a group of apocarotenoids with diverse structures that act as phytohormones and rhizosphere signals. The model legume Lotus japonicus produces both canonical and non-canonical SLs, 5-deoxystrigol (5DS) and lotuslactone (LL), respectively, through oxidation of a common intermediate carlactone by the cytochrome P450 (CYP) enzyme MAX1. However, the pathways downstream of MAX1 and the branching point in the biosyntheses of 5DS and LL have not been elucidated. Here, we identified a CYP and a 2-oxoglutarate-dependent dioxygenase (2OGD) genes responsible, respectively, for the formation of Lotus SLs by transcriptome profiling using RNA-seq and screening of SL-deficient mutants from the Lotus retrotransposon 1 (LORE1) insertion mutant resource. The CYP and 2OGD genes were named DSD and LLD, respectively, after 5DS or LL defective phenotype of the mutants. The involvements of the genes in Lotus SL biosyntheses were confirmed by restoration of the mutant phenotype using Agrobacterium rhizogenes-mediated transformation to generate transgenic roots expressing the coding sequence. The transcript levels of DSD and LLD in roots as well as the levels of 5DS and LL in root exudates were reduced by phosphate fertilization and gibberellin treatment. This study can provide the opportunity to investigate how and why plants produce the two classes of SLs.
ESTHER : Mori_2020_Planta_251_40
PubMedSearch : Mori_2020_Planta_251_40
PubMedID: 31907631

Title : Hydroxyl carlactone derivatives are predominant strigolactones in Arabidopsis - Yoneyama_2020_Plant.Direct_4_e00219
Author(s) : Yoneyama K , Akiyama K , Brewer PB , Mori N , Kawano-Kawada M , Haruta S , Nishiwaki H , Yamauchi S , Xie X , Umehara M , Beveridge CA , Nomura T
Ref : Plant Direct , 4 :e00219 , 2020
Abstract : Strigolactones (SLs) regulate important aspects of plant growth and stress responses. Many diverse types of SL occur in plants, but a complete picture of biosynthesis remains unclear. In Arabidopsis thaliana, we have demonstrated that MAX1, a cytochrome P450 monooxygenase, converts carlactone (CL) into carlactonoic acid (CLA) and that LBO, a 2-oxoglutarate-dependent dioxygenase, can convert methyl carlactonoate (MeCLA) into a metabolite called [MeCLA + 16 Da]. In the present study, feeding experiments with deuterated MeCLAs revealed that [MeCLA + 16 Da] is hydroxymethyl carlactonoate (1'-HO-MeCLA). Importantly, this LBO metabolite was detected in plants. Interestingly, other related compounds, methyl 4-hydroxycarlactonoate (4-HO-MeCLA) and methyl 16-hydroxycarlactonoate (16-HO-MeCLA), were also found to accumulate in lbo mutants. 3-HO-, 4-HO-, and 16-HO-CL were detected in plants, but their expected corresponding metabolites, HO-CLAs, were absent in max1 mutants. These results suggest that HO-CL derivatives may be predominant SLs in Arabidopsis, produced through MAX1 and LBO.
ESTHER : Yoneyama_2020_Plant.Direct_4_e00219
PubMedSearch : Yoneyama_2020_Plant.Direct_4_e00219
PubMedID: 32399509

Title : Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and non-canonical strigolactones in Lotus japonicus - Mori_2020_Phytochemistry_174_112349
Author(s) : Mori N , Sado A , Xie X , Yoneyama K , Asami K , Seto Y , Nomura T , Yamaguchi S , Akiyama K
Ref : Phytochemistry , 174 :112349 , 2020
Abstract : Strigolactones (SLs) are a group of plant apocarotenoids that act as rhizosphere signaling molecules for both arbuscular mycorrhizal fungi and root parasitic plants. They also regulate plant architecture as phytohormones. The model legume Lotus japonicus (synonym of Lotus corniculatus) produces canonical 5-deoxystrigol (5DS) and non-canonical lotuslactone (LL). The biosynthesis pathways of the two SLs remain elusive. In this study, we characterized the L. japonicus MAX1 homolog, LjMAX1, found in the Lotus japonicus genome assembly build 2.5. The L. japonicus max1 LORE1 insertion mutant was deficient in 5DS and LL production. A recombinant LjMAX1 protein expressed in yeast microsomes converted carlactone (CL) to 18-hydroxycarlactonoic acid (18-OH-CLA) via carlactonoic acid (CLA). Identity of 18-OH-CLA was confirmed by comparison of the methyl ester derivative of the MAX1 product with chemically synthesized methyl 18-hydroycarlactonoate (18-OH-MeCLA) using LC-MS/MS. (11R)-CL was detected as an endogenous compound in the root of L. japonicus.(13)C-labeled CL, CLA, and 18-OH-MeCLA were converted to [(13)C]-5DS and LL in plant feeding experiments using L. japonicus WT. These results showed that LjMAX1 is the crucial enzyme in the biosynthesis of Lotus SLs and that 18-hydroxylated carlactonoates are possible precursors for SL biosynthesis in L. japonicus.
ESTHER : Mori_2020_Phytochemistry_174_112349
PubMedSearch : Mori_2020_Phytochemistry_174_112349
PubMedID: 32213359

Title : Lotuslactone, a non-canonical strigolactone from Lotus japonicus - Xie_2019_Phytochemistry_157_200
Author(s) : Xie X , Mori N , Yoneyama K , Nomura T , Uchida K , Akiyama K
Ref : Phytochemistry , 157 :200 , 2019
Abstract : Root exudates from Lotus japonicus were found to contain at least three different hyphal branching-inducing compounds for the arbuscular mycorrhizal (AM) fungus Gigaspora margarita, one of which had been previously identified as (+)-5-deoxystrigol (5DS), a canonical strigolactone (SL). One of the two remaining unknown hyphal branching inducers was purified and named lotuslactone. Its structure was determined as methyl (E)-2-(3-acetoxy-2-hydroxy-7-methyl-1-oxo-1,2,3,4,5,6-hexahydroazulen-2-yl)-3-(((R)-4-methyl-5-oxo-2,5-dihydrofuran-2-yl)oxy)acrylate, by 1D and 2D NMR spectroscopy, and HR-ESI- and EI-MS. Although lotuslactone, a non-canonical SL, contains the AB-ring and the enol ether-bridged D-ring, it lacks the C-ring and has a seven-membered cycloheptadiene in the A-ring part as in medicaol, a major SL of Medicago truncatula. Lotuslactone was much less active than 5DS, but showed comparable activity to methyl carlactonoate (MeCLA) in inducing hyphal branching of G. margarita. Other natural non-canonical SLs including avenaol, heliolactone, and zealactone (methyl zealactonoate) were also found to be moderate to weak inducers of hyphal branching in the AM fungus. Lotuslactone strongly elicited seed germination in Phelipanche ramosa and Orobanche minor, but Striga hermonthica seeds were 100-fold less sensitive to this stimulant.
ESTHER : Xie_2019_Phytochemistry_157_200
PubMedSearch : Xie_2019_Phytochemistry_157_200
PubMedID: 30439621

Title : Regulation of biosynthesis, perception, and functions of strigolactones for promoting arbuscular mycorrhizal symbiosis and managing root parasitic weeds - Yoneyama_2019_Pest.Manag.Sci_75_2353
Author(s) : Yoneyama K , Xie X , Nomura T , Takahashi I , Asami T , Mori N , Akiyama K , Kusajima M , Nakashita H
Ref : Pest Manag Sci , 75 :2353 , 2019
Abstract : Strigolactones (SLs) are carotenoid-derived plant secondary metabolites that play important roles in various aspects of plant growth and development as plant hormones, and in rhizosphere communications with symbiotic microbes and also root parasitic weeds. Therefore, sophisticated regulation of the biosynthesis, perception and functions of SLs is expected to promote symbiosis of beneficial microbes including arbuscular mycorrhizal (AM) fungi and also to retard parasitism by devastating root parasitic weeds. We have developed SL mimics with different skeletons, SL biosynthesis inhibitors acting at different biosynthetic steps, SL perception inhibitors that covalently bind to the SL receptor D14, and SL function inhibitors that bind to the serine residue at the catalytic site. In greenhouse pot tests, TIS108, an azole-type SL biosynthesis inhibitor effectively reduced numbers of attached root parasites Orobanche minor and Striga hermonthica without affecting their host plants; tomato and rice, respectively. AM colonization resulted in weak but distinctly enhanced plant resistance to pathogens. SL mimics can be used to promote AM symbiosis and to reduce the application rate of systemic-acquired resistance inducers which are generally phytotoxic to horticultural crops. (c) 2019 Society of Chemical Industry.
ESTHER : Yoneyama_2019_Pest.Manag.Sci_75_2353
PubMedSearch : Yoneyama_2019_Pest.Manag.Sci_75_2353
PubMedID: 30843315

Title : Genome Sequence of Striga asiatica Provides Insight into the Evolution of Plant Parasitism - Yoshida_2019_Curr.Biol_29_3041
Author(s) : Yoshida S , Kim S , Wafula EK , Tanskanen J , Kim YM , Honaas L , Yang Z , Spallek T , Conn CE , Ichihashi Y , Cheong K , Cui S , Der JP , Gundlach H , Jiao Y , Hori C , Ishida JK , Kasahara H , Kiba T , Kim MS , Koo N , Laohavisit A , Lee YH , Lumba S , McCourt P , Mortimer JC , Mutuku JM , Nomura T , Sasaki-Sekimoto Y , Seto Y , Wang Y , Wakatake T , Sakakibara H , Demura T , Yamaguchi S , Yoneyama K , Manabe RI , Nelson DC , Schulman AH , Timko MP , dePamphilis CW , Choi D , Shirasu K
Ref : Current Biology , 29 :3041 , 2019
Abstract : Parasitic plants in the genus Striga, commonly known as witchweeds, cause major crop losses in sub-Saharan Africa and pose a threat to agriculture worldwide. An understanding of Striga parasite biology, which could lead to agricultural solutions, has been hampered by the lack of genome information. Here, we report the draft genome sequence of Striga asiatica with 34,577 predicted protein-coding genes, which reflects gene family contractions and expansions that are consistent with a three-phase model of parasitic plant genome evolution. Striga seeds germinate in response to host-derived strigolactones (SLs) and then develop a specialized penetration structure, the haustorium, to invade the host root. A family of SL receptors has undergone a striking expansion, suggesting a molecular basis for the evolution of broad host range among Striga spp. We found that genes involved in lateral root development in non-parasitic model species are coordinately induced during haustorium development in Striga, suggesting a pathway that was partly co-opted during the evolution of the haustorium. In addition, we found evidence for horizontal transfer of host genes as well as retrotransposons, indicating gene flow to S. asiatica from hosts. Our results provide valuable insights into the evolution of parasitism and a key resource for the future development of Striga control strategies.
ESTHER : Yoshida_2019_Curr.Biol_29_3041
PubMedSearch : Yoshida_2019_Curr.Biol_29_3041
PubMedID: 31522940
Gene_locus related to this paper: straf-a0a5a7qxe3

Title : Substrate specificity of tuliposide-converting enzyme, a unique non-ester-hydrolyzing carboxylesterase in tulip: Effects of the alcohol moiety of substrate on the enzyme activity - Kato_2019_Bioorg.Med.Chem.Lett_29_664
Author(s) : Kato Y , Futanaga T , Nomura T
Ref : Bioorganic & Medicinal Chemistry Lett , 29 :664 , 2019
Abstract : 6-Tuliposides A (PosA) and B (PosB) are glucose esters accumulated in tulip (Tulipa gesneriana) as major defensive secondary metabolites. Pos-converting enzymes (TgTCEs), which we discovered previously from tulip, catalyze the conversion reactions of PosA and PosB to antimicrobial tulipalins A (PaA) and B (PaB), respectively. The TgTCEs, belonging to the carboxylesterase family, specifically catalyze intramolecular transesterification, but not hydrolysis. In this report, we synthesized analogues of Pos with various alcohol moieties, and measured the TgTCE activity together with a determination of the kinetic parameters for these analogues with a view to probe the substrate recognition mechanism of the unique non-ester-hydrolyzing TgTCEs. It was found that d-glucose-like structure and number of the hydroxyl group in alcohol moiety are important for substrate recognition by TgTCEs. Among the analogues examined, 1,2-dideoxy analogues of PosA and PosB were found to be recognized by the TgTCEs more specifically than the authentic substrates by lowering Km values. The present results will provide a basis for designing simple, stable synthetic substrate analogues for crystallographic analysis of TgTCEs.
ESTHER : Kato_2019_Bioorg.Med.Chem.Lett_29_664
PubMedSearch : Kato_2019_Bioorg.Med.Chem.Lett_29_664
PubMedID: 30595444
Gene_locus related to this paper: tulge-a0a0h5bmx5

Title : Molecular diversity of tuliposide B-converting enzyme in tulip (Tulipa gesneriana): identification of the third isozyme with a distinct expression profile - Nomura_2018_Biosci.Biotechnol.Biochem__1
Author(s) : Nomura T , Kuchida R , Kitaoka N , Kato Y
Ref : Biosci Biotechnol Biochem , :1 , 2018
Abstract : 6-Tuliposide B (PosB), a major secondary metabolite that accumulates in tulip (Tulipa gesneriana), is converted to the antibacterial lactone, tulipalin B (PaB), by PosB-converting enzyme (TCEB). TgTCEB1 and TgTCEB-R, which encode TCEB, are specifically expressed in tulip pollen and roots, respectively, but are hardly expressed in other tissues (e.g. leaves) despite the presence of substantial PosB-converting activity, suggesting the existence of another TCEB isozyme. Here, we describe the identification of TgTCEB-L ("L" for leaf), a paralog of TgTCEB1 and TgTCEB-R, from leaves via native enzyme purification. The enzymatic characters of TgTCEB-L, including catalytic activity and subcellular localization, were substantially the same as those of TgTCEB1 and TgTCEB-R. However, TgTCEB-L did not exhibit tissue-specific expression. Identification of TgTCEB-L explains the PosB-converting activity detected in tissues where TgTCEB1 and TgTCEB-R transcripts could not be detected, indicating that tulip subtilizes the three TgTCEB isozymes depending on the tissue.
ESTHER : Nomura_2018_Biosci.Biotechnol.Biochem__1
PubMedSearch : Nomura_2018_Biosci.Biotechnol.Biochem__1
PubMedID: 29475400

Title : Conversion of carlactone to carlactonoic acid is a conserved function of MAX1 homologs in strigolactone biosynthesis - Yoneyama_2018_New.Phytol_218_1522
Author(s) : Yoneyama K , Mori N , Sato T , Yoda A , Xie X , Okamoto M , Iwanaga M , Ohnishi T , Nishiwaki H , Asami T , Yokota T , Akiyama K , Nomura T
Ref : New Phytol , 218 :1522 , 2018
Abstract : Strigolactones (SLs) are a class of plant hormones which regulate shoot branching and function as host recognition signals for symbionts and parasites in the rhizosphere. However, steps in SL biosynthesis after carlactone (CL) formation remain elusive. This study elucidated the common and diverse functions of MAX1 homologs which catalyze CL oxidation. We have reported previously that ArabidopsisMAX1 converts CL to carlactonoic acid (CLA), whereas a rice MAX1 homolog has been shown to catalyze the conversion of CL to 4-deoxyorobanchol (4DO). To determine which reaction is conserved in the plant kingdom, we investigated the enzymatic function of MAX1 homologs in Arabidopsis, rice, maize, tomato, poplar and Selaginella moellendorffii. The conversion of CL to CLA was found to be a common reaction catalyzed by MAX1 homologs, and MAX1s can be classified into three types: A1-type, converting CL to CLA; A2-type, converting CL to 4DO via CLA; and A3-type, converting CL to CLA and 4DO to orobanchol. CLA was detected in root exudates from poplar and Selaginella, but not ubiquitously in other plants examined in this study, suggesting its role as a species-specific signal in the rhizosphere. This study provides new insights into the roles of MAX1 in endogenous and rhizosphere signaling.
ESTHER : Yoneyama_2018_New.Phytol_218_1522
PubMedSearch : Yoneyama_2018_New.Phytol_218_1522
PubMedID: 29479714

Title : Which are the major players, canonical or non-canonical strigolactones? - Yoneyama_2018_J.Exp.Bot_69_2231
Author(s) : Yoneyama K , Xie X , Kisugi T , Nomura T , Nakatani Y , Akiyama K , McErlean CSP
Ref : J Exp Bot , 69 :2231 , 2018
Abstract : Strigolactones (SLs) can be classified into two structurally distinct groups: canonical and non-canonical SLs. Canonical SLs contain the ABCD ring system, and non-canonical SLs lack the A, B, or C ring but have the enol ether-D ring moiety, which is essential for biological activities. The simplest non-canonical SL is the SL biosynthetic intermediate carlactone. In plants, carlactone and its oxidized metabolites, such as carlactonoic acid and methyl carlactonoate, are present in root and shoot tissues. In some plant species, including black oat (Avena strigosa), sunflower (Helianthus annuus), and maize (Zea mays), non-canonical SLs in the root exudates are major germination stimulants. Various plant species, such as tomato (Solanum lycopersicum), Arabidopsis, and poplar (Populus spp.), release carlactonoic acid into the rhizosphere. These observations suggest that both canonical and non-canonical SLs act as host-recognition signals in the rhizosphere. In contrast, the limited distribution of canonical SLs in the plant kingdom, and the structure-specific and stereospecific transportation of canonical SLs from roots to shoots, suggest that plant hormones inhibiting shoot branching are not canonical SLs but, rather, are non-canonical SLs.
ESTHER : Yoneyama_2018_J.Exp.Bot_69_2231
PubMedSearch : Yoneyama_2018_J.Exp.Bot_69_2231
PubMedID: 29522151

Title : Molecular diversity of tuliposide B-converting enzyme in tulip (Tulipa gesneriana): identification of the root-specific isozyme - Nomura_2017_Biosci.Biotechnol.Biochem_81_1185
Author(s) : Nomura T , Ueno A , Ogita S , Kato Y
Ref : Biosci Biotechnol Biochem , 81 :1185 , 2017
Abstract : 6-Tuliposide B (PosB) is a glucose ester accumulated in tulip (Tulipa gesneriana) as a major secondary metabolite. PosB serves as the precursor of the antimicrobial lactone tulipalin B (PaB), which is formed by PosB-converting enzyme (TCEB). The gene TgTCEB1, encoding a TCEB, is transcribed in tulip pollen but scarcely transcribed in other tissues (e.g. roots) even though those tissues show high TCEB activity. This led to the prediction of the presence of a TCEB isozyme with distinct tissue specificity. Herein, we describe the identification of the TgTCEB-R gene from roots via native enzyme purification; this gene is a paralog of TgTCEB1. Recombinant enzyme characterization verified that TgTCEB-R encodes a TCEB. Moreover, TgTCEB-R was localized in tulip plastids, as found for pollen TgTCEB1. TgTCEB-R is transcribed almost exclusively in roots, indicating a tissue preference for the transcription of TCEB isozyme genes.
ESTHER : Nomura_2017_Biosci.Biotechnol.Biochem_81_1185
PubMedSearch : Nomura_2017_Biosci.Biotechnol.Biochem_81_1185
PubMedID: 28485211

Title : Function and application of a non-ester-hydrolyzing carboxylesterase discovered in tulip - Nomura_2017_Biosci.Biotechnol.Biochem_81_81
Author(s) : Nomura T
Ref : Biosci Biotechnol Biochem , 81 :81 , 2017
Abstract : Plants have evolved secondary metabolite biosynthetic pathways of immense rich diversity. The genes encoding enzymes for secondary metabolite biosynthesis have evolved through gene duplication followed by neofunctionalization, thereby generating functional diversity. Emerging evidence demonstrates that some of those enzymes catalyze reactions entirely different from those usually catalyzed by other members of the same family; e.g. transacylation catalyzed by an enzyme similar to a hydrolytic enzyme. Tuliposide-converting enzyme (TCE), which we recently discovered from tulip, catalyzes the conversion of major defensive secondary metabolites, tuliposides, to antimicrobial tulipalins. The TCEs belong to the carboxylesterase family in the alpha/beta-hydrolase fold superfamily, and specifically catalyze intramolecular transesterification, but not hydrolysis. This non-ester-hydrolyzing carboxylesterase is an example of an enzyme showing catalytic properties that are unpredictable from its primary structure. This review describes the biochemical and physiological aspects of tulipalin biogenesis, and the diverse functions of plant carboxylesterases in the alpha/beta-hydrolase fold superfamily.
ESTHER : Nomura_2017_Biosci.Biotechnol.Biochem_81_81
PubMedSearch : Nomura_2017_Biosci.Biotechnol.Biochem_81_81
PubMedID: 27696958
Gene_locus related to this paper: tulge-a0a0h5bmx5

Title : LATERAL BRANCHING OXIDOREDUCTASE acts in the final stages of strigolactone biosynthesis in Arabidopsis - Brewer_2016_Proc.Natl.Acad.Sci.U.S.A_113_6301
Author(s) : Brewer PB , Yoneyama K , Filardo F , Meyers E , Scaffidi A , Frickey T , Akiyama K , Seto Y , Dun EA , Cremer JE , Kerr SC , Waters MT , Flematti GR , Mason MG , Weiller G , Yamaguchi S , Nomura T , Smith SM , Beveridge CA
Ref : Proc Natl Acad Sci U S A , 113 :6301 , 2016
Abstract : Strigolactones are a group of plant compounds of diverse but related chemical structures. They have similar bioactivity across a broad range of plant species, act to optimize plant growth and development, and promote soil microbe interactions. Carlactone, a common precursor to strigolactones, is produced by conserved enzymes found in a number of diverse species. Versions of the MORE AXILLARY GROWTH1 (MAX1) cytochrome P450 from rice and Arabidopsis thaliana make specific subsets of strigolactones from carlactone. However, the diversity of natural strigolactones suggests that additional enzymes are involved and remain to be discovered. Here, we use an innovative method that has revealed a missing enzyme involved in strigolactone metabolism. By using a transcriptomics approach involving a range of treatments that modify strigolactone biosynthesis gene expression coupled with reverse genetics, we identified LATERAL BRANCHING OXIDOREDUCTASE (LBO), a gene encoding an oxidoreductase-like enzyme of the 2-oxoglutarate and Fe(II)-dependent dioxygenase superfamily. Arabidopsis lbo mutants exhibited increased shoot branching, but the lbo mutation did not enhance the max mutant phenotype. Grafting indicated that LBO is required for a graft-transmissible signal that, in turn, requires a product of MAX1. Mutant lbo backgrounds showed reduced responses to carlactone, the substrate of MAX1, and methyl carlactonoate (MeCLA), a product downstream of MAX1. Furthermore, lbo mutants contained increased amounts of these compounds, and the LBO protein specifically converts MeCLA to an unidentified strigolactone-like compound. Thus, LBO function may be important in the later steps of strigolactone biosynthesis to inhibit shoot branching in Arabidopsis and other seed plants.
ESTHER : Brewer_2016_Proc.Natl.Acad.Sci.U.S.A_113_6301
PubMedSearch : Brewer_2016_Proc.Natl.Acad.Sci.U.S.A_113_6301
PubMedID: 27194725

Title : Molecular identification of tuliposide B-converting enzyme: a lactone-forming carboxylesterase from the pollen of tulip - Nomura_2015_Plant.J_83_252
Author(s) : Nomura T , Murase T , Ogita S , Kato Y
Ref : Plant J , 83 :252 , 2015
Abstract : 6-Tuliposides A (PosA) and B (PosB), which are the major secondary metabolites in tulip (Tulipa gesneriana), are enzymatically converted to the antimicrobial lactonized aglycons, tulipalins A (PaA) and B (PaB), respectively. We recently identified a PosA-converting enzyme (TCEA) as the first reported member of the lactone-forming carboxylesterases. Herein, we describe the identification of another lactone-forming carboxylesterase, PosB-converting enzyme (TCEB), which preferentially reacts with PosB to give PaB. This enzyme was isolated from tulip pollen, which showed high PosB-converting activity. Purified TCEB exhibited greater activity towards PosB than PosA, which was contrary to that of the TCEA. Novel cDNA (TgTCEB1) encoding the TCEB was isolated from tulip pollen. TgTCEB1 belonged to the carboxylesterase family and was approximately 50% identical to the TgTCEA polypeptides. Functional characterization of the recombinant enzyme verified that TgTCEB1 catalyzed the conversion of PosB to PaB with an activity comparable with the native TCEB. RT-qPCR analysis of each part of plant revealed that TgTCEB1 transcripts were limited almost exclusively to the pollen. Furthermore, the immunostaining of the anther cross-section using anti-TgTCEB1 polyclonal antibody verified that TgTCEB1 was specifically expressed in the pollen grains, but not in the anther cells. N-terminal transit peptide of TgTCEB1 was shown to function as plastid-targeted signal. Taken together, these results indicate that mature TgTCEB1 is specifically localized in plastids of pollen grains. Interestingly, PosB, the substrate of TgTCEB1, accumulated on the pollen surface, but not in the intracellular spaces of pollen grains.
ESTHER : Nomura_2015_Plant.J_83_252
PubMedSearch : Nomura_2015_Plant.J_83_252
PubMedID: 25997073
Gene_locus related to this paper: tulge-a0a0h5bmx5

Title : Carlactone is converted to carlactonoic acid by MAX1 in Arabidopsis and its methyl ester can directly interact with AtD14 in vitro - Abe_2014_Proc.Natl.Acad.Sci.U.S.A_111_18084
Author(s) : Abe S , Sado A , Tanaka K , Kisugi T , Asami K , Ota S , Kim HI , Yoneyama K , Xie X , Ohnishi T , Seto Y , Yamaguchi S , Akiyama K , Nomura T
Ref : Proc Natl Acad Sci U S A , 111 :18084 , 2014
Abstract : Strigolactones (SLs) stimulate seed germination of root parasitic plants and induce hyphal branching of arbuscular mycorrhizal fungi in the rhizosphere. In addition, they have been classified as a new group of plant hormones essential for shoot branching inhibition. It has been demonstrated thus far that SLs are derived from carotenoid via a biosynthetic precursor carlactone (CL), which is produced by sequential reactions of DWARF27 (D27) enzyme and two carotenoid cleavage dioxygenases CCD7 and CCD8. We previously found an extreme accumulation of CL in the more axillary growth1 (max1) mutant of Arabidopsis, which exhibits increased lateral inflorescences due to SL deficiency, indicating that CL is a probable substrate for MAX1 (CYP711A1), a cytochrome P450 monooxygenase. To elucidate the enzymatic function of MAX1 in SL biosynthesis, we incubated CL with a recombinant MAX1 protein expressed in yeast microsomes. MAX1 catalyzed consecutive oxidations at C-19 of CL to convert the C-19 methyl group into carboxylic acid, 9-desmethyl-9-carboxy-CL [designated as carlactonoic acid (CLA)]. We also identified endogenous CLA and its methyl ester [methyl carlactonoate (MeCLA)] in Arabidopsis plants using LC-MS/MS. Although an exogenous application of either CLA or MeCLA suppressed the growth of lateral inflorescences of the max1 mutant, MeCLA, but not CLA, interacted with Arabidopsis thaliana DWARF14 (AtD14) protein, a putative SL receptor, as shown by differential scanning fluorimetry and hydrolysis activity tests. These results indicate that not only known SLs but also MeCLA are biologically active in inhibiting shoot branching in Arabidopsis.
ESTHER : Abe_2014_Proc.Natl.Acad.Sci.U.S.A_111_18084
PubMedSearch : Abe_2014_Proc.Natl.Acad.Sci.U.S.A_111_18084
PubMedID: 25425668
Gene_locus related to this paper: arath-AtD14

Title : Confirming stereochemical structures of strigolactones produced by rice and tobacco - Xie_2013_Mol.Plant_6_153
Author(s) : Xie X , Yoneyama K , Kisugi T , Uchida K , Ito S , Akiyama K , Hayashi H , Yokota T , Nomura T
Ref : Mol Plant , 6 :153 , 2013
Abstract : Major strigolactones (SLs) produced by rice (Oryza sativa L. cv. Nipponbare) and tobacco (Nicotiana tabacum L. cv. Michinoku No. 1) were purified and their stereochemical structures were determined by comparing with optically pure synthetic standards for their NMR and CD data and retention times and mass fragmentations in ESI-LC/MS and GC-MS. SLs purified from root exudates of rice plants were orobanchol, orobanchyl acetate, and ent-2'-epi-5-deoxystrigol. In addition to these SLs, 7-oxoorobanchyl acetate and the putative three methoxy-5-deoxystrigol isomers were detected by LC-MS/MS. The production of 7-oxoorobanchyl acetate seemed to occur in the early growth stage, as it was detected only in the root exudates collected during the first week of incubation. The root exudates of tobacco contained at least 11 SLs, including solanacol, solanacyl acetate, orobanchol, ent-2'-epi-orobanchol, orobanchyl acetate, ent-2'-epi-orobanchyl acetate, 5-deoxystrigol, ent-2'-epi-5-deoxystrigol, and three isomers of putative didehydro-orobanchol whose structures remain to be clarified. Furthermore, two sorgolactone isomers but not sorgolactone were detected as minor SLs by LC-MS/MS analysis. It is intriguing to note that rice plants produced only orobanchol-type SLs, derived from ent-2'-epi-5-deoxystrigol, but both orobanchol-type and strigol-type SLs, derived from 5-deoxystrigol were detected in tobacco plants.
ESTHER : Xie_2013_Mol.Plant_6_153
PubMedSearch : Xie_2013_Mol.Plant_6_153
PubMedID: 23204500

Title : Molecular diversity of tuliposide A-converting enzyme in the tulip - Nomura_2013_Biosci.Biotechnol.Biochem_77_1042
Author(s) : Nomura T , Tsuchigami A , Ogita S , Kato Y
Ref : Biosci Biotechnol Biochem , 77 :1042 , 2013
Abstract : Tuliposide A-converting enzyme (TCEA) catalyzes the conversion of 6-tuliposide A to its lactonized aglycon, tulipalin A, in the tulip (Tulipa gesneriana). The TgTCEA gene, isolated previously from petals, was transcribed in all tulip tissues but not in the bulbs despite the presence of TCEA activity, which allowed prediction of the presence of a TgTCEA isozyme gene preferentially expressed in the bulbs. Here, the TgTCEA-b gene, the TgTCEA homolog, was identified in bulbs. TgTCEA-b polypeptides showed approximately 77% identity to the petal TgTCEA. Functional characterization of the recombinant enzyme verified that TgTCEA-b encoded the TCEA. Moreover, the TgTCEA-b was found to be localized to plastids, as found for the petal TgTCEA. Transcript analysis revealed that TgTCEA-b was functionally transcribed in the bulb scales, unlike the TgTCEA gene, whose transcripts were absent there. In contrast, TgTCEA-b transcripts were in the minority in other tissues where TgTCEA transcripts were dominant, indicating a tissue preference for the transcription of those isozyme genes.
ESTHER : Nomura_2013_Biosci.Biotechnol.Biochem_77_1042
PubMedSearch : Nomura_2013_Biosci.Biotechnol.Biochem_77_1042
PubMedID: 23649245
Gene_locus related to this paper: tulge-tcab2 , tulge-tcab1 , tulge-tcab3 , tulge-tcab4

Title : The beta9 loop domain of PA-PLA1alpha has a crucial role in autosomal recessive woolly hair\/hypotrichosis -
Author(s) : Shinkuma S , Inoue A , Aoki J , Nishie W , Natsuga K , Ujiie H , Nomura T , Abe R , Akiyama M , Shimizu H
Ref : Journal of Investigative Dermatology , 132 :2093 , 2012
PubMedID: 22475755
Gene_locus related to this paper: human-LIPH

Title : A novel lactone-forming carboxylesterase: molecular identification of a tuliposide A-converting enzyme in tulip - Nomura_2012_Plant.Physiol_159_565
Author(s) : Nomura T , Ogita S , Kato Y
Ref : Plant Physiol , 159 :565 , 2012
Abstract : Tuliposides, the glucose esters of 4-hydroxy-2-methylenebutanoate and 3,4-dihydroxy-2-methylenebutanoate, are major secondary metabolites in tulip (Tulipa gesneriana). Their lactonized aglycons, tulipalins, function as defensive chemicals due to their biological activities. We recently found that tuliposide-converting enzyme (TCE) purified from tulip bulbs catalyzed the conversion of tuliposides to tulipalins, but the possibility of the presence of several TCE isozymes was raised: TCE in tissues other than bulbs is different from bulb TCE. Here, to prove this hypothesis, TCE was purified from petals, which have the second highest TCE activity after bulbs. The purified enzyme, like the bulb enzyme, preferentially accepted tuliposides as substrates, with 6-tuliposide A the best substrate, which allowed naming the enzyme tuliposide A-converting enzyme (TCEA), but specific activity and molecular mass differed between the petal and bulb enzymes. After peptide sequencing, a novel cDNA (TgTCEA) encoding petal TCEA was isolated, and the functional characterization of the recombinant enzyme verified that TgTCEA catalyzes the conversion of 6-tuliposide A to tulipalin A. TgTCEA was transcribed in all tulip tissues but not in bulbs, indicating the presence of a bulb-specific TgTCEA, as suggested by the distinct enzymatic characters between the petal and bulb enzymes. Plastidial localization of TgTCEA enzyme was revealed, which allowed proposing a cytological mechanism of TgTCE-mediated tulipalin formation in the tulip defensive strategy. Site-directed mutagenesis of TgTCEA suggested that the oxyanion hole and catalytic triad characteristic of typical carboxylesterases are essential for the catalytic process of TgTCEA enzyme. To our knowledge, TgTCEA is the first identified member of the lactone-forming carboxylesterases, specifically catalyzing intramolecular transesterification.
ESTHER : Nomura_2012_Plant.Physiol_159_565
PubMedSearch : Nomura_2012_Plant.Physiol_159_565
PubMedID: 22474185
Gene_locus related to this paper: tulge-tcea1 , tulge-tcea2

Title : Prevalent LIPH founder mutations lead to loss of P2Y5 activation ability of PA-PLA1alpha in autosomal recessive hypotrichosis - Shinkuma_2010_Hum.Mutat_31_602
Author(s) : Shinkuma S , Akiyama M , Inoue A , Aoki J , Natsuga K , Nomura T , Arita K , Abe R , Ito K , Nakamura H , Ujiie H , Shibaki A , Suga H , Tsunemi Y , Nishie W , Shimizu H
Ref : Hum Mutat , 31 :602 , 2010
Abstract : Autosomal recessive hypotrichosis (ARH) is characterized by sparse hair on the scalp without other abnormalities. Three genes, DSG4, LIPH, and LPAR6 (P2RY5), have been reported to underlie ARH. We performed a mutation search for the three candidate genes in five independent Japanese ARH families and identified two LIPH mutations: c.736T>A (p.Cys246Ser) in all five families, and c.742C>A (p.His248Asn) in four of the five families. Out of 200 unrelated control alleles, we detected c.736T>A in three alleles and c.742C>A in one allele. Haplotype analysis revealed each of the two mutant alleles is derived from a respective founder. These results suggest the LIPH mutations are prevalent founder mutations for ARH in the Japanese population. LIPH encodes PA-PLA(1)alpha (LIPH), a membrane-associated phosphatidic acid-preferring phospholipase A(1)alpha. Two residues, altered by these mutations, are conserved among PA-PLA(1)alpha of diverse species. Cys(246) forms intramolecular disulfide bonds on the lid domain, a crucial structure for substrate recognition, and His(248) is one amino acid of the catalytic triad. Both p.Cys246Ser- and p.His248Asn-PA-PLA(1)alpha mutants showed complete abolition of hydrolytic activity and had no P2Y5 activation ability. These results suggest defective activation of P2Y5 due to reduced 2-acyl lysophosphatidic acid production by the mutant PA-PLA(1)alpha is involved in the pathogenesis of ARH.
ESTHER : Shinkuma_2010_Hum.Mutat_31_602
PubMedSearch : Shinkuma_2010_Hum.Mutat_31_602
PubMedID: 20213768
Gene_locus related to this paper: human-LIPH

Title : NO-1886 (ibrolipim), a lipoprotein lipase activator, increases the expression of uncoupling protein 3 in skeletal muscle and suppresses fat accumulation in high-fat diet-induced obesity in rats - Kusunoki_2005_Metabolism_54_1587
Author(s) : Kusunoki M , Tsutsumi K , Iwata K , Yin W , Nakamura T , Ogawa H , Nomura T , Mizutani K , Futenma A , Utsumi K , Miyata T
Ref : Metabolism , 54 :1587 , 2005
Abstract : Although the lipoprotein lipase (LPL) activator NO-1886 shows antiobesity effects in high-fat-induced obese animals, the mechanism remains unclear. To clarify the mechanism, we studied the effects of NO-1886 on the expression of uncoupling protein (UCP) 1, UCP2, and UCP3 in rats. NO-1886 was mixed with a high-fat chow to supply a dose of 100 mg/kg to 8-month-old male Sprague-Dawley rats. The animals were fed the high-fat chow for 8 weeks. At the end of the administration period, brown adipose tissue (BAT), mesenteric fat, and soleus muscle were collected and levels of UCP1, UCP2, and UCP3 messenger RNA (mRNA) were determined. NO-1886 suppressed the body weight increase seen in the high-fat control group after the 8-week administration (585 +/- 39 vs 657 +/- 66 g, P < .05). NO-1886 also suppressed fat accumulation in visceral (46.9 +/- 10.4 vs 73.7 +/- 14.5 g, P < .01) and subcutaneous (43.1 +/- 18.1 vs 68.9 +/- 18.8 g, P < .05) tissues and increased the levels of plasma total cholesterol and high-density lipoprotein cholesterol in comparison to the high-fat control group. In contrast, NO-1886 decreased the levels of plasma triglycerides, nonesterified free fatty acid, glucose, and insulin. NO-1886 increased LPL activity in soleus muscle (0.082 +/- 0.013 vs 0.061 +/- 0.016 mumol of free fatty acid per minute per gram of tissue, P < .05). NO-1886 increased the expression of UCP3 mRNA in soleus muscle 3.14-fold (P < .01) compared with the high-fat control group without affecting the levels of UCP3 in mesenteric adipose tissue and BAT. In addition, NO-1886 did not affect the expression of UCP1 and UCP2 in BAT, mesenteric adipose tissue, and soleus muscle. In conclusion, NO-1886 increased the expression of UCP3 mRNA and LPL activity only in skeletal muscle. Therefore, a possible mechanism for NO-1886's antiobesity effects in rats may be the enhancement of LPL activity in skeletal muscle and the accompanying increase in UCP3 expression.
ESTHER : Kusunoki_2005_Metabolism_54_1587
PubMedSearch : Kusunoki_2005_Metabolism_54_1587
PubMedID: 16311090

Title : Interleukin-6 and erythropoietin act as direct potentiators and inducers of in vitro cytoplasmic process formation on purified mouse megakaryocytes - An_1994_Exp.Hematol_22_149
Author(s) : An E , Ogata K , Kuriya S , Nomura T
Ref : Experimental Hematology , 22 :149 , 1994
Abstract : Mouse megakaryocytes were purified using a rabbit antimouse platelet serum, and magnetic beads were conjugated with an antirabbit IgG antibody. The purified cells were 95.8 +/- 1.2% megakaryocytes, and the recovery and viability of the megakaryocytes were 70 +/- 18.4%, and 80 +/- 13.4%, respectively. The effects of recombinant erythropoietin (Epo), interleukin-6 (IL-6), and IL-1 beta on these purified megakaryocytes were studied. Epo and IL-6 significantly increased DNA synthesis in these cells, but IL-1 beta did not. Similarly, both Epo and IL-6, but not IL-1 beta, increased the acetylcholinesterase (AchE) activity in the megakaryocytes. Epo and IL-6 stimulated the megakaryocytes to form cytoplasmic processes, which are considered to represent in vitro proplatelet formation. This process formation was inhibited by the addition of colchicine to the cultures. It was concluded that Epo and IL-6 are not only direct potentiators of megakaryocytes, but also inducers of in vitro cytoplasmic process formation on megakaryocytes.
ESTHER : An_1994_Exp.Hematol_22_149
PubMedSearch : An_1994_Exp.Hematol_22_149
PubMedID: 8299737