Summary of Research Project Results Under the JSPS FY2000
"Research for the future Program"



1.Research Institution Tokyo University of Agriculture & Technology
 
2.Research Area Life Science
 
3.Research Field Mechanisms of Plant Responses to Environment and Application to Biotechnology
 
4.Term of Project FY1996〜FY2000
 
5.Project Number 96L00605
 
6.Title of Project Molecular Breeding of Woody Plants and Its Fundamental Study

7.Projetct Leader
Name Institution,Department Title of Position
Noriyuki Morohoshi Tokyo University of Agriculture and Technology, BASE Professor

8.Core Members

Names Institution,Department Title of Position
Hiroo Fukuda University of Tokyo, Biological Science Professor
Takao Ito Kyoto University, Wood Research Institute Professor
Toyoki Kozai Chiba University, Horticulture Professor

9.Cooperating Researchers

Names Institution,Department Title of Position
Keishi Takabe Kyoto University, Forest and Biomaterial Associate Professor
Koichi Mizuno Osaka University, Graduate school of Science Associate Professor
Yoshihiro Ozeki Tokyo University of Agriculture and Technology, Technology Associate Professor

10.Summary of Research Results

This project was carried out from two viewpoints which are fundamental and applied fields. In former, the molecular biological and histochemical analyses on the formation of vascular bundle and on the biosynthesis of cellulose and lignin in woody plants were performed. In latter, the following issues were researched the establishment of the techniques, recombination, differentiation and regeneration, and tissue culture in wood, and the development of application techniques to make a useful transgenic trees. These results are as follows:
1) Transcription factors involved in the formation of vascular bundles, the promoter of the hydrolysis enzyme genes expressing when vessels are formed especially, and the factor genes inducing the differentiation were isolated and analyzed. It was clear that these isolated promoters usefully worked in wood.
2) High polymer of cellulose was synthesized in the terminal complexes (TC) on the cell wall. TC was composed of CesA protein, g-tubulin protein and other 5 to 6 kinds of constitutional proteins.
3) PAL, OMT, CAD and PO genes involved in lignin biosynthesis were isolated from wood. Localization of these enzymes on the cell wall was tested using their antibody on the electron microscopic level and the mechanism of lignification was clear on the cell wall.
4) MAT vector system, a recombination technique of woody plants was devised. Using this method, the poplar decreasing 25-45% of lignin content and the Cryptomerea japonica that not bring about pollinosis were made.
5) A mass production system on the bases of the photoautotrophic techniques was developed by the combination of new some techniques on the introduce of adventive embryo, regeneration and tissue culture in sugi, hinoki, mahogany and so on.

11.Key Words

(1)Genes forming vascular bundle、(2)Cellulose、(3)Lignin
(4)Transgenic woody plants、(5)Tissue culture of gymnosperms、(6)Mass-production culture of wood
(7)MAT-vector system、(8)Transgenic sugi without pollinosis、(9)Poplar containing low lignin content

12.References

[Reference Articles]
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Planta   2001 in press.

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Kuriyama, H. Regulation of tracheary element differentiation.
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Tamura, T. Suitability of peroxidase-suppressed transgenic hydrid aspen (Populus Sieboldii x P. gradidentata) for pulping.
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Holzforschung 55 2001 in press

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Nakamura, T. Sedimentable amyloplasts in starch sheath cells of stems of Japanese cherry.
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Advances in Space Research   2001 in press

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Funada, R. Changes in levels of endogenous plant hormones in cambial rcgions of stems of Larix Teptolepis at the onset of cambial activity in springtime.
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Journal of Wood Science   2001 in press

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Yazaki, K. Growth and annual ring structure of Larix sibirica grown at different CO2 concentration and nutrient supply.
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Tree Physiology   2001 in press

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Ebinuma, H. MAT (Multi-Auto-Transformation) vector system. The oncogenes of Agrobacterium as positive markers for regeneration and selection of marker-free transgenic plants.
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In Vitro Cell. Dev. Biol.   2001 in press

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Ebinuma, H. Systems for removal of a selection marker and their combination with a positive marker.
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Plant Cell Rep.   2001 in press

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Endo, S. Inhibition of proteasome activity by the TED4 protein in extracellular space: a novel mechanism for protection of living cells from injury caused by dying cells.
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Plant Cell Physiol. 42 2001 9-19

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Yamamoto, R. Brassinosteroid levels increase drastically prior to morphogenesis of tracheary elements.
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Plant Physiol. 125 2001 556-563

Author Title of Article
Motose, H. An arabinogalactan protein(s) is a key component of a fraction that mediates local intercellular communication involved in tracheary element differentiation of zinnia mesophyll cells.
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Plant Cell Physiol. 42, 2001 129-137

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Obara, K. Direct evidence of active and rapid nuclear degradation triggered by vacuole rupture during programmed cell death in Zinnia.
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Plant Physiol. 125 2001 615-626

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Nakagawa, N. Cell wall integrity controls expression of cndoxyloglucan transferase in Tobacco BY2 cells.
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Plant & Cell Physiol. 42 2001 240-244

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Heo, J. A forced ventilation micropropagation system for photoautotrophic production of sweetpotato plug plantlets in a scaled-up culture Vessel: I. Growth and uniformity.
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HortTechnology 11 2001 90-94

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Kubota C. Mathematical models for planning vegetative propagation undercontrolled environments.
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HortScience 36 2001 15-19

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Wilson, S. B. A forced ventilation micropropagation system for photoautotrophic production of sweetpotato plug plantlets in a scaled-up culture Vessel: II. Carbohydrate status.
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Takeuchi, M. Immunolacalization of O-methyltransferase and peroxidase in differentiating xylem of poplar.
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Holzforschung 55 2001 146-150

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Sato, S. Role of the putative membrane-bound endo-1, 4 β-glucanase KORRIGANin cell elongation and cellulose synthesis in Arabidopsis thaliana.
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Plant Cell Physiol. 42 2001 251-263

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Yasuda, S. Formation and chemical structures of acid-soluble lignin I: sulfuric acid treatment time and acid-soluble lignin content of hardwood.
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Journal of Wood Science 47 2001 69-72

Author Title of Article
Funada, R. Seasonal variations in cndogenous indole-3-acetic acid and abscisic acid in the cambial region of Pinus densiflora stems in relation to earlywood-latewood transition and cessation of tracheid production.
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Holzforschung 55 2001 128-134

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Oribe, Y. Cambial reactivation in the partially heated stem in an evergreen conifer, Abies sachalinensis.
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Planta 212 2001 684-691

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Maruyama, E. Embryogenic cell culture, protoplast regeneration, cryopreservation, biolistic gene transfer and plant regeneration in Japanese Cedar
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Plant Biotechnology 17 2000 281-298

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Hiwatashi, Y. Tissue-specific localization of mRNA for carrot homeobox genes, CHBs.
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Plant Cell Physiol. 41 2000 639-643

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Fukuda, H. Programmed cell death of tracheary elements as a paradigm in plants.
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Plant Mol. Biol. 44 2000 245-253

Author Title of Article
Shinohara, N. Isolation of a vascular cell wall-specific monoclonal antibody recognizing a cell polarity by using a phage display subtraction method.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA. 97 2000 2585-2590

Author Title of Article
Nara-Sato, K. The rates of deceleration of nuclear and organellar DNA syntheses differ in the progenitor cells of the apical meristems during carrot somatic embryogenesis.
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Planta 211 2000 457-466

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Koizumi, K. A series of novel mutants of Arabidopsis thaliana that are defective in the formationof continuous vascular network: Calling the auxin signal flow canalization hypothesis into question.
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Development 217 2000 3197-3204

Author Title of Article
Nakashima, J. Autolysis during in vitro tracheary element differentiation - Formation and location of the perforation.
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Plant Cell Physiol. 41 2000 1267-1271

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Sakamoto, J. Artificial chitin spherulites composed of single crystalline ribbons of chitin via enzymatic polymerization.
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Macromolecules 33 2000 4155-4160

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Kobayashi, S. Formation and structure of artificial cellulose spherulites via enzymatic polymerization.
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Biomacromolecules 1 2000 168-173

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Fujino, T. Characterization of cross-links between cellulose microfibrils, and their occurrence during elongation growth in pea epicotyl.
Journal Volume Year Pages Concerned
Plant & Cell Physiol. 41 2000 486-494

Author Title of Article
Suzuki, K. Highly substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans show a distinct localization pattern in the tissues of Zeamays L.
Journal Volume Year Pages Concerned
Plant & Cell Physiol. 41 2000 948-959

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Afreen-Zobayed, F. A Combination of vermiculite and paper pulp supporting material for the photoautotrophic micropropagation of sweet potato.
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Plant Science 157 2000 225-231

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Kim, H. H. The potential use of photoperiod during transplant production underartificial lighting conditions on floral development and bolting, using Spinach as a model.
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HortScience 35 2000 43-45

Author Title of Article
Zobayed, S. M. A. In vitro propagation of Lagerstroemia spp. from nodal explants and gaseous composition in the culture headspace.
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Environment Control Biology 38 2000 1-11

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Zobayed, S. M. A. Mass propagation of Eucalyptus camaldulensis in a scaled-up vessel under In Vitro photoautotrophic condition.
Journal Volume Year Pages Concerned
Annals of Botany 85 2000 587-592

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Zobayed, S. M. A. Water control and survival of Ipomoea batatas grown photoautotrophically under forced ventilation and photomixotrophically undernatural ventilation.
Journal Volume Year Pages Concerned
Annals of Botany 86 2000 603-610

Author Title of Article
Awano, T. Deposition and localization of glucuronoxylan on the secondary cell wall of Japanese beech as observed using immuno FE-SEM.
Journal Volume Year Pages Concerned
Protoplasma 212 2000 72-79

Author Title of Article
Maeda, Y. Immunolocalization of glucomannans in the cell wall of differentiating tracheids in Chamaecyparis obtusa.
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Protoplasma 213 2000 148-156

Author Title of Article
Nakashima, J. Antolysis during in vitro tracheary element differentiation: Formation and location of the perforation.
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Plant Cell Physiol. 41 2000 1267-1271

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Ozeki, Y. Phenylalanine ammonia-lyase genes involved in anthocyanin synthesis and the regulation of its expression in suspension cultured carrot cells.
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J. Plant Res. 113 2000 319-326

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Yoshimoto, Y. Isolation and Characterization of a cDNA for Phenylalanine Ammonia-lyase (PAL) from Dianthus caryophyllus (carnation).
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Plant Biotechnology 17 2000 321-326

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Nishikubo, N. Specific accumulation of cell wall polysaccharide-linked hydroxycinnamoyl esters in irregularly shaped and collapsed internode parenchyma cells of dwarf mutant rice. Fukei No. 71.
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Plant and Cell Physiology 41 2000 776-784

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Matsui, N. Conversion of syringyl to guaiacyl moieties on the cinnamyl alcohol pathway during the biosynthesis of lignin in angiosperms.
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Planta 210 2000 831-835

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Yasue, K. The effects of tracheid dimensions on variations in maximum density of Picea glehnii and relationships to climatic factors.
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Trees 14 2000 223-229

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Kitin, P. Analysis by confocal microscopy of the structure of cambium in the hardwood Kalopanax pictus.
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Annals of Botany 86 2000 1109-1117

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Hafren, J. Ultrastructural changes in the compound middle lamella of Pinus thunbergii during lignification and lignin removal.
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Holzforschung 54 1999 234-240

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Kimura, S. Different distribution of cellulose synthesizing complexes inbrittle and non-brittle strains of barlcy.
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Plant & Cell Physiol. 40 1999 335-338

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Kimura, S. Immunogold labeing of rosette terminal cellulose synthesizing complexes in a vascular plant (Vigna angularis).
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Plant Cell 11 1999 2075-2085

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Sugiyama, J. Molecular directionality of α-chitin biosynthesis.
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J. Mol. Biol. 286 1999 247-255

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Lloyd, A. Lignin distribution in mild compression wood of Pinus radiata.
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Can J Bot 77 1999 41-50

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Inagaki, Y. Genomic organization of the genes encoding dihydroflavonol 4-reductase for flower pigmentation in the Japanese and common morning glories.
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Gene 226 1999 181-188

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Chen, F. The structural conversion of lignin subunit at cinnamyl alcohol stage in Eucalyptus camaldulensis.
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Journal of Wood Science 45 1999 487-491

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Chen, F. Evidence for a novel biosynthetic pathway that regulates the ratio of syringyl to guaiacyl residues in lignin in the differentiating xylem of Magnolia kobus DC.
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Planta 207 1999 597-603

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Murakami, Y. The differentiation of contact cells and isolation cells in the xylem ray parenchyma of Populus maximowiczii.
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Annals of Botany 84 1999 429-435

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Kitin, P. Variations in the lengths of fusiform cambial cells and vessel elements in Kalopatax pictus.
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Annals of Botany 84 1999 621-632

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Utsumi, Y. The progression of cavitation in earlywood vessels of Fraxinus mandshurica var. japanica during freezing and thawing.
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Plant Physiology 121 1999 897-904

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Sugita, K. Effective selection system for generating marker-free transgenic plants independent of sexual crossing.
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Plant Cell Rep. 18 1999 941-947

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Igarashi, M. Expression of the Zinnia TED3 promoter in developing tracheary elements of transgenic Arabidopsis.
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Kimura, S. A new cellulosic structure, the tunic cord in the ascidian Polyandrocarpa misakiensis
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Protoplasma 204 1998 94-102

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Helbert, W. High resolution electron microscopy on ultrathin section of cellulose microfibrils generated by glomerulocytes in Polyzoa vesculiphora.
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Protoplasma 203 1998 84-90

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Imai, T. Unidirectional action of cellobiohydrolase I on Valonia microcrystals.
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FEBS Lett. 432 1998 113-116

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Imai, T. Nanodomains of I α and I β cellulose in algal microfibrils.
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Macromolecules 31 1998 6275-6279

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Helbert, W. Molecular Imaging of Halocynthia papillosa Cellulose.
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J. Struct. Biol. 124 1998 42-50

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Iwata, K. Effect of ions on the orientation of cortical microtubules in Spirogyra cells.
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Plant & Cell Physiol. 39 1998 1099-1103

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Suzuki, K. Localization of the xyloglucan in cell walls in a suspension culture of tobacco by rapid-freezing and deep-etching techniques coupled with immunogold labelling.
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Plant & Cell Physiol. 39 1998 1003-1009

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Suzuki, K. Cell wall architecture prerequisite for the cell division in the protoplasts of white poplar, Populus alba L.
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Plant & Cell Physiol. 39 1998 632-638

Author Title of Article
Fujino, T. Changes in the three dimensional architecture of the cell wall during lignification of xylem cells in Eucalyptus tereticornis.
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Holzforsch. 52 1998 111-116

Author Title of Article
Kimura, S. A new cellulosic structure, the tunic cord in the ascidian, Polyandrocarpa misakiensis.
Journal Volume Year Pages Concerned
Protoplasma 204 1998 94-102

Author Title of Article
Fujino, T. Change in pectin structure during epidermal cell elongation in pea (Pisum sativum) and its implications for cell wall architecture.
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Plant & Cell Physiol. 39 1998 1315-1323

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Nakagawa, N. Increase in the amount of celA 1 protein in Tobacco BY-2 cells by a cellulose biosynthesis inhibitor, 2, 6-dichlorobenzonitrile.
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Plant & Cell Physiol. 39 1998 779-785

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Aoyagi, S. BEN1 and ZEN1 cDNAsencoding S1-type DNases that are involved in programmed cell death in plants.
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FEBS Lett. 429 1998 134-138

Author Title of Article
Awano, T. Localization of glucuronoxylans in Japanese beech visualized by immunogold labeling.
Journal Volume Year Pages Concerned
Protoplasma 202 1998 213-222

Author Title of Article
Matoh, T. Immunocytochemistry of Rhamnogalacturonan II in Cell Walls of Higher Plants.
Journal Volume Year Pages Concerned
Plant Cell Physiol. 39 1998 483-491

Author Title of Article
Goodell, B. Laccase Immunolabelling and Microanalytical Analysis of Wood Degaded by Lentinus edodes.
Journal Volume Year Pages Concerned
Holzfoschung 52 1998 345-350

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Tanno, F. Antimicrobial TMSAH-Addcd Wood-Inorganic Compostes Prepared by the Sol-Gel Process.
Journal Volume Year Pages Concerned
Holzforschung 52 1998 365-370

Author Title of Article
Goto, M. Histochemistry and UV-microspectrometry of cell walls of untreated and ammonia-reated barley straw.
Journal Volume Year Pages Concerned
Can J Plant Sci. 78 1998 437-443

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Mato, M. Isolation and characterization of a cDNA clone of UDP-galactose: flavonoid 3-O-galactosyltransferase (UF3GaT) expresscd in the seedling of Vigna mungo.
Journal Volume Year Pages Concerned
Plant Cell Physiol. 39 1998 1145-1155

Author Title of Article
Utsumi, Y. Visualization of cavitated vessels in winter and refilled vessels in spring in diffuse-porous trecs by cryo-scanning electron microscopy.
Journal Volume Year Pages Concerned
Plant Physiology 117 1998 1463-1471

Author Title of Article
Hu, W-J. Compartmentalized expression of two structurally and functionally distinct 4-coumarate: CoA ligase genes in aspen (Populus tremuloides).
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 95 1998 5407-5412

Author Title of Article
Sato, Y. Changes in the activity and mRNA of cinnamyl alcohol dehydrogenase during tracheary element differentiation in Zinnia elegans.
Journal Volume Year Pages Concerned
Plant Physiol. 113 1997 425-430

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Ito, M. Cell-cycle-regulated transcription of A-and B-type plant cyclin genes in synchronous cultures.
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Plant J. 11 1997 983-992

Author Title of Article
Fukuda, H. Tracheary element differentiation.
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Plant Cell 9 1997 1147-1156

Author Title of Article
Yamamoto, R. Brassinosteroids induce entry into the final stage of tracheary element differentiation in culturedZinnia cells.
Journal Volume Year Pages Concerned
Plant Cell Physiol. 38 1997 980-983

Author Title of Article
Koyama, M. Parallel-up structure evidences the molecular directionality during biosynthesis of bacterial cellulose.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA. 94 1997 9091-9095

Author Title of Article
Nakashima, J. Immunocylochemical localization of phenylalanine ammonia-lyase and cinnamyl alcohol dehydrogenase in differentiating trachcary elements derived from Zinnia mcsophyll cells.
Journal Volume Year Pages Concerned
Plant Cell Physiol. 38 1997 113-123

Author Title of Article
Nakashima, J. Direct Visualization of lignifying secondary wall thickenings in Zinnia elegans Cells in Culture.
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Plant Cell Physiol. 38 1997 818-827

Author Title of Article
Nakashima, J. Inhibition of phenylalanine ammonia-lyase activity causes the depression of lignin accumulation of secondary wall thickening in isolated Zinnia mesophyll cells.
Journal Volume Year Pages Concerned
Protoplasma 196 1997 99-107

Author Title of Article
Ozeki, Y. Plant cell culture variation during long term subculturing caused by insertion of a transposable element in a phenylalanine ammonia-lyase (PAL) gene.
Journal Volume Year Pages Concerned
Mol. Gen. Genet. 254 1997 407-416

Author Title of Article
Takeda, J. Action spectrum for induction of promoter activity of phenylalanine ammonia-lyase gene by UV in carrot suspension cells.
Journal Volume Year Pages Concerned
Photochem. Photobiol. 66 1997 464-470

Author Title of Article
Sakurai, M. Isolation and characterization of a cDNA for chalcone synthase from cultured cells of the strawberry Fragaria ananassa, cv Shikinari.
Journal Volume Year Pages Concerned
Plant Biotechnol. 14 1997 183-185

Author Title of Article
Zhan, W. C. A new approach based on the leaf disc method for Agrobacterium mediated transformation and regeneration of aspen.
Journal Volume Year Pages Concerned
Plant Science 123 1997 105-112

Author Title of Article
Kajita, S. Structural characterization of modified lignin in transgenic tobacco plants in which the activity of 4-coumarate: coenzyme A ligase is depressed.
Journal Volume Year Pages Concerned
Plant Physiology 114 1997 871-879

Author Title of Article
Kajita, S. Immunological characterization of transgenic tobacco plants with a chimeric gene for 4-coumarate: CoA ligase that have altered lignin in their xylem tissue.
Journal Volume Year Pages Concerned
Plant Science 128 1997 109-118

Author Title of Article
Funada, R. The orientation and localization of cortical microtubules in differentiating conifer tracheids during cell expansion.
Journal Volume Year Pages Concerned
Plant and Cell Physiology 38 1997 210-212

Author Title of Article
Ebinuma, H. Selection of marker-free transgenic plants using the isopentenyl transferase gene as a selectable marker.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 94 1997 2117-2121

Author Title of Article
Ebinuma, H. Germplasm conservation of the tropical forest trees, Cedrela odorata L., Guazumacrinita Mart., and Jacaranda mimosaefolia D. Don., by shoot tip encapsulation in calcium-alginate and storage at 12-25 C.
Journal Volume Year Pages Concerned
Plant Cell Reports 16 1997 393-396

[Reference Books]
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K. Watanabe & A. Komamine Challenge of Plant and Agricultural Sciences to the Crisess of Biospere on the Earth in the 21st Century
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EUREKAH-COM/LANDESBIOSCIENCE 2000 309

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C. Kubota & C. Chun Transplant Production in the 21st Century
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Kluwer AcademicPublishers 2000 290


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