Fundamental study on molecular farming using mulberry plant

1999 Fiscal Year Final Research Report Summary

• Project/Area Number: 09460034
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 蚕糸・昆虫利用学
• Research Institution: KYOTO INSTITUTE OF TECHNOLOGY
• Principal Investigator: SUGIMURA Yukio Faculty of Textile Science, Kyoto Institute of Technology, Associate Professor, 繊維学部, 助教授 (20273542)
• Co-Investigator(Kenkyū-buntansha): KOTANI Eiji Faculty of Textile Science, Kyoto Institute of Technology, Research Associate, 繊維学部, 助手 (10273541) ; FURUSAWA Toshiharu Faculty of Textile Science, Kyoto Institute of Technology, Professor, 繊維学部, 教授 (70127166)
• Project Period (FY): 1997 – 1999
• Keywords: Mulberry / Plantlet regeneration / Gene delivery / Transformant / Molecular farming

Research Abstract

Mulberry plant, a woody feedcrop for the silkworm, is an attractive species for molecular framing since the plant have superior agronomical characteristics. Transgenic mulberry clone, if produced, may provide a farming system for the production of valuable biomolecules. To realize this target, it is essential to establish the methods for (1) plantlet regeneration from explant tissues and (2) gene delivery into target tissue. High frequency of shoot regeneration was achieved by 2 step-culture system consisting of seedling culture for obtaining leaf explants and leaf culture for inducing shoot organogenesis. A critical factor in both cultures was medium composition in which 2, 3, 5-triiodobenzoic acid and cytokinin such as 6-benzylaminopurin and thidiazuron was included. For efficient gene delivery into regenerable leaf tissue, various bombardment conditions were optimized using a particle inflow gun device : gold particle as microprojectile, bombardment frequency, preculture period and conditioning treatment of target tissue. When the optimal conditions were combined, 200-300 gene expressing-cells per cmィイD12ィエD1 tissue were detected. Based on these results, it is possible to generate transgenic mulberry clones.

Research Products

• [Publications] Sugiura, Y. et al.: "Shoot bud formation and plantlet regeneration from the basal tissues of mulberry leaf"J. Seric. Sci. Jpn.. 67. 421-424 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sugimura, Y. et al.: "Efficient induction of shoot organogenesis from leaves of mulberry seedlings using 2, 3, 5, -triiodoben benzoic acid"Plant Biotech.. 16. 123-127 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sugimura, Y. et al.: "Gene transfar by electroporation into protoplasts isolated from mulberry calli"J. Seric. Sci. Jpn.. 68. 49-53 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Adachi, T. et al.: "Adaptability of mulberry plantlets regenerated in vitro to outdoor conditions"J. Seric. Sci. Jpn.. 68. 161-163 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sugimura, Y. et al.: "Transient expression of β-glucuronidase gene transferred Into leaf tissues of mulberry seedings by particle Inflow gun"Sericologia. 39. 33-38 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sugimura, Y. et al.: "Major improvements In gene deliverly Into mulberry leaf cells by particle Inflow gun"J. Seric. Sci. Jpn.. 69(印刷中). (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 1. Sugimura, Y., Adachi, T., Konani, E. and Furusawa, T.: "Shot bud formation and plantlet regeneration from the basal tissues of mulberry leaf."J. Seric. Sci., Jpn.. 67. 421-424 (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 2. Sugimura, Y., Adachi, T., Kotani, E. and Furusawa, T.: "Efficient induction of shoot organogenesis from leaves of mulberry seedlings using 2, 3, 5-triiodobenzoic acid."Plant Biotech.. 16. 123-127 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 3. Sugimura, Y., Miyazaki, J., Yonebayashi, K., Kotani, E. and Furusawa, T.: "Gene transfer by electroporation into protoplasts isolated from mulberry calli."J. Seric. Sci., Jpn.. 68. 49-53 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 4. Sugimura, Y., Adachi, T., Abe, M., Kotani, E. and Furusawa, T.: "Tansient expression of β-glucronidase gene transferred into leaf tissues of mulberry seedlings by the particle inflow gun."Sericologia. 39. 33-38 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 5. Adachi, T., Sugimura, Y., Kotani, E. and Furusawa, T.: "Adaptability of mulberry plantlets regenerated in vitro to outdoor conditions."J. Seric. Sci., Jpn.. 68. 161-163 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 6. Sugimura, Y., Uchida, A., Adachi, T., Hasegawa, H., Kotani, E. and Furusawa, T.: "Major improvements in gene delivery into mulberry leaf cells by particle inflow gun."J. Seric. Sci., Jpn.. 69 (in press). (2000)
  • Description: 「研究成果報告書概要(欧文)」より