| Project/Area Number |
04806024
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
林産学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KURODA Hiroyuki KYOTO UNIV., WOOD RESEARCH INSTITUTE Instructor, 木質科学研究所, 助手 (00115841)
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| Co-Investigator(Kenkyū-buntansha) |
MORIKAWA Hiromichi HIROSHIMA UNIV., SCIENCE Professor, 理学部, 教授 (00089129)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | woody plants / regeneration / aspen / direct gene introduction / パーティクルガン / 形質転換 |
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| Research Abstract |
Sample Preparation of Woody Plants :
First, tissues from woody plants were sterilized. Young shoots were essential for the sterilization while the shoots older than one month failed the decontamination with fungi or bacteria. The sterilized shoots cultured on agar medium were used for the experiments.
Thin layr culture was then examined for introduction of genes into the tissue. Two mm thick discs from the shoots were obtained by a handmade slicer, which was razor blades arranged 2mm interval with spacers. The slicer was able to slice 500-1000 pieces of 2mm-thick discs within several minutes. Another hand-made slicer was also used for the preparation of thin section (500um thick) from aspen shoots.
Before introduction of genes into the discs or thin sections, 2mm thick discs were optimized for regenaration. The discs were cultured on the medium containing various concentration of cytokinin with NAA.After one month culture, 2-5 shoots in an aspen disc were formed with BA(2ppm) and NAA(0.01PPm).
Direct introduction of genes :
pBI221 was used for cytochemical transient assay. Rather low transient expressions were observed in the aspen tissues sliced. The efficiency was 1-40 spot/plate. A pretreatment of the tissues improved the efficiency(200-500 spots/plate). It is high enough to obtain transgenic aspen plants. In order to clarify which factor is concerned with the high transient expression, studies are going on.
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