| Project/Area Number |
07660361
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Zootechnical science/Grassland science
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| Research Institution | MIYAZAKI UNIVERSITY |
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Principal Investigator |
KAWAMURA Osamu MIYAZAKI UNIVERSITY,FACULTY OF AGRICULTURE,ASSOCIATE PROFESSOR, 農学部, 助教授 (00041062)
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| Co-Investigator(Kenkyū-buntansha) |
AKASHI Ryo MIYAZAKI UNIVERSITY,FACULTY OF AGRICULTURE,ASSISTANT PROFESSOR, 農学部, 助手 (20253809)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
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| Keywords | FORAGE / DIGESTIBILITY / LIGNIN / TRANSFORMATION / AGROBACTERIUM / ANTISENSE / CINNAMYL ALCOHOL DEHYDROGENASE / BIRD'S-FOOT TREFOIL |
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| Research Abstract |
In order to produce the high-digestible, as it were, the brown midrib type forage contained less lignin, we carried out the experiments as follows ;
1) Bird's-foot trefoil (Lotus corniculatus L.) are transformed with Agrobacterium tumefaciens carrying the plasmid vector pBI121 contained a kanamycin resistant gene (NTP II) and beta-glucuronidase gene (GUS). Resistant plants were regenerated at high frequency from green calli developed on the inoculated hypocotyl cuttings under the kanamycin selection.
2) The histological GUS assay showed that 7.0% of the resistant shoots also expressed the GUS gene in a variety tissues. The stable integration of this gene was confirmed by polymerase chain reaction (PCR) analysis.
3) A partial gene encoding CAD (Cinnamyl Alcohol Dehydrogenase) was isolated from Aralia cordata using PCR.An 1.2-kb portion of the sequence of the CAD gene obtained by PCR was inserted into an antisense orientation between the 35S promoter and the NOS terminater gene in T-DNA region of pGAH/SL-GUS.This constructed plasmid pGAH/anti-UCAD was introduced into bird's foot trefoil. The transgenic plants were selected by means of the resistance to kanamycin and hygromycin phosphotransferase.
4) The stable integration of the antisense gene was confirmed by PCR in the transgenic plants. The CAD activity of the transgenic plants was lower significantly than that of the control plants, but the lignin and the aldehyde contents and the in vitro digestibility by rumen mincroorganisms were not significantly different between the transgenic and control plants.
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