| Project/Area Number |
13640653
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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 |
植物生理
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| Research Institution | Kagoshima University |
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Principal Investigator |
ABE Mikiko (2003) Kagoshima University, Faculty of Science, Professor, 理学部, 教授 (00107856)
内海 俊樹 (2001-2002) 鹿児島大学, 理学部, 助教授 (20193881)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Akihiro Kagoshima University, Faculty of Science, Assistant Professor, 理学部, 助手 (50305108)
UCHIUMI Toshiki Kagoshima University, Faculty of Science, Associate Professor, 理学部, 助教授 (20193881)
阿部 美紀子 鹿児島大学, 理学部, 教授 (00107856)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | symbiotic hemoglobin / non-symbiotic hemoglobin / leghemoglobin / Lotus japonicus / promoter / transformation / hairly root / GFP / アレイ |
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| Research Abstract |
Lotus japonicus is a promising model legume for studying molecular interactions in symbiotic nitrogen fixation and whole genome sequence of microsymbiont of Lotus japonicus, Mesorhizobium loti MAFF303099, is known. Transcriptional profiles of whole genome of M. loti revealed that the symbiosis island (611 kb region on the chromosome) functions as clustered expression islands to support symbiotic nitrogen fixation. A gene disruption of 1-aminocyclopropane-1-carboxylate deaminase, which is highly up-regulated in bacteroid (symbiotic state of rhizobia), decreased competitiveness on nodule formation. This suggests that disruption of highly expressed genes is a useful strategy for exploring novel gene functions in symbiosis.
A fusion gene of promoter region of a symbiotic hemoglobin gene (LjLb3) of L. japonicus and the structural gene of GFP was constructed and introduced into L. japonicus mediated by Agrobacterum tumefaciens or A. rhizogenes. Inoculation of M. loti to transgenic plants roots and hairy roots resulted in normal nodule formation. The expression of the fusion gene was confirmed by RT-PCR and GFP-expression could be observed by fluorescent microscopy during nodulation process. Interestingly, fluorescent cells were scattered on the surface of the roots after a few hours later of rhizobial inoculation. Using transgenic plants and transgenic hairy roots, quantitative observation of host plant cells that responding to rhizobia could be performed.
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