1998 Fiscal Year Final Research Report Summary
Symbiotic engineering with green manure leguminous plant and nitrogen-fixing rhizobia
| Project/Area Number |
08455380
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Osaka University |
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Principal Investigator |
MUROOKA Yoshikatsu Osaka Univ.Eng.Professor, 大学院・工学研究科, 教授 (60029882)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Takashi Hiroshima Univ.Eng.Professor, 工学部, 教授 (40230461)
YAMASHITA Mitsuo Osaka Univ.Eng.Assoc.Prof., 大学院・工学研究科, 助教授 (40220347)
KANEKO Yoshinobu Osaka Univ.Eng.Assoc.Prof., 大学院・工学研究科, 助教授 (90161182)
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| Project Period (FY) |
1996 – 1998
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| Keywords | nitrogen-fixation / nodule / Mesorhizobium huakuii / symbiont / leguminous plant / Astragallus sinicus / symbiotic engineering |
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| Research Abstract |
1) Physical map of a large symbiotic plasmid in Mesorhizobium huakuii subsp. rengei :
A large plasmid, pRhYM was prepared from M.huakuii subsp. rengei B3, which was isolated from renge-sou (Astragalus sinicus), by pulse field electrophoresis. The gene libraries were constructed from DNA fragments of the plasmid with a super cosmid vector. The libraries were analyzed by hybridization and made contig of about 300 kb.
2) Nucleotide sequence of genes involved in nitrogen fixation and nodulation :
A total about 80 kb nucleotide sequence of the symbiotic plasmid pRhYM was determined. On this sequence, several clusters of the nif and nod genes were located.
3) Development of transformation system of M.huakuii :
By using pBBR plasmid vector, M.huakuji subsp. rengei was transformed at the efficiency of 10^5.
4) Nodule formation on renge-sou with the transgenic Rhizobium :
The transformed strain of M.huakuii subsp. rengei was used to infect renge-sou seeds. The roots formed nodules, and the plasmid pBBR was isolated from the nodules.
Thus, development of symbiotic engineering is now progress both in renge-sou plant and Rhizobium as a model.
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