| Project/Area Number |
18570050
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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・分子
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| Research Institution | Japan Women's University |
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Principal Investigator |
SEKIMOTO Hiroyuki Japan Women's University, Faculty of Sciences, Associate Professor (20281652)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Motomi University of Tokyo, Graduate School of Arts and Sciences, Professor (00193524)
ABE Jun Japan Women's University, Faculty of Sciences, Researcher (10424764)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,020,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Closterium / sexual reproduction / receptor / sex pheromone |
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| Research Abstract |
The desmid Closterium peracerosum-strigosum-littorale complex (C. pslc), which is the closest unicellular sister to land plants, is the best-characterized of the charophycean green algae with respect to the process of sexual reproduction. Two sex pheromones (PR-IP and PR-IP Inducer) involved in the progress of sexual reproduction of the C. pslc have been well characterized. In addition, putative receptor molecules (CpRLK1, CpRLP1) for these pheromones have been identified.
From the results of two hybrid systems, presences of week interactions between PR-IP Inducer and CpRLK1 and between 19-kDa subunit and CpRLPI were confirmed. To identify another receptor molecules for pheromones, two-hybrid screenings from cDNA library for sexual reproductive processes were performed using cDNAs encoding respective pheromones as baits. As a result, two genes showing an increase in expression during sexual reproduction were identified when 19-kDa subunit of PR-IP was used as a bait. In the case of PR-IP Inducer, one gene showing significant similarity with a previously identified gene (Cp-01) was screened at multiple times The expression of Cp-01 was elevated only in mt^+ cells by the addition of PR-IP Inducer.
To further elucidate their sexual reproduction, the transformation system for the C. pslc by using particle bombardment method was constructed. Two GFP constructs driven by endogenous promoters of C. pslc (CpHSP7Op and CpCABp, which are highly and constitutively expressed through the reproduction processes) were prepared and introduced into the cell by particle bombardment. These GFP signals could be transiently detected in the cytoplasm during 48-72 h after bombardment. When a GFP-fusion construct with a nucleus-localized selective marker gene (ble) was introduced, the GFP signal was only visible in the nucleus. These results indicate that this method and strategy are applicable to study cellular localization of unknown gene products during the sexual reproduction in C. pslc.
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