2004 Fiscal Year Final Research Report Summary
Isolation and functional analysis of the self-incompatibility gene in the grasses (Poaceae)
| Project/Area Number |
15580006
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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 |
Breeding science
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| Research Institution | Mie University |
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Principal Investigator |
KAKEDA Katsuyuki Mie University, Faculty of Bioresources, Associate Professor, 生物資源学部, 助教授 (50221867)
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| Co-Investigator(Kenkyū-buntansha) |
KOWYAMA Yasuo Mie University, Faculty of Bioresources, Professor, 生物資源学部, 教授 (80024579)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Self-incompatibility / S gene / Hordeum bulbosum / mRNA fingerprinting / Poaceae / Linkage analysis / Barley / 2D-gel analysis |
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| Research Abstract |
The AFLP-based mRNA, fingerprinting (AME) analysis was conducted to find S genotype-specific transcripts from pistils and anthers at anthesis in Wild barley, Hordeum bulbosum (2x) that possesses the gametophytic self-incompatibility (SI). A detailed genetic linkage map around the S locus was constructed using several clones obtained by the AMF analysis. Comparison of the linkage map with a physical map of barley choromosome 1 indicated that the S locus of H. bulbosum was located in a large chromosomal region across the centromere, where recombination is severely suppressed. In the linkage analysis, we have obtained five clones that showed no recombination with the S locus. Two of those clones showed pistil- or anther-specific expression. From sequence analysis of, the full length cDNA, the pistil-specific clone (HPS10) was shown to encode a small unknown protein that seems to be a putative secreted signaling molecule. Together with a high degree of allelic polymorphism, this clone turned out to be a promising candidate for the pistil S gene. Thus, we have been trying, to transform H. bulbosum calli with its RNAi construct, but so far we have obtained no transformants. For the anther-specific clone (122-7), sequence analysis revealed that it would encode a protein carrying an F-box motif. This finding is notable because the pollen S factor recently identified in the other gametophytic SI species (Solanaceae, etc.) is found to be an F-box protein. Further investigation on allelic polymorphism of the clone is needed. In the final experiment, we conducted 2D-gel analysis of pistil proteins in H. bulbosum and identified an S genotype-specific protein by LC-MS/MS. A cDNA clone coding for the protein was isolated and revealed to be tightly linked to the S locus.
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