| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
1. When expression vectors for efficient cyan fluorescent protein-fused Prunus mumc (Rosaceae) S-RNase (pistil S haplotype determinant) and Venus (variant of Yellow fluorescent protein)-fused P. mumc SLF (pollen S haplotype determinant) were co-introduced into tobacco BY-2 cells, the S-RNase and SLF were co-localized in perinuclear cytoplasmic regions, suggesting the S-RNase and SLF interact with each other. However, the S-RNase-and SLF with either the same or different S haplotypes were co-localized.
2. In order to identify Petunia hybrida (Solanaceae) SLF, expression sequence tag (EST) analysis for pollen tubes of S_<B1> homozygote was carried out. Out of 2,746 ESTs, one sequence homologous to Petunia inflata SLF2 (PiSLF2), a gene known to act as pollen S haplotype determinant, was found, and the gene from S_<B1>homozygote was termed pB1SLE We also acquired genes homologous to PiSLF2 from pollen tube cDNAs of S_<D1> and S_<D2> homozygotes by RT-PCR with primer pair designed from pB1SLF, and the genes from S_<D1> and S_<D2> homozygotes were termed pD1SLF and pD2SLF, respectively.
RT-PCR with primer pair designed from P1SLF2 was also performed and six genes were obtained (termed rB1SLF (linked to S_<B1>haplotype), rB2SLF (linked to S_<B2> haplotype), rD1SLFI and rD1SLF2 (linked to S_<D1>haplotype), and rD2SLF1 and rD2SLF2 (linked to S_<D2> haplotype).
To determine whether these genes act as pollen S haplotype determinants, P. hybrida plants were transformed with these genes and pollen tube behavior of obtained transformants was observed. In obtained transformants, only plants harboring the pD2SLF transgene exhibited breakdown of self-incompatibility, suggesting that the pD2SLF acts as pollen S haplotype determinant.
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