By using OsRacD cDNA as probe to screen the genomic library of photoperiod sensitive genic male sterile rice line Nongken 58S, a positive clone containing 2 kb promoter and 396 bp coding region of OsRacD was obtained....By using OsRacD cDNA as probe to screen the genomic library of photoperiod sensitive genic male sterile rice line Nongken 58S, a positive clone containing 2 kb promoter and 396 bp coding region of OsRacD was obtained. Compared with the promoter of OsRacD cloned by reverse PCR from normal rice variety Nongken 58 (Nongken 58N), the homology was 99.8%, and the different nucleotides were outside the predicted response elements in promoter, suggesting that the fertility between rice varieties Nongken 58S and Nongken 58N under the long-day conditions was not attributed to the difference in the structure of OsRacD upstream regulation sequences, but to the developmental regulation of gene differential expression.展开更多
Using an improved version of mRNA differential display technology, we have obtained a differentially displayed fragment RDP-8. Homologous comparison indicated that the fragment RDP-8 has high homology with the gene en...Using an improved version of mRNA differential display technology, we have obtained a differentially displayed fragment RDP-8. Homologous comparison indicated that the fragment RDP-8 has high homology with the gene encoding maize small GTP-binding protein. By screening cDNA library of the rice Nongken 58N panicle using the newly obtained fragment RDP-8 as probe, we further found the full-length cDNA of osRACD gene that encodes a rice small GTP-binding protein. As compared with maize RACD gene, the osRACD of rice shows remarkable homology in both nucleotide sequence and amino acid sequence, 88% and 97% respectively. Evidence from RT-PCR study indicates that osRACD gene is related to photoperiod fertility conversion of photoperiod sensitive genie male sterility (PSGMS) rice.展开更多
OsRacD, belonging to rice (Oryza sativa L. ssp. japonica) Rho family of the small GTPases, is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice which i...OsRacD, belonging to rice (Oryza sativa L. ssp. japonica) Rho family of the small GTPases, is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice which influences the rice fertility via controlling the pollen tube growth. Using OsRacD as bait, two GDP dissociation inhibitor genes designated as OsRhoGDI1 and OsRhoGDI2 were screened by yeast two-hybrid system. To further study the regulation mechanism of OsRhoGDIs, and also the relationships between OsRhoGDIs and OsRacD, the upstream regulation sequences of OsRhoGDI1 and OsRhoGDI2 were cloned by PCR, and the cis-elements in the promoters were predicted using internet tools in this study. The results showed that there were several kinds of response elements, such as phytohormone response elements, lightregulated elements and adversity induced elements in the promoters of OsRhoGDIs, some of which were similar. Comparing with the promoter of OsRacD, several pollen tube germination related elements were found. On one hand, the expression and regulation mechanism of OsRhoGDIs were complex, they may be regulated by several signaling pathways commonly, and the regulation mechanisms were similar to some extent. On the other hand, it was suggested that the fertility of photoperiod sensitive genic male sterile rice may be controlled by the cooperative expression of OsRhoGDIs and OsRacD.展开更多
文摘By using OsRacD cDNA as probe to screen the genomic library of photoperiod sensitive genic male sterile rice line Nongken 58S, a positive clone containing 2 kb promoter and 396 bp coding region of OsRacD was obtained. Compared with the promoter of OsRacD cloned by reverse PCR from normal rice variety Nongken 58 (Nongken 58N), the homology was 99.8%, and the different nucleotides were outside the predicted response elements in promoter, suggesting that the fertility between rice varieties Nongken 58S and Nongken 58N under the long-day conditions was not attributed to the difference in the structure of OsRacD upstream regulation sequences, but to the developmental regulation of gene differential expression.
基金This work was supported by the Fund of the State "863" and the National Basic Sciences Initiative (Grant No. 2001CB1088).
文摘Using an improved version of mRNA differential display technology, we have obtained a differentially displayed fragment RDP-8. Homologous comparison indicated that the fragment RDP-8 has high homology with the gene encoding maize small GTP-binding protein. By screening cDNA library of the rice Nongken 58N panicle using the newly obtained fragment RDP-8 as probe, we further found the full-length cDNA of osRACD gene that encodes a rice small GTP-binding protein. As compared with maize RACD gene, the osRACD of rice shows remarkable homology in both nucleotide sequence and amino acid sequence, 88% and 97% respectively. Evidence from RT-PCR study indicates that osRACD gene is related to photoperiod fertility conversion of photoperiod sensitive genie male sterility (PSGMS) rice.
文摘OsRacD, belonging to rice (Oryza sativa L. ssp. japonica) Rho family of the small GTPases, is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice which influences the rice fertility via controlling the pollen tube growth. Using OsRacD as bait, two GDP dissociation inhibitor genes designated as OsRhoGDI1 and OsRhoGDI2 were screened by yeast two-hybrid system. To further study the regulation mechanism of OsRhoGDIs, and also the relationships between OsRhoGDIs and OsRacD, the upstream regulation sequences of OsRhoGDI1 and OsRhoGDI2 were cloned by PCR, and the cis-elements in the promoters were predicted using internet tools in this study. The results showed that there were several kinds of response elements, such as phytohormone response elements, lightregulated elements and adversity induced elements in the promoters of OsRhoGDIs, some of which were similar. Comparing with the promoter of OsRacD, several pollen tube germination related elements were found. On one hand, the expression and regulation mechanism of OsRhoGDIs were complex, they may be regulated by several signaling pathways commonly, and the regulation mechanisms were similar to some extent. On the other hand, it was suggested that the fertility of photoperiod sensitive genic male sterile rice may be controlled by the cooperative expression of OsRhoGDIs and OsRacD.