벼의 c/DRE Binding Factor 4 유전자를 이용한 내염성 벼 형질전환 계통 개발
Development of Salt-Tolerant Transgenic Rice using Rice c/DRE Binding Factor 4 cDNA
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초록
This study was conducted to isolate a salt tolerant gene and to develop salt tolerant rice for reclaimed-saline areasthrough genetic transformation. A rice c/DRE binding factor 4 (OsCBF4) cDNA was isolated from rice (cv. Nipponbare) usingRT-PCR. The full-length cDNA of the CBF4 gene consists of 1,429 nucleotides and 274 amino acid residues. The OsCBF4 sharesfrom 33 to 49% identity of deduced amino acid sequence with other CBFs of rice. In order to develop salt tolerant rice, transgenicrice plants containing the OsCBF4 gene were obtained via Agrobacterium-mediated transformation. The stable incorporation ofthe OsCBF4 gene into rice genome was confirmed by PCR and Southern analysis. The stable expression of introduced gene wasalso validated by RT-PCR analysis in T2 plants. Biological assay of T3 progeny of the transgenic plants in Yoshida solutioncontaining 120 mM Nacl for 2 weeks, confirmed that the OsCBF4 confers salt tolerance to transgenic rice plants. OsCBF4transgene in the transgenic line CBF4-10 was markedly expressed up to over three-fold in the leaf by 120 mM NaCl treatment. Real-time PCR analysis revealed that the levels of the transgene expression were markedly increased under salt treatment. Thetransgenic line CBF4-10 which showed highest ability to recover from the saline stress could be used as a potential source forsalt tolerance in rice breeding programs.
Abstract
This study was conducted to isolate a salt tolerant gene and to develop salt tolerant rice for reclaimed-saline areasthrough genetic transformation. A rice c/DRE binding factor 4 (OsCBF4) cDNA was isolated from rice (cv. Nipponbare) usingRT-PCR. The full-length cDNA of the CBF4 gene consists of 1,429 nucleotides and 274 amino acid residues. The OsCBF4 sharesfrom 33 to 49% identity of deduced amino acid sequence with other CBFs of rice. In order to develop salt tolerant rice, transgenicrice plants containing the OsCBF4 gene were obtained via Agrobacterium-mediated transformation. The stable incorporation ofthe OsCBF4 gene into rice genome was confirmed by PCR and Southern analysis. The stable expression of introduced gene wasalso validated by RT-PCR analysis in T2 plants. Biological assay of T3 progeny of the transgenic plants in Yoshida solutioncontaining 120 mM Nacl for 2 weeks, confirmed that the OsCBF4 confers salt tolerance to transgenic rice plants. OsCBF4transgene in the transgenic line CBF4-10 was markedly expressed up to over three-fold in the leaf by 120 mM NaCl treatment. Real-time PCR analysis revealed that the levels of the transgene expression were markedly increased under salt treatment. Thetransgenic line CBF4-10 which showed highest ability to recover from the saline stress could be used as a potential source forsalt tolerance in rice breeding programs.
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