| Chilling injure in seedling period is one of major limiting factors for productivity of rice (Oryza sativa L.). To improve rice cold tolerance in seedling period, it is important to exploit novel cold-resistant genes, elucidate the underlying molecular mechanism and utilize the genes in cold-resistant molecular breeding. A novel cold-inducible gene encoding a putative C2H2-type zinc finger protein, designated OsCOI (Oryza sativa Cold-Inducible), has been identified in our former research. In this study, the gene was cloned by PCR method from rice. Plant binary over-expression and RNAi-expression vectors were construted and transformed into rice by Agrobacterium-mediated, and obtained some transgenic rice plant lines. The work established a basis for further studying the function and application potential of the gene in rice seedling cold tolerance. The main results were as following:1. The cDNA of OsCOI gene was cloned by PCR method from rice. The sequence analysis showed that the total length of cDNA was 1235 bp with an open reading frame encoding 269 amino acids. Homology analysis of the nucleotide sequence showed that the identity was 100% to the published sequence AK072942. The results from bioinformatics software analysis showed that the gene wae a zinc finger protein with two C2H2domains.2. Over-expression and RNAi-expression vectors were constructed and identificated by the enzymes digesting, and the different vectors were transformed into Agrobacterium tumefaciens by electroporation, which were proved by PCR assay with two pairs of specific primers of the gene.3. The OsCOI gene was transformed into japonica rice variety Nipponbare using the two plant expression vectors by Agrobacterium-mediated approach. Three over-expression transgenic plants and twelve RNAi transgenic plants were obtained by screening in the media contained hygromycin. The results of PCR assay showed that the gene had been integrated into rice genome of T0 plants.
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