Cloning And Characterization Of Arabidopsis Thaliana NCED4Gene And Medicago Sativa L. MsFTa Gene

In this study, we characterized the MsFTa gene from Medicago sativa (alfalfa) and NCED4gene from Arabidopsis thaliana by gene genetic engineering, transgenic technology and phenotype analysis. The results are showed below:1. A1785-bp genomic DNA sequence named NCED4was obtained by PCR. It encoding595amino acids and does not contain introns, NCED4protein isolated from arabidopsis thaliana compared with homologous protein of Arab is lyrata L., the gene have high homology and similar conservative area, there is no signal peptide.2. NCED4gene was inserted into plasmid pBI121with DNA recombination technology, resulting a plant expressing vector pBI-NCED4. Transgenic Arabidopsis plants were obtained by the floral dip method with Agrobacterium tumefaciens GV3101harboring pBI-NCED4plasmid and seeds of transgenic Arabidopsis were selected on MS plates containing50mg/L Kanamycin. PCR and Real Time RT-PCR analysis showed that the target gene was inserted into the genomes of the Arabidopsis thaliana and NCED4gene was over-expressed in transgenic Arabidopsis successfully. Gene function is relate to the regulation and control of abscisic acid in plant.3. Real-time quantitative experiments have shown that NCED4gene in different tissue of Arabidopsis have significant differences in expression, laminae levels were strongest and the flower was weakest; NCED4gene expression quantity is obviously influenced by hormone of extraneous sources:under the action of abscisic acid, amount of gene expression reduced after rising first as time accumulation, while under the effect of a certain amount of GA3, amount of gene expression reduced accumulated over time.4. A1528-bp genomic DNA sequence named MsFTa was obtained by PCR. It encoding176amino acids, MsFTa protein isolated from a Medicago saliva L. compared with other species of homologous protein, the gene have high homology and similar conservative area, there is no signal peptide.5. MsFTa gene was inserted into plasmid pBIl21with DNA recombination technology, resulting a plant expressing vector pBI-MsFTa. Transgenic Arabidopsis plants were obtained by the floral dip method with Agrobacterium tumefaciens GV3101 harboring pBI-MsFTa plasmid and seeds of transgenic Arabidopsis were selected on MS plates containing50mg/L Kanamycin. PCR and Real Time RT-PCR analysis showed that the target gene was inserted into the genomes of the Arabidopsis thaliana and MsFTa gene was over-expressed in transgenic Arabidopsis successfully. Gene function is relate to the regulation and control of flowering in plant.6. MsFTa gene in different tissue of Arabidopsis have significant differences in expression, flower levels were strongest and the root was weakest; By the treatment of extraneous source GA3, the expression levels of MsFTa gene is obviously increased, and as extend the time of processing, MsFTa showed a trend of rising the level of gene expression Under the condition of low temperature processing, MsFTa appeared rising trend.In a ward, MsFTa gene function is relate to the regulation and control of flowering and abscisic acid in plant.