| The flowering transformation process in higher plants is strictly controlled by both the inherent regulation and the exterior environmental signals. The flowering process is a significant one in the transition from vegetative growth to reproductive growth, which plays a crucial role in guarantying the successful reproduction and deciding the time of flowering. The investigation to the molecular mechanism regulated by flowering time in plants would make a significant contribution to regulating plant reproduction and growing, raising crop production and quality. In the earlier stage of experiment, an early flowers mutant, named as efs-1 (early flowering in short mutant 1) was achieved by screening from the Arabidopsis T-DNA insertion mutant library. By Genome Walking technology and bioinformatics methods, a novel Arabidopsis flowering regulating related gene EFS1 was identified. By complementary reply and over-expression test, the EFS1 gene was confirmed to be Arabidopsis flowering control gene. By RT-PCR and GUS histochemistal staining technique, the expressional level and localization of EFS1 gene was clarified. Through RT-PCR and Real-Time PCR technology, the localization in the flowering control pathway and the model regulating the flowering program was preliminarily clarified. In order to give further and deeper clarification of the function of EFS1 gene, the further experiment mentioned blow was preceded.1. The wild type, EFS1 mutant, revertant and over-expression mutant were disposed with gibberellin, vernalization and cultivated under short-day conditions, meanwhile, the plants without any treatment were treated as the control group. By observing the flowering time, calculating the rosette leaf number, and preceding the RT-PCR analyzing expression level, it can draw a conclusion that primarily the EFS1 gene completes its function of regulating the plant flowering in photoperiod pathway and gibberellin pathway.2. By using RNAi technology and anti-RNA technology, the EFS1 function- absence mutant was achieved. Furthermore, the EFS1 gene anti-RNA and RNAi vectors, pCAMBIA1300-EFS1A and pCAMBIA1300-EFS1i, were constructed. The pCAMBIA1300-EFS1A and pCAMBIA1300-EFS1i vectors were induced by Agrobactrium to transform the Arabidopsis. By hygromycin screening, 11 anti-RNA transformants and 10 RNAi transformants were achieved. Through the PCR test, it can be verified that exogenous gene was successfully induced into the genome of the transgenic plants. The RT-PCR consequence demonstrated that the expression level of EFS1 gene in transgenic plant was suppressed differently, which can identify that the achieved transgenic plants all were EFS1 gene mutants.3. By using the CO and FLC gene that play important roles in flowering pathway, the over-expression vectors pCAMBIA1300-CO and pCAMBIA1300-FLC were constructed. The pCAMBIA1300-CO and pCAMBIA1300-FLC vectors were induced by Agrobactrium to transform the Arabidopsis. By hygromycin screening, 9 CO over-expression transformants and 8 FLC over-expression transformants were achieved. Through the PCR test, exogenous gene was successfully induced into the genome of the transgenic plants. The RT-PCR results illustrated that the expression level of relevant genes were enhanced differently, which proved that the achieved transgenic plants were the relevant gene mutants in order to make preparation for the further double over-expression experiment.4. 5 seeds of mutants achieving from SALK mutants library, i.e. AT3G11540,AT5G15840,AT1G01060,AT2G19520,AT1G22770,AT4G36680, were reproduced and a great amount of mutants were achieved. These mutants were tested by homozygous validation and the relevant homozygous mutants were achieved, which make preparation for the further double mutant test.5. Through yeast double hybrid, we have constructed the bait expression vector pAS1--EFS-CDS.
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