Rice Transformation Of OsMADS59、OsDDL Genes And Analysis Of Expression Of OsMADS59in OsTFL2RNAi Transgenic Rice

The process of growth and development of plants were effected by many genes. The gene of flowering suppressor played a role in the flowering regulatory networks of Arabidopsis. The FLC was a MADS-box gene. In the rice genome, the OsMADS59was the highest homology with Arabidopsis FLC. In view of the conservative of the biological evolution and important functions of Arabidopsis FLC, researching the functions of OsMADS59in rice and the relations with other flowering regulatory genes was greatly significant. In this study, OsMADS59overexpression and RNAi vector were transformed into rice plants by Agrobacterium tumefaciens mediated transformation, which provided material for researching the biological functions of OsMADS59. In order to investigate the regulatory relations between OsMADS59and OsTFL2, in situ hybridization was used to study the expression of OsMADS59in OsTFL2RNAi transgenic rice. In addition, this study constructed RNAi vector of a new gene that was OsDDL, and the gene laid basis for finally proving biological function because it obtained OsDDL RNAi transgenic plants through involving in the regulation of rice flowering. The main results of this study were as follows:1、Obtained OsMADS59overexpression and OsMADS59RNAi transgenic rice. In order to filter resistant callus under50μg/mL concentration of hygromycin, the embryo callus of Rice NONGKEN58(58N) were inducted and pre-group materials were infected by Agrobacterium EHA105which was transformed by OsMADS59overexpression vector and RNAi vector through agrobacterium tumefaciens mediated transformation.15resistant strains with OsMADS59overexpression were obtained and13positive strains were obtained with PCR detection. In addition,33resistant strains which with OsMADS59were obtained and22positive strains were obtained with PCR detection.2、In this study, leaves and shoot tips were took on27days and54days as materials which were used to do RNA in situ hybridization experiment with the TFL2RNAi transgenic rice58N as research background and58N rice as blank. The results showed that the OsMADS59expression of meristems of leaves and shoot tips in58N CK materials significantly increased with extending the growth time of rice. But the OsMADS59expression of meristems of leaves and shoot tips in58N TRI significantly declined.3、RNAi vector of OsDDL was successfully constructed. Building RNA interference structure was completed with the RNAi fragments which were amplified out from genomic DNA of Nipponbare rice and cloned on the pUCm-T vector and reverse connected in the176vector. Finally RNAi vector of OsDDL was constructed by connecting RNA interference structure to pICAMBIA1300, the matching results were completely correct through the evaluation of sequences.4、Through callus materials were infected by EHA105Agrobacterium which was transformed by OsDDL RNAi vector.14hygromycin resistant strains were differentiate from callus and11transgenic positive strains were obtained with PCR detection.