棣棠花器官决定的AGAMOUS同源基因克隆和功能分析-生物化学与分子生物学专业论文.docxVIP

三峡大学硕士学位论文 三 峡 大 学 硕 士 学 位 论 文 PAGE VI万方数据 PAGE VI 万方数据 Abstract Kerria japonica (Rosaceae) was deciduous shrubs, it had two varieties with single and double flower under natural conditions. The variation of flower morphologies has made Kerria japonica（Rosaceae）as an excellent model for studying floral organs morphogenesis. The C-class MADS-box gene AGAMOUS (AG) plays crucial roles in Arabidopsis thaliana development by regulating the organ identity of stamens and carpels, the repression of A-class genes, and floral meristem determinacy. Although there are extensive reports on AGAMOUS homologous genes involved in flower organogenesis in model and economically important flower plants, there are few reports on this research area in woody plants. In this paper, we took Homology-based cloning and Rapid-amplification of cDNA ends (RACE) to isolate AGAMOUS homologous genes, which regulate stamen and carpels development, combining with the recover study of the gene on Arabidopsis ag-1 mutant indicated the main function in controlling Kerria japonica flower development. The main results are as followings: （1）cloning and sequence analysis of KjAG from Kerria japonica A MADS-box gene KjAG involved in flower development had been isolated from Kerria japonica via the way of homology cloning combined with RACE technique.Protein sequence alignment and molecular phylogeny analysis suggested that, KjAG was an AG homology in Arabidopsis, at the transcriptional activation domain of C-terminal existed two conserved motifs: AGⅠ and AGⅡ. （2）Functional analysis of KjAG from Kerria japonica A sense expression vector of KjAG had been introduced into Arabidopsis ag-1 mutant via agrobacterium-mediated method then screen positive transformation plants,and proceeded phenotypic analysis of transgenic arabidopsis plants.The results showed that, compared with ag-1 mutant, 35S::KjAG transgenic plants indicated that the gene recovered flower morphology of ag-1, such as producing stamens organs in