Studies On Cloning And Transformation Of Gibberellin Receptor Gene In Arabidopsis

There are four pathways that Controlling the flowering time in plants, including Photoperiod pathway, Gibberellin pathway, Autonomous pathway and Venalization pathway. So far, some reports suggested the possible interaction between different signal pathway during flowering evocation. GA regulates a series of growth and development processes, as well as flowering time control. In order to further investigate the interaction between these pathways, we have cloned genes encoding GA receptors, and made overexpression constructs. These genes were transformed into Arabidopsis Colombia wild-type, and good transgenic lines with high expression level were screened.Our work will provide a basis for studying the Gibberellin pathway in flowering time control.In this experiment, the total RNA was extracted and the first chain of cDNA was synthesized with RT-PCR. GID1 genes were cloned by PCR technology. Then the genes were cloned into the entry vector with Gateway cloning system. After sequencing of the the proper gene fragment, these genes were cloned into the destination expression vector of PK2GW7-HA.Transformation into Colombia wild-type plants was done with Agrobacterium, and transgenic lines were screened for singe insertion of transgene. Good transgenic lines were obtained after expression level was analyzed with Real-time PCR.Those transgenic lines showed a typical phenotype with high level GA biosythesis, which suggest GA signal pathway was enhanced.