| Torenia fournieri L is an important flower that grows in semi-tropics and tropics, with special and naked megaspore, and it is also a model plant which can be used to study flower apparatus's development and the progress of impregnation. Chimeric gene Psag12-IPT is composed of SAG12 cloned from Arabidopsis thaliana and IPT corned from the tmr region of plasmid of Agrobacterium tumefaciensand encoding isopentenyl transferase. This chimeric gene is always used to be transformed to plant in order to improve its resistance.In this study,The vector pCAMBIA1301 and the vector pSG516 were cut by Xba I ,and then were linked .After transformed to EcoliDH5a, we got the expression vector pHQIPT. This vector has the target gene, the reporter gene GUS, the selectiong gene Hygromycin.Using freezing-melting method , the expression vector pHQIPT was transformed to Agrobacterium tumefaciens.Leaf di scs of Torenia fournieri L and Agrobacterium tumefaciens coculture on coculture: medium after 4 days.Ninety percent of transient expression of GUS gene of explants after coculture are positive. The explant was selected on shoot-induced medium (Cef500mg/L, Hygl5mg/L)after 20 days ,then was transfered to rooting medium(Cef500mg/L, Hyg0mg/L) and stayed 15 days.The explant then was transferred to rooting medium(Cef500mg/L, Hygl5mg/L)20 days .After this disposal ,we got nine resistant shoot. Expression of GUS gene of these resistant shoot are positive. PCR .Southern blot and Northern blot analysis confirmed that Psag12-IPT gene was already integrated into the genome of Torenia fournieri L.But the morphology of these shoot were different to that of wild types.These lateral bud seem limitless propagation,these leaves are small and crisp, the roots of these plant are shoirt and little. Phapas there are some special trans-acting factors in Torenia fournieri L that can active SAG12 promoter, and the IPT gene is overexpressed. |
