|
|
|
| Transformation of Blue Gene in Narcissus tazetta var. chinensis Mediated by Agrobacterium tumefaciens |
|
|
|
|
Abstract Hf2 gene(1527bp) encoding flavonoid 3’,5’-hydroxylase (F3’5’H) and dihydroflavonol 4-reductase (dfr) (1212 bp) gene were isolated from Petunia hybrida and Ipomoea nil petals respectively. Hf2 and anti sense dfr genes were subcloned into pBRLys vector together generating pUbi-Hf2-dfr vector which contained Ubiquitin(Ubi) promoter, and then the pUbi-Hf2-dfr was transferred into Agrobacterium tumefaciens strain EHA105. Result of PCR amplification with transformant colony showed that the expression vector pUbi-Hf2-dfr was successfully transferred into Agrobacterium. Hf2 and anti sense dfr genes were transformed into callus of Narcissus tazetta var. chinensis mediated by Agrobacterium. Extracted the genomic DNA of 130 regenerated plants with CTAB method, and then performed PCR with specific primers and electrophoresis results showed that there were 3 regenerated plants with the intergration of Hpt and blue genes. PCR-Southern blotting indicated the same result.
|
|
Received: 24 August 2009
|
|
|
|
|
|
|
