Construction And Function Analysis Of The Expression Vector Carrying Blue Genes Which Is Driven By Flower-Specific Promoter

With the graceful posture, tall and straight flower stalk, Lily(LiLium spp.)is suitable for using in the garden. As one of the most famous cut flowers in the world, it is widely favored by the peoples from various countries. In the nature, Lily owns various colors such as white, pink, red and yellow, but lacks blue flowers. Breeding blue Lily flower is the research emphasis of flower molecular breeding via genetic engineering. As the most influential factors in the color formation of higher plants, anthocyanin biosynthetic pathway has been studied clearly, a large number of related genes have been cloned and identified.For seeking breeding way of blue lily flower, in the previous work, we cloned chalcone synthase promoter sequences from petals of orential Lilium 'Sorbonne' and verified its functions initially; cloned DFR gene sequences coding dihydroflavonol 4-reductase from petals of Hyacinthus orientalis; cloned Hf 1gene sequences coding flavonoid 3 ', 5'- hydroxylase from royal purple petals of Petunia hybrida; cloned DifF gene sequences coding B5 protein from royal purple petals of Petunia hybrida. On this basis, our experiment have constructed two multi-gene expression vector, and transformed model plant petunia via Agrobacterium in order to reference for blue Lily flower breeding. Main results were obtained as follows:1.Construction of multi-genes expression vectorsInosculating flower specific promoter PchsA and Hf1 gene, we constructed intermediate plant expression vector pCAM-Hf1, after that constructed DFR and DifF genes into pCAM-Hf1, respectively. Finally, we gained two multi-genes expression vectors pCAM -Hf1-DFR and pCAM-Hf1-DifF;2. Transformation of petunia and PCR detection of hygromycin positive plantsWe transformed model plant petunia via Agrobacterium. After hygromycin resistance screening, gained some hygromycin positive plants. Extracted DNA of them using CTAB method and were carried out PCR detection using corresponding genes specific primer, transgenic petunia DNA amplified expected peculiar PCR fragments, respectively. We obtained transgenic petunia plants initially.