F3'5'H Gene Influenced Flower Color In Transgenic Tobacco

Flower color is an important one of qualitative characters and economic parameters for the ornamental plants. Because of the colors for many focal flowers are limited, for example, the roses, carnations, tulips and so on, which flowers are lack of the color for blue and violet. And the reforms for the ornamental plants colors are provided with the advantages such as the visual arts, and the large economic potential. Gene engineering technology is superior to the Cross breeding and Directive breeding technology with its short cycle , low cost and high benefit. Though traditional breeding technology has been used for a long time. In this dissertation, the author introduced F3'5'H into tobacco. In order to research the function of F3'5'H was observed the influence of floral color. The main experiment and results of this paper are such as follows:1. Construction of plant expressional vector p^{BinF3' 5'H} and use it to transffere into LBA4404P^{ucm-F3' 5'H} p^BinAR digested with KpnI BamHI was constructed for plant expressional vector p^{BinF3' 5'H} of sense F3'5'H gene. The plant expressional vector p^{BinF3' 5'H} was belong to the Lab, and used it to transffere into LBA4404, respectively.2. Introduction of F3'5'H gene into tobacco by Agrobacterium tumefaciensYoung leaves of were excised from sterile seedlings of pure deep red petunia, cut into5 X 5mm², dipped into Agrobaterium (OD₆00 adjusted to 0.2) for 15min. All the explants were washed in liquid MSO medium for 2 hours. After washed with sterile water for 5 times and patted dry on the filter paper again, patted dry on the filter paper. Transferred to MS1 medium and co-cultured for 3-4 days in darkness. Subsequently, the explants were transferred td MS2 upplemented for 3-6 days for selection culture. The resistant shoots were rooted on MS3 and then transferred the plantlets with normal roots to field for further analysis. Statistic results showed, that the resistant shoots Y^{F3' 5'H} has 20 lines.3. The identification of transformed plants by PCR and the mensuration of anthocyanin intransgenic tobaccoHfl of F3'5'H were amplified, respectively, by common PCR to identify transformed petunia. The PCR positve results suggested that transgenic YF3'5H has 20 lines (YF3 5 H n, n=l-20.The anthocyanin was extracted from various developmental stages of YF3 5 H which were PCR positve results and changed flower color. Y ° are controls . Their contents were measured by colorimetry. The results showed that the anthocyanin of Y° were less than YF3 5 H .4. The analysis of the changes of flower colors and the mutations of flower organs by transformed sense F3' 5' H geneWe recorded the color changes of YF3 5 H, especially the color, distribution and veins of the flower petals that growing in V period and would open in 2 days. Results showed that the flower petals color of Y° are lighter than YF3 5 H. In YF3 5 H lines, like deep red color brushing the petals from corollar crackle to both sides. The color of YF3 5 H lines are more than Y°. There are pale pink veins or sectors on the edge or middle part of corolla.The variational criterion of flower organs is the phenotype of bud, the number of sepal, the diametical sizes of corollar tube and corolla, the number of corollar crackle, and the number of pistal and stamen, etc. Results showed that there are no variants in YF3 5 H linesIn TG-YF3 5 H lines, we detected a diffrent varinant with particular color phenotype.