The Gene Silencing Vector Constrction Of CHSJ Gene Using Artificial MiRNA And The Selection For The Suitable Petunia Lines In Transformation

Petunia have many advantages, including easy to regeneration, easy to manage, the longer of growth cycle, the longer of flowering period, not strictly for the environ-mental requirements and so on. So, petunia is not only the important ornamental flow ers, but also the important model plant in the research of gene. Genotype is the impor-tant factors In the Agro bacterium-mediated transformation. In this study, we selected the suitable petunia lines for the Agro bacterium-mediated transformation. We carried out optimization for the selected petunia lines. The results are as following:1. We successfully constructed a petunia pMADS9 miRNA gene silencing vector for plant expression vector by overlaping PCR method, Using Tools carrier RS300, the intermediate vector pMF604 and plant expression vector pC2301,2. After the Chlorine disinfection in 4h, the seed has the highest seed germination rate and the lowest pollution; to compared with the seed or the explants by the mercuric disinfection, the seed by the Chlorine disinfection have many advantages, as short to germination time and easy to operate.In agronomic traits, we seleced 26 petunia lines. Compared with the petunia lines A01, we selected 6 petunia lines (C01, C02, C03, C04, C05 and C06).2. After the infection by agrobacterium, we gathered statistics of the selected 6 petunia lines (C01, C02, C03, C04, C05 and C06) in GUS transient expression. Compared with the petunia lines A01, we selected the petunia lines C01, C04 and C06.3. For the three petunia lines C01, C04 and C06, we optimized the transformation conditions, the results show that:In a certain ranges of the concentration of Agro bacterium (OD600=0.3-0.4), time of inoculation(5-10 min) and time of co-culture(2-3 d); we would have higher conversion and lower contamination rate.