Studies On Agrobacterium-Mediated Genetic Transformation Of Different Blossom Gene Of London Plane(Platanus Acerifolia Willd.)

Platanus acerifolia was widely grown in our country and has become China’s major cities street trees and landscaping trees. At yellow river and yangtze basin, more than70%of the trees are Platanus acerifolia in Wuhan,Nanjing, Shanghai and other cities. However, Platanus acerifolia fruit is synthesized by the majority of small nut which set synthesis of spherical poly fruit,the long villi surrounded the base of small nuts.And in every year between April and June,with aging fruits abscission,many seed hairs and pollen from neonatal male flowers distribute in the air.Those questions not only pollute the environment, induce allergic symptom but also affect people’s lives and health.The the development of plant genetic engineering provides a new way in breeding of no-seed varieties.Many varieties mediated by Agrobacterium Tumefaciens and putting exogenous genes into the recipient shorten the breeding time and directly cultivate new varieties.Based on many years of breeding no-seed varieties,this study uses GUS reporter gene in condition of influence the transformative adventitious using zygotic embryos as explants.Using the optimization genetic transformation system transformates the carrier with flowering-related genes. Utilizing the PCR tests results. The main findings are as follows:1. By preliminary exploration of the storage conditions, firstly considering the survival rate, budding rate, and finally considering the growing post-emergence determined the optimal combination. Processing conditions for the selected paper bags, placed72h, and stored at-20℃vinyl bag, placed72h, stored at-20℃, moisture content of about6%were the best combinations.2.In the optimization of regeneration system of zygotic embryos,the seedling still can differentiated in the100mg/L Km concentration.But the number of differentiation decreased.In the Km concentration of75mg/L with long time selection,the growth of non-transformed seedlings can be completely inhibited.In the rooting experiment,the75mg/L Km concentration can completely inhibited root growth.In the study of the concentration of Tm influencing the growth of zygotic embryos,300mg/L was the most suitable concentration of bacteriostasis.3.By the analysis of GUS transient expression rates and other relevant data,the results showed optimum conversion conditions were the infection time of1h,mannitol treatment of4h, pre-training of6-7t,co-cultured of3t in the AS concentration of100mg/L. Through the bacterium Agrobacterium tumefaciens concentration and type of analysis, EHA105and OD=0.6-0.8made the Agrobacterium strongest infection.4.After more than six months screening, the conversions of five kinds of target gene as follows:resistant plants of LFY-FUli got two positive plants, A-114、A-303、A-306were the positive plants using the PCR detection, only one strain of TFLi resistant plants was the positive of plant, LFY did not get positive strains,PI-Barnase resistant plants have been tested and four were positive plants.