Agrobacterium Tumefactions-mediated Genetic Transformation Of Apical Meristem And Tillring Node In Bread Wheat (Triticum Aestivum L.)

In wheat genetic transformation, there existed some problems including low transformation rate, genetically poor stability, etc. In the present study, Agrobacterium tumefactions- mediated transformation techniques of wheat apical meristem and tillering node were investigated with young embryos of four bread wheat as cultivars, i.e. Henong825, Hnong827, Henong 58-3 and and Zhoumai 1 as receptor, and OsDREB 2.2 gene from rice （Oryza sativa） as donor gene. The main results are shown as follows:1. After 5h germination, wheat bud growing point as the receptor was sufferred 3 and 4 degree wounds. The rates of regeneration bud were 69.35% and 54.66%, respectively. The transformation rate of 4 degree wound was higher, which was 3.64%. Wheat coleoptile was removed after 15h germination, and bud growing point was sufferred 1, 2 and 5 degree wounds. After infection the transformation rates were 37.06%, 30.02% and 19.64%, respectively. Early transformation rate of 2 degree wound was higher , which was 2.65%.2. The Survival rate of T₀ with tillering node-infection is 100%, detected by PCR, positive plants occupied 3.25%. Monoclonal testing to future generations of four positive strain lines, the genetic transfer rate of the two lines from Henong 827 respectively was 35.71% and 30.43%, and the two lines of Henong 58-3 respectively was 13.63%and 16.33%.3. 58 transgenic plants were selected out in this study, and 54 plants were obtained by apical merited infection method, 4 plants were obtained by tillring node infection meristem.4. The result of wheat apical meristem infection experiment showed that the appropriate density of agrobacterium was 0.5 OD600, 300mg/L carbenicillin was good as a antibiotic, and the most appropriate density of kanamycin was 90mg/L as a resistant plants selection agent.