| Mature embryos of 27 wheat genotypes were cultured to compare the efficiency of induction, differentiation and regeneration of mature embryos callus. The factors which affect the induction, differentiation and regeneration of mature embryos callus were studied. The wheat genotypes with high efficiency of induction and regeneration of mature embryos callus were screened. The mature embryos callus of genotypes screened were transformed by Agrobacterium tumefacienens. The factors (A. tumefaciens cell density, inoculation duration, pre-cultivation duration, treatment of sonication and vacuum infiltration) influencing the efficiency of transformation were researched. The results showed that1. The genotype and concentration of 2,4-D and NAA were significantly factor influencing the induction and regeneration of wheat mature embryos callus. The best suitable concentration of 2,4-D and NAA were 2.0 mg l-1 and 0.5 mg l-1. 12 wheat genotypes with high efficiency of induction and regeneration of mature embryos callus were selected out of 27 wheat genotypes. The optimal method was established.2. The A. tumefaciens cell density, inoculation duration, pre-cultivation duration and treatment of sonication and vacuum infiltration, were important influence factors for transformation of wheat mature embryos. At the condition of cell density OD600=0.8 and inoculation duration 60min, the rate of resistant embryo was 4.21%. The treatment of sonication(7.5min) and vacuum infiltration(7.5min) in infection could increase the ratio of transformation from 3.09%to 8.16%. The system optimized relatively of genetic transformation of mature embryo of wheat seed mediated by A. tumefacienens was preliminarily established.3. The transgenic wheat of improved resistance to the powdery mildew was production,which has better resistance than non-transformation wheat . Independent plantlets were obtained from the kanamycin-resistant calli after screening. PCR analysis indicated that they were transgenic plants. |
PDF Full Text Request