关键词: 葡萄, 器官发生途径, 不定芽, 组织培养

Abstract: Establishing an efficient regeneration and transformation system is a fundamental role to the transgenic study. For Centennial Seedless, the effects of growth regulators, explants and ablastin to the generation of adventitious shoots were studied, based on which the optimal condition for regeneration and transformation was determined. Our experiments showed that, when being used alone or accompanied with auxins, the cytokinin TDZ is always better than BA and KT in inducing adventitious shoots. The adventitious shoots of Centennial Seedless could be efficiently induced in the media of MS + TDZ 2.0 mg·L^-1, MS + TDZ 4.0 mg·L^-1 or MS + TDZ 4.0 mg·L^-1 + IAA0.1 mg·L^-1and adventitious shoot regeneration rates were 37.4%,40.2% and 50.0%, respectively. For fixed medium, adventitious shoots can be generated with minor difference in efficiency from leaf disc, stem and petiole. When this regeneration system was used in genetic transformation mediated by Agrobaterium tumefaciens, the optimal concentration of screening resistance medium was 20 mg·L^-1 for kanamycin, and for the control of agrobacterium growth, the best concentration was 400 mg·L^-1 for cephaloridine or 300 mg·L^-1 for carbencillin . Transformants were obtained 45. PCR and Southern blotting confirmed that foreign gene was integrated genome of grape.

Key words: grape, organogenesis approach, adventitious shoot, tissue culture