Abstract:

The efficient acceptors for maize transgenic engineering are insufficient in China now. Seed production by male sterility is the best method for advancing authenticity of maize hybrid. Maize inbred line 18-599 (white) is an antivirus high quality maize self-line in China, which have been used for lots of maize hybrid cultivars. Establishment of a good efficiency transgenic acceptors is necessary for the advance of transgenic efficiency in maize transformation work. In this study, the efficient transgenic acceptors were established and optimized. 18-599 (white) was studied in estate, types of culture mediums, times of callus regeneration and concentration of screening reagent Basta. The result showed that N6-4 medium was the best one in 8 types of mediums for immature embryo of 18-599 (white), 1.6 mm length was the feasible length of immature embryos for tissue culture in establishing transgenic acceptor system, it was within 5 times for suitable callus subculture. With the optimized transgenic acceptors, barnase gene was translated triumphantly into 18-599 (white) by particle gun using bar as a marker gene. Basta was used as the screening reagent, its lethal concentration was 8 mg·L^-1 and its working concentration for screening was 6, 8, and 6 mg L^-1 in turn in 3 times for callus regeneration. In this study, a transgenic plant with male sterility was gained through molecule detecting and observing in the field. The result has an important regnificance for creation of new male sterility self-lines on maize in the future.

Key words: Maize, Inbred line, Transgenic acceptor, Particle gun, Molecular detection