玉米自交系18-599(白)转化受体体系的建立和转barnase基因植株的获得

以抗病优质综合性状优良的玉米自交系18-599（白）为试材，优化其遗传转化受体体系，并进行了barnase基因的转化。结果表明，在基于N6和MS培养基的8个方案中，N6-4培养基是转化受体系统建立的最佳培养基；幼胚的最佳接种长度为1.6 mm；用于转化的幼胚愈伤组织继代次数最好控制在5次之内。利用18-599（白）优化受体体系进行了barnase基因的基因枪转化，用除草剂Basta作筛选剂，其致死浓度为8 mg L^-1，3轮筛选的Basta浓度依次为6、8和6 mg L^-1。对转化植株的分子检测和大田不育性状观察表明，获得了转barnase基因的雄性不育18-599（白）转化植株。

Establishment of Transgenic Acceptor and Transformation of barnase Gene by Particle Gun in Maize Inbred Line 18-599 (White)

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.