作　　者： ; ; ; ; ; ; ; ;

机构地区： 广东省农业科学院

出　　处： 《分子植物育种》 2014年第6期1195-1200,共6页

摘　　要： 本研究拟采用香蕉多芽体薄片为转化受体材料,建立起根癌农杆菌介导的香蕉遗传转化体系。以巴西蕉组培苗的球茎横切薄片为起始材料,诱导获得多芽体,利用多芽体薄片为受体,通过其对潮霉素的敏感性和GUS基因的瞬时表达率试验,以期找到适合的遗传转化条件。结果表明:巴西蕉球茎横切薄片在多芽体诱导培养基(MS+6-BA 10 mg/L+PP3331 mg/L+NAA 0.21 mg/L)上,经过4至5个月的培养,形成了花椰菜结构的多芽体。巴西蕉多芽体横切薄片对潮霉素敏感,最佳的潮霉素素筛选浓度是25 mg/L,转化的最佳共培时间为4 d。本研究从3次转化中的共300个薄片中获得抗性芽18个,经GUS检测和PCR鉴定,获得的抗性芽全部为阳性。本研究建立的以多芽体薄片为转化受体的转化体系,为今后将具有重要经济性状的基因导入香蕉提供了技术参考。 The objective of the study is to establish an efficient Agrobacterium-mediated genetic transformation protocol for Baxi banana （Musa acuminate cv. Baxi） using multiple bud clumps （MBC） as receptor materials. The experiment initiated from the transverse corn slice of in vitro plantlets of Baxi banana, from which the MBC was induced. Using MBC slice as starting transformation receptor, the optimal transformation conditions were obtained through the hygromycin sensibility selection and GUS gene transient expression test. The results showed on the inducing medium （MS＋6-BA 10 mg/L＋PP333 1 mg/L＋NAA 0.21 mg/L）, the transverse corn slices of Baxi banana were formed cauliflower shaped MBC 4 or 5 months later. The MBC slices were sensitive to to hygromycin, the suitable selection concentration of hygromycin was 25 mg/L and the best co-culture time was 4 days. Using the optimal transformation protocol, a total of 18 hygromycin-resistant shoots were regenerated from the initial 300 MBC slices in 3 separative experiments. All these shoots were confirmed to be positive transformed shoots using histochemical GUS assay and PCR methods. These results suggest that an efficient Agrobacterium-mediated genetic transformation protocol has been developed by using MBC slices for stable integration of foreign genes into banana, this transformation system will provide an important technique for future studies on transferring economically important genes into banana.

关 键 词： 香蕉 遗传转化 多芽体

领　　域： [农业科学]