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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。
In this study, we plan to use banana multi-buds as transformation receptor materials to establish Agrobacterium tumefaciens mediated transformation system of banana. In this study, the multi-buds were induced by the cross-section of the corm of B. brasiliensis seedlings and the multi-buds were used as the receptor. Through the sensitivity to hygromycin and the transient expression rate of GUS gene, Find suitable genetic transformation conditions. The results showed that the transplanted bulbs of Brazilian banana bulbs were formed in the multi-bud induction medium (MS + 6-BA 10 mg / L + PP3331 mg / L + NAA 0.21 mg / L) after 4-5 months of culture The multi-buds of cauliflower structure. The cross-cut thin slices of Bradyrhizobium japonicus were sensitive to hygromycin. The optimum concentration of hygromycin was 25 mg / L, and the best co-cultivation time was 4 d. In this study, 18 resistant shoots were obtained from a total of 300 pieces of 3 transformations. All the resistant shoots obtained by GUS assay and PCR were positive. The transformation system established by this study using multi-buds as the transformation receptor provides a technical reference for the future introduction of genes with important economic traits into bananas.