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为提高辽宁棉花品种的耐盐性,利用农杆菌介导上胚轴外植体转化法,将来源于盐生植物獐茅的Na~+/H~+逆向转运蛋白基因(AlNHX1)导入棉花辽棉19号中,分析影响棉花上胚轴外植体农杆菌转化的几个重要因素,建立并优化了该品种的转化体系,结果表明:1)农杆菌侵染的最佳时间为90s,共培养过程中乙酰丁香酮最适浓度为100 mg/L,筛选培养基中潮霉素最适浓度为30 mg/L,生根培养基中吲哚乙酸IBA的最适浓度为10mg/L;2)对转化植株进行PCR检测,表明耐盐基因AlNHX1已导入辽棉19号中;3)在盐胁迫下,转基因植株的电导率,渗透势都显著低于未转化植株。结果表明,通过筛选并优化转化体系,辽棉19号品种的耐盐性有较明显的提升,为沿海滩涂地区种植棉花提供了一定的参考。
In order to improve the salt tolerance of cotton varieties in Liaoning, the Na ~ + / H ~ + antiporter gene (AlNHX1) derived from Haliotis discus hannai was introduced into cotton Liaodong Cotton 19, the paper analyzed several important factors that affect the transformation of cotton epicotyl explants Agrobacterium, established and optimized the transformation system of the varieties, the results showed that: 1) the best time for Agrobacterium infection was 90s, a total of The optimum concentration of acetosyringone was 100 mg / L, the optimal concentration of hygromycin in the culture medium was 30 mg / L, the optimal IBA concentration in rooting medium was 10 mg / L; 2) Transformed plants were detected by PCR, indicating that salt-tolerant gene AlNHX1 was introduced into Liaomian 19; 3) Under salt stress, the conductivity and osmotic potential of transgenic plants were significantly lower than those of untransformed plants. The results showed that by screening and optimizing the transformation system, the salt tolerance of Liaodian 19 varieties increased obviously, which provided some references for planting cotton in coastal tidal flat areas.