辽棉19号转化耐盐基因AlNHX1的研究  

Research on transforming salt-tolerant gene AlNHX1 to cotton Liao19

作　　者：丰明[1] 王旭达[1] 王鹤[1] 张高华[1] 

机构地区：[1]辽宁省农业科学院大连生物技术研究所,辽宁大连116029

出　　处：《中国农业大学学报》2017年第10期36-44,共9页Journal of China Agricultural University

基　　金：辽宁省自然科学基金(2014027023)

摘　　要：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。Using Agrobacterium transformation method, the epicotyl explants of cotton Liao19 were transformed with Aefuropus vacuolar Na＋/H＋ antiporter gene AINHX1 to improve its salt resistance. We have established and optimized a transformation system was established and optimized to study several important factors on cotton transformation. The results showed that： 1） The optimum Agrobacterium infection time was 90 s. In the co-culture medium,the optimum concentration of AS was 100 mg/L. The hygromycin optimal concentration of the cotton filter medium was 30 mg/L. The IBA optimal concentration of rooting medium was 10 mg/L; 2） For all the transformation cottons, PCR test results showed that the salt tolerance gene AINHX1 was successfully transformed into cotton Liao19;3） Physiological studies showed that the relative conductivity and osmotic potential of transgenic plants were all significantly lower than non- transgenic ones under the salt stress. This study confirmed that the cotton Liao19 obtained certain salt resistance by screening and optimizing the transformation system and provided reference for the cultivation of cotton in coastal beach area.

关 键 词：棉花 NA+/H+逆向转运蛋白 农杆菌转化 上胚轴外植体 相对电导率 细胞渗透势 

分 类 号：S562[农业科学—作物学] Q945.79[生物学—植物学]