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通过农杆菌介导的方式将獐茅液泡膜Na~+/H~+逆向转运蛋白基因(AlNHX)表达在马铃薯中,并对影响马铃薯转化的几种因素(抗生素浓度、农杆菌菌液浓度、共培养时间等)进行优化。结果表明:最适农杆菌菌液浓度是OD600=0.6,最适侵染时间是5min,最适共培养时间是2d,马铃薯转化中最适头孢霉素浓度是400mg/L;经PCR检测确认的转基因马铃薯植株在含0.7%NaCl的培养基中可以生长,而野生型马铃薯无法在此培养基中正常生长。在盐胁迫条件下,转基因马铃薯的Na~+、K~+含量及K~+/Na~+的比值均高于野生型马铃薯。研究证实马铃薯植物的的耐盐性可以通过农杆菌转化导入獐茅液泡膜Na~+/H~+逆向转运蛋白基因而得到提高。
Through Agrobacterium-mediated transformation, the Na ~ + / H ~ + antiporter gene (AlNHX) of Solanum nigrum var. Solani was expressed in potato and several factors affecting the potato transformation (antibiotic concentration, Agrobacterium tumefaciens concentration, Co-cultivation time, etc.) to optimize. The results showed that the optimal concentration of Agrobacterium was OD600 = 0.6, the optimal infection time was 5 min, the optimum co-culture time was 2d, and the optimal concentration of cefotaxime in potato was 400 mg / L. Transgenic potato plants grew in medium containing 0.7% NaCl, whereas wild type potatoes did not grow normally in this medium. Under salt stress, the contents of Na ~ +, K ~ + and K ~ + / Na ~ + in transgenic potato were higher than those in wild type. Studies have confirmed that the salt tolerance of potato plants can be transformed into Agrobacterium tumefaciens Na + / H + antiporter gene and improved.