摘要
[Objective] The aim was to investigate AtNHX1 gene transformation in Brassica napus L. mediated by Agrobacterium tumefaciens. [Method] By using Agrobacterium-mediated method and cre/lox plant expression vector,the transformation of AtNHX1 gene of Na＋/H＋ antiporter in Brassica napus was studied. [Result] The regeneration rate of cotyledon with petiole was much higher than that of hypocotyl,thus,the cotyledon with petiole was selected as the recipient for transformation. After the cotyledon with petiole was soaked in bacterial solution （OD600=0.4） for 8-10 min,kanamycin-resistant green seeding percentage could reach 3.75%. [Conclusion] The PCR detection of kanamycin-resistant plants proved that NHX1 gene had been inserted into Brassica napus genome. And this research could provide a new way to improve the salt tolerance of Brassica napus.
[目的]研究根癌农杆菌介导AtNHX1基本对甘蓝型油菜的转化。[方法]采用根癌农杆菌介导法,应用cre/lox植物表达载体,研究Na+/H+逆向转运蛋白AtNHX1基因对甘蓝型油菜的转化。[结果]甘蓝型油菜带柄子叶的再生频率远大于下胚轴的再生频率,因此选择带柄子叶作为转化受体。带柄子叶经浓度OD600为0.4的菌液中侵染8～10min,卡那霉素抗性绿苗率达3.75%。[结论]对抗性植株的PCR检测证明,外源基因已经整合到油菜基因组中,为提高甘蓝型油菜耐盐能力研究提供了新的途径。
基金
Supported by Key Project of Nanjing Xiaozhuang University(2007NXY01)
Natural Science Foundation of the Higher Education Institutions of Jiangsu Province (08KJD180011)
College Student Practice and Innovation Training Program in Jiangsu Province(2009-2011)~~
