本研究建立了以甘蓝型油菜下胚轴为外植体的一步不定芽再生培养和遗传转化体系。首先,以甘蓝型油菜中双11号含部分子叶节的下胚轴为外植体,从6-BA和NAA配比、Ag NO3以及无菌苗苗龄等方面对影响油菜组织培养的因素进行了研究,建立了甘蓝型油菜快速高频一步不定芽再生培养技术体系。该技术体系为,切取5 d苗龄含部分子叶节的下胚轴置于添加4 mg/L 6-BA的MS基本培养基中培养,5 d再生出不定芽,再生频率为100%。在此基础上,进行了甘蓝型油菜的遗传转化,成功地将用p FGC5941双元载体构建的Bn TFL1基因干扰载体转入中双11号中。整个转化过程中,芽的诱导生成只需5 d,接着完成抗性芽的PPT筛选需要30 d,整个转化周期从播种到得到生根抗性苗仅需大约70 d,而传统转化方法抗性芽的诱导筛选需要90 d,整个转化周期需要130 d左右,大大缩短了转化周期,简化了转化过程,提高了转化效率。 In this study, we established a one-step adventitious bud regeneration culture and genetic transformation system using the hypocotyls of Brassica napus as explants. First of all, taking the hypocotyls of some cotyledon nodes of double 11 in Brassica napus as explants, the factors influencing the tissue culture of canola were studied from the aspects of 6-BA and NAA ratio, AgNO3, and seedling age of sterile germ The research established a rapid high-frequency adventitious bud regeneration culture system of rapeseed. The technical system is that the hypocotyls of 5-day-old seedlings with some cotyledonary nodes are cut and cultured in MS basic medium supplemented with 4 mg / L 6-BA, and adventitious buds are regenerated on the 5th day with a regeneration frequency of 100%. On this basis, the genetic transformation of Brassica napus was carried out and the Bn TFL1 gene interference vector constructed with the pFGC5941 binary vector was successfully transferred into Zhongshuang11. In the whole transformation process, bud formation was only 5 days, followed by PPT screening of resistant buds for 30 days. Only about 70 days had elapsed between sowing and obtaining resistant seedlings, while the traditional transformation method resistant shoots Of the screening needs 90 d, the entire conversion cycle takes about 130 d, greatly reducing the conversion cycle, simplifying the conversion process and improving the conversion efficiency.