甘蓝型油菜带节子叶再生体系的建立及遗传转化应用

优良再生体系的建立是农杆菌介导植物遗传转化的基础。本研究建立以甘蓝型油菜含部分子叶节的子叶为外植体的再生体系,该再生体系为:切取5日苗龄甘蓝型油菜含部分子叶节的带柄子叶为外植体,置于添加3 mg/L6-BA的MS基本培养基中培养5天,外植体再生出丛生不定芽,不定芽再生率为100%。基于新建立的带节子叶再生体系,通过农杆菌介导,成功地将用p FGC5941载体构建的Bn TFL1基因干扰载体转入甘蓝型油菜中,从播种到得到生根抗性苗,整个转化周期只需要60~70天,较传统甘蓝型油菜以不带节子叶为外植体100~130天的转化周期大大缩短。

Establishment of Regeneration System of Cotyledon With Cotyledonary Node From Brassica napus L. And Transformation Application

The establishment of a good regeneration system is the foundation of agrobacterium- mediated genetic transformation. With cotyledons including part cotyledonary node fromBrassica napus as explants, a regeneration system was established. The content of this system was that cotyledons including part cotyledonary node from five days age of rapeseed seedlings were cut as explant and then were cultured in the MS media supplemented with 3 mg/L 6-BA, and five days later, shoot regenerated from the explant. The frequency of shoot regeneration was 100% . Based on the new regeneration system of cotyledon with cotyledonary node, aBnTFL1 RNAi vector constructed from pFGC5941 vector was transformed intoBrassica napus successfully by agrobacterium mediation in the research. The whole transformation period from seeding to getting rooted resistant seedlings was 60- 70 days. Compared with 100- 130 days period of traditional transformation based on regeneration culture of cotyledon without cotyledonary node, this new transformation method obviously shortened transformation period.