农杆菌介导反义油酸脱饱和酶基因转化甘蓝型油菜

Introduction of Trans Desaturase Gene into Brassica napus via Agrobacterium Mediated Transformation

以甘蓝型油菜花油3号的子叶柄作为转化受体,利用农杆菌介导法,将反义油酸脱饱和酶基因导入甘蓝型油菜,获得了卡那霉素抗性植株.对再生植株进行PCR-Southem blotting检测,证明外源基因已整合到油菜基因组中.本文还进一步分析了影响遗传转化的一些因素.结果表明:对子叶柄进行适当的预培养,可以提高转化效率.延迟筛选也有利于提高转化效率.将处于对数生长期的农杆菌菌液稀释10倍后,外植体浸泡5～7min的转化效果最佳.

Using the cotyledon petioles of cultivar Huayou No.3 Brassica napus as materials, trans desaturase gene was introduced into rapeseed mediated by Agrobacterium. The regenerated seedlings with Kanamycin resistant was obtained. The results of PCR-Southern blotting indicates that trans desaturase gene had been integrated into the genome of rapeseed. Factors which affected the transformation of cotyledon petiole were studied. Results indicated that suitable pre-culture could increase transformation frequency, and delayed selection was also advantageous to increasing it. Additionally, 5~7 min was the most suitable detaching time under the condition that the germ liquid which was in log phase had been diluted 10 times.