摘要
采用农杆菌介导的大豆子叶节转化系统成功地将Bt基因 (cryIA)导入大豆。从发芽 5 - 7天的大豆无菌苗切取子叶节外植体 ,经农杆菌感染和共培养后 ,在选择培养基上 4周左右出现抗性不定芽。将不定芽转移到芽伸长培养基上 ,4 - 6周后再生苗长至 2 .5 - 3cm高。再将再生苗切下转入生根培养基 ,2周左右生根。生根后的再生植株经逐步锻炼移入盆中 ,所有植株均能正常开花结荚。在移栽成活的 8株T0 植株中有 7株PCR检测呈阳性反应 ;在 7个T1株系中有 4个株系存在PCR阳性植株。取 4个稳定遗传的T1代株系内的阳性植株的叶片提取DNA ,用地高辛标记的Bt基因探针进行Southern杂交分析 ,结果 4个株系均呈现阳性 ,证明Bt基因已整合到受体大豆的基因组内并能传递给后代。
Bacillus thuringiensis cryIA gene was introduced into soybean successfully with Agrobacterium-cotyledonary node transformation system. Cotyledonary node explants were prepared from 5-7 day old seedlings, infected and co-cultivated with Agrobacterium tumefaciens. Resistant adventitious buds emerged after 4 weeks on selective medium. The buds were transferred onto elongation medium and regenerated shoots grew to 2.5-3 cm high after 4-6 weeks. The shoots were cut off and transferred onto rooting medium . After about 3 weeks, the rooted plantlets were hardened off and transferred into pots. All the plants could flower and set seeds normally. Among the 8 transplanted survival T 0 plants, seven gave positive PCR reaction; and 4 of the 7 T 1 plant lines had PCR positive plants. DNA was extracted from the leaves of positive plants in the 4 T 1 plant lines, and southern blot was conducted with probe of Bt gene. All the 4 lines are positive, indicating that the Bt gene has integrated into the receiver's genome and can be passed to progenies.
出处
《大豆科学》
CAS
CSCD
北大核心
2001年第3期157-162,F003,共7页
Soybean Science
