摘要： 利用农杆菌〔GUS基因（β-葡萄糖苷酸酶基因）为标记基因，NPTⅡ为选择基因〕，用共培法对棉花下胚轴切段直接转化。在0.1mg/L2,4-D和0.1mg/LKT的MS培养基上共培48h,转移到加头孢霉素500mg/L和50~100mg/L卡那霉素的上述培养基中诱导和筛选抗性愈伤组织，70~80天计算愈伤组织的诱导频率，并用组织化学定位法检测GUS基因的表达，统计转化频率。结果表明：平均出愈率为40%左右，最高达60%以上。愈伤组织转化率平均为30%左右，最高达70%.再生植株转化频率一般在30%左右，最高达60%.中国农科院生物技术中心和中国科学院遗传所对DNA分子杂交检测结果，证实BT基因已整合到转化棉株的基因组中，且遗传给后代植株

关键词: 棉花, 下胚轴, 出愈率, 转化率

Abstract: Hypocotyl segments from 5 6 day old seedling of Gossypium hirsutum were co-cultured with Agrobacterium tumefacience,GUS gene as marker gene,NPT as selecting gene.After the cultures were co cultured in MS medium contained 0.1 mg/L 2,4 D and 0 1mg/L KT for 48h,then they were transferred into the medium described above suplemented with 500mg/l cefotaxime and 50 100mg/L kanamycin to induce and select resistant calli.After 70 80d the induction frequence of calli was calculated.The expression of Gus gene was tested with histochemistry location method and the transformation frequency was recorded.The experimental result indicated that the average rate of callus was 40% or so, and the highest was over 60%.The average rate of callus transformation was 30% or so,and the highest was 70%.The frequency of regenerated plant transformation was 30% or so,and the highest was over 60%.According to the result of DNA molecular hybridization,the presence of introduced gene in the genome of the transgenic plants was confirmed. It was also confirmed that BT gene had been integrated into genome of transgenic plants and was inherited into its progenies.

Key words: Cotton, Hypocotyl, Callus rate, Transformation rate

中图分类号: 

• S562.035