摘要
利用表面活性剂silwet-77辅助渗透进行floral-dip转化甘蓝型油菜的结果表明,0.1%以上的高浓度表面活性剂silwet-77处理对植株的结实率存在明显的抑制作用,且转化不稳定。0.05%浓度的silwet-77处理获得的转化率较高,未用silwet-77处理的转化频率明显下降。同一浓度的表面活性剂处理下,6d内每隔1d进行连续3次浸渍处理转化频率优于1、2次处理。对13170粒T1代种子进行Km抗性筛选获得了103株抗性植株,对抗性植株进行GUS组织化学检测表明,目的基因已经导入到甘蓝型油菜中,抗性植株进一步的分子检测和田间性状鉴定正在进行中。
Using surfactant, silwet-77, as infiltrating assistant, B.napus floral-dip transformation mediated by Agrobacterium was carried out in this research. The results indicated that high concentration (≥0.1%) of surfactant silwet-77 can obviously effect the seed setting rate of B.napus and the stability of transformation. The results showed that 0.05% silwet-77 was up to high transformation frequency and low transformation frequency without adding silwet-77. Transformation with floral-dip treated by 3 times day after day, would be more superior to that with 1 or 2 treatment under the condition of the same silwet-77 concentration and during in six days. 103 Km resistant transgenic plants were identified from the 13170 T1 seeds. GUS assay of the Km resistant plants indicated that the foreign gene had been integrated into the rape genome. Further molecular assay is being in progress.
出处
《分子植物育种》
CAS
CSCD
2004年第5期661-666,共6页
Molecular Plant Breeding
基金
四川省科技厅应用基础项目
四川省教育厅油菜育种与生物技术应用研究项目资助
