摘要
目的:对农杆菌介导的水稻遗传转化实施优化。方法:以籼稻品种9311成熟胚、明恢86以及粳稻品种日本晴为水稻遗传转化优化的研究材料。试验中使用的农杆菌菌株为LBA4404,采用的质粒为pCAMBIA1301A-At MYB44。配制多种溶液后,制备农杆菌感受态细胞。使用制备好的农杆菌感受态细胞将质粒pCAMBIA1301A-At MYB44导入农杆菌菌株LBA4404中。应用导入后的菌液对三种水稻实施侵染,实现其遗传转化。提取遗传转化后水稻植株的DNA分子并实施PCR检测。结果:优化后DNA提取量、DNA纯度、DNA完整性均更高。优化后对于籼稻品种的遗传转化效果更好。优化后PCR条带更加清晰,同时其出现非特异性条带的情况不明显PCR检测阳性率更高。结论:在研究中成功实现了对于农杆菌介导的水稻遗传转化的优化,具有重大现实意义。
Objective:To optimize the genetic transformation of rice mediated by Agrobacterium.Method:Mature embryos of indica rice variety 9311,Minghui-86,and japonica rice variety Nihon Haruna were used as research materials for genetic transformation optimization of rice.The Agrobacterium strain used in the experiment was LBA4404,and the plasmid used was pCAMBIA1301A At MYB44.After preparing multiple solutions,prepare Agrobacterium competent cells.Transfer plasmid pCAMBIA1301A At MYB44 into Agrobacterium strain LBA4404 using prepared Agrobacterium competent cells.Apply the imported bacterial solution to infect three types of rice and achieve their genetic transformation.Extract DNA molecules from rice plants after genetic transformation and perform PCR detection.Result:After optimization,the DNA extraction amount,DNA purity,and DNA integrity were all higher.After optimization,the genetic transformation effect of indica rice varieties is better.After optimization,the PCR bands are clearer,and the occurrence of non-specific bands is less obvious,resulting in a higher positive rate of PCR detection.Conclusion:The optimization of Agrobacterium mediated rice genetic transformation has been successfully achieved in the study,which has significant practical significance.
作者
周晓佟
ZHOU Xiao-tong(Northeast Forestry University,Harbin 150000,China)
出处
《北方水稻》
2025年第5期18-23,共6页
Northern Rice
关键词
农杆菌
质粒
水稻遗传转化
DNA分子提取
PCR检测
Agrobacterium
Plasmid
Rice genetic transformation
DNA molecule extraction
PCR detection
