提高水稻基因枪转化效率的研究

Some Improvements of the Biolistic Transformation System for Oryza sativa

在转化前将受体材料转移到高渗培养基上，即用含有一定浓度的甘露醇和山梨醇的培养基对受体材料进行处理。基因枪轰击后把受体材料转出，结果表明，这样做明显提高了瞬时表达（ＧＵＳ基因的表达）效果，用可筛选的标记基因（潮霉素磷酸转移酶基因，ＨＰＴ）试验表明，经高渗处理后，稳定转化的效率也有提高。用基因枪将带有ＨＴＰ基因的粒子轰击其盾片，经过筛选及植株再生，获得了转基因植株。

Osmotic treatment of the target tissues during particle bombardment significantly influenced the efficiencies of transient β glucuronidase (GUS) expression and stable transformation of a selectable gene (hygromycin phosphotransferase (HPT) gene).Among different treatments,the supplementation of 0.5～0.75mol/L of sorbitol and mannitol in media resulted in more than 100 fold increase in transient expression and more than 5 fold increase in stable transformation.Mature embryos from seeds of two japonica rice varieties were bombarded with plasmid DNA which contained HPT gene and GUS gene.After selection for hygromycin B resistance and plant regeneration,transgenic plants were obtained.Integration of introduced HPT gene into plant genome was demonstrated by molecular analysis.Mature embryos provide the most convenient materials for biolistic transformation of rice.